BACKGROUND: The p27, one of the cyclin-dependent kinase inhibitors in renal disease, regulates cell growth through affecting cellular events. Abundant evidence has suggested that p27 has great role in regulating re nal cell proliferation, hypertrophy, differentiation and apoptosis, etc. OBJECTIVE: To investigate the relationship of protein 27 expressions in renal tissue with cell proliferation and apoptosis. DESIGN: Completely randomized grouping design, control animal experi ment. SETTING: Nephrology Laboratory, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: This experiment was carried out at Nephrology Laboratory of Internal Medicine, Tongji Hospital and Micro-Pathology Center in Tongji Medical College from September 2004 to January 2005. A total of 48 adult male SD rats were enrolled, all belonged to cleaning grade. METHODS: ①Establishing diabetic models: diabetic rat models were es tablished with streptozotocin, 60 mg/Kg, given through intraperitoneal in. Jection in the experimental group. Blood glucose and urine glucose from caudal vein were measured after 48 hours. Animals in experimental group whose random serum glucose at least 16.7 mmol/L and urine glucose ()- ()-() were regarded as successful models. ②Six rats in experimental group and the control group, respectively were sacrificed in the experimental pe riod of the 3rd, 7th, 14th and 28th days. Before sacrificed, creatinine clearance (Ccr) was measured. Concentration of Retinal binding protein (Rbp) in urine was measured. Proteinuric elements were analyzed by sodium dodecyl sulfate-polyacrlamide gelelectrophoresis (SDS-PAGE). ③Renal tissues were observed pathologic changes through light microscopy and electronic mi croscopy. ④Expression of p27 and proliferation cellnuclear antigen (PC NA) were detected by immunohistochemical staining method, and apoptosis was determined by Terminal deoxynucleotidy 1 transferase-mediated dUTP nick end labeling (TUNEL). We use positive glomerular cells / total glomerular cells to present positive rate and apoptosis rate. ⑤The data were analyzed for statistically significant group difference using unpaired t test and Spearman correlative-analysis. MAIN OUTCOME MEASURES: Renal function, urine protein, p27 ex pression in glomerular cells, positive rate and apoptosis rate of proliferating cell nuclear antigen (PCNA) expression and compared with the control group. RESULTS: Totally 48 rats were involved in the result analysis. ①Changes of pathology: At the 7th day after establishing models, glomerular hypertro phy was found through light microscopy and focal fusion of epithelial foot processes through electron microscopy. At the 14th day, mesangial region, which kept expanding through electronic microscopy, was seen under electron microscope. At the 28th day, slight segmental hypercellularity be gan to show in light microscopy. ②Ccr and Rbp levels of diabetic rats were higher significantly than those in the control group at the 7th, 14th and 28th days (t=2.143-3.004,P ＜ 0.05 ). The results of SDS-PAGE showed that middle and low molecular protein could be observed in every group from the 3rd day. ③The level of p27 expression in glomerular cells were lower obviously at the 3rd day(t=2.536,P ＜ 0.05), but higher at the 7th, 14th and 28th days in diabetic rats, which was higher remarkably than that in the control group (t=2.704-2.823,P ＜ 0.05). The number of PCNA positive cell in diabetic glomerulus became the largest at the 3rd day, then decreased continuously, it was still higher than that in the control group at the 7th dan 14th days (t=2.681,2.525,P ＜ 0.05), but became normal at day 28. The number of TUNEL positive cells in glomerulus also increased significantly at the 3rd day (t=2.764,P ＜ 0.05), but it was smaller than that in the control group at the 7th, 14th and 28th days(t=2.096-2.627,P ＜ 0.05). ④The level of p27 expression and the number of PCNA positive cells in glomerulus were negative correlated (r=-0.446, P ＜ 0.05). The level of p27 expression and the number of TUNEL positive cells in glomerulus were also negative correlated (r=-0.517, P ＜ 0.05).CONCLUSION: Functional impairment of reabsorption induced by proximal convoluted renal tubule and high filtering of glomerulus in early diabetics occurs synchronously. Protein p27 may be participated in the occurring and developing of diabetic nephropathy by checking of proliferation and apoptosis of renal cell in the early stage of diabetes.

The effects of mycophenolic acid (MPA) on high glucose-induced expression of transforming growth factor-beta (TGF-beta) and connective tissue growth factor (CTGF) in mesangial cells (MC) were investigated. Rat MC were cultured in the presence of different concentrations of MPA (1.0 and 10.0 micromol/L) or MPA plus high glucose for 72 h. The expression of TGF-beta and CTGF was detected by Western blot. The results showed that high glucose could induce the expression of TGF-beta and CTGF in MC, but MPA could inhibit this effects. MPA did not influence the expression of TGF-beta and CTGF in normal glucose. It was concluded that MPA might prevent the progression of diabetic nephropathy by inhibiting the expression of TGF-beta and CTGF in MC.

Objective: To study the effects of soybean phospholipids on fatty acids and phospholipids content in erythrocyte membranes of mice. Methods:The mice were fed with soybean phospholipids of 2 . 5, 5 . 0 , 10.0 g/kg bw respectively while control group with distilled water for 30 days. Then the mice were killed and contents of fatty acids and phospholipids in erythrocyte membranes were detected by HPLC. Results: (1)The contents of linoleic acid (C18 ∶2) and linolenic acid ( C18 ∶3), the composition of polyunsaturated fatty acid (PUFA) were increased obviously (P

Objective:To study the effects of soybean phospholipids on learning and memory abilities and contents of fatty acids in hippocampus of mice and its mechanism. Method:Mice were randomly divided into four groups:low dose(2.5g/kg) ,middle dose(5.0g/kg) and high dose(10.0g/kg) groups of soybean phospholipids and control group. The treatment groups were fed with soybean phospholipids every day while the control group with distilled water for 30 d.After learning and memory abilities were measured,the mice were killed and contents of fatty acids in hippocampus of mice were determined by high performance liquid chromatography(HPLC) . Results:(1) The learning and memory abilities of mice in middle and high dose groups were significantly improved,but only memory abilities in low dose groups were stronger than control group.(2) The contents of protein in hippocampus in middle and high dose groups were increased extremely.(3) The contents of oleic acid(C18:1) and docosahexaenoic acid(DHA,C22:6 ) were obviously enhanced in hippocampus of high dose group;but decreased in eicosapentaenoic acid(EPA,C20:5) and linoleic acid(C18:2) . The content of DHA was increased and EPA decreased significantly in middle dose group.The increase of arachidonic acid(C20:4) was not significant.Conclusion:Soybean phospholipids may improve learning and memory abilities,which might contribute to change of fatty acids contents in hippocampus of mice.

Objective To study the expression of interleukin-18(IL-18)during the progression of renal interstitial fibrosis in rat kidney after unilateral ureteral obstruction(UUO)and the effect of Shenfu injection.Methods The obstructive nephropathy model was established by unilateral ureteral obstruction(UUO).Fifty。Six rats were randomly assigned into shame operation group,operation group(UUOgroup)and treatment group(UUO+Shenfu).After 7 and 14 days,the renal function and histopathological changes were evaluated.Immunohistochemistry was used to examine the expression of IL-18 in renal tissue.Results In comparison with the shame opeartion group,the operation group showed obvious renal interstitial fibrosis.And the expression of IL-18 increased signifieantly(P<0.05).Compared with the operation group,the degree of interstitial fibrosis was obviously ameliorated in the treatment group,and the expression of IL-18 decreased significandy after treatment for 7 days(P<0.05),and decreased more after treatment for 14 days(P<0.05).Conclusions Shenfu injection may protect renal function by decreasing the expression of IL-18 in the progression of renal interstitial fibrosis.

Objective To investigate the dynamic changes of MDA, NO, SOD and pathologic changes of the lung and kidneyduring repefusion after haemorrhagic shock in rabbits, and to study the protective effects of edaravone during thecourse.Method Totally 29 beparinized (3 mg/kg) rabbits were randomly divided into three groups:tho sham-operatedcontrol group (group C, n = 7), the haemorrhagic shock group (group I/R, n = 10), and the haemorrhagicshock group with edaravone infusion (group I/R-edaravone, n = 12). Rabbits in the latter two groups were bledfrom left arteria cmralis in 10 minutes with MAP maintained at 40 mmHg for 60 minutes, and then group I/R-edar-avone was given edaravone intravenously. After that, resuscitation began:all blood loss was replaced with normalsaline within 60 minutes with MAP at the end ≥ 70% MAP before haemorrhagic shock. Edaravone was reinjectedat 10 hours after shock.All rabbits were killed at 20 h after reperfusion.Plasma nitric oxide(NO), malonyldialde-hyde (MDA) and superoxide dismutase(SOD) in every group were measured before shock,60 minutes after shockaad 1 h, 5 h and 20 h after reperfusien. Part of the right lung and the right kidney tissues were taken from everyrabbit for pathologic examnation after sacrifice.Results There was no significant difference in MDA,NO aad SOD among three groups before shock. A higherlevel of MDA (5.35±0.29 μmol/L), NO(27.75 ±2.88 μmol/L)and lower serum concentration of SOD(194.58±14.42U/ml)could be found in group I/R during haemorrhagic shock,as compared to group C(4.44±0.59 μmol/L,25.01±4.95μmol/L,210.86±24.54U/ml,respectively,P<0.01).At 20 hours after resuscitation,MDA and NO contents continued to increase(5.69±0.24 μmol/L and 28.01±3.10 μmol/L respectively,P<0.05)while SOD contents kept decreasing(151.83±9.36 U/ml,P<0.05)in group I/R.Comparing to group I/R,group I/R-edaravone had significant lower level of MDA(3.48±0.23 μmol/L,P<0.01)and higher concentration of SOD(195.10±11.87U/ml,P<0.01).Edaravone attenuated the pathologic changes in the lung and kidney.Conclusions Edaravone could effectively protect vital organs from reperfusion injury caused by free radicals following haemorrhagic shock by reducing plasma levels of MDA,NO and increasing levels of SOD.

The apoptosis and the expression of tumor suppressor gene p53 in hypercholesterolemia (HC)-induced renal injury were investigated in rats. A high cholesterol diet (HCD)-induced HC rat model was made and serum lipid, urinary protein excretion (UPE) and N-aceto-β-D-glucosidase (NAG) were measured. The levels of malondialdehyde (MDA), as an index of lipid peroxidation, in renal cortex and serum were compared between the two diet groups. Apoptosis and p53 expression were determined by TUNEL and immunohistochemistry, respectively. In the HCD-induced HC group, serum total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) as well astriglyceride (TG) were significantly increased, while the level of high density lipoprotein-cholesterol (HDL-C) decreased. Meanwhile, increased excretions of UPE and NAG in urine were observed, which were accompanied with a decrease in urinary creatinine clearance (Ccr) and indicated both glomerular and tubular damages. In addition, apoptotic cell death coexisted in the kidney, as revealed by increased TUNEL positive cells. Finally, an increase in p53 expression was observed in tubuli, but not in glomeruli. Both TUNEL positive cells and p53 expression were found to be correlated to the level of renal cortical MDA (r=0.817, P<0.01 and r=0.547, P<0.01, respectively). The major manifestation of HCD-induced renal injury is apoptosis. The lipid peroxidation is a critical event to induce DNA damage and p53 is involved in the pathogenesis of lipid-induced renal injury.

The effects of mycophenolic acid (MPA) on high glucose-induced expression of transforming growth factor-β (TGF-β) and connective tissue growth factor (CTGF) in mesangial cells (MC) were investigated. Rat MC were cultured in the presence of different concentrations of MPA (1.0 and 10.0 μmol/L) or MPA plus high glucose for 72 h. The expression of TGF-β and CTGF was detected by Western blot. The results showed that high glucose could induce the expression of TGF-β and CTGF in MC, but MPA could inhibit this effects. MPA did not influence the expression of TGF-β and CTGF in normal glucose. It was concluded that MPA might prevent the progression of diabetic nephropathy by inhibiting the expression of TGF-β and CTGF in MC.

Objective To identify the significance of serum phospholipase A2 receptor antibody (PLA2R-Ab) in idiopathic membranous nephropathy (IMN) patients.Methods A total of 108 patients diagnosed as IMN by medical history,physical examination,laboratory examination and renal biopsy in Tongji Hospital affiliated to Tongji Medical College,Huazhong University of Science and Technology between Dec 1,2014 and Aug 31,2017 were enrolled,and all related data were recorded.According to the results of serum PLA2R-Ab test,patients were divided to positive group and negative group,and the data were compared with the independent sample t test and the chi-square test.Kaplan-Meier survival analysis was performed to compare remission rates between groups,and the Logrank method was used to evaluate the significance of differences.Univariate and multivariate Cox regression analysis were used to verify predicting factors for achieving remission.Results Overall,67.6％(73/108) patients had detectable serum PLA2R-Ab.Compared with patients in negative group,patients in positive group exhibited higher proportion of male patients (P=0.002),lower level of serum albumin (P ＜ 0.001),higher level of cholesterol (P ＜ 0.001),lower level of immunoglobulin G (P ＜0.001),higher level of proteinuria (P=0.003),a lower of chance of remission (P=0.049),longer time needed to achieve partial remission (P=0.001) and complete remission (P=0.002).The 1-and 2-year cumulative renal partial remission rates were 72.4％,86.1％,and the cumulative renal complete remission rates were 43.8％,54.0％,respectively.Patients in negative group had higher partial remission (x2=9.84,P=0.002) and complete remission (x2=15.50,P＜0.001) than those in positive group.Multivariate Cox regression model indicated that serum positive PLA2R-Ab was a significant independent risk factor.Conclusions IMN patients with serum PLA2R-Ab show more severe condition and lower remission rates than those without serum PLA2R-Ab.Serum positive PLA2R-Ab is an independent remission-related predictor for IMN patients.

Objective To investigate the clinical characteristics and risk factors of secondary infection in patients with anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV).Methods One hundred and eighteen patients newly diagnosed with AAV at the institute of nephrology,Tongji hospital affiliated to Huazhong university of science and technology,from 2012 to 2017,were analyzed retrospectively.Induction therapy included single corticosteroids,combination of corticosteroids with cyclophosphamide and combination of corticosteroids with other immunosuppressive agents.End point was defined as moderate to severe infection which was diagnosed by the clinical and radiological manifestation as well as microbiological evidences.The infection-related survival curve was drawn to reflect the time when the infection occurred.The clinical baseline variables in patients with and without infection were compared.Multivariate Logistic regression model was used to determine the independent predictors of infection.Receiver-operating characteristic curve (ROC) was plotted for evaluating the predictive value of lymphocyte on moderate to severe infection.Results During followup of median 3 months (1-30 months),88 infection episodes were found in 63 (53.4％) patients,of which 54 times (61.4％) occurred within 6 months after treatment,46 times (52.3％) happened within 3 months after treatment.The most common organ of infection was lung (62.5％),and the most common pathogen was bacteria (51.1％).Multivariate Logistic regression model showed that lung involvement (OR=4.44,95％ CI 1.59-12.41),moderate reduction of lymphocyte in follow-up (OR=5.69,95％ CI 2.05-15.85) and severe lymphocyte reduction (OR=36.28,95％CI 3.45-381.17) were independent risk factors of secondary infection in AAV patients (all P ＜ 0.05).ROC curve showed that the area under the curve of lymphocyte as a predictor of severe infection was 0.767 (95％ CI 0.64-0.89,P ＜ 0.05).Based on lymphocyte less than 0.49× 109/L which was the cut-off value for predicting severe infection,the sensitivity and the specificity were 83.9％ and 71.9％,respectively.Conclusions Lung involvement and moderate-severe lymphopenia during follow-up are independent risk factors of secondary infection in AAV patients.Hence,physician should pay more attention to those patients,and adjust treatment in time to avoid the occurrence of infection.