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Objective:To compare the effect of various antigen retrieval methods in paraffin sections of renal biopsy tissue, and explore the best antigen retrieval method.Methods:Forty-five renal biopsy specimens were collected from the First Affiliated Hospital of Zhengzhou University, including lupus nephritis ( n=10), membranous nephropathy ( n=10), IgA nephropathy ( n=10) and amyloidosis glomerulopathy ( n=15). Five retrieval methods (including high pressure thermal retrieval combined with trypsin retrieval, microwave thermal retrieval combined with trypsin retrieval, high pressure thermal retrieval, microwave thermal retrieval and gastroprotease retrieval) were used for immunofluorescence staining of paraffin sections. Renal tissue specimens were divided into six groups according to different antigen retrieval methods, frozen section specimens used as a control group. The immunofluorescence semi-quantitative scores of paraffin sections of the five heat-repairing antigen methods and frozen sections were compared. Results:Immunofluorescence staining of hyperbaric thermal retrieval combined with trypsin retrieval group and microwave thermal retrieval combined with trypsin retrieval group were similar with those of frozen sections. Compared with the control group, there were no significant difference in the semi-quantitative immunofluorescence scores between the two groups (all P>0.05). However, Immunofluorescence staining of hyperbaric thermal retrieval, microwave thermal retrieval, pepsin digestion had significantly higher false negative rate than those of frozen sections. Compared with the control group, the difference in semi-quantitative immunofluorescence score was statistically significant (all P<0.05). Conclusion:High pressure heat retrieval combined with trypsin retrieval or microwave heat retrieval combined with trypsin retrieval is the first choice of antigen retrieval methods.
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Objective To study the role of alternative complement pathway overactivation in malignant nephrosclerosis.Methods (1) Fifty patients with confirmed malignant nephrosclerosis by renal needle biopsy were enrolled.Meanwhile,twenty-five cases of time-zero renal needle biopsy were enrolled as control subjects.Enzyme linked immunosorbent assay (ELISA) was used to detect alternative complement pathway of the complement initiation factor B,positive regulation factor P,negative regulation factor H,and the complement end products C3a and C5a in the plasma and urine.(2) Immunohistochemistry was used to detect the deposition of the complement end product C5b-9,C4d and mannan binding lectin (MBL) of lectin pathway in the renal biopsies.Double immunofluorescence labeling method was used to assay the deposition of C5b-9 and CD34 (endothelial cell marker) in the arteriolar endothelium and glomerular capillary endothelium.Results (1) The plasma and urine levels of complement factor B,factor P,C3a and C5a in malignant nephrosclerosis patients were significantly higher than those in control subjects (all P ＜ 0.05),while the plasma and urine levels of complement factor H in malignant nephrosclerosis patients were lower than those in control subjects (all P ＜ 0.05).(2) The plasma level of factor P was positively correlated with 24 h urine protein (rs=0.465,P=0.001).Urinary factor B/urinary creatinine,urinary factor P/urinary creatinine and urinary C3a/urinary creatinine were positively correlated with serum creatinine in malignant nephrosclerosis patients (rs=0.483,P ＜ 0.001;rs=0.352,P=0.012;rs=0.319,P=0.024),while urinary factor H/urinary creatinine was negatively correlated with serum creatinine and 24 h urine protein (rs=-0.299,P=0.035;rs=-0.342,P=0.015).Urinary C5a/urinary creatinine was positively correlated with serum creatinine and 24 h urine protein (rs=0.525,P ＜ 0.001;rs=0.496,P ＜ 0.001).(3) Immunohistochemical results showed that there were C5b-9 deposited in the arterioles and glomerular capillary wall in malignant nephrosclerosis patients,and no deposition in control renal tissues.Meanwhile,the semi-quantitative scores showed that C5b-9 deposition intensity was positively correlated with serum creatinine and 24 h urine protein (rs=0.791,P＜ 0.001;rs=0.345,P=0.014).The double immunofluorescence labeling analysis showed that the C5b-9 and CD34 deposited in the arteriolar endothelium and glomerular capillary endothelium.(4) Plasma level of factor B in malignant nephrosclerosis patients was positively correlated with plasma C3a level (r=0.331,P=0.022).Plasma level of factor P was positively correlated with C5b-9 score (rs=0.300,P=0.034).Urinary B was positively correlated with urinary C3a,C5a and C5b-9 score (rs=0.311,P=0.028;rs=0.465,P=0.001;rs=0.428,P=0.002).Urinary factor P was also positively correlated with urinary C3a and C5a (rs=0.307,P=0.030;rs=0.442,P=0.001).Immunohistochemical result showed that there were C4d deposited in the arterioles and glomerular,and no deposition of MBL.Conclusion Complement activation via the alternative pathway may be involved in malignant nephrosclerosis and related to the severity of the disease.
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Objective@#To study the role of alternative complement pathway overactivation in malignant nephrosclerosis.@*Methods@#(1) Fifty patients with confirmed malignant nephrosclerosis by renal needle biopsy were enrolled. Meanwhile, twenty-five cases of time-zero renal needle biopsy were enrolled as control subjects. Enzyme linked immunosorbent assay (ELISA) was used to detect alternative complement pathway of the complement initiation factor B, positive regulation factor P, negative regulation factor H, and the complement end products C3a and C5a in the plasma and urine. (2) Immunohistochemistry was used to detect the deposition of the complement end product C5b-9, C4d and mannan binding lectin (MBL) of lectin pathway in the renal biopsies. Double immunofluorescence labeling method was used to assay the deposition of C5b-9 and CD34 (endothelial cell marker) in the arteriolar endothelium and glomerular capillary endothelium.@*Results@#(1) The plasma and urine levels of complement factor B, factor P, C3a and C5a in malignant nephrosclerosis patients were significantly higher than those in control subjects (all P<0.05), while the plasma and urine levels of complement factor H in malignant nephrosclerosis patients were lower than those in control subjects (all P<0.05). (2) The plasma level of factor P was positively correlated with 24 h urine protein (rs=0.465, P=0.001). Urinary factor B/urinary creatinine, urinary factor P/urinary creatinine and urinary C3a/urinary creatinine were positively correlated with serum creatinine in malignant nephrosclerosis patients (rs=0.483, P<0.001; rs=0.352, P=0.012; rs=0.319, P=0.024), while urinary factor H/urinary creatinine was negatively correlated with serum creatinine and 24 h urine protein (rs=-0.299, P=0.035; rs=-0.342, P=0.015). Urinary C5a/urinary creatinine was positively correlated with serum creatinine and 24 h urine protein (rs=0.525, P<0.001; rs=0.496, P<0.001). (3) Immunohistochemical results showed that there were C5b-9 deposited in the arterioles and glomerular capillary wall in malignant nephrosclerosis patients, and no deposition in control renal tissues. Meanwhile, the semi-quantitative scores showed that C5b-9 deposition intensity was positively correlated with serum creatinine and 24 h urine protein (rs=0.791, P<0.001; rs=0.345, P=0.014). The double immunofluorescence labeling analysis showed that the C5b-9 and CD34 deposited in the arteriolar endothelium and glomerular capillary endothelium. (4) Plasma level of factor B in malignant nephrosclerosis patients was positively correlated with plasma C3a level (r=0.331, P=0.022). Plasma level of factor P was positively correlated with C5b-9 score (rs=0.300, P=0.034). Urinary B was positively correlated with urinary C3a, C5a and C5b-9 score (rs=0.311, P=0.028; rs=0.465, P=0.001; rs=0.428, P=0.002). Urinary factor P was also positively correlated with urinary C3a and C5a (rs=0.307, P=0.030; rs=0.442, P=0.001). Immunohistochemical result showed that there were C4d deposited in the arterioles and glomerular, and no deposition of MBL.@*Conclusion@#Complement activation via the alternative pathway may be involved in malignant nephrosclerosis and related to the severity of the disease.
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Objective To study the role of C3a and C5a in focal segmental glomerulosclerosis (FSGS) patients. Methods (1) A total of 66 patients with FSGS confirmed by renal biopsy were selected, including 18 cases of tip lesion, 11 cases of perihilar, 22 cases of not otherwise specified (NOS), 10 cases of cellular, and 5 cases of collapsing FSGS. The normal renal tissue resected from patients with kidney tumor was taken as a negative control. The expression of C3a and C5a in renal tissues was detected by immunohistochemistry. (2) Serum and urine samples from these 66 FSGS patients were collected, and serum and urine samples from 10 healthy adult selected from the same physical examination center in the same term were used as normal controls. The levels of C3a and C5a in serum and urine were detected by enzyme - linked immunosorbent assay (ELISA). Results (1) Immunohistochemical results showed that C3a and C5a were deposited in glomerulus of FSGS patients, and no deposition in normal renal tissues. The semi - quantitative score showed that kidney C3a score was significantly correlated with serum creatinine (r=0.547, P＜0.001) and 24 h urine protein (r=0.329, P=0.007) in FSGS patients, and kidney C5a score was also significantly correlated with serum creatinine (r=0.415, P＜0.001) and 24 h urine protein (r=0.414, P＜0.001) in FSGS patients. (2) The levels of serum C3a and C5a in FSGS patients were higher than those in healthy adults (both P＜0.05), but there was no significant difference among the five pathological types (P＞0.05). The levels of urinary C3a/urinary creatinine, urinary C5a/urinary creatinine were higher in FSGS patients than those in healthy adults (all P＜0.05). The levels of urine C3a/urinary creatinine and urinary C5a/urinary creatinine in collapsing FSGS were higher than other FSGS types (all P＜0.01), but there was no significant difference among the tip lesion, the perihilar, the not otherwise specified and the cellular (P＞0.05). (3) Urinary C3a/urinary creatinine levels were significantly correlated with serum creatinine (r=0.774, P＜0.001) and 24 h urine protein (r=0.430, P＜0.001) in FSGS patients, and urinary C5a/urinary creatinine levels were also significantly correlated with serum creatinine (r=0.677, P＜0.001) and 24 h urine protein (r=0.333, P=0.007) in FSGS patients. Conclusion Complement C3a and C5a may be involved in the pathogenesis of FSGS and may be related to the severity of FSGS.
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Objective:To investigate the role of C3a,C5a and their receptors in the pathogenesis of IgA nephropathy (IgAN). Methods:A total of twenty-eight 6-8 weeks old female BALB/c mice were investigated.And they were negative control group , WT group,C3aR-/-group,C5aR-/-group(the latter three groups were named as experimental groups ),seven mice in each group.All the mice were infected through respiratory tract with infectious SV (experimental groups) or PBS(negative control group),combined with tail vein challenge to make IgAN animal model.Testing 24 h total urinary protein , serum urea nitrogen ( BUN ) and creatinine ( Cr ) , using direct immunofluorescence to test the renal deposition of IgA and C 3,observing renal pathologic lesion under PAS staining with light microscopy.RT-qPCR was used to test the relative mRNA expression of TNF-α,TGF-β,IL-1β,IL-6,MCP-1.Results: After 15 weeks,the level of UTP in experimental group was significantly higher than negative control group ,and the same results as WT group than C3aR-/-group and C5aR-/-group.There was no significant difference among groups for BUN and Cr.Combined with negative control group , experimental groups had significant renal pathological lesions , and the changes of WT group was more severe than C3aR-/-group and C5aR-/-group.The results of relative mRNA expression of TNF-α,TGF-β,IL-1β,IL-6,MCP-1 was the same as the level of 24 h UTP,at the same time,the relative mRNA expression of IL-1β,IL-6,MCP-1 in C3aR-/-group was significantly less than C5aR-/-group.Conclusion:The deficiency of C3a/C5a receptors can protect kidney from injury in IgAN ,and C3a receptor has more significant role in protect kidney from injury in IgAN.
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<p><b>BACKGROUND</b>Mycophenolate mofetil (MMF) and cyclophosphamide (CTX) are widely used in treating various kidney diseases. However, whether they are effective and which one is better for treating IgA nephropathy patients with proliferative pathological phenotype in renal diseases, such as endocapillary proliferation, cellular crescents, and/or capillary loops fibrinoid necrosis is still unknown. We, therefore, initiated a study to compare the effects of MMF and CTX in treating IgA nephropathy with the above pathological lesions.</p><p><b>METHODS</b>One hundred and nineteen patients with IgA nephropathy who had at least one of the three aforementioned lesions were enrolled. All patients were treated with prednisone; 48 patients received prednisone only (Pred group), 40 received MMF and prednisone (MMF + Pred group), and 31 were treated with CTX and prednisone (CTX + Pred group). The median time of follow-up was 30 months (maximum: 96 months). The primary endpoint was defined as renal survival. The incidence of remission of proteinuria was the secondary endpoint.</p><p><b>RESULTS</b>Serum creatinine in all groups declined significantly at different follow-up times (P = 0.002), and the differences among the three groups were significant (P < 0.001). At 24 months of follow-up, the decline rates were 12.35%, 32.95%, and 24.14% in the Pred, MMF + Pred, and CTX + Pred groups respectively. For urine protein excretion, the decline rates were 49.12% (Pred), 73.67% (MMF + Pred), and 63.53% (CTX + Pred) respectively at 24 months of follow-up. The differences among the three groups were not significant (P = 0.714). Renal survival (the primary endpoint) was significantly different (P = 0.027); however, the sencondary endpoint was similar for all the three groups (P = 0.100).</p><p><b>CONCLUSIONS</b>For IgA nephropathy patients with endocapillary proliferation, cellular crescents, and/or fibrinoid necrosis of capillary loops, prednisone combined with MMF was more effective in lowering the serum creatinine than with CTX. Combined MMF and prednisone treatment led to a better renal survival compared to that of prednisone with CTX.</p>
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Adult , Female , Humans , Male , Young Adult , Glomerulonephritis, IGA , Drug Therapy , Pathology , Immunosuppressive Agents , Therapeutic Uses , Mycophenolic Acid , Therapeutic Uses , Retrospective StudiesABSTRACT
Objective To investigate the protective effects of Wnt-7a protein on renal interstitial fibrosis in mice of unilateral ureteral obstruction (UUO)model.Methods Eighteen male C57BL/6 mice were randomly divided into 3 groups:sham-operation group,the UUO model group and Wnt-7a treatment group.The body weight of mice was measured everyday.All the mice were sacrificed at thc seventh day after the operation.The left kidney was taken for histology evaluation and molecular biology assay.Masson's stain was performed as a main indicator of interstitial fibrosis.The expression of vimentin,α-smooth muscle actin,and E-adherin in renal tissue was detected by immunohistochemistry staining and the expression of α-smooth muscle actin and E-cadhe(nn) in renal tissue was detected by Western blotting.Results Compared with sham-operation group,body weight of the (,)odel group was significantly lower (P＜0.05),and the relative area of interstitial fibrosis was significantly larger (P＜0.05).Furthermore,the expression of vimentin and α-SMA was significantly up-regulated (P＜0.05),and the expression of E-cadherin was significantly down-regulated (P＜0.05).Compared with model group,all the above-mentioned abnormalities were restored to some extent and showed significant differences (P＜0.05) in Wnt-7a treatment group.Conclusion Wnt-7a protein can decrease the interstitial fibrosis by inhibiting epithelial to mesenchymal transition in UUO mice.
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Objective To explore the clinicopathological characteristics of lupus nephritis (LN) with antinucleosome antibody (AnuA).Methods Data of 481 patients with biopsy-proven LN in the First Affiliated Hospital of Zhengzhou University from 2004 to 2011 were analyzed retrospectively.The patients were divided into two groups:AnuA-positive group (76 patients) and AnuA-negative group (405 patients).The clinical manifestations,laboratory examinations,histopathologic classes of LN,disease activity measured by SLE disease activity index (SLEDAI) of two groups were investigated and compared.Results There were 15 male patients in positive group (15/76,19.74％) with mean age of (27.99±10.88) years and 45 patients in negative group (45/405,11.11％) with mean age of (31.15±12.15) years respectively,which showed that male patients were more common in positive group (P＜0.05).Incidences of oral ulcer,fever,anemia,low complement and positive anti-dsDNA antibody were higher in positive group (P＜0.05).Percentage of diffuse proliferative lupus nephritis (class Ⅳ ) and pathological activity index (AI) in positive group were higher compared to negative group (all P＜0.05),while no significant differences of other pathological types,chronic index (CI) and SLEDAI were found between two groups.Conclusion LN patients with positive AnuA have special clinicopathological characteristics and AnuA may be used as a promising biomarker for the proliferative LN.
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Objective To investigate the effect of soluble Tie2 fusion protein(sTie2/Fc)on the angiogenesis of peritoneal vessels in uremic peritoneal dialysis (PD) rats.Methods Rats were randomly divided into 6 groups:normal rats as control group (group1),rats with sham operation (group2),uremic rats without PD (group3),uremic rats dialyzed with 4:25％ PD solution (group4),uremic rats dialyzed with 4.25％ PD solution and treated by subcutaneous injection of 2.5 μg/kg sTie2/Fc (group5),uremic rats dialyzed with 4.25％ PD solution and treated by subcutaneous injection of 5.0 μg/kg sTie2/Fc (group6).sTie2/Fc was given every other day during peritoneal dialysis period,total 14 doses.After regular PD for 28 days,RT-PCR and tissue immunohistochemical staining were used to detect the mRNA and protein expressions of Angpt-2 in peritoneal tissues in each group of rats.Microvessel density (MVD) of peritoneum was detected and quantified with immunohistochemical staining by using anti-CD34 antibody.Results The expression of Angpt-2 mRNA and protein was found in each group.There was no significant difference of.Angpt-2 expression both in mRNA and protein level between group1 and group2.Compared with group1,the mRNA and protein expression of Angpt-2 were significantly increased in group3 and group4 (all P＜0.05).Compared with group3,the mRNA and protein expression of Angpt-2 were significantly increased in group4 (all P＜0.05).Compared with group 4,the mRNA and protein expression of Angpt-2 were significantly decreased in group5 and group6 (all P＜0.05).Compared with group5,the mRNA and protein expression of Angpt-2 were significantly decreased in group6 (all P＜0.05).Only few new microvessel was found in group1 and group2.Compared with group1,MVD was significantly up-regulated in group3 and group4 (all P＜0.05).Compared with group4,MVD was significantly down-regulated in group5 and group6 (all P＜0.05).Conclusions Peritoneum neoangiogensis can be effectively inhibited by sTie2/Fc in uremic rat treated with PD.Blocking of signal transduction may be involved in the mechanism of sTie2/Fc inhibiting peritoneal angiogenesis.
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Objective To explore the prevalence of chronic kidney disease(CKD) and its related risk factors in the people aged 60 years or over in Zhengzhou.Methods One thousand two hundred and ninety-one residents aged 60 years or over were selected with stratified-multi-steps-cluster sampling method in 4 communities from 4 different districts in Zhengzhou city.The indicators of renal damage and its related factors were collected with questionnaires.Results Eligible data of 1247 subjects were collected.After adjustment for age and gender, the prevalence of albuminuria,hematuria and reduced eGFR were 13.3% ,6.0% and 2.0% respectively, and the prevalence of CKD was 17.5%.Multiple logistic regression analysis showed that hypertension, diabetes and overweight were independent risk factors for CKD.Conclusions The prevalence of chronic kidney disease in the urban people aged 60 years or over in Zhengzhou is higher than in the non-elderly people and preventive measures should be adopted.
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Objective To study the prevalence and risk factors of chronic kidney disease (CKD) among adults in Zhengzhou. Methods One thousand eight hundred and fifty five residents (≥ 20 years) from 4 communities in 4 districts of Zhengzhou city were randomly selected by using a stratified,multistage sampling. They were interviewed, and received physical examination and measurements of urine and blood for renal damage as well as risk factors. Results Eligible data of 1752 subjects were included in the study. After the adjustment of age and gender component, albuminuria was found in 5.78% of the subjects, hematuria in 8.19%, and reduced renal function in 1.58%. Male had lower prevalence of albuminuria and hematuria (4.37% vs 7.29%, X2=6.252, P=0.012; 5.08% vs 11.51%, X2=24.499, P＜0.01), but higher prevalence of reduced eGFR(2.26% vs 0.86%, X2=5.830, P=0.016) as compared with female. The prevalence of albuminuria and reduced eGFR increased with age. The crude prevalence of CKD was 14.50%, while the standardized rate was 13.57%. The prevalence of female was higher than that of male (17.83% vs 9.59%, X2=23.132, P＜0.O1), which also increased with age. The most common manifestations of CKD were hematuria and albuminuria. Gender, age, smoking, hypertension,diabetes mellitus, obesity and hyperuricaemia were independently associated with CKD. The awareness rate of CKD was 8.27% and only 7.09% of the subjects received treatment. Conclusions The prevalence of CKD is 13.57% and the recognition is 8.27% in urban adult population of Zhengzhou.lndependent risk factors associated with kidney damage are gender, age, smoking, hypertension, diabetes mellitus, obesity and hyperuricaemia.
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Objective To observe the clinical curative effect and side-effect of MMF plus corticosteroid hormone compared with cyclophosphamide(CTX)plus corticosteroid hormone for idio-focal segmental glomerular sclerosis(I-FSGS).Methods Thirty patients with I-FSGS confirmed by renal biopsy in the Nephrology Department,the First Affiliated Hospital of Zhengzhou University from 2004 to 2006 were randomly divided into two groups-15 patients in each group:MMF combined with corticosteroid hormone as therapy group,and CTX combined with corticosteroid hormone as control group.The two groups were compared on urine protein in 24 hours,serum albumin and creatinine clearance rate(Ccr).Results There were significant differences(P
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Objective To evaluate prognostic value of electroencephalogram (EEG) in neonatal with hypoxic ischemic encephalopathy (HIE). Methods Sixty two infants with HIE was studied by doing physical examination, intelligence test, CT and EEG. Their clinical outcome was compared to their EEG in the first month after birth. Results Incidence of sequelae in normal or mild abnormal EEG and severe abnormal EEG were 3%, 29%, respectively. Among three infants of EEG cases, two dead and one developed cerebral palsy infant. Among four of burst suppression found in the EEGs, two cases dead, one developed cerebral palsy, one was normal. In five of hypoactive EEG, one dead, one developed cerebral palsy, and the other was with low IQ. Conlusion The prognosis is related to the background activity of EEG. The presence of a burst suppression EEG pattern and a hypoactive/flat EEG are negative prognostic criteria.