Objective To investigate the efficacy differences of different doses of cyclophosphamide (CTX) among subcategories of diffuse proliferative lupus nephritis (LN). Methods Clinical data of 133 LN patients diagnosed by renal biopsy with class Ⅳ or class Ⅳ+Ⅴ who were treated with corticosteroid plus CTX were analyzed retrospectively. The baseline Scr, 24 h urine protein, CTX dosages and prognosis were compared among different dosages for each subcategory. Results The average cumulative dose of CTX within 6 months was (11.1±4.1) g. The high dose group was >12 g, the medium dose group was >6-12 g and the low dose group was ≤6 g. Compared to low dose group, high dose CTX increased the remission rate of class Ⅳ+Ⅴ (67% vs 40%, P=0.314) and chronic renal lesion (43% vs 0%, P=0.212), but such enhancement was not obvious in class Ⅳ (Ⅳ-S: 67% vs 50%, P=0.548, Ⅳ-G: 65% vs 70%, P=0.560). Difference of overall adverse reactions was not significant between high dose group and low dose group (51% vs 37%，P=0.224). Conclusion Corticosteroid plus high dose CTX may improve the remission rate of patients with class Ⅳ+Ⅴ and chronic renal lesions.

Objective To investigate the impact of metabolic syndrome (MS) on clinicopathology of IgA nephropathy (IgAN). Methods A total of 118 IgAN patients complicated with MS were enrolled in the study as IgAN-MS group. Then 118 IgAN patients of same age arrange without MS were randomly selected as IgAN-non-MS group. A comparative analysis of clinical and pathological data between these two groups was performed. Results The urine protein, serum creatinine, body mass index, mean arterial pressure, serum triglyceride, fasting blood glucose and serum uric acid in IgAN-MS group were all significantly higher than those in IgAN-non-MS group (P＜0.05 or P＜0.01). The serum HDL-C level in IgAN-MS group was significantly lower than that in IgAN-non-MS group (P＜0.01). The percentages of patient with hypertension, abnormal glucose metabolism or abnormal lipid metabolism in IgAN-MS group were also significantly higher than those in IgAN-non-MS group (P＜0.01). The glomerular and tubulointerstitial pathological changes in IgAN-MS group were significantly more severe than those in IgAN-non-MS group (P＜0.01). There were significantly positive correlations between MS and urinary protein quantity, serum creatinine level, and glomerular damage index or tubulointerstitial damage index (P＜0.01) by Spearman rank correlation analysis. Conclusion MS may be an important risk factor of IgAN progression.

Objective To investigate the clinicopathologic characteristics, classification and outcome in lupus nephritis(LN) patients with thrombotic microangiopathy（TMA）. Methods LN patients with TMA proven by renal biopsy, from January 2000 to February 2009 in our hospital were enrolled. They were classified as poly-immunocomplex deposit group (n=35) and pauci-immunocomplex deposit group (n=25). Their clinicopathologic features and outcome were analyzed retrospectively. Results (1)The incidence of TMA in lupus nephritis was 9.2% (n=62), which presented severe hypertension, prominently elevated serum creatinine, anemia, thrombocytopenia, and was also the poorest prognosis of all the vascular lesion types. The incidence of death/end stage renal disease (ESRD) was 25.0%, with a mortality rate of 13.6%. (2) According to immunocomplex deposit in renal tissue, LN complicated with TMA could be classified as “poly-immunocomplex deposit subtype” and “pauci-immunocomplex deposit subtype”. The former presented better response to steroid and immunosuppressant therapy, in spite of more active clinicopathologic manifestations. The incidences of death/ESRD of poly subtype and pauci subtype were 8.8% and 32.0% respectively. Conclusions TMA presenting severe manifestations and the poorest prognosis is not rare in LN. LN with TMA may be classified as poly-immunocomplex deposit subtype and pauci-immunocomplex deposit subtype. This classification may be helpful in prognosis because the latter shows bad response to steroid- immunosuppressant therapy.

Objective To examine the correlation of urinary podocyte number and giomerular podocalyxin expression with clinicopathology in IgA nephropathy (IgAN) patients. Methods Morning urinary specimens (100 ml) 3 days before renal biopsy from 50 patients with IgAN diagnosed by renal biopsy and from 20 healthy volunteers as control were collected. After centrifugation, 300 μl sediment was used for smear. Immunohistochemical staining with monoclonal anti-podocalyxin antibody was performed to detect urinary podocytes and the number of podocyte was counted under optical microscope. Computer image analysis system was used to examine glomerular PCX expression. Renal pathology and classification were investigated based on Lee’s grading and Katafuchi semi-quantitative integration method. Relevance analysis was carried out on urinary podocyte number, glomerular PCX expression with pathological score and clinical data. Results The amount of urinary podocytes in IgAN was obviously higher than that in healthy controls (P<0.01). Significant differences were found in multiple comparison of the median of urinary podocytes among Lee’s grade groups. Ⅰ-Ⅱ group was lower as compared to Ⅲ, Ⅳ, Ⅴ groups (all P<0.05). Ⅲ group was lower as compared to Ⅴ group (P<0.05). The positive rate of urinary podocyte was the highest in Ⅳ and Ⅴgroups (100%), while the lowest in Ⅰ-Ⅱ group (55%). Glomerular PCX expression in IgAN decreased with the aggravation of renal pathology. Significant differences were found in multiple comparison of the glomerular PCX expression with the pathological score. Lee’sⅠ-Ⅱ group was higher as compared to Ⅲ, Ⅳ, Ⅴ groups (all P<0.05). Ⅲ and Ⅳ groups were higher as compared to Ⅴ group (P<0.05). In IgAN, urinary podocyte excretion was negatively correlated with glomerular PCX expression (r=-0.702, P<0.01), positively correlated with 24-hour urinary protein (r=0.465, P<0.01) and positively correlated with glomerular and tubular scores (r=0.233, 0.307, P<0.05). Glomerular PCX expression was negatively correlated with 24-hour urinary protein (r=-0.367，P<0.05) and negatively correlated with glomerular and tubular scores (r=-0.560,-0.377, P<0.05). Conclusions Injury and desquamation of glomerular podocytes may involve in the development of IgAN. The number of urinary podocyte can reflect the loss of podocytes in renal tissue, which may be used as a marker of disease progression of IgAN.

Objective To study the change of serum insulin-like growth factor 1 (IGF-1) in primary nephrotic syndrome (PNS) patients and its relationship with bone metabolism, and to investigate the clinical significance of IGF-1 in the mechanism of bone metabolic disorders in PNS patients. Methods A total of 30 PNS patients with chronic kidney disease(CKD) stage 1 and 2 were randomly selected from 2008.1 to 2009.5 in our hospital. Serum IGF-1, albumin, calcium, phosphorus, PTH，25 hydroxy vitamin D3, bone gla protein (BGP), degradation products of C-terminal telopeptides of type I collagen (CTx), 24-hour urinary protein excretion, and ratio of urinary calcium to creatinine (UCa/Cr) were measured. Healthy control group of 61 persons were randomly selected from our medical examination center at the same time. Results Serum levels of calcium, 25 hydroxy vitamin D3 and BGP were significantly lower; CTx and UCa/Cr were significantly higher in PNS patients (P<0.05) as compared to healthy control group. BMD of PNS patients was lower but without significant difference compared with healthy control group [(1.078±0.090) g/cm2 vs (1.090±0.062) g/cm2, P>0.05]. Serum level of IGF-1 was significantly lower in PNS patients and was positively correlated with BMD and BGP, and negatively correlated with 24-hour urinary protein excretion and CTx. Conclusions Bone metabolic disorder exists in PNS patients with the appearance of decreased bone formation and increased bone absorption. Serum level of IGF-1 has good correlations with bone biochemical markers, which may be used as a new bone biochemical marker of bone metabolism in kidney disease.

Objective To investigate the underlying mutation in a late-onset Chinese patient with classic Bartter syndrome. Methods The mutation analysis of CLCNKB gene was performed by the PCR direct sequencing. The patient’s parents and siblings were studied as well. Fifty normal volunteers were analyzed as control group. Results The heterozygous deletion mutation cDNA 753delG and heterozygous missense mutation G433E were detected in the patient. Her father was found to carry heterozygous G433E and her mother to carry cDNA 753delG mutation respectively. Her brother carried heterozygous G433E and her sister was normal. Conclusions Two mutations of the CLCNKB gene in this Chinese patient with late-onset classic Bartter syndrome are identified. The cDNA 753delG mutation has not been reported previously.

Objective To study the role of PI3K-Akt-GSK-3β signaling in the apoptosis of renal tubular cells after ischemia-reperfusion injury and the protective mechanism of recombinant human erythropoietin (rHuEPO). Methods The human kidney tubular epithelial cells (HK-2) were cultured in vitro in different conditions as control group with serum, ischemia-reperfusion (IR)group, LY294002 group with LY294002 (AKT inhibitor) 10 μmol/L 30 minutes before IR treatment, LiCl group with LiCl (GSK-3β inhibitor) 20 μmol/L 30 minutes before IR treatment, rHuEPO group with EPO 20 U/ml 30 minutes before IR treatment, rHuEPO+LY294002 group with EPO 20 U/ml and in the presence of LY294002 (10 μmol/L) 30 minutes before IR treatment, rHuEPO +LiCl group with EPO 20 U/ml and in the presence of LiCl (20 μmol/L) 30 minutes before IR treatment. Akt, GSK-3β and caspase-3 activation were measured by Western blotting. The apoptotic ratio of HK-2 cells was measured by flow cytometry. Cell viability was detected by MTT. Results In comparison with the control group, the apoptotic ratio raised up to 15.20%±1.43％, the expression of Akt activity decreased, GSK-3β activity and caspase-3 activity markedly elevated in IR group (P<0.05). LY294002 group up-regulated the apoptotic ratio (18.20%±2.06％), decreased the expression of Akt activity, increased GSK-3β activity and caspase-3 activity, however, LiCl group down-regulated the apoptotic ratio (12.30%±0.85％), increased the expression of Akt activity, decreased GSK-3β activity and caspase-3 activity compared with IR group (P<0.05). rHuEPO group remarkably decreased the apoptotic ratio (11.10%±1.62％), increased the expression of Akt activity, decreased GSK-3β activity and caspase-3 activity compared with IR group (P<0.05). rHuEPO+LY294002 group elevated the apoptotic ratio (13.40%±1.94％), decreased the expression of Akt activity, increased GSK-3β activity and caspase-3 activity, meanwhile, rHuEPO +LiCl group down-regulated the apoptotic ratio (7.50%±1.31％), increased the expression of Akt activity, decreased GSK-3β activity and caspase-3 activity compared with rHuEPO group (P<0.05). Conclusions PI3K-Akt-GSK-3β signaling pathway is involved in HK-2 cells apoptosis induced by ischemia-reperfusion injury and rHuEPO may be used as a new therapy.

Objective To investigate the effect of surfactant protein D (SP-D) overexpression on lipopolysaccharide(LPS)-induced monocyte chemoattractant protein-1(MCP-1) expression in human renal proximal tubular epithelial cells (HK-2) and its mechanism. Methods HK-2 cells were treated with LPS at various concentrations(0, 0.1, 1, 2, 5, 10 mg/L) for 8 h and at 5 mg/L for various time points (0, 2, 4, 8, 16, 24 h). Expression of SP-D was detected by Western blotting and real-time PCR. Expression of MCP-1 was determined by ELISA and real-time PCR. Human SP-D cDNA eukaryotic expression vector pEE14-hSP-D was transfected to HK-2 cells. The changes in transfected cells of SP-D protein were observed by Western blotting. Expression of MCP-1 was detected by ELISA and real-time PCR. Results SP-D was expressed in HK-2 cells. The levels of SP-D protein and mRNA in HK-2 cells were significantly decreased after treatment with LPS (P<0.05). Expression of MCP-1 protein and mRNA was increased remarkably after treatment with LPS (P<0.05). HK-2 cells transfected with pEE14-hSP-D showed up-regulated expression of SP-D. The overexpression of SP-D inhibited the LPS-induced expression of MCP-1(P<0.01). Conclusions SP-D inhibits LPS-induced expression of MCP-1 in HK-2 cells. SP-D may play an important role in the modulation of renal inflammation.

Objective To investigate the protective effect of intermedin(IMD) on renal ischemia reperfusion injury(IRI) and its mechanism. Methods A total of twenty-four male Wistar rats were randomly divided into four groups: control group, IRI group, empty plasmid group and IMD group. After remove of right kidney, plasmid was transfected into the kidney by ultrasonic microbubbles technology, and IRI model was made after 1 week. Renal pathology was observed by PAS staining. Renal tissue superoxide dismutase (SOD), myeloperoxidase (MPO), caspase-3 activity, and malondialdehyde (MDA) content were detected by colorimetric method. The intercellular cell adhesion molecule-1(ICAM-1), endothelin 1(ET-1) and P-selection expression of renal tissue were detected by immunohistochemical method. Apoptosis of renal tubular cell was detected by TUNEL. Results Compared with control group, tubulointerstitial pathological injury was significant aggravated in IRI group(P<0.01); compared with IRI group, IMD pretreatment significantly alleviated the degree of renal injury(P<0.01). Compared with control group, in IRI group, SOD activity was significantly decreased (P<0.05), MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 were increased significantly (all P< 0.01). Compared with IRI group, IMD pretreatment significantly increased SOD activity (P<0.05), decreased the MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1(all P<0.01). The apoptosis rate of renal tubular epithelial cells in IRI group was significantly higher than that in control group (34.83%±8.75% vs 3.33%±0.47%, P<0.01), while the apoptosis rate of IMD group (20.67%±7.71%) was significantly lower than that of IRI group. There was no difference of above indexes between empty plasmid group and IRI group. Conclusions IMD pretreatment protects against renal IRI. The mechanism may be at least partly related to the clearance of oxygen free radicals, the improvement of lipid peroxidation, inflammatory cell infiltration and cell apoptosis, leading to the decrease of the production of reactive oxygen species caused by oxidative stress.

Objective To investigate the possible correlation between chronic periodontitis (CP) and chronic renal failure(CRF) by establishing chronic periodontitis and chronic renal failure model in SD rats. Methods Forty health male SD rats were divided into four groups: control group (A), CP group (B), CRF group (C), CP accompany with CRF group (D). Ten rats were sacrificed in every group at the end of week 8. The periodontal index, levels of serum Scr and BUN, the concentration of IL-1β and TNF-α were examined. The severity CP and CRF was quantified by histopathology. The date was statistically analyzed. Results Animal models were established successfully. Scr and BUN in group D, BUN were higher than that in group C [Scr (120.54±21.29) μmol/L vs (93.63±18.82) μmol/L, BUN (34.20±14.44) mmol/L vs (17.77±4.15) mmol/L, P<0.05]. The kidney change of inflammation was observed in group B, the grade of PAS and Masson in group C and D were higher than that in group A(P<0.01), and that in group D was higher than group C(P<0.05). Obvious inflammation of periodontal tissue was observed in group B and D. Attachment loss level (AL) in group D was higher than that in group B [(173.60±16.75) μm vs (124.00±23.87) μm, P<0.05]. The level of IL-1β and TNF-α in group B and C and D were higher than that in group A(P<0.05), and IL-1β in group D was higher than that in group B and C (P<0.05), TNF-α in group D was higher than that in group B(P<0.05). 2×2 factorial design revealed that there were interactions between CP and CRF on the numerus of Scr and BUN and AL P<0.05), and the influence of each factor on that was significant (P<0.05), no interactions were noted between CP and CRF on IL-1β and TNF-α(P>0.05), but the influence of each factor on that was significant (P<0.05). Conclusions The SD rat models can appear chronic periodontitis and chronic renal failure at the same time. There is correlation between chronic periodontitis and chronic renal failure. Chronic periodontitis can aggravate chronic renal failure throngh the role of inflammation.

Objective To explore the effects of ethyl-3,4 dihydroxybenzoate (EDHB), a prolyl hydroxylase inhibitor, pretreatment on the tubular epithelial cells apoptosis induced by albumin and hypoxia. Methods To investigate the effects of albumin and hypoxia on cells, rat tubular epithelial cells (NRK-52E) were incubated for 24 h in: (1) normoxia (5％CO2); (2) hypoxia (1% O2); (3) albumin (30 g/L) under normoxia; (4) albumin (30 g/L) under hypoxia. To investigate the effects of EDHB pretreatment on cells apoptosis, NRK-52E were incubated in hypoxia for 24 h in: (1) normoxia; (2) hypoxia; (3) hypoxia+albumin (30 g/L); (4) hypoxia+EDHB（500 μmol/L）; (5) EDHB pretreatment (albumin 30 min after EDHB). Apoptosis was measured by flow cytometry (AnnexinV-FITC-PI). bcl-2, bax and vascular epithelial growth factor (VEGF) mRNA expression were detected by RT-PCR. VEGF protein expression was detected by Western blotting. Results NRK-52E apoptosis was not significantly different between hypoxia and normoxia groups (P>0.05), but increased significantly in albumin (30 g/L) under hypoxia group compared with albumin (30 g/L) under normoxia group (37.36%±4.95% vs 25.59%±3.32%, P< 0.05). There was an increase in bax mRNA expression and a decrease in bcl-2 mRNA expression in albumin (30 g/L) under hypoxia group compared with albumin (30 g/L) under normoxia group (P< 0.05). EDHB pretreatment improved these impairments of albumin (30 g/L) under hypoxia on NRK-52E (P<0.05). VEGF expression elevated in hypoxia compared with normoxia (P<0.05), decreased in albumin (30 g/L) under hypoxia groups compared with that without albumin groups (P<0.05). EDHB pretreatment significantly improved VEGF expression compared with albumin (30 g/L) under hypoxia group (P<0.05). Conclusion NRK-52E cells apoptosis induced by albumin is accelerated by hypoxia, however partially improved by EDHB pretreatment, probably through the up-regulation of VEGF expression.

Objective To investigate the renoprective effect and its possible mechanism of Niaoduqing on the adriamycin(ADR)-induced nephropathy rats. Methods Forty eight male Sprague-Dawley rats were randomly divided into normal control (n=12), ADR-induced nephropathy(model, n=12), Benazepril-treated ADR nephropathy(Benazepril, n=12) and Niaoduqing-treated ADR nephropathy (Niaoduqing, n=12) groups. The rat nephropathy model was established by adriamycin injection and unilateral nephrectomy. The rats were sacrifficed per batch at the 4th and 8th weekend. The pathological change of nephridial tissue, the 24-hour urinary protein excretion and renal function were examined. Immunohistochemistry was used to meassure the expression of fibronection(FN), collagenⅣ(COLⅣ), osteopontin (OPN). Results The 24-hour urinary protein excretion, BUN, Scr, triglyeride (TG), cholesterol (Cho) in model group were significantly higher than those in normal group(P<0.01）, as well as more server glomerulosclerosis in kidney were observed in model group than those in control group(P<0.01), while the albumin (Alb) was lower (P<0.01). Compared with model group, the 24-hour urinary protein excretion, BUN, Scr, TG, Cho were significantly reduced and renal glomerulosclerosis was improved in Niaoduqing group (P<0.01）, while the Alb was higher (P<0.01). Conclusion Niaoduqing palys an important role in the prevention and treatment for nephropathy.