Objective:To investigate the effect of HMGB1 and TLR-4 in the renal injury of SLE.Methods:The level of HMGB1 in serum from 16 patients suffering from SLE without kidney damage,18 patient with lupus nephritis (LN),and 12 healthy individuals were measured by ELISA.The fresh PBMCs were isolated and the total RNA was extracted.Then the mRNA expression of HMGB1 was amplified by RT-PCR.Flow cytometric analysis(FCM) was performed to study cell surface markers and the expression of TLR-4.Results:RT-PCR and ELISA results showed that the expressions of mRNA and protein were higher in patients with LN than in SLE without kindey damage and healthy people.The expression of TLR-4 in CD14+ monocytes of patients with LN was higher either,while there were no significant changes in CD3+ T cells among LN,SLE and healthy control.Conclusion:PBMCs in patients with LN could synthesize and secrete HMGB1 initiatively,which are correlated with serum HMGB1 level.HMGB1 might play a role in autoimmunity of lupus nephritis partly by activation of monocytes through its binding to TLR-4.

Objective:To investigate the effects of TLR/STAT pathway in the proliferation of mesangial cell induced by HMGB1.Methods:Human mesangial cells were inoculated in the dose of 1?104 ml-1.After 24 h,cells were cultured with standard medium as control group or with medium supplement with 10 ?g/L human recombinant protein HMGB1 as trial group in vitro.Then the cells were collected in 6,12 and 24 h respectively,as well as control group cells.Immunocytochemical staining was adopted to examine the expressions of PCNA proteins on mesangial cells in different groups.Immunocytochemical staining and FCM were performed to detect the changes of TLR2 protein expression.STAT1 and STAT3 mRNA were examined by RT-PCR technique.Results:Immunocytochemical staining indicated that the mesangial cells could multiply after they were induced by human recombinant protein HMGB1.Immunocytochemical staining showed that the level of TLR2 protein in trial groups were higher than those in control groups.FCM indicated that HMGB1 could significantly up-regulate the expression of TLR2 protein time-dependently.The STAT1 and STAT3 mRNA in HMGB1 groups were higher than those in control groups.The expression of TLR2 protein was positively correlated with those of STAT1 and STAT mRNA respectively.The positive rate of PCNA was remarkably correlated with the expression of STAT1 and STAT3 mRNA.Conclusion:HMGB1 could activate STAT1/STAT3 through combining with its cell-surface receptor TLR2,which may play an important role in promoting the proliferation of mesangial cells and then damaging the renal of lupus nephritis.

Objective:To investigate the expression and mechanism of NF-κB signal pathway in murine lupus nephritis.Methods:The BXSB mice as well as C57BL/6 of 16 weeks were used.Transmission electron microscope and PAS were used to detect the pathological change of renal tissue.RT-PCR and ELISA were used to detect the expression of HMGB1 mRNA and protein.The expression of HMGB1,p- NF-κB,RAGE,IκB and PCNA protein was detected by immunohistochemical stain,FCM and Western blot.Results:The level of BUN in serum and Micro-albumin in urine of BXSB mice was higher than that in C57BL/6 mice.The expression of HMGB1 mRNA and HMGB1 protein level in peripheral blood increased significantly in BXSB group.Compared with those in control group,electron microscopy and PAS revealed the thickness of glomerular basement membrane(GBM),fusion of foot processes partly of epithelial dell and subepithelial electron-dense deposits in the renal tissue of BXSBA mice.Compared with that of control group,expression of PCNA was higher in glomeruli of BXSB mouse.HMGB1 protein over-expression localized in cytoplasm and extracellular milieu,especially in proliferative glomeruli in BXSB group,while the HMGB1 protein primarily confined to the nuclear of tubule in control group.In BXSB group,the expression of p-NF-κB and RAGE increased,while the expression of IκB decreased.There were positive correlation between the expression of HMGB1,RAGE and p-NF-κB protein (r=0.833,0.621,0.848,P＜0.01),while the expression of p-NF-κB protein negatively correlated with that of IκB.Conclusion:HMGB1 could activate NF-κB through combining with its receptor-RAGE,induce the form of proliferative glomerulonephritis by promoting the proliferation of inherent cell of glomeruli,which may play an important role in the murine lupus nephritis.

Objective To investigate the relationship between the effect of high mobility group protein box 1 (HMGBI) on the renal injure of systemic lupus erythematosus (SLE) and the expression of Toll-like receptor 4(TLR4). Methods The level of HMGB1, MMP-2 and TIMP-2 in serum from 16 pa-tients with SLE, 18 patients with lupus nephritis(LN) and 12 healthy people were measured by ELISA. The fresh peripheral blood mononuelear cell (PBMC) were isolated and the total RNA was extracted. Then the mRNA expression of HMGB1 was amplified by RT-PCR. Flow cytometry analysis was performed to study cell surface markers and the expression of TLR4. Results RT-PCR and ELISA results showed that the expres-sions of mRNA and level of HMGB1 protein in serum were higher in patients with LN than those in SLE and healthy people. The expression of TLR4 in CD14+ monecytes of patients with LN was higher than that with SLE and healthy people, while there were no significance in CD3+ T cells among LN, SLE and healthy peo-ple. The expressions of MMP-2 and TIMP-2 in serum of LN was lower than that in SLE and healthy people, at the same time the ratio of MMP-2/TIMP-2 decreased in LN group. HMGB1 mRNA and CD14+/TLR4+ was negatively correlated with the ratio of MMP-2/TIMP-2, and the level of HMGB1 in serum was positively correlated with proteinuria, while negatively correlated with the ratio of MMP-2/TIMP-2 in LN. Conclusion HMGB1 is one of the important cytokine in the pathogenesis of lupus nephritis. HMGBI might play a role in proteinuria of lupus nephritis in part via TLR4 pathway to activate monocytes and decrease the expression of MMP-2/TIMP-2.

Objective Based on the theoretical connotation of"Dredge Du meridian remodeling god"intervention in PSD,two rounds of expert questionnaire were conducted in combination with Delphi survey method to optimize the diagnosis and treatment scheme of massage intervention in PSD,and to develop an optimal operation mode of Tuina for PSD.Methods Experts with rich experience in Tuina intervention in different provinces and cities in China were selected to participate in two rounds of questionnaires.Items were screened after statistical analysis of experts′ basic information,positive degree,coordination coefficient and authority coefficient of the two rounds of questionnaires.Results In the two rounds of Delphi survey,34 and 37 experts were selected for questionnaire survey,and the positive coefficients of experts were 94.44%and 94.47%,respectively.The average expert authority coefficients of the two rounds were 0.65 and 0.74.The expert coordination degree of the two rounds of questionnaires was medium,and Kendall coefficient W>0.4.Conclusion After two rounds of expert research in Delphi,the specific techniques,techniques and treatment frequency were screened and demarcated,and the optimized PSD diagnosis and treatment plan of"Dredge Du meridian remodeling god"Tuina intervention with high expert authority and good consensus was formed,which can be promoted and applied to PSD patients clinically.

Nonalcoholic fatty liver disease （NAFLD） is an abnormal lipid metabolic disorder of the liver characterized by accumulation of a large amount of lipids in the liver， and it is currently the most common liver disease around the world. Mendelian randomization （MR） incorporates genomic data into traditional epidemiological study designs to infer the causal relationship between exposure factors and disease risk. In recent years， MR has been widely used in studies on inference of the etiology of NAFLD. This article systematically summarizes the advances in the application of MR in NAFLD research， so as to provide new ideas for understanding the nature of the disease and scientific interventions.

Objective: To observe the effect of atorvastatin combined with insulin glargine on renal function in patients with early diabetic nephropathy. Methods: From January 2016 to March 2019, 100 patients with early diabetic nephropathy admitted to Hanzhong 3201 Hospital Affiliated with Xi′an Jiaotong University Medical School were selected as subjects. According to the random number table, patients were divided into control group and observation group, with 50 cases in each group. All patients underwent diet control, blood pressure control and symptomatic treatment. Patients in the control group were treated with insulin glargine to control blood glucose. Patients in observation group were given atorvastatin on this basis. After 16 weeks of treatment, the therapeutic effects of the two groups were observed, as well as the change in urinary albumin excretion rate (UAER), serum creatinine (Scr), C-reactive protein (CRP), total cholesterol (TC), and triglyceride (TG). Adverse reactions were observed during treatment in both groups. Results: After treatment, the levels of UAER, Scr, CRP, TC and TG of the two groups were lower than those before treatment, and the above indexes of the observation group were lower than those of the control group. The difference were statistically significant (P<0.05). During the treatment period, the incidence of adverse reactions in control group and observation group was 4.00%(2/50) and 12.00%(6/50), and there was no significant difference (P>0.05). Conclusions Atorvastatin combined with insulin glargine in the treatment of early diabetic nephropathy can effectively reduce the levels of UAER, Scr, CRP, TC and TG, and has good safety.