Objective  To describe the clinical characteristics, and to analyze the AGXT gene mutation in three siblings with primary hyperoxaluria type I (PHI).  Methods  AGXT gene mutation was analyzed by direct sequencing analysis in this family, and the minor allele status was also tested. One hundred unrelated healthy subjects were also analyzed as controls.  Results  Three mutations in AGXT were identified in each of three patients including two novel heterozygous missense mutations and one previously reported variant. One mutation was a methionine to leucine substitution at position 49 (p. M49L, c.145A＞C) in exon 1, one was an asparagine to isoleucine transition at codon 72 (p.N72I, c.215A＞T) in exon 2, and another was a heterozygous nonsense mutation at codon 333 (p.R333*). Both p.M49L and p.R333* occured in cis configuration with the minor allele IVS1+74 bp.  Conclusions  Two novel mutations are identified probably in association with PHI, however their pathogenicity and potential molecular mechanisms should be explored by further investigations. This is the first investigation on mutant gene analysis of PHI in China.

Objective  To compare the characteristics of renal disease based on renal biopsy data between the Tibetan plateaus and the plain.  Methods  160 chronic kidney diseases patients underwent renal biopsy from the plain and 80 cases from Tibet plateau were compared by parallel controlled manner. The relationship of renal pathology and clinical signs were also compared.  Results  (1) The male to female ratio was quite different between Tibet plateau and plain groups (0.60∶1 vs 1.11∶1, P＜0.01). (2) The distribution of pathological types between two groups was quite different. The main reason of primary renal disease at Tibet plateau was minimal change disease (51.39% vs 14.53%, P＜ 0.01), but at plain it was IgA nephrology (49.57% vs 6.94%, P＜0.01). Meanwhile, the LN ratio in the secondary glomerulonephritis at the plateau region was significantly lower than those in the plain region (12.6% vs 34.9%, P＜0.05).  Conclusions  The most common reason of primary glomerular disease in plateau region is minimal change, and the most common clinical manifestation is nephrotic syndrome. IgA nephropathy in the plain is the most frequent primary renal disease. In terms of the secondary renal diseases, allergic purpura nephritis is dominated in the plateau region, whereas LN is frequently found in the plain.

Objective  To finding out the characters of vascular remolding after the establishment of native arteriovenous fistula on the wrist, and exploring the influential factors. Methods  Doppler ultrasound was used to monitor the diameter of cephalic vein, brachial artery, radial artery and ulnar artery at the time before the surgery and one day, one week, two weeks, four weeks and eight weeks after the surgery respectively. The tendency of the diameter change was analyzed.  Results  Twenty eight patients completed the whole monitor session, in which eleven were female. The average age of those patients was (53.68 ± 2.61) years old. Twelve of them were diabetic nephropathy. The diameters of all vessel were increased more rapidly at the first day than any other days after surgery(all P＜0.01). The patients were divided into two groups depending on whether diabetic nephropathy. No significant difference was found between the two groups on the tendency of diameter change in cephalic vein and brachial artery (all P ﹥ 0.05). However, the tendency of diameter change in radial artery and ulnar artery was statistically significant difference between the two groups (all P＜0.05).  Conclusions  Cephalic vein, brachial artery, radial artery and ulnar artery are all apparently dilated on the first day after the surgery. The vascular dilation and diameter increasing become much slower after the period, the diameter tend to be stable. The primary diseases may affect the tendency of the diameter change in radial artery as well as ulnar artery.

Objective  To clarify whether the NADPH oxidases (NOXs) family contributed to the reactive oxygen species (ROS) production and subsequent interstitial fibrosis in unilateral ureter obstruction (UUO) rats.  Methods  Male Wistar rats were randomly divided into sham operation group (n＝8), sham operation + apocynin treatment group (n＝8), UUO operation group (n＝8) and UUO operation+apocynin treatment group (n＝8). Either vehicle or apocynin (100 mg/kg per day) were given by gavage for 7 days after surgery. Rats were sacrificed at 7^th day. ELISA was used to detect the activity of superoxide dismutase (SOD) and catalase (CAT), and the level of 8-iso-prostaglandin F2alpha (8-iso- PGF2α) in renal tissue. Western blotting was used to detect the protein expressions of NADPH oxidase subunit NOX2 and NOX4, α- smooth muscle actin(α-SMA), collagen I (COL-I) and the level of ERK1/ 2 phosphorylation (p-ERK1/2).  Results  UUO rats with vehicle displayed increased oxidative stress, as measured by renal tissue 8-iso-PGF2α, accompanied with increased renal expression of NADPH oxidases (NOX2, 1.5-fold and NOX4, 1.7-fold, respectively), compared with sham-operated rats (P＜ 0.05). Furthermore, vehicle treated UUO rats showed increased renal COL - I and α - SMA levels, compared with sham-operated rats (P＜0.05). ERK1/2 was also activated as detected by p-ERK1/2 expression in UUO rats with vehicle (P＜0.05). Apocynin treatment significantly decreased renal tissue 8-iso-PGF2α level and expressions of NOX2 (-28.7%) and NOX4 (-31.0%) in UUO rats, respectively, compared with vehicle treated rats (P＜0.05). And significant decrease of COL-I (-26.4%) and α-SMA expression (-80.0%) were also observed (P＜0.05). The activation of ERK1/2 in UUO rats was greatly inhibited by apocynin treatment (P＜0.01). Despite the pronounced dysregulation of pro – oxidative NOXs family, no compensatory increase of antioxidative enzyme activities occurred.  Conclusion  The NOXs family contributes largely to the production of ROS and subsequent interstitial fibrosis after ureter ligation, and inhibition of the NOXs family may be a choice for preventing interstitial fibrosis.

Objective  To explore the effects of physiological intrauterine hypoxia on the mammal kidney development in ex vivo model.  Methods  Kidney rudiments were micro-dissected from embryonic day 13.5 SD rat embryos. They were pooled and assigned randomly to control group (normoxia, 21% O₂) and experimental groups(hypoxia, 3% - 5% O₂). For conventional culture, the rudiments were placed on 0.4 μm pore-size polycarbonate filters at the bottom of a well insert in a six well plate and incubated at 5% CO₂ at 37℃ . The hypoxia micro - environment was created in an incubator through injecting N2. Immunostaining was carried out to label the structure of developing kidney and pictures were taken by a laser confocal microscope. TdT-mediated dUTP nick end labeling was used to detect cell apoptosis while EdU (5-Ethynyl-2'- deoxyuridine) was added to the medium for cell proliferation detection during kidney development. After culture kidneys of both groups were processed and the changes of genes expression were measured by real time PCR.  Results  Kidney development was significantly suppressed under hypoxia condition with decreased ureteric buds as well as nephrons compared with that under normoxia condition (P＜0.05). Hypoxia inhibited the proliferation of cultured kidneys while reduced the apoptosis of their cells (P＜0.05). Six2 – positive progenitors were better maintained within kidneys cultured under low O₂ (P＜0.05). Moreover, genes regulating metanephric mesenchymal to epithelial transformation such as Wnt9b and Wnt4 were down- regulated while genes functioning to maintain metanephric mesenchymal progenitors such as Six2, Wt-1 and Pax2 were up - regulated in hypoxia.  Conclusion  Physiological intrauterine hypoxia as an important microenvironment factor may suppress mammal kidney development through inhibiting the proliferation and differentiation of embryonic kidney progenitors while maintain their undifferentiated status and self-renewal, thus contributing to the balance between differentiation and maintenance of a mesenchymal progenitor cell pool which determines the final number of nephrons in adult kidneys.

Objective  To observe the effects of metformin on expression of Adenosine 5’- monophosphate (AMP)-activated protein kinase (AMPK), nuclear factor-κB (NF-κB) and transforming growth factor β1 (TGF - β1) in cultured rat glomerular mesangial cells (MCs), and explore its reno - protective mechanisms.  Methods   MCs were cultured in the medium with normal glucose (group NG, 5.6 mmol/L), high glucose (group HG, 25mmol/L) and different concentrations of metformin (group M1, M2, M3). After 48 h exposure, the supernatants and MCs were collected. The expression of NF-κB and TGF-β1 mRNA was analyzed by real time-PCR. Total-AMPK, phospho-Thr-172 AMPK (p-AMPK), NF -κB p65 and TGF-β1 were visualized by Western blot.  Results  The real time-PCR and Western blot result showed MCs could express AMPK, NF-κB and TGF-β1 mRNA and protein. After stimulated by HG, the levels of intracellular NF - κB and TGF - β1 expressions were significantly increased compared with group NG (P＜0.05); The levels of NF-κB and TGF-β1 were significantly decreased in group M1, M2 and group M3 compared with group HG in a dose-dependent manner. After stimulated by HG, the level of intracellular p-AMPK were down-regulated compared with group NG(all P＜0.05); The expression of p-AMPK increased with the rising of metformin concentration, presenting the opposite trend (P＜0.05), while the level of total-AMPK protein was unchanged with exposure to HG or different concentrations of metformin(P＞0.05).  Conclusion  Metformin can suppress the expression of NF- κB and TGF-β1 of glomerular MCs induced by HG via AMPK activation, which may partly contribute to its reno-protection.

Objective  To investigate the effect of fluvastatin on TGF-β1 expression in a rat model of peritoneal dialysis(PD).  Methods  A rat model of PD was built by intraperitoneal injection of 2.5% or 4.25% peritoneal dialysate. SD rats were randomly divided into 7 groups: (1) Normal control group; (2)Saline control group: saline 100 ml/kg intraperitoneal injection(IP) each day; (3) Fluvastatin treatment group: fluvastatin intragastrically administration 10 mg/kg each day; (4) 2.5% PD group: 2.5% peritoneal dialysate IP 100 ml/kg everyday; (5)4.25% PD group: 4.25% peritoneal dialysate IP 100 ml/kg everyday; (6)2.5% PD plus fluvastatin treatment group: 2.5% peritoneal dialysate IP 100 ml/kg plus fluvastatin 10 mg/kg everyday; (7)4.25% PD plus fluvastatin treatment group: 4.25% peritoneal dialysate IP 100 ml/kg plus fluvastatin 10 mg/kg everyday. The rats were sacrificed at 6 weeks and peritoneal tissues were dissected. The expressions of TGF-β1 and FN were examined by RT-PCR and immunohistochemical analysis. Masson staining was used for histological examination.  Results  Masson staining showed that the peritoneum thickened in 2.5% and 4.25% PD group than in normal control group and saline control group. The fluvastatin treatment ameliorated the thickening of peritoneum induced by PD. RT-PCR and immunohistochemical analysis showed that the mRNA and protein expression of TGF- β1 and FN increased in 2.5% and 4.25% PD group than in normal and saline control group (all P＜0.05). The fluvastatin treatment ameliorated the increased expression of TGF-β1 and FN induced by PD. There was no statistically significant difference among normal control group, saline control group and fluvastatin treatment group in both peritoneal thickness and the expression of TGF-β1 and FN.  Conclusion  Fluvastatin can reduce the increased expressions of TGF -β1 and FN in rat peritoneum and ameliorate the thickening of peritoneum induced by PD.