Objective To study the direct effect of parathyroid hormone(PTH) on osteoclasts (OCs) differentiation and bone resorption in vitro and to explore the relationship between PTH and expression of RANKL (ligand of receptor activator of nuclear factor kappa B)gene and OPG (osteoprotegerin) gene in osteoblasts (OBs) in order to provide some theoretic evidence for understanding the pathogenesis of high-transferred renal osteodyslrophy (ROD). Methods PTH was directly used to induce OCs differentiation from whole bone marrow of C3h mouse by using OCs formation assay and then inspected the bone resorption of these OCs in vitro by using pits assay. Multiple RT-PCR assay was used to examine the expression of RANKL-mRNA and OPG-mRNA in different PTH concentrations and different treating times by PTH. Results (1) PTH in vitro could induce OCs formation form whole bone marrow of wild-type C3h mice.In a certain concentration scope, as the concentration of PTH increased, the number of differentiated OCs and the degree of bone destruction of OCs increased. (2) Expression of RANKL-mRNA and OPG-mRNA was in dose-dependent and time-dependent manner within a certain range of concentration and time. Conclusion PTH in vitro mediates OCs differentiation and bone resorption probably through the up-regulated expression of RANKL-mRNA and OPG-mRNA.

Objective To investigate the effect of purified urinary IgG from patients with minimal change nephrotic syndrome and membranous nephropathy on the expression of macrophage migration inhibitory factor (MIF) in human proximal tubular epithelial cells (HK-2). Methods Proteins were precipitated from urine with (NH4)2SO4, and urinary IgG was purified by protein G column chromatography. SDS-PAGE and Western blotting were performed to analyze the urinary IgG. HK-2 cells were incubated with urinary IgG (0,0.5,1.0,2.5,5.0,10.0 mg/ml) from either minimal change nephrotic or membranous nephropathy patients. The expression of MIF mRNA was assessed by RT-PCR, and MIF protein was assessed by Western blotting. Results SDS-PAGE showed that there were four bands, which were all IgG fractions confirmed by Western blotting. The urinary IgG up-regulated the expression of MIF mRNA and protein in HK-2 cells. The expression increased in a dose-dependent manner. The expression of MIF mRNA and protein in HK-2 cells increased significantly when they were incubated with urinary IgG from membranous nephropathy patients at 1mg/ml (P

AIM: To investigate the expression of osteopontin (OPN) and CD44 in human renal proximal tubular epithelial cells stimulated by human serum albumin (HSA). METHODS: Proximal tubular epithelial HK-2 cells were stimulated by HSA at different concentrations for different time, then OPN mRNA production was detected by RT-PCR, and OPN protein was detected by Western blotting. The expression of OPN and CD44 in HK-2 cells after stimulation for 24 h or 48 h were detected by immunofluorescence with confocal laser scanning microscope. RESULTS: Osteopontin mRNA in HK-2 cells showed a highest expression at 3 h and 48 h after HSA stimulation. However, the expression of OPN protein in HK-2 cells reached the maximum at 24 h after HSA stimulation. OPN mRNA and protein showed a strong dose-dependence relation with the concentration of HSA. HSA also stimulated HK-2 cells to express CD44 protein, the fluorescence of CD44 was most prominent at 48 h after HSA stimulation. CONCLUSION: HSA stimulates human renal proximal tubular epithelial cells to express OPN and CD44.

Objective To study the role of JAK-STAT singal transduction pathway in the interstitial fibrosis of unilateral ureter obstruction (UUO)mice. Methods Mice UUO model was established and the phosphorylation of JAK-STAT was examined at day 1,4,7 and 14 after ligation of the ureter.Mice in the treatment group were treated with daily injection of selective JAK2 inhibitor AG490 starting 2 h before ureter ligation until sacrifice while vehicle alone was given to mice in the model control group.Mice were sacrificed at day 14 after the establishment of model.Renal tubular lesion and interstitial fibrosis were assessed on paraffin section.Immunohistochemistry was used to detect renal macrophage infihration and α-SMA expression.The expression of collagen Ⅲ and MCP-1 mRNA was measured by RT-PCR.Phosphorylation of JAK2and STAT1 was examined by Western blotting. Results JAK2-STAT1 signaling transduction pathway was activated in UUO model.The activation of JAK2-STAT1 was closely correlated with the progression of renal injury,tubular histological lesions and interstitial fibrosis.AG490 treatment significantly inhibited the phosphorylation of JAK2 and STAT1 (P＜0.01).AG490 treatment also significantly reduced tubular lesions[(21.7 ±1.7)% vs (49.4±1.0)%]and interstitial fibrosis(1.0±0.1 vs 2.3±0.2),α-SMA expression(0.9±0.1 vs 2.1±0.2)and maerophage accumulation[(13.3±1.6)cells/HPF vs (34.4±1.0)cells/HPF](all P＜0.01).In addition,AG490 significantly inhibited the expression of collagen Ⅲ and MCP-1 mRNA. Conclusion JAK-STAT signaling plays an important role in renal tubulointerstitial inflammation and fibrosis.

Objective To investigate the relationship between the biochemical markers of bone turnover and renal osteodystrophy (ROD) in patients with primary nephrotic syndrome (PNS).Methods A total of 30 patients with PNS and 50 healthy subjects (controls) were included in the study.The BMD of lumbar vertebrae and femur was measured by dual-energy X-ray absorptiometry.The levels of total procollagen type Ⅰ amino-terminal propeptide (TP Ⅰ NP),β-isomerized carboxyterminal propeptide (β-CTx),intact PTH (iPTH),serum calcium,serum phosphorus,ALP,25-OH-Vitamin D3 (25-OH-VD3),β2-micro-globulin(β2-MG),and ratio of urinary to creatinine (UA/Cr) were measured and calculated.The risk factors related to ROD in PNS patients were analyzed.Two-sample t test,multiple linear regression and partial correlation analysis were used to analyze data.Results The BMD values of lumbar vertebrae and femur in the PNS group were significantly decreased compared with those in controls (t =6.162,5.583,3.891 (＜40 years),5.923,5.324,3.129 (≥40 years),all P＜0.05) and the serum levels of TPⅠNP,β-CTx,iPTH,ALP,β2-MG and UA/Cr in the PNS group were significantly higher than those in controls (t:2.738-10.129(＜40 years),3.226-12.581 (≥40 years),all P＜0.05),and the levels of serum calcium and 25-OH-VD3 in the PNS group were significantly lower than those in controls (t =3.624,7.223 (＜40 years),2.011,2.564 (≥40 years),all P＜0.05).But there was no significant difference for serum phosphorus between the 2 groups (t=0.811,0.513,both P＞0.05).TP Ⅰ NP was positively correlated with β-CTx,iPTH,ALP,UA/Cr,β2-MG(r:0.512-0.682,all P＜0.01),and TP Ⅰ NP was negatively correlated with serum calcium and 25-OH-VD3(r=-0.314,-0.562,both P＜0.01)in the PNS group.β-CTx was positively correlated with iPTH,ALP,UA./Cr,β2-MG(r:0.459-0.618,all P＜0.01),and negatively correlated with serum calcium and 25-OH-VD3(r=-0.212,-0.589,both P＜0.01).The iPTH was positively correlated with ALP,UA/Cr and β2-MG (r =0.410,0.606,0.508,all P＜0.05),and negatively correlated with serum calcium and 25-OH-VD3(r=-0.315,-0.516,both P＜0.05).Conclusions The BMD in PNS patients is lower than that in healthy subjects.Combined measurement of TP Ⅰ NP,β-CTx and the BMD is helpful for the diagnosis of ROD in PNS patients.

Objective To study the relationships between patients'personality traits,coping styles, perceived social support and the doctor?patient relationship,try to build the relationship model of patients’ psychological factors and the doctor?patient relationship,and to understand the patients psychological factors influence on the doctor?patient relationship,to provide a basis for improving the doctor?patient relationship. Methods The doctor patient relationship evaluation questionnaire, Eysenck personality questionnaire re?vised,short scale for Chinese( EPQ?RSC) ,simplified coping style questionnaire( SCSQ) and perceived social support scale( PSSS) were used to survey 400 outpatients,using SPSS 22.0 and AMOS 17.0 for data analysis. Results There was statistically significant difference in psychoticism between the high and low doctor?pa?tient relationship evaluation score groups( t=-4.537, P<0.01).There was statistically significant difference in positive coping,family support,friend support,other support and perceived social support between the high and low score groups(P<0.05).Psychoticism had a direct positive effect on doctor?patient relationship( r=0.228, P=0.000) ,and perceived social support had a direct negative effect on the doctor?patient relationship ( r=-0.256, P=0.000).Psychoticism,neuroticism,positive coping and negative coping styles could affect the doctor?patient relationship indirectly by perceived social support. Conclusion Patient personality can affect the doctor?patient relationship directly,meanwhile,it can affect the doctor?patient relationship indirectly by perceived social support,and coping styles can only affect the doctor?patient relationship indirectly by per?ceived social support.

Objective To investigate the survival rate in elderly patients with long-term catheters hemodialysis.Methods A total of 127 elderly patients receiving maintained hemodialysis were included in this study.Patients were divided into two groups according to catheter location:long term indwelling internal jugular vein catheter group and long term indwelling vena iliac external catheter group.The baseline anthropometric and laboratory parameters were measured.The catheter dysfunction and catheter related complications were observed.Results The incidence of catheter dysfunction was higher in iliac vein catheter group than in internal jugular vein catheter group (51.5 ％ vs.24.5％,x2 =8.459,P=0.015),and there were no significant differences in the catheter related infection rate and cardiovascular (CVD) event rate between the two groups.Kaplan-Meier survival curves showed that mean survival time was lower in vena iliac external catheter group than in internal jugular vein catheter group (1.9 years vs.2.5 years,x2 =10.847,P=0.001),and still was lower after controlling the catheter dysfunction and cardiovascular events (x2 =9.915,P=0.002;x2=4.368,P =0.037).Multivariate logistic regression analysis showed that levels of hemoglobin,creatinine,systolic blood pressure,parathormone and age were the independent risk factors for mortality in elderly patients with long term catheter hemodialysis.Conclusions There is a high incidence of CVD events in elderly patients with long-term catheter hemodialysis.The incidence of catheter dysfunction is higher and the survival rate is lower in elderly patients with long-term indwelling vena iliac external catheter hemodialysis.

Objective To investigate the survival of older patients with iliac vein tunneled cuffed catheters on maintenance hemodialysis. Methods A total of 70 older patients with external iliac vein tunneled cuffed catheters on maintenace hemodialysis were included in this study, there were 94 patients with internal jugular vein tunneled cuffed catheters as control group. The baseline anthropometric and 1aboratory parameters were measured. The catheter dysfunction and catheter related complications were documented. Results There were lower survival rate and catheter survival in the external iliac vein group compared with internal jugular vein group(χ2=13.714, P<0.01; χ2=13.093, P<0.01). Compared with internal jugular vein group, there was lower rate of catheter infection in external iliac vein group(χ2=9.416, P<0.01); In addition, there were higher rate of cardiovascular disease(CVD) events and catheter dysfunction among patients in external iliac vein group (χ2=7.492, P<0.01; χ2=5.912, P<0.05). Furthermore, the incidence of catheter dysfunction and cardiovascular disease events were the independent risk factors of mortality for older patients with iliac vein tunneled cuffed catheters on maintenance hemodialysis by Cox regression model. Conclusions Patients with iliac vein tunneled cuffed catheters have a shorter survival time. Those with catheter dysfunction or cardiovascular disease events are in higher risk of mortality.

To understand the function of Mayven and investigate the pathogenesis of multiple sclerosis, the gene sequences of different truncated Mayven were amplified from the gene library of human brain. These truncated fragments, including fragment P1 (1-902 bp), fragment P2 (1-523 bp), fragment P3 (507-182 bp) and fragment P4 (887-1782 bp), were cloned into pGEX-4T-2 vector to construct recombinant plasmids. The recombinant plasmids were transformed into E. coli BL21(DE3) and induced to express by IPTG. The expressed proteins were detected by SDS-PAGE and Western blot, and were purified by GST purifying system. The results showed that recombinant express vectors of different truncated GST-Mayven were successfully constructed and were expressed in soluble form protein induced by IPTG. The fusion proteins have good reactivity to GST antibody. The construction of recombinant express vectors of different truncated GST-Mayven lays a basis for further function study on Mayven.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Microfilament Proteins ; genetics ; metabolism ; Multiple Sclerosis ; genetics ; Nerve Tissue Proteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism

BACKGROUND: A large number of experiments in vivo and in vitro have shown that mesenchymal stem cells may obviously inhibit the lymphocytes and other immunocytes.OBJECTIVE: To investigate the effect of human umbilical cord mesenchymal stem cell transplantation on the immune function and prognosis of patients suffering decompensated liver cirrhosisn due to hepatitis B.METHODS: 118 patients with decompensated cirrhosis due to hepatitis B were randomly divided into control group (n=59) and observation group (n=59). The two groups all received normal medical treatment, and in addition, the observation group also received human umbilical cord mesenchymal stem cell transplantation. (4.0-4.5)×108 stem cells were transplanted twice by intervention via proper hepatic artery (10 mL) and intravenous infusion (10 mL) within 1 week after admission. The levels of serum interleukin-6, tumor necrosis factor-α, interleukin-10, transforming growth factor-β and the percentage of lymphocyte subsets in the peripheral blood were determined in the two groups before and 1, 4 weeks after treatment. The model for end-stage liver disease (MELD) score and Child-Pugh score of 118 patients after treatment for 12 weeks were observed and recorded, and liver failure, complications and survival during follow-up period in the two groups were observed. RESULTS AND CONCLUSION: After treatment for 1 and 4 weeks, the levels of serum interleukin-6 and tumor necrosis factor-α in the observation group were significantly lower than those in the control group (P < 0.05 or P <0.001), but the levels of serum interleukin-10 and transforming growth factor-β in the observation group were significantly higher than those in the control group (P < 0.05 or P < 0.001). After treatment for 1 week, the percentagesof CD3+CD4+T cell and CD4+CD25+Treg cells in the observation group were significantly higher than those in the control group (P < 0.001), but the percentages of CD3+CD8+ T cells and CD3-CD19+ B cells were significantly lower than those in the control group (P < 0.05 or P < 0.001). After treatment for 4 weeks, the percentages of CD3+ T cell ,CD3+CD4+ T cells and CD4+CD25+ Treg cells in the observation group were significantly higher than those in the control group (P < 0.05 or P < 0.001), but the percentages of CD3+CD8+ T cell and CD3-CD19+ B cells were significantly lower than those in the control group (P < 0.05 or P < 0.001). After treatment for 12 weeks, the MELD and Child-Pugh scores in the observation group were significantly lower than those in the control group (P < 0.05).During the follow-up period, none of the cases in the observation group developed liver failure, but five cases in the control group did. In addition, the incidence of complications and cumulative mortality in the observation group were significantly lower than those in the control group (P < 0.05). These results show that the human umbilical cord mesenchymal stem cell transplantation may alleviate liver inflammation and improve liver function, and then may reduce the incidence of hepatic failure and mortality for patients with decompensated cirrhosis due to hepatitis B.