20 February 2019, Volume 33 Issue 1
    

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  • Editorial
    C.E. Gallenga, F. Pandolfi, A. Caraffa, S.K. Kritas, G. Ronconi, E. Toniato, S. Martinotti, P. Conti
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 1-6.
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    Activated mast cells (MCs) secrete a number of compounds including pro-inflammatory and anti-inflammatory cytokines. MCs are a potential source of cytokines and chemokines which participate in allergic reactions and inflammation. MCs can be activated by IgE through its receptor FceRI, but also by Toll-like receptors and/or interleukin (IL)-1. MCs can be a target for both pro-inflammatory and anti-inflammatory cytokines. IL-1 activates MCs to release inflammatory chemical mediators, and cytokines/chemokines, an effect which can be potentially inhibited by IL-37. In addition, IL-36 is also a powerful cytokine with a pro-inflammatory activity. IL-38 binds IL-36R and inhibits the pro-inflammatory activity of IL-36, thus performing a therapeutic action. In this article we review the role of MCs in relation to pro-inflammatory and anti-inflammatory IL-1 family member cytokines and a possible therapeutic effect in inflammatory disorders.

  • Article
    J. Liao, B. Liu, W. Zhong, G-D. Wang, Y-L. Xu, X. Chen
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 7-17.
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    This study aimed to explore the protective effect of Lycium barbarum polysaccharides (LBPs) against hyperlipidemia and lipid-induced renal injury in a rat model. Male Sprague-Dawley rats (n=30) were randomly divided into three equal groups: a control group (fed a regular diet) and two experimental groups (fed a high-fat diet). By feeding rats a high-fat diet for 12 weeks, an animal model of hyperlipidemia was established, after which one experimental group received oral LBPs at a dose of 300 mg/kg per day. Blood lipids, renal function, and urinary proteins were measured after 12 weeks. Changes in renal pathology and expression levels of sterol regulatory element binding transcription factor 1 (SREBP-1), interleukin-6 (IL- 6), tumor necrosis factor-alpha (TNF-α), AMP-activated protein kinase (AMPK) were determined. Rats with hyperlipidemia induced by a high-fat diet showed increases in blood lipids and blood urea nitrogen, serum creatinine, and urinary protein, as well as increases in renal levels of SREBP-1, TNF-α, and IL-6 and decreases in renal levels of adiponectin and AMPK. Administration of LBPs restored blood lipid, blood urea nitrogen, serum creatinine, and urinary protein levels, downregulated renal levels of SREBP-1, TNF-α, and IL-6, and upregulated renal levels of adiponectin and AMPK. These results indicate that LBPs may mediate lipid metabolism, enhance anti-inflammatory responses, and ameliorate renal injury caused by lipid metabolism isorders in a rat model of hyperlipidemia.

  • Article
    B. Bobrowska-Korczak, D. Skrajnowska, K. Seidel, W. Bielecki, B. Gadzinski
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 19-28.
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    The aim of the present study was to assess the effect of zinc and copper, separately or in combination with resveratrol, on the concentration of 15-hydroxyeicosatetraenoic acid (15-HETE) in urine of rats with mammary cancer (adenocarcinoma) induced with 7,12- dimethyl-1,2-benz[a]anthracene (+ DMBA). The research focused on the kinetics of alterations in urinary 15-HETE at the early stage of carcinogenesis, as well as on the influence of dietary factors on the process. The content of 15-hydroxyeicosatetraenoic acid in the rats' urine was determined by enzyme-linked immunosorbent assay (ELISA). The 15-HETE level was standardized by conversion to the creatinine level. Regardless of the diet (standard, Zn, Zn+resveratrol, Cu, Cu+resveratrol), DMBA-induced breast carcinogenesis was not inhibited. On the contrary, in the Zn+resveratrol supplemented group, tumorigenesis developed at a considerably faster rate. It was found that the supplementation of the rats' diet with zinc only or zinc plus resveratrol resulted in a reduction of the 15-HETE level in urine of rats with mammry cancer (+ DMBA) as compared with control rats (- DMBA). A statistically lower concentration of 15-HETE was found in urine of rats supplemented with zinc or copper with resveratrol when compared with the animals that received only zinc or copper (p=0.02; p=0.0001). Summing up the obtained results, it can be concluded that the level of 15-HETE in urine of the examined animals depended on the applied supplementation.

  • Article
    M. Oštrić, M. Kukuljan, D. Markić, A. Gršković, A. Ivančić, D. Bobinac, J. Španjol, J. Maroević, I. Šoša, T. Ćelić
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 29-38.
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    Since vascular calcification is considered a process regulated similar to that of bone tissue mineralization, we investigated the participation of bone formation proteins. We analyzed the correlation of serum circulating bone markers, osteoprotegerin (OPG) and receptor activator of nuclear factor ĸB ligand (RANKL) in chronic kidney disease (CKD) patients, to coronary artery calcification score. We also considered the effect of inorganic phosphate on pro- and anti-calcifying tissue factors. We confirmed that circulating OPG is an independent calcium score predictor with its high serum concentration favoring high coronary artery calcification. In tissue samples of non-diseased human renal arteries, the expression of OPG and receptor activator of nuclear factor ĸB (RANK) was positive, while expression of RANKL was absent. In atherosclerotic specimens and arteries with medial calcification, the most upregulated was expression of bone morphogenetic proteins, BMP-2 and BMP-7, as well as expression of RANK and RANKL. In the diseased arteries, OPG expression was present only in areas where bone structures were formed. In atherosclerotic and medial calcification arteries, loss of alpha-smooth muscle actin (α-SMA) expression was observed. These data suggest a possible regulatory role of the examined proteins, especially OPG and RANKL, in vascular calcification, as well as their possible clinical significance as circulating predictors of vascular calcification.

  • Article
    M. Brązert, D. Iżycki, W. Kranc, B. Borowiec, M. Popis, P. Celichowski, K. Ożegowska, M. Jankowski, M. Jeseta, L. Pawelczyk, A. Bręborowicz, D. Rachoń, M. Nowicki, B. Kempisty
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 39-51.
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    The ovarian granulosa cells (GCs) that form the structure of follicle undergo substantial modification during the various stages of human folliculogenesis. These modifications include morphological changes, accompanied by differential expression of genes, encoding proteins which are mainly involved in cell growth, proliferation and differentiation. Recent data bring a new insight into the aspects of GCs' stem-like specificity and plasticity, enabling their prolonged proliferation and differentiation into other cell types. This manuscript focuses attention on emerging alterations during GC cell cycle - a series of biochemical and biophysical changes within the cell. Human GCs were collected from follicles of women set to undergo intracytoplasmic sperm injection procedure, as a part of remnant follicular fluid. The cells were primarily cultured for 30 days. Throughout this time, we observed the prominent change in cell morphology from epithelial-like to fibroblast-like, suggesting differentiation to other cell types. Additionally, at days 1, 7, 15 and 30, the RNA was isolated for molecular assays. Using Affymetrix® Human Genome U219 Array, we found 2579 human transcripts that were differentially expressed in GCs. From these genes, we extracted 582 Gene Ontology Biological Process (GO BP) Terms and 45 KEGG pathways, among which we investigated transcripts belonging to four GO BPs associated with cell proliferation: "cell cycle phase transition", "G1/S phase transition", G2/M phase transition" and "cell cycle checkpoint". Microarray results were validated by RT-qPCR. Increased expression of all the genes studied indicated that increase in GC proliferation during long-term in vitro culture is orchestrated by the up-regulation of genes related to cell cycle control. Furthermore, observed changes in cell morphology may be regulated by a presented set of genes, leading to the induction of pathways specific for stemness plasticity and transdifferentiation in vitro.

  • Article
    JC. Zhang, WX. Li, L. Wu, X. Liu, L. Zhang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 53-62.
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    The role of Grb2-associated binder 1 (Gab1) in bFGF-activated PI3K-AKT pathway of endothelial cells remains largely unknown. To elucidate this role, a set of studies with siRNA knockdown of Gab1 was performed. Knockdown of Gab1 using siRNA was performed in fused endothelial cell line EA.hy926 and the low level of Gab1 was confirmed with quantitative R-T PCR and Western blotting. Effects of Gab1 down-regulation were examined on several aspects: bFGF-induced AKT phosphorylation, proliferation, migration and vessel tubing formation of EA.hy926 cells. The bFGF-induced AKT phosphorylation of wild-type EA.hy926 cells was both dose-dependent and time dependent with a peak at 10 ng/ml and about 30 min after bFGF treatment. The AKT activation was significantly reduced in Gab1 siRNA-treated EA.hy926 cells. The blocking of Gab1-AKT path resulted in a set of biological alterations of EA.hy926 cells: (i) reduced proliferation; (ii) impaired migration; (iii) decreased vessel tubing formation in both 2D and 3D culture. All data support that Gab1 is associated with angiogenesis function of EA.hy926 endothelium cells via PI3K-Akt signaling pathway.

  • Article
    X. Yuan, YS. Fan, L. Xu, GQ. Xie, XH. Feng, K. Qian
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 63-71.
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    Gout is a common metabolic disease and acute gouty arthritis (AGA) is one of the important complications. Jia-Wei-Si-Miao-Wan is a newly developed drug for treating acute gouty arthritis, but the molecular mechanism has not been completely clarified. Thus, this study was aimed to explore the regulation of Jia-Wei-Si-Miao-Wan on NLRP3 inflammasome and TLR/NF-κB signaling, which are two important signaling pathways in inflammation. AGA rat model was established by injecting monosodium urate into the right knee. Colchicine and Jia-Wei-Si-Miao-Wan were administrated by gavage. The circumference of the knee was measured. IL-1β and IL-18 level in the flushing fluid was detected by enzyme linked immunosorbent assay (ELISA). Western blot, immunohistochemistry and quantitative real-time RT-PCR were used to detect the protein and mRNA expression of TLR4, NLRP3, ASC, caspase-1, NF-κB and p-NF-κB. The results showed that IL-1β and IL-18 level in the flushing fluid was increased and TLR4, NLRP3, ASC, caspase-1, NF-κB and p-NF-κB expressions were up-regulated after the establishment of AGA rat model. Colchicine and Jia-Wei-Si-Miao-Wan administration could alleviate the inflammation in the knee by inhibiting NLRP3 inflammasome and TLR/NF-κB signaling. In vivo data showed that the therapeutic effect of Jia-Wei-Si-Miao-Wan could be comparable with colchicine but had lower hepatic and renal toxicity. In conclusion, Targeting NLRP3 inflammasome and TLR/NF-κB signaling by Jia-Wei-Si-Miao-Wan could be effective in treating AGA.

  • Article
    D. Zhu, R. Ding, LJ. Li, YM. Zheng, H. Wang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 73-80.
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    This study aimed to investigate the effect of T cell subsets with different proportions on renal function and blood lipids in patients with preeclampsia (PE). Ninety pregnant women were included in this experiment and classified into 3 groups: group A (30 with severe PE), group B (30 with mild PE) and group C (30 healthy pregnant women). Clinical characteristics were examined. The ratio of T cell subsets in the three groups was detected by flow cytometry. The relationship between PE and T cell subsets was determined. The results showed that no significant difference was found in age or gestational age (P>0.05). The number of CD4+ T cells (CD4 is mainly expressed in helper T cells, referred to as TH) in group A increased significantly compared with group B and group C (P less than 0.05), while the number of CD8+ T cells (CD8+ T cell, also known as cytotoxic T cell (CTL) in group A decreased noticeably in comparison to group C (P less than 0.05). The ratio of CD4+ cell number to CD8+ cell number (TH/CTL) in group A was elevated significantly compared with group B and group C (P less than 0.05). The uric acid (UA) concentration of group A was noticeably elevated compared to group C (P less than 0.05), which differed insignificantly between group B and C (P>0.05). Glomerular filtration rate (GFR) of group A was obviously impaired in comparison to group B and group C (P less than 0.05), which differed insignificantly between group B and C (P>0.05). Total cholesterol (TC) concentrations in group A, C and B were examined. TC concentrations in the former two groups were slightly higher compared to the latter (P less than 0.05), and those in the former two groups were also higher than the normal range (P less than 0.05). Mean triglyceride (TG) concentrations in all 3 groups were above the normal range, and those in group A and B were significantly higher compared to group C (P less than 0.05). TG concentrations differed insignificantly between group B and C (P>0.05). Serum TG, UA and TC in PE patients were positively related to TH/CTL (P less than 0.05). In conclusion, PE is related to T cell subsets, and T cell subsets are closely related to kidney injury and dyslipidemia.

  • Letter
    H-H. Meng, C. Shu, X-Y. Mo
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 81-84.
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    To investigate the pathogenesis, diagnosis and treatment of heterotopic pregnancy (HP), a retrospective analysis of the clinical data for in vitro fertilizations and embryo transfer procedures (IVF-ET) was performed at the Hospital of Jilin University. In this case study, the subject was admitted to the hospital for monitoring the IVF-ET results by color Doppler ultrasonography and found to have an intrauterine pregnancy combined with the income of the annex area. Postoperative pathology confirmed the HP. The patient was provided with appropriate expectant management, and finally cesarean section was performed for delivery of a full-term infant. IVF-ET patients should be alerted to the occurrence of HP. If HP is diagnosed, a comprehensive assessment based on the patient’s clinical and extrauterine pregnancy characteristics should be undertaken to determine the treatment plan. Expectant management may also lead to a good outcome for mother and infant.

  • Letter
    B. Zhang, J. Zhao, H.J. Jing, R.M. Yu, Y. Song, A.H. Hu
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 85-89.
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    Acute kidney injury (AKI) is often associated with adverse clinical outcomes and is difficult to diagnose in the early stages in septic patients. This study aims to investigate the correlation between serum C-reactive protein (CRP) levels and AKI severity diagnosis in the patients with sepsis. This retrospective study included 60 patients, who were divided into an AKI group and a non-AKI group. CRP levels were compared between the two groups. CRP was evaluated by analysis of the area under the receiver operating characteristics curve (ROC, AUC=0.648, Z=1.965, P=0.0494, 95%CI 0.514-0.767). CRP levels were significantly different between the AKI group, 99.79(49.26-172.59) mg/L, and non-AKI group, 23.13(5.80-65.49), p=0.001, Z=-3.0409. CRP had a sensitivity of 55.26% and a specificity of 81.82% at the optimal cut-off value. Subsequent multivariate logistic regression analysis indicated that CRP could predict AKI severity in septic patients (OR=1.01; 95%CI=1.00-1.02; P=0.001). High level of CRP correlates with more severe AKI in patients with sepsis. In conclusion, CRP could be an attractive predictor in early diagnosis of AKI in septic patients.

  • Letter
    HJ. Wang, JF. Ding, YL. Yu, LJ. Zhu, ZM. Ying, DH. Cao, MC. Wang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 91-96.
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    The aim of this study was to investigate the effect of oxycodone on the immune function and biochemical parameters in puerpera undergoing cesarean section. The puerperas undergoing cesarean section were randomly divided into two groups, namely, the oxycodone hydrochloride group (group A) and the morphine group (group B). After the operation, patient controlled intravenous analgesia (PCIA) was used for medication. The analgesic effects at different time points, after 1 h, 2 h, 6 h, 12h, 24 h and 48 h, were recorded. The score of the numerical rating scale (NRS) of uterine contraction pain was evaluated. At the same time, peripheral blood was taken at different time points after analgesia (immediately after surgery, then 24 h, 48 h and 72 h after surgery). Th1 and Th2 cell levels and biochemical parameters were determined. Compared with the morphine group, the Th1 cell level in the oxycodone hydrochloride group was significantly increased at 24 h and 48 h after surgery (P<0.05). Th2 level was significantly increased compared to that of morphine at 24 h, 48 h and 72 h after surgery (P<0.05). The alanine aminotransferase (ALT) level and aspartate aminotransferase (AST) level in the oxycodone hydrochloride group were significantly decreased compared to that of the morphine group at 24 h after surgery. AST level in the oxycodone hydrochloride group was significantly increased compared to that of the morphine group at 72 h after surgery (P<0.05). The total bilirubin (TB) level was significantly increased in the oxycodone hydrochloride group compared to the morphine group at 24 h, 48 h and 72 h post-surgery (P<0.05). Compared with morphine, oxycodone can increase the level of Th1 and Th2 immune cells in peripheral blood in puerperas undergoing cesarean section, and has an increased effect on liver function, without causing abnormal liver function.

  • Letter
    J. Hu, Y. Zeng, C. Shen, H. Sun, X. Lian
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 97-103.
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    The aim of work was to study the mechanism of platelet-rich plasma (PRP) in promoting the repair of lacunar bone defects in the lower extremities of rabbits. PRP was first prepared, and PRP and artificial bone were mixed into PRP gel at a ratio of 2:1. Forty-nine healthy New Zealand white rabbits were selected, and unified standard PRP was prepared using the blood collected from one of these rabbits. The other rabbits were randomly divided into two groups, a PRP group (the bone defect area was filled with PRP gel) and an artificial group (the bone defect area was filled with only artificial bone). At weeks 3, 5, 7 and 9 post-operation, the animals were sacrificed, and the bone defect repair was observed by gross observation, X-ray, pathological and immunohistochemical methods. The bone repair of each group was scored, and the barrier density value in the bone defect area and the proportion of positive staining of the immunohistochemical sections were recorded. The results showed that the wound healing of the two groups was good, and the radiological score and the value of refractive density were significantly different between the two groups at weeks 3, 5 and 7 post-surgery (P < 0.05). Both gross observation and HE staining showed that healing of the bone defect in the PRP group was better than that in the artificial group, and the proportion of positive staining of the immunohistochemical sections in the PRP group was higher than that in the artificial group at weeks 3 and 5 post-operation. In conclusion, PRP can promote repair and healing of bone defects. Vascular endothelial growth factor (VEGF) is an important stimulating factor for early bone regeneration.

  • Letter
    S. Manzoor, S. Younis, M. Hussain, F. Ehsan, N. Sheikh, M.H. Abbasi
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 105-108.
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    The success of interferon-free regimens using new direct-acting antiviral (DAA) is a revolution and major breakthrough in the development of new therapeutic options against hepatitis C virus (HCV). Accumulating evidence suggest sustained virological response (SVR) with DAA in 95% of patients. To date, however, there are very few data related to efficacy of DAA in the Pakistani population. We aimed to investigate the efficacy of sofosbuvir-based regimen among Pakistani population. A total of 1,913 patients who attained SVR24 after being treated with sofosbuvir and ribavirin from August 2015 to March 2017 were enrolled in this study. We analyzed the demographic, clinical and virological data and screened all patients for HCV in March 2017 to evaluate the response rate. We found an overall response rate of 92.8%. In addition, we also observed lower response rates among older patients. It can be inferred that a large proportion of patients achieved SVR after treatment with sofosbuvir-based regimen.

  • Letter
    J. Wu, Q. Zhang, S. Qian, L. Jin
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 109-117.
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    The aim of this study was to investigate the effect of active vitamin D deficiency on bone marrow ablation (BMX) repair in mice. Six-week-old α(OH)ase knockout [1α(OH)ase-/-] mice and control wild type mice were used to generate the BMX model. Serum Ca, P and PTH (parathyroid hormone) concentration were determined at 0 week (before operation), 1 week (1W), 2 weeks (2W) and 3 weeks (3W) post-operation. Callus formation and absorption 1W, 2W and 3W post-operation were observed by imaging and histochemistry. The morphological changes and osteoblast/osteoclast content in callus were determined by immunohistochemical analysis. The expression of gene and protein associated with bone resorption were determined in callus tissue to evaluate the impact of vitamin D on bone formation. The results showed that 1α(OH)ase-/- mice suffered from low Ca, low P and hyper PTH due to deficiency of active vitamin D, and the symptoms persisted in the process of BMX and subsequent bone repair. In control mice, new calluses were observed in the medullary cavity at the first week, then part of the callus was absorbed in the second week, and most of the callus was absorbed in the third week. In the experimental group, new calluses were observed in the first week and reached its peak in the second week, and most of these calluses were mineralized in the third week. Meanwhile, the cortical bone became thinner and callus mineralization was decreased in 1α(OH)ase-/- mice compared to the control mice. In conclusion, active vitamin D deficiency causes hyper PTH in mice. Hyper PTH promotes trabecular bone formation and decreases bone turnover; increases cortical bone formation and bone resorption while improving bone turnover.

  • Letter
    S. Borys-Wójcik, M. Brązert, M. Jankowski, K. Ożegowska, B. Chermuła, H. Piotrowska-Kempisty, D. Bukowska, P. Antosik, L. Pawelczyk, M. Nowicki, M. Jeseta, B. Kempisty
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 119-123.
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    Human Wharton’s jelly mesenchymal stem cells (WJ-MSCs) exhibit CD29, CD79 and CD105 markers, characteristic for mesenchymal cell lines. Under the influence of the appropriate factors, WJ-MSCs can be dedifferentiated to osteoblasts, chondrocytes, adipocytes, myocytes, cardiomyocytes, glial cells and dopaminergic neurons. Wharton’s jelly (WJ) is one of the potential sources of mesenchymal stem cells (MSCs) - obtaining these cells does not raise moral or ethical objections, because the umbilical cord (UC) is a regular waste material. The expression of the OCT-4 and Nanog proteins, which are characteristic for WJ-MSCs may indicate that these cells have retained some embryonic character. The collected data suggests that WJMSCs show increased division and telomerase activity compared to bone marrow MSCs (BM-MSCs). The published results showed no human leucocyte antigen (HLA) class II expression, with the possibility of HLA class I modification by WJ-MSCs, allowing for the transplantation of these cells both within the same and other species - which allows the use of human cells in animal models. The results of selected studies indicate that WJ-MSCs can be an essential element of regenerative medicine of the 21st century.

  • Letter
    S. Këlliçi, J. Mihani, M. Miraçi, M. Fida, T. Backa
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 125-131.
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    Non-adherence in patients with rheumatoid arthritis (RA) using disease-modifying antirheumatic drugs (DMARDs) may lead to increased disease activity and symptoms, poorer quality of life and increased costs related to the disease treatment. The aim of this study is to determine the level of adherence to the treatment protocols of a chronic disease such as rheumatoid arthritis in the Albanian population and to investigate the predictors of adherence to medication. This is a single-center cross-sectional study in which included 125 patients diagnosed with RA recruited from the University Hospital “Mother Teresa” in Tirana, Albania from January to December 2013. The study was based on self-reporting by patients through a standard questionnaire divided into two parts. The first part consisted of a face-to-face interview with the patients in the hospital setting and the second part aimed at evaluation of the adherence to the prescribed therapy and was filled in via a telephone interview 4 weeks after the patient’s discharge from the hospital (117 patients participated in the second part of the interview). The Morisky Medication Adherence 4-item Scale (MMAS-4) was used to measure patients’ self-reported adherence to their medications. Factors affecting adherence are evaluated through the values of χ2 (chi-squared test of Pearson) for categorical variables and p (Student’s t-test) for continuous variables; 30.8% of full adherence to DMARDs was found in this study. Adherence to the medication was associated with education, income, number of comorbidities, use of biologic DMARDs, use of traditional medication and patients neglectful behavior toward medication. Three of the most important reasons reported by the patients as reasons for non-adherence to the medication were: the cost of the treatment; forgetfulness in taking the drug; and side effects of the treatment. The findings emphasize the need for intervention of the health authorities to improve the access to medicines for RA. Health professionals have a crucial role to inform and educate patients in order to support them in improving their adherence.

  • Letter
    C. Jin, L. Yu, S. Fang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 133-138.
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    The aim of this study was to investigate the effect of glucagon-like peptide-1 (GLP-1) on islet β-cell autophagy in mice with type 2 diabetes induced by high-fat diet and low-dose streptozotocin (STZ). Twenty-four 6-week-old male C57BL/6J mice were divided into four groups: group A (control group; 8-week low-fat diet and 8-week normal saline treatment), group B (8-week high fat diet and streptozotocin intervention as well as 8-week normal saline treatment), group C (8-week high fat diet and streptozotocin intervention as well as 2-week GLP-1 intervention), and D (8-week high fat diet and streptozotocin intervention as well as 8-week GLP-1 intervention), with six mice in each group. At the end of 16-weeks, the mice were sacrificed, and blood and tissues were collected. The pathophysiological and morphological changes were evaluated by H&E staining. Immunohistochemical staining of proliferating cell nuclear antigen (PCNA) was performed. Autophagy in islet cells was accessed by immunofluorescence staining of LC3 and P62. Glucose tolerance test (GTT) showed that blood glucose was lower in group C than in group B at 60 min after fasting and glucose load (P < 0.05). Insulin tolerance test (ITT) showed that blood glucose in group C was lower than that in group B at 30 min, 60 min, and 120 min, respectively (P < 0.05). H&E staining showed that the hepatic steatosis, necrosis and inflammatory cell infiltration were significantly reduced in group D compared with group A, B and C (P < 0.05); mice in group B had the highest degree of islet morphological changes, cell necrosis, and inflammatory cell infiltration among the four groups (P < 0.05). The level of LC3 was higher in group D than in group B (P < 0.05). The P62 level was reduced in group D compared with that in group B (P < 0.05). In conclusion, GLP-1 intervention can change the proliferation and autophagy status of islet β cells in mice, and thus protect the function of islet β cells.

  • Letter
    PC. Li, XS. Yang, WY. Li
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 139-143.
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    Invasive fungal disease (IFV) has a high incidence among autoimmune disease patients who have poor immunity. Diagnosis of IFD is difficult in the early stage. Conventional imaging technologies are poor in sensitivity, specificity and detection rate, therefore, an effective method is needed for improving the diagnosis of autoimmune disease complicated with IFD. To study the benefits of galactomannan testing in the diagnosis of IFD, 256 patients who were diagnosed with autoimmune disease in our hospital between December 2014 and December 2016, and who developed pulmonary infection in the process of immunosuppressant treatment, were selected. They were classified into 2 groups: autoimmune disease patients with IFD and those with non-IFD, based on the diagnostic criteria and treatment principles of invasive pulmonary fungal infection. Among the patients with autoimmune disease in combination with IFD, there were 12 confirmed cases, 24 clinically diagnosed cases and 49 suspected cases. The 36 confirmed and clinically diagnosed cases were used as the observation group, while 171 patients with autoimmune disease in combination with non-IFD were placed in the control group. Galactomannan testing was performed on both groups. The results demonstrated that the level of galactomannan in the patients in the observation group were significantly higher than those in the control group (P<0.05). Receiver operating characteristic (ROC) curves revealed that galactomannan testing had high diagnostic value for autoimmune disease combined with IFD. The area under ROC curve was the largest when galactomannan value higher than 0.7 was taken as the positive breakpoint, and the sensitivity and specificity at this critical value were 87.1 and 92.6%, respectively. Therefore it can be concluded that galactomannan testing has important application values in the diagnosis of autoimmune disease in combination with IFD.

  • Article
    M. Kulus, M. Brązer, M. Popis, B. Borowiec, D. Bukowska, M. Jeseta, H. Piotrowska-Kempisty, M. Nowicki, B. Kempisty, P. Antosik
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 145-149.
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    Shuttling proteins are molecules that can facilitate transport through the nuclear envelope. A very large number of proteins are involved in this process that includes nuclear pore buildup, signal, receptor and enzyme proteins. There are many examples of proteins whose biological activity depends on nucleocytoplasmic transport. Very often they are largely responsible for the proper occurrence of cell division, maturation, development and differentiation. Thanks to the well mastered methods of in vitro cell culture, it is possible to trace the levels of protein expression and their distribution in cells. Advanced molecular techniques allow for precise determination of their displacement in time. Several studies are still being carried out, using primary cultures, to identify the factors that determine the maturation, development and differentiation of cells. In understanding of the detailed mechanisms controlling cell life, the key is not the level of expression of a specific protein, but its distribution in individual cellular compartments.

  • Letter
    Z. Ahmad, M.H. Abbasi, N. Sheikh, S.R. Ahmad
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 151-155.
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    Mitochondrial gene, COI has been identified as the standard barcode for molecular identification of many species. To date, no attempt has been reported for the molecular characterization of bats in Pakistan. The current study is therefore planned to design primers for the molecular marker COI through multi-species sequence alignment approach for Pakistani bat species. The COI sequences from major bat families were retrieved from NCBI and aligned on Mega 7 to find primers from most conserved regions. Based on the alignment results three primers with degeneracy were designed with one forward and two sets of reverse primers (PBCOIdF, PBCOIdR1 and PBCOIdR2). Touchdown PCR was carried out to amplify the COI gene from two major bats families i.e., Vespertilionidae, and Emballonuridae from the Punjab region. Primer pair (PBCOIdF and PBCOIdR1) yielded clean amplicons of size 579 and 576 bp for the bat specimens belonging to the Vespertilionidae family, whereas second primer pair (PBCOIdF and PBCOIdR2) gave good quality bands of size 673 and 664 for the Emballonuridae family. NCBI blast sequence analysis for specimens from Vespertilionidae and Emballonuridae revealed the identity score of 95% (Pipistrellus tenuis) and 97% (genus Taphozous), respectively. Group specific primers designed in the current study gave successfully satisfying results when applied to two large bat families. This work will certainly help to identify bats species across Pakistan and generate data regarding genetic diversity of this taxon parallel with other countries.

  • Letter
    S.B. Shah, A.C. Kaushik, F. Ali, L. Huang, X. Lu, L Sartaj, P. Xu, H. Tang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 157-162.
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    Hexabromocyclododecanes (HBCD), an additive brominated flame retardant, is causing great concern as a result of its extensive use and increasing volumes in different environmental systems. In this study, the HBCD degrading gene dehHZ1 was identified from strain HBCD-sjtu by homology modelling of enzyme DehHZ1 sequence with LinB from strain Sphingobium indicum B90A, an HBCD degrading enzyme. The three dimensional (3D) model structures of enzyme DehHZ1 were obtained. Structure and functional relationship of the enzyme DehHZ1 with compound HBCD was performed using active sites. The HBCD degrading ability of the transformant containing the gene dehHZ1 was observed at 30ºC, pH 7.0 and 200 RPM, in mineral salt medium (MSM) with 0.1 mM HBCD. An intermediate 1, 5, 9-cyclododecatriene (CDT) was recognized based on gas chromatography mass spectrometry analyses after HBCD degradation. These findings may provide new insights into the environment friendly route of HBCD degradation and its conversion into new metabolite.

  • Letter
    XL. Qiao, Y. Dong, XY. Feng, HQ. Wang, CM. Jiang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 163-167.
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    Nonalcoholic fatty liver disease (NAFLD) is gradually becoming one of the most frequent causes of liver diseases. The aim of this study was to establish a mouse model of nonalcoholic fatty liver disease induced by high fat diet, which could be used for studying the pathogenesis of NAFLD. Forty male C57BL/6J mice were randomly divided into normal and high-fat model groups. Compared with the normal group, after 5 weeks of high-fat diet, the model group showed significantly elevated serum TG, TC, fasting glucose, transaminase, hepatic TG, TC and TNF-α levels (P <0.05). In contrast, hepatic adiponectin content was significantly reduced (P <0.05) in the model group compared to the normal group. The morphological alterations determined by HE and Sudan III further showed diffuse hepatic steatosis and massive inflammatory cell infiltration in the model group. Immunohistochemistry staining further revealed significantly elevated CYP2E1 and TGF-β1 expression and markedly decreased catalase and adiponectin expression in the model group compared to the control group (P <0.05). This study confirmed that high-fat diet significantly increased liver CYP2E1 and TGF-β1 expressions and decreased adiponectin and catalase expressions in NAFLD mouse model.

  • Letter
    DL. Guo, ZW. Gong, T. Zhang, L. Xue, JL. Tong, XQ. Li
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 169-174.
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    To evaluate the practical value of delayed enhancement 320 row volume multidetector computed tomography (DE-320 row volume MDCT) for reperfused acute and old myocardial infarction (MI) in a porcine model, 14 pigs underwent balloon-induced occlusion of the middle segment of the left circumflex coronary artery (LCX) for 90 min. The balloons were removed and reperfusion was performed. DE-320 row volume MDCT was performed 15 min after the injection of iodinated contrast. Delayed-enhancement magnetic resonance imaging (DE-MRI) was completed 15 min after injection of gadolinium-DTPA. The pigs were then divided into acute vs old MI groups. Six pigs in the acute MI group were sacrificed immediately and their hearts were harvested for triphenyltetrazolium chloride (TTC) pathology. The pigs in the old MI group were reared for another 6 months before undergoing repeat DE-320 row volume MDCT and DE-MRI prior to being sacrificed. On DE-320 row volume MDCT, the size of the acute and old myocardial infarction was 23.95±9.8% and 16.93%±7.04%, respectively, which had a high consistency and correlation with DE-MRI and TTC pathology. The CT values of the delayed enhanced area were different between acute MI and old MI and were 132.5±30.5 HU and 91.2±18.3 HU, respectively (P <0.001). The CT values of the delayed enhanced areas were different from that of viable myocardium (acute, P <0.001, old, P <0.001). The diagnosis of the infarcted segments by DE-320 row volume MDCT had good consistency with the results of TTC pathology (acute, kappa = 0.76; old, kappa = 0.64). DE-320 row volume MDCT can be an effective method for assessing MI.

  • Letter
    T-M. Guan, S. Xiao, D-CH. Liu, X-H. Liao, W-D. Li, K-F. Wu, R. Huang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 175-181.
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    The fruits of wampee have been used traditionally as stomachic and vermifuge herbs. Previous research has shown that wampee fruits possess antioxidant and antialcoholism effects. Drug-induced liver injury is very common when using anti-tubercular drugs, and is considered to be linked to oxidative stress. This study was designed to investigate the hepatoprotective activity of wampee fruits against antituberculosis drug-induced hepatotoxicity. Hepatotoxicity was induced in rats by oral administration of a combination of isoniazid and rifampicin. An aqueous extract from the fruits of wampee (AEFW) at doses of 1.5 and 3.0 g/kg/d were evaluated for their potential hepatoprotective effects. The serum levels of hepatic alanine transaminase (ALT) and aspartate transaminase (AST) were estimated at the end of the experiments. An analysis of hepatic histopathology was carried out to assess injury to the liver. Western blot and immunostaining were used to determine the levels of protein expression. The results showed that AEFW (1.5 and 3.0 g/kg/d) was found to significantly decrease the levels of ALT and AST and reduce hepatocellular necrosis and liver fibrosis in treated rats, compared with the control group. Furthermore, Western blot and immunostaining analysis revealed that the extract can down-regulate the expression of MyD88 and NF-κB p65, and inhibit NF-κB p65 phosphorylation. Therefore AEFW can alleviate anti-tubercular drug-induced liver damage by enhancing anti-inflammatory effects.

  • Letter
    J. Gil, D. Ramsey, P. Pawlowski, E. Szmida, P. Leszczynski, M. Bebenek, M.M. Sasiadek
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 183-185.
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    Despite great progress in colorectal cancer (CRC) prevention, early recognition and treatment, a high frequency of morbidity and mortality of CRC patients is still observed even in developed countries. Molecular characterization of this tumor becomes a standard procedure allowing for application of personalized therapy. However, searching for new biomarkers and for new individual therapeutic strategies is increasingly desirable. In this study on molecular background of CRC, we focused on analyses of mRNA levels of autophagy gene ATG9A and pro-apoptotic gene BAX. Genes involved in autophagy, that is the catabolic and conservative cellular process, have been revealed as a promising new cancer biomarker as well as anti-cancer target. Under normal circumstances autophagy occurs at a low, basal level in most human cells, however in cancer cells its expression varies from down- to upregulation. Despite the fact that the complex link between autophagy and apoptosis is well documented, these interrelations have not yet been fully uncovered. The relative expression of mRNA values of ATG9A and BAX genes in colorectal cancer samples vs normal adjacent tissues was assessed by Real-time PCR with Universal Probe Library. The results of our study revealed a statistically significant correlation between the expression of BAX and ATG9A genes, showing that in CRC a higher expression of BAX gene is connected with lower expression of ATG9A.

  • Letter
    G. Koracević, M. Stojanović, D. Lović
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 187-189.
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    There is no reason to believe that excessive alcohol consumption during weekends substantially increases the risk of atrial fibrillation (“Holiday heart syndrome”) but it is not the case with the risk of blood pressure (BP) elevation. Practical implication of this “Weekend - Masked arterial hypertension (MAHT)” is to record 24-h ambulatory BP monitoring also during the weekend, in patients at high risk of MAHT, if recordings during workdays are normal.

  • Letter
    X. Wang, X. Duan, X. He
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 191-196.
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    The aim of this study was to explore the interaction of immune factors in acquired immune deficiency syndrome (AIDS) patients with hepatitis C infection and its effects on immune and liver function. Fasting venous blood was taken from hepatitis C virus (HCV) patients (HCV group), human immunodeficiency virus (HIV)-treated AIDS patients (HIV group), HIV-infected HCV patients (HIV/HCV group), and healthy controls (control group). Enzyme linked immunosorbent assay (ELISA) was used to detect immune factors (IL-10, IL-6, TNF-α) closely related to disease and intestinal microbial translocation products [Lipopolysaccharide (LPS), bacterial lipopolysaccharide binding protein LPS-binding) Protein (LBP), soluble CD14 molecule (soluble CD14, sCD14), intestinal fatty acid binding protein (IFABP), and anti-endotoxin core antibody (EndoCAb)]. Flow cytometry was used to detect the number of peripheral blood CD4+ T cells. The immune factors closely related to the disease after HIV and HCV infection change in microbial translocation products, and correlations between peripheral blood CD4+ T cell counts and immunization, microbial translocation are compared. The results show that the concentrations of immune factors and intestinal microbial translocation products in the peripheral blood of the HIV/HCV group were higher than those in the HIV and HCV groups. HIV and HCV interact with the intestinal microbial translocation products LPS, LBP, sCD14, and IFABP. There were no significant correlations between immune factor and microbial translocation products in the HIV/HCV group and the number of CD4+ T cells in peripheral blood. In conclusion, after HIV is co-infected with HCV, the systemic immune system is activated, releasing a large number of immunostimulatory factors, further damaging the body. Due to the combined effect of HIV and HCV virus, the liver is further damaged, and the degree of damage mainly depends on the degree of HCV infection and microbial displacement.

  • Letter
    H. Miao, B. Cao, W. Ge, W. Zhao
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 197-203.
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    This study aimed to investigate the expression of p16 and p27 protein in cervical exfoliative cells and explore the relationship between the expression of HR-HPV (High-risk human papilloma virus) and cervical lesions. One hundred-twenty cervical cytology specimens were selected as cases from patient cervical cancer screening. According to the double screening, results were divided into a double negative group, a single positive group, a double positive group [including Thinprep cytologic test (TCT) ≤ Low-grade squamous intraepithelial lesion (LSIL)] and an HPV positive group, [TCT ≥ High-grade squamous intraepithelial lesion and HPV positive group (HSIL)]. Flow cytometry was used to detect the expression of p16 and p27 protein in the specimens of the exfoliated cells of the cervix. The result showed that the expression of p16 in the TCT ≥ HSIL group was higher than other groups (P<0.05), and the expression of p27 was lower than the other groups (P<0.05). In group CIN3, the expression of p16 and p27 was P<0.01 compared to the other groups. The positive rate of the A9 group in group CIN3 and the inflammatory and CIN1 group was statistically different (P<0.05). Moreover, in the single positive groups (group A9, group A5/6, A7 group, and mixed-infection group), the infection rate of group A9 was significantly higher than that of the other groups. With the progression of cervical lesions, the expression of p16 increased and the expression of p27 decreased, and both expressions were associated with HR-HPV infection.

  • Letter
    J. Zhou, T. Li, K. Cai, X. Xu, S. Zhang, L. Yan, X. Sun, D. Lu
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 205-211.
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    The study was aimed to elucidate the molecular mechanism of Farnesyl X receptor (FXR), Bile salt export pump (BSEP) and sodium-taurocholate cotransporting polypeptide (NTCP) in the metabolism of bile acid and cholesterol in hyperlipoidemia rats. Fifty male Wistar rats weighing 140.6±9.5g were randomly divided into two groups: the control group was given normal diet while the experimental group was given high-fat diet. Body weight was measured on a regular basis and liver weight was measured after sacrifice. Plasma glucose and lipids (total cholesterol, triglycerides, high density lipoprotein, low density lipoprotein) were determined by commercially available kits. Hepatic FXR, BSEP and NTCP gene expression and protein distribution were determined by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and immunohistochemistry. The results showed that the expression of FXR and BSEP mRNA in the experimental group was higher than that in the control group (P < 0.05). The expression of NTCP mRNA in the control group was higher than that in the experimental group (P < 0.05). Immuno-histological results showed that the positive rates of FXR, BSEP and NTCP expression in the experimental group were significantly different from those in the control group (P < 0.05). In conclusion, FXR and Bsep and Ntcp gene changes played an important protective role in lipid metabolism, especially for the rats with high-fat diet.

  • Letter
    GH. Li, DH. Ren, S. Yu, J. Chen, K. Fang, ZH. Li, L. Cai, ZL. Shi, JY. Zhang, JI. Ma
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 213-218.
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    Sepsis is a common complication in surgical patients and is the first cause of death in Intensive Care Unit (ICU) patients. The aim of this study was to identify the effect of cholecalciferol on the expression level of T helper 17 cells (Th17) and regulatory T cells (Tregs) in patients with severe sepsis, and to explore their role in immune function in severe sepsis. Forty patients with severe sepsis from Intensive Care Unit (ICU) were selected as the experimental group. Blood samples were collected from all patients on day 1 after their admission to ICU. CD3+CD25+ cells were isolated from blood specimens and used to measure the differentiation of CD3+CD25+ to Th17 cells and Treg cells under different concentrations of cholecalciferol: 0 nM, 1 nM, 5 nM and 10 nM. Apache II scores, Sofa Scores, blood pressure and platelet count decreased in the group with low cholecalciferol levels compared to the group with normal cholecalciferol level. The percentage of CD4+IL-17+ and CD8+IL-17+ cells decreased significantly in the 4 groups under cholecalciferol treatment in a dose-dependent manner. The percentages of CD4+CD25+Foxp3+ and CD8+CD25+Foxp3+ cells increased significantly in the 4 subgroups under cholecalciferol treatment in a dose-dependent manner. Cholecalciferol modulates the differentiation of CD3+CD25+ T cells into Th17 cells and Treg cells by decreasing the Th17 cells levels and increasing the Treg cell levels in a dose-dependent manner, suggesting that cholecalciferol level can affect the progression of severe sepsis patients.

  • Letter
    T. Batool, T. Akhtar, A. Asghar, M.B. Khawar, N. Sheikh
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 219-224.
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    Obesity is an epidemic issue associated with low-grade inflammation and in most of the cases with poor iron condition that influences nearly every community. The present research experiment was designed to inspect the consequences of obesity on the micro architecture of heart and kidney tissues as well as on the expression of mRNA of some hepatic genes like Hepcidin, Transferrin and TNF-α in association with their role in iron regulation and proinflammatory responses in the body. Two tentative groups of Rattus norvegicus (each supplied with different proportions of fat in their food) and one control group were analysed. After a period of sixteen weeks, the rats were dissected and their organs, liver, heart and kidneys, were excised and preserved accordingly. Histological analysis of heart tissues of obese rats showed hemorrhagic condition along with interstitium widening and interstitial inflammatory condition (myocarditis) and also necrosis of some myocardial fibers. Microanalysis of kidney tissues of obese rats showed glomerular distortion, glomerular basement membrane thickening and enlargement of Bowman’s capsule along with mesangial expansion. mRNA expressional studies of some hepatic genes of the obese rats showed significant elevation in the expression of hepicidin (P-value < 0.001) and TNF-α (P-value < 0.001) and at the same time significant decline in the expression of transferrin (P-value < 0.001) was also observed as compared to the control group. Taken together, from these findings it can be concluded that fatty diet induces many changes in the cellular architecture of heart and kidney as well as in the expression of hepatic genes involved in iron regulation (Hepcidin and transferrin) and proinflammatory responses (TNF-α).

  • Letter
    SW. Chen, HX. Xia, QQ. Fan, AW. Xie, Y. Yu
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 225-230.
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    Primary liver cancer is a commonly seen tumor, and early surgical resection can achieve a favourable effect, however traditional laparotomy has the shortcomings of great trauma and multiple postoperative complications, which are not beneficial to the recovery of patients. To explore the characteristics and effectiveness of precise hepatectomy, 116 patients with primary liver cancer who were admitted to Chenzou No. 1 People’s Hospital, Hunan, China, between August 2015 and December 2016 were selected. They were randomly divided into a conventional hepatectomy group and a precise hepatectomy group, 58 in each group. The surgical condition, changes of liver function indexes, prognosis and incidence of postoperative complications were observed and compared between the two groups. The results suggested that patients who underwent precise hepatectomy had less average intraoperative bleeding and shorter hospital stay compared to those who underwent conventional hepatectomy, but the duration of the surgery was longer (P < 0.05). Compared to pre-surgery, the content of total bilirubin in serum (TBIL), alanine transaminase (ALT) and aspartate transaminase (AST) of the two groups decreased, and the content of albumin (ALB) increased after treatment. The changes of the indexes of the precise hepatectomy group were more obvious, and the liver function of was superior to that of the conventional hepatectomy group; the comparison within group and between groups had statistical significance (P<0.05). The recurrence rate of tumor of the precise hepatectomy group was significantly lower than that of the conventional hepatectomy group (P<0.05). The lesion removal rate, one-year survival rate, and two-year survival rate of the former were obviously higher than that of the latter (P<0.05). The difference of the incidence of complications between the two groups had statistical significance (P<0.05). Precise hepatectomy has a favourable effect in the treatment of primary liver cancer as it can effectively control cancer recurrence and improve prognosis; hence it is worth promotion.

  • Letter
    LR. Qiu, LJ. Zhou, BY. Wang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 231-235.
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    Nursing intervention is a positive method for the recovery of patients’ health. To study the clinical effect of intraoperative nursing intervention in patients with cholecystolithiasis who underwent laparoscopic cholecystectomy, 100 patients with cholecystolithiasis were randomly divided into two groups: an experimental group that was given nursing intervention, and a control group given conventional nursing care. The average length of stay, average hospitalization expenditure, postoperative recovery and nursing satisfaction of the two groups were compared. We found that there were significant differences in the average hospitalization time, average hospitalization expenses, postoperative recovery and nursing satisfaction between the two groups. The average hospitalization time of the experimental group was 4.4±0.7 days, and the cost was 7985±561 yuan, which were lower than those in the control group. Postoperative recovery and nursing satisfaction of the experimental group were better than those of the control group, indicating that nursing intervention achieved a favorable effect.

  • Letter
    LQ. Huang, W. Shen, T. Qiu, YH. Zhi, LC. Wang, SH. Mao, X. Xing, SH. Shen, RL. Jiang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 237-244.
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    This study aimed to prepare an animal model of qi-deficiency gastrointestinal failure, and verify feasibility of the model. One hundred and twenty healthy adult male ICR mice were randomly divided into a control group (24 mice), a placebo group (24 mice), and an experimental group (72 mice). Mice in the control group were given a normal diet for 10 days, those in the placebo group were additionally given normal saline solution at 4°C for 10 days, and mice in the experimental group were additionally given 4°C supersaturated sodium sulfate solution for 10 days. After 10 days, mice in the experimental group were randomly divided into three subgroups: a model group, a self-healing group and a Sijunzi decoction group, with 24 mice in each group. Mice in the model group were immediately sacrificed, those in the Sijunzi decoction group were fed with Sijunzi decoction for 7 days, and mice in the self-healing group were given the same amount of normal saline for 7 days. Mice in each group were randomly divided into three subgroups, namely, pathological group, immunohistochemistry group and toner propelling test group, with 8 mice in each subgroup. The general condition of mice in the toner propelling test group was observed. The changes of intestinal villi, intestinal gland and intestinal epithelial cells in the intestinal mucosa of mice in the pathological group were observed. In the immunohistochemical group, apoptosis and 5-HT of the intestinal epithelial cells were detected. All indicators were found to be significantly different from those of the normal control group. After 7 days of traditional Chinese medicine treatment, mice in the Sijunzi decoction group had gained weight, reduced stool, and increased gastrointestinal propulsion index, and the pathological damage scores of intestinal mucosa were decreased. In conclusion, the mouse model of qi-deficiency gastrointestinal failure was successfully established, which is a reliable, stable and simple method to investigate the disease.

  • Letter
    W. Deng, KY. Chen, WY. Liu, ZM. Lv, C. Zhang
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 245-250.
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    The aim of this study was to investigate the effect of Vitamin D on pancreatic islet β cell injury induced by low concentration cadmium in mice. The islet β cell line NIT-1 cells were divided into normal control group, Cdcl2 group and Cdcl2+vitamin D3 (VD) group. Cell viability was detected by CCK-8 assay. Insulin content in the supernatant was detected by enzyme-linked immunoassay (ELISA). Reactive oxygen species (ROS) content was detected by flow cytometry, and changes of insulin-related genes were detected by real-time fluorescence quantitative PCR. Changes of insulin receptor substrate-1 (IRS-1) and insulin receptor substrate-2 (IRS-2) were detected by Western blotting. The results showed that the concentration of Cdcl2 (0.05 mM) which had no significant effect on the viability of NIT-1 cells was screened by CCK-8 method, where the concentration of active vitamin D (1 nM) for the cytotoxicity induced by a low concentration of cadmium could be reduced. The insulin secretion ability of NIT-1 cells was detected. The results showed that under 4-day exposure to a low concentration of cadmium, the insulin secretion of NIT-1 cells increased at first (P<0.05), and then decreased with the prolongation of cadmium exposure (P<0.05). The intervention of active vitamin D eventually increased the insulin secretion of the cadmium-exposed cells (P<0.05). At the gene expression level, long-term exposure of a low concentration of cadmium did not significantly change the expression of insulin gene, but significantly increased the ROS content in NIT-1 cells (P<0.05), while the ROS content in NIT-1 cells was decreased by the intervention of active vitamin D (P<0.05). In conclusion, the insulin secretion of islet β cells increased at the early stage of long-term low concentration cadmium exposure, and then decreased with the prolongation of cadmium exposure. This effect was affected by active vitamin D intervention.

  • Letter
    LJ. Liu, XJ. Lu, JM. Gao, RJ. Wang, GX. Cheng
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 251-257.
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    This study aimed to investigate the effect of the APP17 peptide on diabetic encephalopathy. Streptozotocin (STZ)-induced type 2 diabetes mellitus rats were used to obtain a model of diabetic encephalopathy with cognitive impairment. After treatment with APP17 peptide, the changes in blood glucose levels, body weight, and memory ability in the model rats were observed. Glucose metabolism was detected by Positron Emission Tomography/Computerized Tomography (PET/CT) in brain of rats from each group. After 12 weeks of STZ-induced diabetes, the escape latency of the diabetic encephalopathy (DE) group was significantly prolonged compared with the normal control (NC) group (p<0.05) and the uptake of glucose in the cerebral cortex and hippocampus was decreased compared with the NC group. The glucose uptake was significantly increased in the group treated with APP17 (APP17+DE), compared with the DE groups and the level of glucose metabolism in brain tissue was greatly improved (p<0.05). In conclusion, it was confirmed that APP17 peptide could improve the cognitive function and glucose metabolism in diabetic encephalopathy rats.

  • Letter
    F. Di Berardino, D Zanetti
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 259-262.
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    The nasopharyngeal cavity is the crucial point of the upper airway; its obstruction is associated with allergic rhinitis, upper airway respiratory infections and is the basis of middle ear diseases. The purpose of this preliminary study was to evaluate the ability of a new device to empty the nasopharyngeal cavity from mucus and restore free nasal breathing. The present study evaluated the effects of nasopharyngeal drainage using a new device (Atomix wave®) on nasal expiratory flow rates in 20 volunteers (12 males and 8 females, mean age 44±18 years) with nasal obstruction. Nasal spirometry was performed before and after nasopharyngeal cavity treatment with the new device. The nasal maximum mid-expiratory flow rate (n-MEF 25/75) was significantly increased after treatment (baseline value 0.92±0.43 and 0.67±0.33; post-treatment 1.31±0.56 and 1.25±0.44; p less than 0.05). The new device proved effective in clearing the nasopharyngeal cavity.

  • Letter
    A. Ballini, S. Cantore, E.A. Fotopoulou, I.P. Georgakopoulos, E. Athanasiou, D. Bellos, G. Paduanelli, R. Saini, G. Dipalma, F. Inchingolo
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 263-268.
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    Orthodontic appliances protect the plaque from the actions of brushing, mastication, and salivary flow. Plaque control is a very difficult task for patients with fixed orthodontic appliances. It has been demonstrated that chemical agents are useful additions in plaque control for these patients. To improve mechanical plaque removal, studies in the field suggest to add a chemo–therapeutic agent, such as an antibacterial mouth rinse, to the oral hygiene regimen. Noteworthy evidence from clinical trials has shown that oral hygiene status is significantly improved when antibacterial mouth rinses are added to daily oral hygiene measures (tooth brushing and flossing) compared with tooth brushing and flossing alone. The purpose of this study was to evaluate the effect of a combined rinsing solution (sea salt, xylitol and lysozyme) on plaque and gingival index in orthodontic patients with fixed appliances.

  • Letter
    S. Capodiferro, L. Calabrese, F. Maffini, E. Cascardi, G Favia, E. Maiorano
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 269-273.
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    Oral squamous cell carcinoma in patients under 20 years of age is extremely rare, the alveolar gingiva being the most frequently affected site. The metachronous occurrence (eight years later) of a dentinogenic ghost cell tumour of the anterior maxilla in the same patient has never been previously reported. The patient had been diagnosed with a poorly differentiated squamous cell carcinoma of the ventral tongue when 12 years of age and was treated by en-block resection with functional neck dissection. Eight years later, he showed a dentinogenic ghost cell tumour in the anterior maxilla, associated with an impacted central incisor, which was treated by conservative surgery with wide margins; the patient has remained free of disease, without recurrence, at the one-year follow-up. Though extremely rare, malignancies should be considered in the diagnostic work-up of paediatric/adolescent patients with head and neck neoplasms. Additional difficulties arise when dealing with multiple tumours, both under a differential diagnostic point of view and for the surgical management, due to the lack of unanimously accepted guidelines for treatment and follow-up. To the best of our knowledge, oral squamous cell carcinoma and dentinogenic ghost cell tumour are unrelated to each other and may represent a casual combination, which raises additional questions as to the clinical management.

  • Letter
    S. Bernardi, S. Mummolo, G. Varvara, E. Marchetti, M.A. Continenza, G. Marzo, G. Macchiarelli
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 275-280.
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    Regenerative dentistry aims to restore the volumes and tissues lost during traumas or periodontal disease. In the past, research focused on the development of new materials which could constitute the best graft for tissue regeneration. Dentinal matrix has been considered one of the materials that can be used for this purpose. The aim of this study is to evaluate the in vitro biocompatibility of a mineralized dentinal graft material. Human periodontal ligament fibroblast (HPLF) cell line was used for subculture, according to the manufacturer’s instructions. XTT essay was performed to evaluate the cell survival, and histological and scanning electron microscopy (SEM) analyses were performed to evaluate the morphological changes in response to the new material. The dentinal particles were obtained according to the Smart Grinder protocol and Bio-Oss material was used as positive control. The response was evaluated after 24 hours, 72 hours and 7 days from the seeding. The 24-h results showed a cell survival of 96.7% in the sample tested with the dentin and of 98.9% in the sample tested with the Bio-Oss. The 72-h results showed a cell survival of 105% in the sample tested with the dentin and of 140% in the sample tested with the Bio-Oss. The 7-day results confirmed the trends of the cellular growth: the cell survival rate of the sample tested with the dentin was 152% and of 186% in the sample tested with the Bio-Oss. The morphological data confirmed the biological essays, showing the perfect biocompatibility and the “cell-friendly” properties of the tested material. HPLF cells showed a very promising growth reaction to the tested material, comparable to the considered standard material (Bio-Oss). In vivo studies, RCTs and systematic reviews are necessary to validate the use of this material for regenerative periodontal interventions in clinical practice.

  • Letter
    G. Spoto, L. D'Alessandro, C. Rubini, M. Fioroni, A.W.S. Rutjes, S. Spoto, M. Ferrante, M. Della Malva, M. Petrini
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 281-285.
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    The aim of this study was to evaluate whether or not the expression of cAMP-phosphodiesterases (cAMP-PDE) varies in different thyroid pathologies and to elucidate the relationship between the expression of cAMP-PDE and clinic-pathologic factors. Forty thyroid biopsy samples, excised to perform the biopsy, were microscopically examined and classified. Then cAMP-PDE expression was assessed using high performance liquid chromatography. The expression of cAMP-PDE showed different patterns according to the histological features of the samples (p <0.001). A strong expression of cAMP-PDE was observed more frequently in papillar and follicular carcinoma than in controls and benign pathologies. We did not detect any level of endogenous cAMP. cAMP could be used as additional diagnostic or prognostic biomarker in patients suspected of thyroid disease, however further studies are needed to confirm these results.

  • Letter
    L. Giannetti, E. Spinas, A. Murri Dello Diago, U. Consolo, L. Generali
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 287-290.
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    The aim of this work was to describe the rehabilitation of a superior monoedentulia by positioning a conometric connection implant and a prosthetic crown using CAD/CAM technique. Surgical steps are explained and the biological aspects of this particular morse conometrical connection are evaluated, with particular attention to the possibility of removing the crown by means of an innovative attachment hole in the abutment. This type of monoedentulia can now be solved by implant-prosthetic replacement, and the dentist must choose the most suitable implant for each individual case, the most favorable connection and the most appropriate prosthetic material with regard to bone and soft tissue biology.

  • Letter
    F. Melfa, R. Rauso
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 291-292.
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    Botulinum toxin type A injection is a well established procedure for the treatment of frontal lines. In the present letter the authors propose to hyperdiluite the toxin and to perform a gentle massage after the injections to let the drug to easily spread into the muscle, reducing the amount of BoNTA units required and achieving a natural result.

  • Letter
    G.M. Nardi, R. Grassi, F.R. Grassi, S.E. Aragona, B. Rapone, F. Della Vella, S. Sabatini
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 293-297.
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    The aim of the study was to evaluate the efficacy of a polarized, polychromatic, non-coherent light (Bioptron), in addition to full mouth scaling and root planing, in chronic periodontitis. Forty adult patients affected by chronic periodontitis were enrolled in the study and were randomly divided into two groups. The control group received only full mouth scaling and root planing (FMSRP), while in the test group, Bioptron light treatment was added to FMSRP. The Full Mouth Plaque Score (FMPS), Full Mouth Bleeding Score (FMBS), Gingival Index (GI) and Probing Pockets Depth (PPD) were recorded in both groups at the first examination, after 1 month (T1) and after 3 months (T2). Both groups showed a significant reduction of FMPS at T1 and T2 (P < 0.05). Both groups showed a significant reduction of FMBS at T1 and T2 (P < 0.05), but a higher improvement (P < 0.05) was recorded in the study group at T1. Study group GI was significantly reduced at T2 (P < 0.05), differently from the control group (p>0.05). PPD showed a reduction in both groups with no statistically significant differences (p>0.05) between them. Polarized, polychromatic non-coherent light seems to be an effective additional therapy in the management of adult chronic periodontitis, reducing inflammatory clinical indexes.

  • Letter
    B. Defraia, R. Piumelli, AM. Buccoliero, G. Cannella, V. Bugelli, R. Grifoni, M. Focardi, V. Pinchi
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 299-302.
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    Congenital adrenal hyperplasia (CAH) is a group of autosomal recessive disorders due to decreased activity of enzymes necessary for cortisol biosynthesis. Each of the CAH disorders represents a clinical spectrum which reflects the consequence of a specific mutation. The Salt Wasting (SW) form of CAH due to 21-hydroxylase deficiency (21-OHD) is a life-threatening disease when untreated. It is easily diagnosed with newborn screening. Early detection of the defect prevents severe complications and an adrenal crisis that often leads to death. The authors report a case of a 20-day-old male infant, seemingly in good health, who died suddenly despite early cardiopulmonary resuscitation. The forensic autopsy showed the presence of enlarged adrenal glands. Analysis of a blood spot sample using tandem mass spectrometry revealed high serum levels of 17 hydroxyprogesterone (17-OHP), consistent of 21-OHD. The cause of death was a hypovolemic shock heart failure due to dehydration, caused by 21-OHD CAH. The report highlights the importance of extensive newborn screenings that would have prevented this death. The case evokes two relevant medico-legal issues, firstly for the hospital where the child was born, due to the lack of adequate information about the limited and inhomogeneous screening panels routinely adopted by different hospitals. Secondly, for suspicious clinical signs appearing after the discharge from hospital that could have been neglected by the pediatrician.

  • Letter
    F. Ricciardiello, F. Oliva, M. Mesolella, G. Sequino, G. Motta, G. Petruzzi, E. CANTONE
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 303-308.
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    The aim of the study was to evaluate the postoperative endoscopic score and the quality of life (QoL) in two groups of adults undergoing septoplasty with turbinoplasty and who were treated after surgery with a nasal spray containing silver vitellinate, hyaluronic acid and sodium benzoate, or saline solution alone. This single-blind randomized study was carried out on 54 patients (30 males, and 24 females, mean age 33.5±2.7 years) undergoing septoplasty and volumetric tissue reduction of inferior turbinates for nasal obstruction. All subjects underwent the Visual Analogue Scale (VAS) questionnaire and the nasal endoscopy score by Lund and Kennedy (LK) at baseline before surgery (T0), 15 days (T15) and 21 days (T21) after treatment. All subjects were randomized into 2 groups, the experimental arm, group I (GI), and the control arm, group II (GII). Patients were given the treatment, a nasal spray containing silver vitellinate, hyaluronic acid and sodium benzoate for group I and a saline solution alone (sodium chloride 0.9%) for group II. After therapy, better VAS and LK scores were found (P < 0.05) in the GI in GII. No patients reported adverse reactions or complications. Our findings suggest that silver vitellinate, hyaluronic acid and sodium benzoate nasal spray improved not only the endoscopic score, but also the quality of life (QoL) in the early postoperative period after septoplasty. In addition, the absence of adverse reactions or complications pointed out the tolerability and safety of the treatment.

  • Letter
    A. Lucchese, R. Botta, C. Lucchese, S. Croce, M. Marcolina, M. Manuelli
    Journal of Biological Regulators and Homeostatic Agents. 2019, 33(1): 309-313.
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    The objective of this work was to provide a review of the literature on studies of stem cells and their potential applications in dental practice. Research included every article published between 2000 and 2018 in PubMed PMC, Scopus, Cochrane Trial Library, and Web of Science databases featuring the keywords: regenerative dentistry, stem cells, tissue engineering. Papers that did not directly address the subject were excluded. Emphasis was given to scientific studies that showed possibilities of stem cell therapy in the oral and maxillofacial region. The methodological quality of selected papers was scored using the “Swedish Council on Technology Assessment in Health Care Criteria for Grading Assessed Studies” (SBU) method. Two review authors independently assessed eligibility, extracted data and verified the quality of the studies.