1.Effect of fosinopril on the expression of NADPH oxidase p22phox mRNA in the kidneys of diabetic rats
Yanqiang PENG ; Yulian JI ; Zongpei JIANG ; Xueqing YU ; Haiyan ZHANG
Chinese Journal of Nephrology 2008;24(5):343-348
Objective To study the effect of fosinopril on the expression of NADPH oxidase subunit p22phox mRNA and the extracellular matrix (ECM) accumulation in the kidneys of rats with diabetes mellitus . Methods Diabetic rats induced by streptozotocin were randomly divided into control group(DM group) and fosinopril group (fosinopril 10 mg'kg-1·d-1) (DM+Fosin group) and treated for 12 weeks . Expression of p22phox mRNA of NADPH oxidase in kidneys was measured by RT-PCR . The expression of fibroneetin was studied by immunohistochemistry and matrix metalloproteinases 9 activity was detected by Zymography . Meanwhile, the kidney hypertrophy index, serum creatinine level and 24-hour urinary protein excretion were evaluated . Results The expression level of p22phox mRNA in the kidneys of DM+Fosin group rats was decreased by 45% than that of DM group at week 4 (P<0 .05) . At week 8 fosinopril significantly decreased the expression of glomerular and tubulointerstitial fibronectin by 52,5% and 42 .9% respectively (P<0 .05), while increased MMP-9 activity by 29 .6% (P<0 .05) compared with DM group . Fosinopril significantly decreased 24-hour urinary protein excretion of diabetic rats from week 8 . Serum creatinine level, 24-hour urinary protein excretion and kidney hypertrophy index were significantly decreased by 35 .9%, 50 .2% and 17 .2% in rats of DM+Fosin group than those of DM group at week 12 (P<0 .05) . Fosinopril did not affect blood sugar significantly . Conclusion Fosinopril has beneficial effect on diabetic nephropathy partly through inhibiting the expression of NADPH oxidase p22 phox mRNA .
2.Results analysis of urine cellular components tested by Urit-1500 urine analyzer
Houlong LUO ; Xingchao LIU ; Xueqing ZHOU ; Fen PENG ; Donghong LIU
International Journal of Laboratory Medicine 2014;(16):2212-2213,2215
Objective To explore the clinical accuracy of urine cellular components of RBC,WBC,etc.detected by the Urit-1500 urine analyzer.Methods 1 085 urine samples in this hospital from June to October 2013 were selected and detected by the Urit-1500 urine analyzer and the microscope detection respectively.The results of urine cellular components detected by the urine analy-zer were analyzed.Results In 1 085 urine samples,the coincidence rate of two detection methods in detecting RBC and WBC was 93.8%(1 018/1 085)and 94.0%(1 020/1 085)respectively.In 295 RBC positive urine samples tested by the urine analyzer,the number of negative results tested by microscopy was 49,accounting for 16.6%.In 790 RBC negative urine samples tested by the u-rine analyzer,the number of negative results tested by microscopy was 18,accounting for 2.3%.In 197 WBC positive urine samples tested by urine analyzer,the number of negative results tested by microscopy was 28,accounting for 14.2%.In 888 WBC negative urine samples by urine analyzer,the number of negative results tested by microscopy was 37,accounting for 4.2%.Conclusion De-tecting urine RBC and WBC by using the Urit-1500 urine analyzer should combined with the microscopy and other test indexes and even clinical data to conduct the comprehensive analysis for reducing the misdiagnosis and missed diagnosis as far as possible.
3.The expression and clinical significance of miR-98 in peripheral blood mononuclear cells in asthmatic children
Xueqing XIAO ; Wanhong XIA ; Hong ZHAI ; Jun PENG
Chinese Pediatric Emergency Medicine 2017;24(3):201-204
Objective To explore the role of miR-98 in peripheral blood mononuclear cells (PBMC) in the pathogenesis and development of childhood asthma.Methods A total of 43 cases of asthmatic children and 30 cases of healthy controls were enrolled in the study.Peripheral blood mononuclear cells were isolated in both healthy subjects and asthmatic children in acute attack and remission stages.The expressions of miR-98 and interleukin-4(IL-4) and IL-13 mRNA were detected by real-time quantitative PCR.Results The miR-98 levels of asthmatic children in attack stage were significantly lower than those in remission stage and control group (P<0.01).The IL-4 and IL-13 mRNA levels of asthmatic children in attack stage were significantly higher than those in remission stage and control group (P<0.01).There was no significant difference of miR-98,IL-4 and IL-13 mRNA between asthmatic children in remission stage and the controls (P>0.05).Furthermore,a negative correlation was found between the expression of miR-98 and IL-5(r=-0.794,P<0.01) and between the expression of miR-98 and IL-13 mRNA (r=-0.804,P<0.01) in asthmatic children in attack stage.A positive correlation was also found between IL-4 and IL-13 mRNA in asthmatic children in attack stage (r=0.853,P<0.01).Conclusion The expression of miR-98 decreased in asthmatic children,and miR-98 might be involved in the pathogenesis and development of asthma.
4.Effect of dialysate on the peritoneal membrane function and the mesothelial cell morphology in chronic peritoneal dialysis rats
Qiongqiong YANG ; Rengao YE ; Xiao YANG ; Wenxing PENG ; Xueqing YU
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To investigate the influence of dialysate on the cell morphology and the peritoneal membrane function in chronic peritoneal dialysis rats. METHODS: 40 SD rats were divided randomly into 4 groups. Except control group, other groups daily received infusion of 20 mL dialysate (4.25% dialysate(HG), 1.50% dialysate(LG), Riger's solution(RG)) respectively for 8 weeks. The peritoneal membrane function was investigated by peritoneal transport test, and the rat peritoneal mesothelial cells(PMCs)morphology was analyzed by the cell imprints. RESULTS: The intraperitoneal volume and net ultrafiltration in HG and LG groups were significantly lower, with D/P_(urea) significantly higher, and C_(urea) after 4 h of dialysis significantly lower than that in RG and control groups. The density of cell population from analysis of cell imprints in HG and LG groups was significantly lower, but the mean surface area were significantly larger than that in RG and control groups. These change had no difference between HG and LG group. Using the high glucose dialysate for 8 weeks significantly decreased the ultrafiltration volume ,which was significantly relate to the increasing of cell surface area (r=-0.896,P
5.Effect of urinary IgG from nephrotic patients on expression of macrophage migration inhibitory factor in human proximal tubular epithelial cells
Weiying CHEN ; Fang FANG ; Hui PENG ; Xueqing YU
Chinese Journal of Nephrology 2005;0(07):-
Objective To investigate the effect of purified urinary IgG from patients with minimal change nephrotic syndrome and membranous nephropathy on the expression of macrophage migration inhibitory factor (MIF) in human proximal tubular epithelial cells (HK-2). Methods Proteins were precipitated from urine with (NH4)2SO4, and urinary IgG was purified by protein G column chromatography. SDS-PAGE and Western blotting were performed to analyze the urinary IgG. HK-2 cells were incubated with urinary IgG (0,0.5,1.0,2.5,5.0,10.0 mg/ml) from either minimal change nephrotic or membranous nephropathy patients. The expression of MIF mRNA was assessed by RT-PCR, and MIF protein was assessed by Western blotting. Results SDS-PAGE showed that there were four bands, which were all IgG fractions confirmed by Western blotting. The urinary IgG up-regulated the expression of MIF mRNA and protein in HK-2 cells. The expression increased in a dose-dependent manner. The expression of MIF mRNA and protein in HK-2 cells increased significantly when they were incubated with urinary IgG from membranous nephropathy patients at 1mg/ml (P
6.Intestinal permeability and injury change in chronic renal failure rats
Fangfei LI ; Zongpei JIANG ; Wenxue HU ; Yu ZHANG ; Yafang LI ; Wenxing PENG ; Xuchu HU ; Xueqing YU
Chinese Journal of Nephrology 2011;27(5):369-373
Objective To observe the histopathologic injury of small intestine and intestinal permeability in chronic renal failure (CRF) rats. Methods Twenty male Sprague-Dawley rats were randomly assigned to CRF group (n=10) and control group (n=10). 5/6 nephrectomy was used to establish CRF rats, while sham operation for control. Blood biochemistry was regularly monitored until CRF model was successfully established. The model rats were fed with lactulose (L) and mannitol (M) through intragastric administration. Urine was collected after 6 hours, and the concentration of lactulose and mannitol in urine was measured using high pressure liquid chromatograph with refractive index detector (HPLC-RID), and the ratio of urinary excretion of L/M was calculated to evaluate intestinal permeability. Small intestinal mucosa were stained by hematoxylin-eosin (HE) and observed with light microscope (villus height, thickness of muscle layer and villus count), histological damage score was used to evaluate intestinal injury. Results The L/M ratio of CRF group was higher than that of control group (1.75±0.11 vs 1.20±0.06, P<0.01). The small intestinal mucosal villus height and thickness of muscle layer in CRF group were higher (P<0.01), and the number of villi was lower compared to control group (P<0.01). The score of histopathologic intestine damage of CRF group was higher than that of control group (1.00±0.71 vs 0, P<0.01). Conclusion The intestinal permeability of CRF rats is increased with varying degrees of intestinal damage.
7.Expression of PPAR-γ,TLR4 and activation of STAT1 signal protein in rats with peritoneal dialysis related acute peritonitis induced by LIPS
Xunliang ZOU ; Xiao YANG ; Funfang ZHANG ; Xiuqing DONG ; Wenxing PENG ; Changyun WANG ; Xueqing YU
Chinese Journal of Microbiology and Immunology 2009;29(8):716-722
ated acute peritonitis induced by LPS in rats.
8.Effects of PPARγ, agonist on the expression of PPARγ. toll-like receptor 4 and STAT1 signal protein activation in rats with peritoneal dialysis-related acute peritonitis
Xunliang ZOU ; Xiao YANG ; Yunfang ZHANG ; Xiuqing DONG ; Wenxing PENG ; Changyun WANG ; Xueqing YU
Chinese Journal of Nephrology 2008;24(7):476-481
Objective To explore the effects of peroxisome proliferator-activated receptorγ (PPARγ) agonist rosiglitazone and 15-deoxy-delta-12,14-prostaglandin J2 (15d-PGJ2) on the expression of PPARγ, toll-like receptor 4 (TLR4) and the activation of STAT1 as well as the local inflammation reaction of abdominal cavity in sprague dawley (SD) rats with peritoneal dialysis- related acute peritonitis induced by lipopolysaccharide (LPS). Methods Twenty-four male SD rats were equally randomized to four groups(n=6 each): control group, injected with 4.25% dextrose peritoneal dialysate (PDF) via abdominal cavity(90 ml/kg); LPS group, injected with LPS(1 mg/kg) via abdominal cavity 4 hours later follewed by PDF injection; rosiglitazone plus LPS group (Rosi group), preconditioned with rosiglitazone (20 mg·kg-1·d-1) by intragastric way for 3 days, then injected with LPS and PDF via abdominal cavity; 15d-PGJ2 plus LPS group (15d-PGJ2 group), preconditioned with 15d-PGJ2 (0.3 mg·kg-1·d-1)via abdominal cavity injection for 3 days, then injected with LPS and PDF via abdominal cavity. The rats were killed 4 hours after PDF injection, IL-6 level in abdominal dropsy was determined by ELISA. Peritoneum tissue was stained by Masson. Leucocyte count in abdominal dropsy was performed. The mRNA expression of PPARγ and TLR4 in peritoneum tissue was determined by RT-PCR; the protein expression of PPARγ, TLR4, p-STAT1 and STAT1 in peritoneum tissue was analyzed by Western blot. Results IL-6 level of abdominal dropsy in LPS group [median 268.53 (range 201.87-335.19) ng/L] was significantly higher than that of control group [median 147.62 (range 130.60-164.64) ng/L] (P<0.01). The IL-6 level of abdominal dropsy in Rosi group [median 110.20 (range 77.60-142.80) ng/L] was significantly lower than that of LPS group (P<0.05). Compared to that of control group, the edematous degree of peritoneum in LPS group was significantly severer, meanwhile, mRNA and proteins expression of PPARγ and TLR4 in rat peritoneum were also significantly higher (P<0.05, P<0.01). Compared to that of LPS group, the edematous degree of peritoneum in Rosi group was lighter, the expression of PPARγ and TLR4 mRNA was significantly up-regulated (P<0.05), meanwhile their proteins expression was down-regulated (P<0.05); and in 15d-PGJ2 group, the edematous degree of peritoneum, the expression of PPARγ mRNA and protein was also decreased (P<0.05), but TLR4 mRNA expression was up-regulated (P<0.01), however, its protein expression was down-regulated (P<0.05). There were no significant differences in leucocyte count of abdominal dropsy among the four groups. The p-STAT1 expression in the rats peritoneum induced by LPS was markedly increased by both rosiglitazone and 15d-PGJ2 (P<0.01). Conclusions Both rosiglitazone and 15d-PGJ2 can down-regnlate the inflammatory reaction in rat peritonitis induced by LIPS, which may be involved in modulating the expression of associated functional protein during LPS signal pathway.
9.Supernatant of cultured mesangial cells with IgA1 from IgA nephropathy induces apoptosis of podocyte
Cheng WANG ; Ying TANG ; Hui PENG ; Zengchun YE ; Zhujiang CHEN ; Xueqing YU ; Tanqi LOU
Chinese Journal of Nephrology 2008;24(6):387-391
Objective To investigate the effects of supernatant of cultured mesangial vcells with serum IgA1 from [gA nephropathy patients on apoptosis of podocyte. Methods Jacalin affinity chromatography and Sephacryl S-200 molecular sieve chromatography were used to isolate IgA1. Apoptosis rate of podocyte was assessed by flow cytometer. Monomeric IgA1 (mIgA1) was transformed to aggregated IgA1(aIgA1) by heating. IgA-mesangial cell supernatant was prepared by collecting spent medium in which growth-arrested mesangial cells were incubated with different aIgA1, then the medium with RPMI 1640 containing 0.5%FBS was cultured with growth-arrested podocyte. Real time PCR was used to detect the mRNA expression of Bcl-2, Bax, Fas and Fas-L. Results Apoptosis rate of podocyte by supematant of cultured mesangial cell with algal from IgAN patients was higher than that from healthy and control groups [(28.5±5.9 ) % vs (22.5± 5.8)%, (20.5±4.5)%, all P<0.05]. Fas mRNA expression of podocyte exposed to supematant of cultured mesangial cells with aIgA1 from IgAN patients increased significantly and was 1.89 folds of control (P<0.05), while Bcl-2 mRNA expression significantly decreased and was 72% of control (P<0.05). The concentrations of Ang Ⅱ and TGF-β in supernatant of cultured mesangial cells with IgA1 from IgA nephropathy were significantly higher than those from healthy control [(13.2±3.4) ng/L vs (8.2±2.3) ng/L, /'<0.05; (15.4±3.4) ng/L vs (10.8±3.2) ng/L, P<0.05]. Conclusion Supernatant of cultured mesangial cells with IgA1 from IgA nephroapthy patients can induce apoptosis of podocyte, which may play a role in the progression of IgAN.
10.Renal lesion associated with Schimke immuno-osseous dysplasia: a case report and review of the literature
Wenfang CHEN ; Xueqing YU ; Shicong YANG ; Weiming GUAN ; Wenxing PENG ; Yu DONG ; Jinlang WU
Chinese Journal of Nephrology 2009;25(2):97-100
Objective To report a Chinese boy suffering from nephrotic syndrome associated with Schimke immuno-osseous dysplasia (SIOD). Methods The clnical data and pathological changes of renal biopsy were analyzed and associated literatures were reviewed. The clinicopathological features and diagnosis of SIOD were discussed. Results The first symptom of the patient was recurrent infections. Growth retardation, spondyloepiphyseal dysplasia accompanied by nephrotic syndrome and defective cellular immunity were seen as clinical features in this patient. Renal pathology showed focal segmental glomerulosclerosis. Conclusion Combining the clinical manifestation with renal pathology, the case is diagnosed as Schimke immuno-osseous dysplasia.