Objective To summarize the clinical, pathological features and inheritance mode of familial collagen type Ⅲ glomerulopathy. Methods The clinical manifestations and pathological findings of 2 affected brothers and their family information were collected. Results Two affected brothers, one was 33-year old and the other was 34-year old. Both of them had great amount of protein excretion in urine (3.1 g, 6.38 g respectively), and one of them had nephrotic syndrome. Both presented hypertension and renal insufficiency (serum creatinine 128μmol/L, 313 μmol/L respectively). Neither hematuria nor abnormalities of nail and bone was found. Their serum concentrations of procollagen Ⅲ peptide were elevated (>50 ng/L). Renal biopsy revealed that massive buddle fibrils were deposited in mesangium and glomerular basement membrane subendothelial area by electron microscopy. Strong staining of type Ⅲ collagen was observed in the mesangial area and along the glomerular capillary loops. Family survey showed their parents’^‘marriage was consanguineous. The concentration of procollagen Ⅲ peptide was also obviously elevated (>50 ng/L) in their younger sister but no proteinuria, hematuria, nor hypertension was detected and her renal function was normal. Conclusion Familial collagen type Ⅲ glomerulopathy is rare. Our findings supported an autosomal recessive pattern of inheritance. It is the first familial case reported in Chinese population.

Objective To screen for the COL4A3 and COL4A4 mutations in a Chinese consanguineous family with autosomal recessive Alport syndrome (ARAS). Methods Using PCR and direct sequencing, all 52 coding exons of the COL4A3 gene and 46 exons, except exon-1, of the COL4A4 gene were analyzed to detect mutations in the pedigree with ARAS. Furthermore, mutation was identified by restriction endonuclease AvaII in all other 20 members. Results A novel missense mutation (3725 G>A, G1242D ) in exon 42 of the COL4A3 gene was identified in homozygous form. This pathogenic mutation was demonstrated in heterozygous forms in all carriers in this family, whereas it was detected neither in the other normal members of the family nor in the 50 controls. In addition, 10 polymorphisms, including one nonglycine missense variants and 9 neutral polymorphisms, were detected in COL4A3/COL4A4. Conclusion The novel pathogenic mutation (3725 G>A, G1242D) of the COL4A3 gene may be the underlying pathogen in this family and it is the first reported case in ARAS.

Objective To investigate the mutations in Chinese families with Fabry disease. Methods Genomic DNA was extracted from peripheral blood cells of three probands diagnosed as Fabry disease and some family members. Seventy genomic DNA samples extracted from 70 unrelated normal persons were used as control. By PCR and direct sequencing, all 7 exons and their neighboring intronic sequences of the GLA gene of the probands were analyzed. Results Three mutations were identified in 3 probands: (1) deletion of 1 bp at nucleotide 1142 in exon 7 (1142DelG), leading to premature termination of protein translation at codon 390. (2) 902 G to A transition in exon 6 (codon 301), resulting in replacement of an arginine residue by glutamine (902G >A, R301Q). (3) 484 T to C transition in exon 3 (codon 142), resulting in replacement of a cysteine residue by arginine (484T >C, C142R). Mutation of GLA gene in 13 relatives of 3 probands was also screened and 6 cases with the same mutation as the relevant proband were found, including 5 heterozygotes and 1 hemizygote. Conclusion Three mutations including one novel mutation (1142DelG) are found in 3 Chinese families with Fabry disease by PCR-DNA sequencing.

Objective To investigate the effect of NPHS2 gene mutation of both V165X(467-468insT) and R168H (503G>A) on the expression and distribution of podocins in HEK293 cells. Methods The wild-type, V165X and R168H mutant cDNAs in the expression plasmids were transfected into HEK293 cells, and the subcellular localization of the wild-type, mutant podocins was studied by immunofluorescence staining with a specific podocin N-terminal antibody (P21) and a specific podocin C-terminal antibody (P35), immunolabeling and confocal microscopy. Results The fluorescence stainings of the wild-type, V165X and R168H mutant podocins with antibody P21 were positive. The fluorescence stainings of the wild-type and R168H mutant podocins with antibody P35 were also positive, whereas the staining of V165X mutant podocin with P35 was negative. The staining for wild-type podocin was distributed around nuclei and mainly on the cell membrane surface in a filamentous pattern, whereas V165X and R168H mutant podocins staining localized predominantly around nuclei with a loss of surface expression. Conclusions Both the molecular structure and the subcellular localization of V165X mutant podocin are changed evidently, so is the subcellular localization of R168H mutant podocin, whereas the molecular structure of R168H mutant podocin is little changed. The normal biological functions of the wild podocin rely on its normal molecular structure and subcellular localization as well.

Objective To investigate the effects of polycystin-1 N-terminal peptide (PC-1NTP) on proliferation, cell cycle and apoptosis of cystic-lining epithelial cells in human autosomal dominant polycystic kidney disease (ADPKD). Methods Cystic-lining epithelial cells were treated with PC-1NTP in vitro. MTT assay was used to detect the PC-1NTP effects on cells proliferation. Cell cycle and apoptosis were analyzed by flow cytometry. The mRNA expression of cyclinD1, p21^WAF1, bax, bcl-2 and MCM-2 were measured by fluorescence quantitative PCR. Results The growth and apoptosis of cystic-lining epithelial cells were significantly inhibited by PC-1NTP. The percentage of cells in G0/G1 phase increased, while decreased in S phase remarkably. The mRNA expression of p21^WAF1 and bcl-2 was significantly higher than that in the control group(P < 0.01), while the mRNA expression of cyclinD1,bax and MCM-2 was significantly lower than that in the control group(P < 0.01 or P < 0.05). Conclusions PC-1NTP resulted in G0/G1 phase arrest and elicited an attenuating effect on the proliferation and apoptosis of cystic-lining epithelial cells, which may be realized through regulating the expression of G1/S checkpoint regulation factor cyclinD1/p21^WAF1 and apoptosis regulating protein bcl-2/bax. PC-1NTP may be a new prospective maneuver in the treatment of ADPKD.

Objective To study renal involvement in hepatic glycogen storage disease(GSD) in childhood. Methods One hundred and eight patients aged less than 21 years old with typeⅠ a GSD (54 cases), typeⅢ (29 cases) and uncertain type hepatic GSD (25 cases). Urine analysis, urine albumin, urine protein of 24 h, urine β2-MG, BUN, creatinine, Ccr were evaluated. Results Of 108 patients with hepatic GSD, 16 patients (20.8%) had proteinuria proven by urine albumin or urine protein of 24 h, their ages first found proteinuria were 8~15 years. Two 15-year-old patients had proteinuria over 1.0 g/24 h. Among 72 patients, urine β2-MG of 51 cases (70.8%) increased (175~10 623 mg/L), and the mean urine β2-MG of typeⅠa GSD was much higher than that of typeⅢ GSD, 4138.2 and 1790.1 mg/L respectively. Of 91 patients, 10 had renal insufficiency, 1/10 (15-year-old girl) had heavy proteinuria (3.5 g/24 h), elevated BUN (9.3 mmol/L) and Scr (1061 μmol/L). Five elder patients (11~21 years old) had hematuria with renal colic caused by renal calculus. Conclusions Persistent protenuria, increased urine β2-MG, decreased Ccr, and renal stones are common complications of hepatic GSD in childhood. Renal function should be thoroughly evaluated during follow-up.

Objective To investigate the possibility of using serum and urinary endothelin-1 (ET-1), interleukin-6 (IL-6) assay as screening tools for atherosclerotic renal artery stenosis (ARAS). Methods Serum and urinary samples from 49 patients with ARAS, 32 cases with ≥2 risk factors for atherosclerosis and 30 normal controls were detected for ET-1 and IL-6 by RIA. The receiver operating characteristic (ROC) curves were then generated to assess their accuracy in screening ARAS by using selective renal angiography as golden standard. Results Urinary ET-1, urinary IL-6 and urinary-serum ratio of ET-1 in ARAS cases were higher than those in two control groups, and were all correlated with degree of renal artery stenosis. The area under the ROC curve of urinary ET-1, urinary IL-6 and urinary-serum ratio of ET-1 was 0.792, 0.756 and 0.779, respectively. The sensitivity and specificity of urinary ET-1 to distinguish ARAS (≥50%) was 80.0% and 72.8% respectively using 6.72 ng/mmol creatinine as the cut-off point. The cut-off value of ET-1 urinary-serum ratio was 12.59 with a sensitivity of 66.7% and a specificity of 61.7%. The sensitivity and specificity of urinary IL-6 were 73.3% and 70.4% respectively using 23.85 ng/mmol creatinine as the cut-off point. The sensitivity and specificity were improved to 80.0% and 77.8% respectively when using 12.60 ng/mmol creatinine as the cut-off point and combined with hypertension to perform series test. Conclusion Urinary ET-1 and urinary IL-6 could be used as screening tools for ARAS.

Objective To analyze the relationship between pathological diagnosis and clinical manifestation, and to investigate the importance of renal biopsy on the diagnosis of type 2 diabetic patients complicated with renal diseases. Methods The clinical and pathological data of 52 type 2 diabetic patients with abnormal urinalysis or increased serum creatinine who had renal biopsy performed in our department were studied. Results Among 52 patients, 32 cases were distinctly diagnosed as diabetic nephropathy (DN), including 3 presenting with diabetic nephropathy complicated with non-diabetic renal disease (NDRD), and 20 with the NDRD only. The preoperative diagnosis of 24 cases (46.15%) was accordant with the diagnosis after renal biopsy, while 10 cases (19.23%) were misdiagnosed. Between the two groups, except the differences in blood urea nitrogen, serum creatinine, diabetic duration and whether the patients were complicated with diabetic retinopathy, other clinical manifestations and laboratory findings were not significantly different. Conclusions Type 2 diabetes complicated with proteinuria may result from NDRD. It is difficult to differentiate the renal pathological changes merely by analyzing the clinical data. Therefore, renal biopsy is an important tool to confirm renal pathological changes.

Objective　To investigate the relationship between TGF-β1-induced fibronectin (FN) expression and up-regulation of integrin-link kinase (ILK) in human kidney tubular epithelial cells (HKC). Methods　Using cell culture and Western blot, the TGF-β1-induced expression of FN and ILK was tested. The ILK expression plasmid (pCMV-wtILK) containing wild-type full-length human ILK cDNA, or kinase dead ILK cDNA were transfered to HKC. The effect of overexpression of ILK on FN expression, and the blockage of ILK activation on the action of TGF-β1 on HKC were studied. Results　TGF-β1 induced FN expression of HKC in a dose-depended manner. In the time-course study, TGF-β1 upregulated ILK expression of HKC as early as 8 hours, meanwhile it induced FN expression. Overexpression of ILK in HKC tansfered with pCMV-wtILK could induce FN expression. Blockage of ILK activation abrogated TGF-β1-induced FN expression. Conclusions　A close relationship exists between TGF-β1-induced FN expression and upregulation of ILK in HKC. Blockage of ILK activation obliterates TGF-β1-induced FN expression.

Objective To investigate the role of caspase 3 in ATP depleted apoptosis in renal tubular epithelial cells(RTEC). Methods To induce apoptosis, REC were subjected to 60 min ATP depletion followed by recovery. Wild-type bcl-2 was overexpressed by infecting opossum kidney(OK) cells with adenovirus containing wild-type human bcl-2. Apoptotic cells were detected with Hoechst 33342 dye. Flow cytometry was used to quantify apoptosis and necrosis. Active caspase 3 and bcl-2 degradation were assessed by Western blot. Results ATP depletion and recovery resulted in activation of caspase 3 and the progressive accumulation of bcl-2 cleavage products and apoptosis. Overexpression of bcl-2 ameliorated apoptosis in the ATP-depleted RTEC followed by recovery. Caspase 3 reproduced the effect of caspase 3 on bcl-2 cleavage, whereas caspase 3 specific inhibitor DVED prevented bcl-2 cleavage in vitro. Conclusion Caspase 3 activition and mediated-bcl-2 degradation are likely to contribute to ATP depletion-induced apoptosis in RTEC.

Objective To investigate the signalling events follow the activation of RAGE in podocytes. Methods AGE and CML generated dichloroflurescin-sensitive intracellular ROS were measured by confocal microscopy. The activation of MAP kinases family were studied using Western blotting. MCP-1 mRNA expression was detected by semi-quantitative RT-PCR. Results Basal ROS was located in nucleus of starvation podocytes. AGE and CML rapidly generated intracellular ROS in podocytes. NAC pre-treated podocytes suppressed basal and inducible ROS generation and antibody for RAGE suppressed the inducible ROS. Blockage of ROS induced by NAC suppressed the expression of CML and H2O2-induced MCP-1. Phosphorylated extracellular signal-regulated kinase (ERK) was found in CML incubated podocytes at 10 min and was prevented by NAC or AFC. PD98058 Pre-treated podocytes partially inhibited the expression of CML-induced MCP-1 mRNA. No evidence were showed that p38 MAPK, SAPK/JNK, PI3K and PKC were involved in the signal transduction. Conclusion Activation of RAGE induces MCP-1 expression in podocytes via ROS-ERK signalling pathway.