Objective To initially map the gene responsible for autosomal dominant familial IgA nephropathy of a Chinese family by exclusive the five loci that had been reported with linkage analysis. Methods The genetic pattern of the familial IgA nephropathy was identified and the genomic DNA was extracted from the blood samples collected from the family members. Short tandem repeat (STR) inside the loci that had been reported was selected, such as 2q36, 3p23-24, 4q26-31, 6q22-23, 17q12-22, and the data with two-point linkage analysis were performed. Results Autosomal dominant inheritance pattern was demonstrated in phenotypes of the family and there was no linkage relationship in the above five loci of chromosomes because the maximum two-point LOD score was 0.39 at D17S1868. Conclusion Following exclusion of the loci which had been reported, there are other new pathopoiesis loci of FIgAN and it reveals that FIgAN has the genetic heterogeneity according to initial result at the same time.

Objective To observe the change of insulin-like growth factor 1 (IGF-1) before and after glucocorticoid (GC) therapy and to explore the effect of its change on bone metabolism in primary nephrotic syndrome (PNS) patients. Methods A total of 39 PNS patients with mean age of (36.73±12.15) years received GC therapy were selected from January 2008 to August 2009 in our hospital. Serum IGF-1, albumin, calcium, phosphorus, parathormone (PTH), 25 hydroxy vitamin D3, bone gla protein (BGP), degradation products of C-terminal telopeptides of type I collagen (CTx), 24-hour urinary protein excretion and the ratio of urinary calcium to creatinine were measured at five time points—before GC therapy, 4 weeks, 8 weeks, 12 weeks and 24 weeks after the use of GC. BMD was also detected at the same time points. Correlations among indexes were analyzed by Pearson. Results Thirty-six PNS patients fulfilled the follow-up and had complete clinical data, while other 3 patients lost. After GC treatment, serum calcium and 25 hydroxy vitamin D3 were significantly increased in a time-dependent manner and were negatively correlated with 24-hour urinary protein excretion (r=-0.749, r=-0.831, P<0.05, respectively). Serum BGP and IGF-1 were decreased after GC therapy in a time-dependent manner while CTx was significantly increased until week 12 after treatment (P<0.05). Compared with pre-treatment, BMD of various parts had no significant difference at week 4; BMD of lumbar spine (L1-L4) was significantly decreased until week 8 (P<0.05); BMD of femoral neck and femoral shaft was significantly decreased at week 24 (P<0.05). IGF-1 was positively correlated with BGP and BMD (r=0.896, r=0.495, P<0.05) and negatively correlated with serum CTx (r=-0.697, P<0.05 ). Conclusions Serum IGF-1 level decreases in a time-dependent manner after GC treatment, which is correlated to BGP, CTx and BMD. Glucocorticoid treatment affects bone metabolism through IGF-1 pathway possably in patients with PNS. IGF-1 may be used as a new bone biochemical marker of glucocoritcoid-induced osteoporosis.

Objective To study the prevalence of chronic kidney disease (CKD) among patients with bronchial asthma in Zhengzhou. Methods A total of 655 patients (older than 14 years) were interviewed and received physical, uria and blood examination. Bronchial asthma and CKD were diagnosed according to related definition. Results The prevalence of albuminuria, hematuria, reduced eGFR and CKD was 9.41%,10.37%, 3.03% and 17.38% respectively in the above special patients, which was higher than that of the general population. Women had higher prevalence of CKD than men (21.47% vs 13.99%, χ2=6.060, P=0.014). Prevalence of 1 to 5 stage of CKD was 8.61%, 5.10%, 3.03%, 0.48% and 0.16% respectively. In the acute phase of asthma, prevalence of CKD (24.42%) was significantly higher than that in chronic duration and clinical remission period (χ2=12.445, P=0.002) and the same significant differences were found in albuminuria and reduced eGFR (χ2=19.619, P＜0.01 and χ2=9.305, P=0.010). Conclusion The bronchial asthma patients have higher incidence of renal damage and should be estimated, especially in the acute phase.

Objective To explore the effect of MICA-Ab expression on the prognosis of sensitized renal transplantation recipients. Methods A total of 51 sensitized recipients (PRA more than 20%) in our hospital from August 2007 to April 2010 were enrolled in the study. In these patients，29 cases received protein A immunoadsorption and detection of MICA-Ab was performed before and after protein A immunoadsorption. Other 22 patients received MICA-Ab detection when they were hospitalized. Associations of PRA, HLA-matches, acute rejection, and serum creatinine of postoperative week 1 and week 4 with MICA-Ab were analyzed retrospectively. Results Sixteen recipients (31.4%) had positive MICA-Ab expression but their acute rejection rate was not higher as compared to the patients with negative MICA-Ab expression. Recipients with PRA>40% showed higher expression level of MICA-Ab than recipients with PRA≤40% (P≤0.05). HLA-match did not show influence on MICA-Ab expression. MICA-Ab positive group had no higher serum creatinine level than negative group in postoperative week 4. MICA-Ab level decreased significantly after protein A immunoadsorption. Conclusions MICA-Ab expression increases in the sensitive recipients but does not influence the prognosis. Protein A immunoadsorption can eliminate MICA-Ab effectively in sensitized recipients.

Objective To retrospectively evaluate the relevant factors for hepatitis B virus-associated glomerulonephritis (HBV-GN). Methods A total of 86 patients with pathology-proven HBV-GN and 135 HBV carriers with non-HBV-GN were included in this retrospective case-control study. Logistic regression analysis was used to detect the relevant factors for HBV-GN. Results On univariate analysis, the factors associated with HBV-GN were as follows: male (OR 2.79, 95％CI 1.48-5.25, P=0.001), HBeAg positivity (OR 2.60, 95％CI 1.49-4.53, P=0.001), HBV replication (OR 3.63, 95％CI 1.80-7.33, P<0.01), liver cirrhosis (OR 4.58, 95％CI 1.41-14.91, P=0.011), and elevated alanine aminotransferase (ALT) (OR 2.53, 95％CI 1.42-4.51, P=0.002). On multivariate analysis, the associations remained significant for male (OR 2.21, 95%CI 1.12-4.33, P=0.022), HBV replication (OR 2.77, 95%CI 1.28-5.97, P=0.01), liver cirrhosis (OR 4.55, 95%CI 1.29-16.10, P=0.019) and elevated ALT (OR 1.96, 95%CI 1.04-3.69, P=0.037). Compared with HBV-associated IgA nephritis (HBV-IgAN) in multivariate model, HBV-associated membranous nephropathy (HBV-MN) or membranoproliferative glomerulonephritis (HBV-MPGN) was significantly associated with male (OR 6.51, 95%CI 1.76-24.11, P=0.005) and HBV replication (OR 7.22, 95%CI 1.68-30.97, P=0.008). Conclusions Male, HBV replication, liver cirrhosis and elevated ALT may be predictive factors for HBV-GN. Compared with HBV-IgAN, HBV-MN or HBV-MPGN is significantly associated with male and HBV replication.

Objective To investigate the expression of metabotropic glutamate receptor (mGluR) in murine podocytes. Methods Conditional immortalized podocytes were used in the research. RT-PCR was used to estimate the mRNA expression. Western blotting, immunofluorescence staining and immunoelectron microscopy were employed to determine the protein production. EIA, EMSA and Western blotting were used to examine the cAMP generation and cAMP response element-binding protein(CREB) activation. Intracellular calcium was investigated using confocal microscopy. Results mGluR1 and 5 mRNA and protein were expressed in murine brain and podocytes. In glomeruli, most of mGluR1 expression located in podocytes and was expressed in the submembrane space of the podocytes. Podocytes treated with (S)-3,5-dihydroxyphenylglycine (DHPG, an agonist for mGluR1/5) rapidly generated cAMP and activated CREB. (RS)-1-Aminoindan-1,5-dicarboxylic acid (AIDA, a selective antagonist of mGluR1/5) and SQ22536 (an adenylate cyclase inhibitor), but not 2-aminoethoxydiphenyl borate (2-APB an antagonist of canonical transient receptor potential) blocked DHPG-induced cAMP generation and CREB activation. Following DHPG treatment, intracellular calcium level rose and was prevented by pre-treatment with AIDA and 2-APB. DHPG-induced calcium influx was also prevented by incubation with calcium-free medium. Conclusion Podocytes express functional mGluR1 and mGluR5.

Objective To study the role of endoplasmic reticulum stress in phenotypic change of cultured human glomerular mesangial cells induced by high glucose. Methods Cultured human glomerular mesangial cells were divided into three groups: control group, high glucose group and high glucose+ 4-phenylbutyric acid(4-PBA) group. Cell number of proliferation was assessed by MTT assay. Cell cycle was measured by flow cytometric analysis. Expression of α-SMA was assessed by immunohistochemistry and was observed by laser scanning confocal microscope. Involved mRNA and protein expression were measured by real-time PCR and Western blotting. Results (1)Cell number of proliferation and S transition proportion in high glucose group significantly increased than that in control group(P < 0.05). High glucose could induce α-SMA expression significantly (P<0.05). 4-PBA could significantly inhibit human glomerular mesangial cells proliferation (P<0.05), S transition arrest(P<0.05) and expression of α-SMA (P<0.05) induced by high glucose. (2) Compared with control group, high glucose could significantly increase the expression of glucose-regulated protein78（Grp78） mRNA and protein (P< 0.05), which could be inhibited by 4-PBA treatment (P<0.05). (3)High glucose could induce the mRNA and protein expression of TGF-β1 and FN significantly, which could be inhibited by 4-PBA treatment (P<0.05). Conclusion Endoplasmic reticulum stress plays an important role in phenotypic change of cultured human glomerular mesangial cells induced by high glucose.

Objective To investigate the effect of erythropoietin(EPO) on mesenchymal stem cells (mMSCs) proliferation under acute kidney injury (AKI) microenvironment,and to study its possible mechanism. Methods C57BL/6 mice’s MSCs (mMSCs) were isolated by Percoll density gradient centrifugation and adherence cultivation. Surface markers were identified by flow cytometry. AKI mice models were made by clamping bilateral renal pedicles for 30 minutes and reopening for 30 minutes. Then both renal cortex was drew immediately to make IR kidney homogenate supernatant. P3-mMSCs were divided into different groups: Group A: low glucose DMEM medium with 10% fetal bovine serum; Group B: low glucose DMEM medium with 10% fetal bovine serum plus IR kidney homogenate supernatant; Group C: low glucose DMEM medium with 10% fetal bovine serum plus IR kidney homogenate supernatant and different concentrations of EPO (1, 5, 10, 50 U/ml). Each group was incubated for 1 d, 3 d, 5 d, 7 d. Proliferation of mMSCs was detected by CCK-8, and apoptosis was detected by TUNEL. The protein expression of erythropoietin receptor(EPOR) and the proteins of proliferation/apoptosis related signal pathway were examined by Western blotting. Results Under IR kidney homogenate supernatant, the proliferation ability of mMSCs decreased significantly (P＜0.01), while the apoptoic percentage was significantly higher than that of Group A(P＜0.01). After intervention of EPO, mMSCs proliferation enhanced, at the same time, the apoptoic percentage decreased, in a dose-dependent manner. EPOR was positive in P3-mMSCs by Western blotting. EPO decreased the expression of caspase-3 in mMSCs under AKI microenvironment in a dose- and time-dependent manner, but increased the expression of Bcl-2. Cultured for 5 d, the expression of phosphor-Janus kinase2(p-JAK2) [(0.641±0.028) vs(0.456±0.012)] and phosphor-signal transducer and activator of transcription(p-STAT5) [(0.398±0.016) vs(0.209±0.020)] was significantly higher in 10 U/ml EPO group compared to group B. Conclusion Erythropoietin can promote proliferation of mMSCs in vitro under AKI microenvironment, which is mediated by EPOR and related with proliferation/apoptosis signal pathway.

Objective To observe the expression changes of renal tissue Bax, Bcl-2 and caspase-3, to wxamine the correlation between the ratio of Bax to Bcl-2, caspase-3 and renal tubular cells apoptosis, and to investigate the role of caspase-related signal pathway. Methods Forty-eight male Wistar rats were randomly divided into three groups: control (CN, n=8), exhaustive swimming (ES, n=24) and inula britannica (IB, n=16) group. The rats of CN were quiet without swimming. The rats of ES swam to exhaustive state and were sacrificed at immediately(ESI), 6 hour(ES 6 h) and 24 hour(ES 24 h) after exhaustive swimming respectively. The rats of IB took orally inula britannica at the dose of 25 ml/kg body weight at 24 h before swimming and then swam to exhaustive state. The rats of IB group were sacrificed at 6 hour (IB 6 h) and 24 hour (IB 24 h) after exhaustive swimming. The animal model of overtraining-induced acute kidney injury was developed by exhaustive swimming. The renal cell apoptosis was measured by the method of TUNEL. The expressions of Bax, Bcl-2, caspase-3 in renal tissue were observed by immunohistochemistry. The expression of caspase-3 protein was examined by Western blotting. The correlation between the ratio of Bax to Bcl-2 and caspase-3 was analysed by Pearson method, and the correlation between the ratio of Bax to Bcl-2, caspase-3 and renal tubular cell apoptosis was analysed by Spearman method. Results The number of renal tubular apoptotic cells was increased progressively in ESI to ES 24 h rats by TUNEL (P<0.05). Immunohistochemistry staining showed that the ratio of Bax to Bcl-2 and caspase-3 in renal tubular cells were increased progressively at 0 h, 6 h and 24 h after exhaustive swimming compared with control group (P<0.05). The change of renal tissue caspase-3 was also revealved by Western blotting analysis. The ratio of Bax to Bcl-2 and caspase-3 in renal tubular cell was correlated positively (r=0.865, P<0.05), The ratio of Bax to Bcl-2, and caspase-3 was also correlated positively to renal tubular cell apoptosis (r=0.674, r=0.837, P<0.05) in ES rats. Pretreatment with inula britannica inhibited the up-regulation of the ratio of Bax to Bcl-2, caspase-3 and cell apoptosis in renal tubular cell induced by exhaustive swimming. Conclusion Overtraining can induce renal tubular cells apoptosis through activating caspase-related signal pathway by impairing the balance of Bax and Bcl-2, which may be one of the important molecular mechanisms of overtraining-induceed renal tubular cells apoptosis.

Objective To study the effect of continuous venovenous haemodiafiltration (CVVHDF) on the clearances of various solutes in multiple organ dysfunction syndrome (MODS) dogs. Methods Dogs were subjected to hemorrhagic shock plus resuscitation and endotoxemia to set up MODS model, then they were randomly divided into 2 groups: MODS group (M group) and MODS+CVVHDF group(M+C group). A PRISMA pre-dilution system with AN-69 filter was applied. Solute clearance of nitric oxide(NO), urea nitrogen(UN), creatinine (Cr), interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-α(TNF-α) and endotoxin (LP) was determined with same dialysate and ultrafiltrate volume during CVVHDF. The clearance of various solutes (Kd) was calculated by the concentration of the cleared solutes in effluents (E) based on the formula Kd=(E×QE)/P. Results Clearances of NO, UN, Cr were greater than those of IL-6, IL-10, TNF-α, LP during the treatment of CVVHDF. Conclusion CVVHDF could effectively reduce the levels of various solutes in the development of MODS by convection and absorption.