Objective To investigate the regulatory effect of adrenomedullin (AM) on the cell-cell contact formation of podocytes and the possible mechanism.Methods Podocytes were treated with AM (10-7 mol/L),AM combined with a PKA inhibitor H89 (10-4 mol/L),and forskolin (10-5 mol/L) as positive control respectively for 12 hours.Immunofluorescent staining was applied to observe the distribution of cell adhesion molecules and actin-associated proteins.Western blotting assay was used to assess their protein levels.Rho GTPases activity was analyzed by GST-pull down assay and their protein levels were tested by Western blotting.Results AM induced the redistribution of adhesion molecules,actin-associated proteins as well as the F-actin at cell-cell contacts between podocytes.This effect was similar to that of forskolin and could be blocked by H89.The levels of those proteins did not change significantly (P ＞ 0.05).AM up-regulated the activities of RhoA,Rac1 and Cdc42 (P ＜ 0.05),which were partially blocked by H89.The protein levels of Rho GTPases showed no difference compared with the control (P ＞ 0.05).Conclusions AM may promote cell-cell contact formation of podocytes,probably through enhancing the activity of Rho GTPases and then resulting in the redistribution of adhesion molecules,actin-associated proteins and F-actin,which is partially mediated through cAMP-PKA signaling pathway.

To explore the serum miR-338-5p expression characteristics in renal transplant recipients ,and the role of regulating BAFF signal ,then investigate its biological significance.Methods:Healthy volunteers were enrolled as control group.Serum miR-338-5p was detected by real-time PCR;soluble BAFF was detected by ELISA;anti-HLA-Ⅰantibody,anti-HLA-Ⅱ antibody and anti-MICA antibody were detected by liquid chip technology.SPSS17.0 software was applied.t-test was used to compare the means of two independent samples;Paired samples t-test was used to compare the means of two paired samples;Spearman method and Pearson method were used to analyse the correlation;P<0.05 was considered to be statistically significant.Results: Compared with healthy controls,serum miR-338-5p in renal transplant recipients decreased significantly (P<0.001),while serum BAFF increased significantly (P<0.01).Serum miR-338-5p levels within 1 year post-transplantation were significantly lower than that of more than 1 year post-transplantation (P<0.01);Serum miR-338-5p levels within 3 years post-transplantation were significantly lower than that of more than 3 years post-transplantation (P<0.01);To all research objects,serum miR-338-5p was significantly negatively correlated with serum BAFF (r=-0.51,P<0.001),and serum miR-338-5p was significantly negatively correlated with anti-HLA-Ⅱ antibody(r=-0.322, P<0.05);Serum miR-338-5p within 3 years was significantly negatively correlated with anti-HLA-Ⅱantibody (r=-0.423,P<0.05), and serum miR-338-5p within 3 years was significantly negatively correlated with anti-MICA antibody(r=-0.411,P<0.05);Serum miR-338-5p more than 3 years was significantly positively correlated with anti-MICA antibody(r=0.486,P<0.05),and Serum miR-338-5p more than 3 years was significantly positively correlated with anti-HLA & MICA antibody(r=0.578,P<0.01).Conclusion:miR-338-5p may directly or indirectly target BAFF signal ,and participate antibody mediated immune response by regulating its target genes and interfere with the long-term survival of transplanted renal.

Objective To explore the significantly differentially.expressed microRNAs during antibody-mediated renal allograft rejection.Method MicroRNA array assay was used,and the obtained data were analyzed by bioinformatics analysis.The obtained significant microRNAs were further analyzed to forecast the targeted genes in the common database,then experimental means were used to testify the targeted genes.Result During the antibody-mediated renal allograft rejection,the significantly over-expressed microRNAs were miR-200c,miR-200b,miR-30c,miR-30b and miR-30e+,etc.The significantly down-expressed microRNA was miR-338-5p.The bioinformatics analysis results indicated that TRAF3 was the targeted gene of miR-338-5p,which was testified by real time PCR,immunohistochemical assay and fluorescence reporter assay.Conclusion miR-338-5p anticipated in the antibody-mediated renal allograft rejection by targeting TRAF3.

ObjectiveTo explore the relationship between the negative co-inhibitor programmed death-1 ( PD-1 ) and the pathogenesis of myasthenia gravis ( MG), by detecting the expression of PD-1 and programmed death ligand-1 ( PD-L1 ) on peripheral blood mononuclear cells (PBMCs) and soluble PD-1 (sPD-1) in plasma from myasthenia gravis patients. MethodsPeripheral blood samples were collected from 45 MG patients and 33 healthy persons without prednisone or other immunodepressant treatment during the half year ahead of withdrawal.The expression of PD-1 and PD-L1 on PBMCs were detected using immuno-fluorescence labeling and flow cytometry, and the concentrations of sPD-1 in plasma were measured using an ELISA kit. Results(1) The proportion of CD4+ PD-1 + T cells, as well as CD14+ PD-L1 +monocytes of the MG group was higher than that of the control group. There were no significant differences in the proportion of CD4+ PD-1 + T cells or CD14+ PD-L1 + monocytes in the MG sub-groups between different genders or MG types. While the proportion of CD4+ PD-1 + T cells of the late-onset MG (age ≥40) group was higher than that of the early-onset MG group (age ＜40). And it was higher in the MG patients with thymoma or thymus hyperplasia than that from the MG patients with normal thymus. The proportion of CD14+ PD-L1 +monocytes from the MG patients with thymoma or thymus hyperplasia group decreased obviously compared with that of the patients with normal thymus group; but no difference could be found between the late-onset group and early-onset group. (2)The concentration of sPD-1 in the plasma from the group of MG patients was(6. 92 ±0. 72) ng/ml,which was higher than that of the healthy control group ( (3.28 ±0. 42) ng/ml),even more, it was significantly higher in the early-onset MG group than that of the late-onset MG group,there was a negative correlation( r =-0. 526, P =0. 000) between the age of onset and the concentration of sPD-1. ConclusionsThe increased expressions of PD-1 on CD4+ T cells and PD-L1 on CD14+ monocytes in MG patients suggested the involvement of the couple of molecules in the pathogenesis of MG.Higher concentration of soluble PD-1 in the plasma of patients with MG suggested that it might disturb the ligation of PD-1 and PD-L1 on T cells and antigen presenting cells, which might result in the abnormal transportation of the negative modulating signal, and accelerate the pathological progress of MG.

Objective To observe the effectiveness of injecting mouse nerve growth factor ( mNGF) on the recovery of hand motor function among patients with cubital tunnel syndrome. Methods A total of 138 patients with moderate to severe cubital tunnel syndrome were randomly divided into groups designated as A, B and C, each of 46. Twenty micrograms of mNGF was injected daily 1 mm from the ulnar nerve at the cubital tunnel for the patients of group A and the injection site was moved 1 mm distally everyday along the nerve , but injected intramuscularly for those in group B. Those in group C received 500 μg of mecobalamin injected intramuscularly 3 times a week. The whole intervention consisted of two 4-week phases, with an interval of 2 months. Before and after the intervention, the function of internal hand muscles, hand function recovery rates and any electrophysiological changes in the ulnar nerve were measured and compared between the two groups. Results All of the patients showed significant improve-ment in hand muscle function and neuroelectrophysiology. The incidence of had muscle atrophy, Tinel′s sign, posi-tiveness in the paper clamping test and claw hand all significantly improved compared with before the treatment in all three groups. The average Disability of Arm Shoulder and Hand score in group A after the treatment was significantly higher than the group B and group C averages. The average ulnar nerve conduction velocity, incubation period and amplitude of group A after the treatment were all significantly better than before the treatment and better than the other groups′averages. After the treatment, the average hand function recovery in group A reached 91％, significantly high-er than in groups B ( 76％) and C ( 59％) . Conclusion Injecting mNGF next to the ulnar nerve is superior to in-jecting it intramuscularly in promoting the recovery of the ulnar nerve and hand function for patients with moderate to severe cubital tunnel syndrome.

Objective :To evaluate therapeutic effect of allisartan isoproxil tablet on carotid atherosclerosis (CAS) le‐sion in patients with mild to moderate essential hypertension (EH).Methods :A total of 120 patients with mild to moderate EH complicated CAS treated in our hospital were enrolled .They were randomly divided into valsartan group (n＝60 ,received valsartan 80mg／d) and allisartan isoproxil group (n＝60 ,received allisartan isoproxil tablet 240mg／d) ,both groups received measure by carotid vascular ultrasound before treatment ,24 weeks and 48 weeks af‐ter treatment and its result were compared between two groups .Results :Compared with before treatment ,there were significant reductions in IMT ,size ,thickness and number of carotid atherosclerotic plaques on 24 weeks and 48 weeks after treatment in two groups ,and above indexes of 48 weeks were significantly lower than those of 24 weeks , P＜0.05 or ＜0.01. The decreased value of IMT in allisartan isoproxil group was significantly higher than that of valsartan group ,and there were no significant difference between two groups in time point of size ,thickness and number of carotid atherosclerotic plaques , P＞0.05 all.Conclusion :Therapeutic effect of two drugs on size ,thick‐ness and number of carotid atherosclerotic plaques are similar ;but therapeutic effect of allisartan isoproxil tablet on IMT is significantly better than that of valsartan .

Objective To evaluate the diffusion distribution of bone cement in the vertebral body by quadrant method,and to analyze and evaluate the correlation between the diffusion distribution type of bone cement and new vertebral fractures after vertebral augmentation.Methods A total of 170 subjects who met the conditions from January 2020 to December 2021 were collected.According to the anteroposterior and lateral view of the spine,the injured vertebra was divided into four quadrants,and divided into homogeneous diffusion group and uneven diffusion group according to the postoperative diffusion distribution of bone cement in the injured vertebra.The incidence and types of refracture were followed up,and the VAS score and Cobb angle were compared between the two groups.Results 170 patients were followed up for at least 12 months,including 90 patients in homogeneous diffusion group and 80 patients in heterogeneous diffusion group.There were 33 cases of refracture(19.41％),12 cases of refracture(13.33％)in the diffuse homogeneous group,and 21 cases of refracture(26.25％)in the diffuse heterogeneous group,and the difference between the groups was statistically significant(P＜0.05).The site of refracture in the diffuse homogeneous group was mainly the clinical vertebral fracture,while the probability of refracture in the diffuse heterogeneous clinical vertebra and the operated vertebra was similar.The incidence of postoperative bone cement leakage in the diffuse homogeneous group was significantly lower than that in diffuse heterogeneous group(P＜0.05).The VAS score and Cobb angle were significantly improved in both groups after surgery and at the last follow-up compared with those before surgery,but there was no significant difference between groups.Conclusion The incidence of new vertebral fractures after vertebroplasty is closely related to the type of cement diffusion,and the risk of refracture defined as uneven cement diffusion by quadrant method is high.

Objective To explore the immunopathological mechanism for the imbalance between the positive signal mediated by inducible costimulator (ICOS) and the negative signal mediated by programmed death-1 (PD-1) in patients with myasthenia gravis (MG).Methods Eighty-two patients with MG,56 healthy controls (HC) and 20 non-MG (NMG) patients,collected in the First Affiliated Hospital of Suzhou University from February 2014 to December 2016,were chosen to participate in the study.The expression of ICOS and PD-1 on peripheral blood mononuclear cells was detected by immuno-fluorescence staining and flow cytometry.The levels of soluble programmed death-1 (sPD-1),soluble programmed death ligand 1 (sPD-L1),IL-4 and other cytokines were detected by enzyme-linked immunosorbent assay.Results (1) Flow cytometry analysis:The co-expression of PD-1,ICOS on CD4 + T cells from MG group (9.64％ (8.82％)) was higher than in HC (1.81％ (2.10％),Z =-7.389,P ＜0.05) and NMG group (2.86％ (1.49％),Z =-4.636,P ＜ 0.05).The expression of ICOS on CD4 + T cells,ICOS ligand (ICOSL) on CD14+ monocytes and CD19+ B cells were increased in MG group comparing with that of the control groups.The proportion of PD-1 + CD4 + T cells (MG group 16.82％ (10.66％),HC 9.34％ (9.18％),Z =-4.345,P＜0.05;NMG group 7.07％ (3.40％),Z=-4.594,P＜0.05) and PD-1 Ligand (PD-L1) + CD14+ monocytes was higher in MG patients.All of these were detected by flow cytometry.(2) ELISA analysis:Serum sPD-1 expression significantly increased in MG group compared with that in the control groups (MG group (1.87 ± 0.64) ng/ml,NMG group (1.49 ± 0.70) ng/ml,t =2.04,P ＜ 0.05;HC (1.05 ± 0.50)ng/ml,t =2.08,P ＜ 0.05),while for serum sPD-L1,there was no significant difference between MG and control groups.(3) Serum cytokines detection:The expression of IL-4 was increased in MG patients (MG group (61.88 ±5.15) pg/ml,HC (32.03 ±1.84) pg/ml,t=2.50,P＜0.05;NMG group (42.62± 3.31) pg/ml,t =2.34,P ＜0.05),and there was a negative correlation between the expression of sPD-1 and the concentration of IL-4.Conclusions The increased expression of PD-1 + ICOS + CD4 + T cells suggested the subset involved in the pathological progress of MG.sPD-1 might disturb the ligation of PD-1 on T cells and PD-L1 on antigen presenting cells,while the ligation of ICOS and ICOSL passed positive signal,leading to over activity of the subsets and the progression of disease.