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ObjectivesTo evaluate renal expression of C4d, a complement component in the classical/mannose binding lectin (MBL) pathway, in patients with primary Sjögren’s syndrome (pSS)-associated renal impairments.MethodsWe retrospectively reviewed the clinical and pathological data from 39 patients with pSS presenting with renal impairments. C4d was examined in paraffin-embedded biopsy tissues using immunohistochemistry. Glomerular C4d positive was defined when >75% glomeruli were globally stained. Tubulointerstitial C4d (TI-C4d) were scored semi-quantitatively as 0 (absent), 1 (spotty or weak), 2 (patchy) and 3 (diffuse). A TI-C4d score ≥2 was considered TI-C4d positive and included in the TI-C4d+ group and vice versa. Peritubular capillary (PTC) C4d was scored as 0 (absent), 1 (0∼10%, minimal), 2 (10%∼50%, focal), and 3 (>50%, diffuse).ResultsGlomerular C4d deposition was observed in all 8 patients with pSS-related membranous nephropathy (MN) without obvious C1q deposition. Two of 5 patients with mesangial proliferative glomerulonephritis and 1 of 2 patients with IgA nephropathy had mild mesangial C4d deposition. Sixteen patients (6 glomerular dominant and 10 tubulointerstitial dominant) presented TI-C4d score ≥2. Patients in the TI-C4d+ group exhibited a higher serum creatinine level at the time of renal biopsy (TI-C4d+ 132.5 [89.7, 165.5] vs. TI-C4d- 83.0 [70.7, 102.0] μmol/L, P=0.008). PTC C4d was observed in 12 patients, with each of minimal, focal and diffuse staining being noted in 4 patients.ConclusionsThe MBL pathway of complement activation was potentially involved in pSS-related MN. Tubulointerstitial C4d might be a pathological marker of severe renal injury in patients with pSS-related renal impairments.

Aim To investigate the effect of low-dosage steroid therapy in patients with severe aristolochic acid nephropathy (AAN). Methods Forty-three chronic AAN patients in the Peking Union Medical College Hospital and the First Affiliated Hospital of Xinxiang Medical College were included in this study from November 1998 to October 2013. According to the treatment method, the patients were divided into a steroid group (SG, n = 25) and a control group (CG, n = 18). The serum biochemical indicators at the basement in the two groups exhibited no obvious statistical differences. In comparison with the baseline data, the levels of serum creatinine at 3, 6, 9, and 12 months were analyzed. The blood pressure, haemoglobin, serum biochemical indicators, and the side-effects of steroid application were also observed. Urinary macrophage chemoattractant protein-1 (MCP-1) and transforming growth factor-1 (TGF-1) amounts were measured as well. Results (i) The serum creatinine content in the CG group was significantly higher than the baseline level during the follow-up(6, 9, and 12 months later), whereas in the SG group it decreased during the 3-6 month period and remained stable within 1 year. (ii) The biochemical indicators, blood pressure, and haemoglobin persisted stable. (iii) The side-effects of low-dosage steroid therapy were not severe and were tolerated by the AAN patients. (4) Urinary MCP-1 and TGF-1 concentrations were positively correlated with serum creatinine and decreased in the SG group. Conclusion Low-dosage steroid therapy reversed or delayed the renal failure progression in severe chronic AAN patients, which may be associated with the suppression of MCP-1 and TGF-β1 activities.

Transforming growth factor-β1 (TGF-β1)‑induced epithelial‑mesenchymal transition (EMT) is one of the important cellular and molecular mechanisms involved in renal fibrosis. Smad3 and miR-192 (a Smad3-dependent microRNA) are involved in TGF-β1-mediated EMT. Vascular endothelial growth factor (VEGF) is a renal tubular epithelial survival factor. Therefore, in the present study, we investigated the role of Smad3 and miR‑192 in the effects of VEGF on TGF‑β1‑mediated tubular EMT. A human kidney cortex (HKC) cell line stably overexpressing VEGF (HKC-SOEV) was established. The normal HKC cells and HKC‑SOEV cells were treated with TGF-β1 (5 µg/l) or/and LY294002 (20 µmol/l) for 24 and 48 h (LY294002 blocks the effect of VEGF). The protein expression of Smad2, Smad3, Smad4 and phosphorylated Smad3 (p‑Smad3) were measured by western blot analysis. The expression of Smad3 and miR-192 was determined by real‑time PCR. E-cadherin and α-smooth muscle actin (α-SMA) expression was detected by western blot analysis and laser scanning confocal microscopy (LSCM). TGF-β1 was found to induce the expression of α-SMA in the HKC cells. TGF-β1 also induced Smad3, miR-192 and p-Smad3 expression, but suppressed E‑cadherin expression. However, in the HKC-SOEV cells, the expression levels of α-SMA, Smad3, miR-192 and p‑Smad3 upon TGF-β1 stimulation were significantly reduced. In these cells, the suppressive effect of TGF-β1 on E‑cadherin was also reduced. Importantly, treatment with LY294002 significantly diminished the effect of VEGF. VEGF suppressed Smad3 and miR‑192, and subsequently inhibited EMT induced by TGF-β1.

Recent information indicates that pentoxifylline (PTX) has the ability to suppress inflammation and profibrotic cell proliferation. In this study, we investigated the effect of PTX on tubulointerstitial fibrosis and the expression of vascular endothelial growth factor (VEGF) in a rat model of obstructive nephropathy.Wistar rats with left ureteral ligation were divided into control and PTX-treated groups. The histopathologic degree of tubulointerstitial fibrosis was scored with PAS and Masson-stained sections. The protein and mRNA for vascular endothelial growth factor (VEGF) were semiquantitatively measured with immunohistochemistry and RT-PCR. The protein for transforming growth factor beta1 (TGFbeta1) and hypoxia-induced factor 1 alpha (HIF-1alpha) was determined by Western blot.Compared with the control group, PTX treatment reduced fibrosis scores at d 7 and d 14 (P0.05). The reduction was accompanied by inhibited expression of transforming growth factor-beta 1 (TGFbeta1), a key cytokine in tubulointerstitial fibrogenesis (P0.01). Meanwhile, VEGF protein and mRNA in the kidney were increased in the PTX-treated group compared with the control group (P0.01). PTX up-regulated expression of VEGF mRNA in a dose- and time-dependent manner in cultured HK-2 cells (P0.01). However, expression of HIF-1alpha (a key transcription factor for VEGF gene expression) was unchanged by PTX treatment. PTX prolonged the half-life of VEGF mRNA by a 1.07-fold increase.PTX inhibited tubulointerstitial fibrosis in a rat model of obstructive nephropathy while preventing loss of VEGF. PTX up-regulated expression of VEGF mRNA through stabilization of its mRNA in cultured renal tubular epithelial cells.

To investigate the effect of low-dosage steroid therapy in patients with severe aristolochic acid nephropathy (AAN).Forty-three chronic AAN patients in the Peking Union Medical College Hospital and the First Affiliated Hospital of Xinxiang Medical College were included in this study from November 1998 to October 2013. According to the treatment method, the patients were divided into a steroid group (SG, n = 25) and a control group (CG, n = 18). The serum biochemical indicators at the basement in the two groups exhibited no obvious statistical differences. In comparison with the baseline data, the levels of serum creatinine at 3, 6, 9, and 12 months were analyzed. The blood pressure, haemoglobin, serum biochemical indicators, and the side-effects of steroid application were also observed. Urinary macrophage chemoattractant protein-1 (MCP-1) and transforming growth factor-1 (TGF-1) amounts were measured as well.(i) The serum creatinine content in the CG group was significantly higher than the baseline level during the follow-up(6, 9, and 12 months later), whereas in the SG group it decreased during the 3-6 month period and remained stable within 1 year. (ii) The biochemical indicators, blood pressure, and haemoglobin persisted stable. (iii) The side-effects of low-dosage steroid therapy were not severe and were tolerated by the AAN patients. (4) Urinary MCP-1 and TGF-1 concentrations were positively correlated with serum creatinine and decreased in the SG group.Low-dosage steroid therapy reversed or delayed the renal failure progression in severe chronic AAN patients, which may be associated with the suppression of MCP-1 and TGF-β1 activities.

Vascular endothelial growth factor (VEGF) has been shown to be a survival factor for renal tubular epithelial cells. In the present study, we investigated whether administration of VEGF ameliorates tubulointerstitial fibrosis in a mouse model of unilateral ureteral obstruction (UUO).Thirty-six male CD-1 mice were randomly divided into three groups: sham-operation, UUO and UUO+VEGF group. VEGF (50 μg/kg) was subcutaneously injected twice daily from d 1 to d 14. Mice in each group were killed at d 3, 7, or 14 after the operation, and the tubulointerstitial fibrosis was histopathologically evaluated. Human proximal tubular epithelial cells (HK-2) were used for in vitro study. The expression levels of α-SMA, E-cadherin, TGF-β1, CTGF, and BMP-7 in the kidney were determined using Western blot and RT-PCR.In the UUO mice, the degree of interstitial fibrosis was dramatically increased in a time-dependent manner. At d 3, 7, and 14, both the mRNA and protein expression levels for α-SMA, TGF-β1, and CTGF were significantly upregulated, whereas those for E-cadherin and BMP-7 were significantly downregulated. At d 3 and 7, VEGF treatment significantly reduced interstitial fibrosis and the expression levels for α-SMA, TGF-β1, and CTGF, while significantly increased the expression of E-cadherin and BMP-7, as compared with the UUO mice. At d 14 after operation, no significant differences were observed in the expression of the examined markers between VEGF-treated mice and UUO mice, with the exception of CTGF. In HK-2 cells, VEGF blocked TGF-β1-induced α-SMA and vimentin expression and restored E-cadherin expression in a dose-dependent manner.VEGF may ameliorate renal tubulointerstitial fibrosis at the early stage in UUO mice. This effect may be related to inhibition of VEGF on renal tubular epithelial-mesenchymal transition (EMT).

To examine the relationship between effect of vascular endothelial growth factor (VEGF) on epithelial-myofibroblast transition (EMT) of HK-2 cells and changes in expressions of bone morphogenetic protein-7 (BMP-7) and inhibitor of DNA binding/differentiation (Id) 2, Id3.The cultured HK-2 cells were co-treated with transforming growth factor-beta1 (TGF-beta1) (5 ng/ml) and VEGF165 (0.1, 1, 10, 100 ng/ml), or with TGF-beta1 (5 ng/ml) and VEGF receptor-1 neutralized antibody (10 microg/ ml), and were also co-treated with TGF-beta1 (5 ng/ml) and VEGF165 (100 ng/ml) with or without activin receptor-like kinase 6 (Alk6)/Fc Chimera (2 microg/ml, to neutralize endogenous BMP-7) for 48 hours. mRNA and protein expressions of alpha-smooth muscle actin (alpha-SMA), E-cadherin, BMP-7, Id2 and Id3 of HK-2 cells were assessed with double-stain immunocytochemistry, real-time PCR and Western blot respectively.Compared with normal controls, alpha-SMA expression significantly increased, while E-cadherin, BMP-7, Id2, and Id3 mRNA and protein expressions markedly decreased in HK-2 cells treated with TGF-beta1 (5 ng/ml) (P0.05). VEGF165 interrupted TGF-beta1 induced alpha-SMA expression in a dose-dependent manner and upregulated BMP-7, Id2 mRNA and protein expressions of the cells (P0.05). alpha-SMA expression increased, while E-cadherin, BMP-7, and Id2 expressions decreased further in HK-2 cells co-treated with TGF-beta1 and VEGFR1 antibody compared with normal controls (P0.05). When endogenous BMP-7 was neutralized with Alk6/Fc Chimera in the cells co-treated with TGF-beta1 and VEGF165, alpha-SMA expression upregulated (P0.05), while Id2 was not changed.VEGF165 may partially inhibit TGF-beta1-induced EMT of HK-2 cells in vitro. This effect is related to the upregulated expressions of BMP-7 and Id2. Id2 may be upregulated directly by VEGF165, but not related to BMP-7.

The aim of this study is to examine the effect of mycophenolate mofetil (MMF) on epithelial-myofibroblast translation (EMT) in adenine-induced chronic renal failure (CRF) rat model and the role of vascular endothelial growth factor(VEGF) and inhibitor of differentiation (Id2 and Id3) in EMT in the rat kidney.Sixty-four male Wistar rats were randomly assigned to the following groups: normal control (n = 16), CRF (n = 24) and MMF(n = 24). CRF was induced by gastric gavage of adenine (125 mg kg(-1) d(-1)) to rats for eight weeks. CRF rats were treated with MMF (15 mg kg(-1) d(-1)) as "MMF" group. The rats were sacrificed at week 2, 4, 6 and 8, respectively. Urinary protein and serum creatinine levels were measured, and the histopathologic degrees of interstitial fibrosis were evaluated in Masson-stained sections. Expressions of alpha-smooth muscle actin (alpha-SMA), transforming growth factor beta1 (TGFbeta1), VEGF and Id (Id2 and Id3) in the kidney tissue were assessed by immunohistochemistry, RT-PCR and/or Western blot methods.The urinary protein level in MMF group was evidently lower than that in CRF group (P0.01), whereas no statistically significant difference was observed in serum creatinine level between the two groups. Renal interstitial fibrosis was reduced significantly with MMF treatment (P0.01). Expression of alpha-SMA in MMF group was lower than that in CRF rats at week 6, 8 (P0.01), while expression of TGFbeta1 was decreased markedly at week 2, 4, 6 (P0.01). The expressions of VEGF in MMF rats were increased significantly at week 6, 8 (P0.01), and Id2, Id3 in MMF rats were increased significantly at week 4, 6 (P0.05).MMF may ameliorate chronic renal fibrosis and EMT in adenine-induced CRF rats. This effect of MMF on EMT is probably related to upregulation of VEGF, Id2 and Id3 expressions and suppressing overexpression of TGFbeta1 in renal tissue. The exact mechanism needs to be studied further.

To examine the effects of monocyte chemoattractant protein-1 (MCP-1) on epithelial-myofibroblast transition (EMT) of renal proximal tubular epithelial cells and the possible mechanism involved.Human renal proximal tubular epithelial cells of the line HK-2 were cultured and divided into three groups: negative control group; positive control group, treated with transforming growth factor (TGF)-beta1 5 microg/L, and MPC-1 group, treated with MCP-1 0.1, 1, 10, and 100 microg/L for 24 h, 36 h, 48 h, and 72 h respectively. The expressions of alpha-smooth muscle actin (alpha-SMA) mRNA and protein of cells were detected by s. Western blotting and RT-PCR were used to detect the expression of P-Erk1/2, Erk1/2, P-P38MAPK, P38MAPK, and RhoA protein levels of the HK-2 cells, and RT-PCR was used to detect the expression of RhoC and Snail mRNA. Specific inhibitors of the Erk, P38MAPK, and Rho signal transduction pathways PD98059, SB203580, and Y27632 were added into the culture fluid of HK-2 cells respectively, 1 h later MCP-1 microg/L was added for 48 h, and Western blotting was used to detect the protein expression of alpha-small muscle actin (SMA).The expression levels of alpha-SMA protein and mRNA significantly increased in the MCP-1 treated HK-2 cells,and these expression levels were the highest in the HK-2 cells treated with MCP-1 1 microg/L for 48 h. The ratios of (P-Erk1/2)/ (Erk1/2) and P-P38MAPK/P38MAPK were significantly increased (all P0.05) in the MCP-1 treated HK-2 cells with the highest ratios seen in the HK-2 cells treated by 100 microg/L of MCP-1. The expression levels of RhoA protein and RhoC mRNA of the HK-2 cells stimulated with MCP-1 of different concentrations were not significantly different (all P0.05). MCP-1 induced expression of a-SMA was only partly inhibited by PD98059 but not by SB203587 or Y27632. MCP-1 dose-dependently increased the expression of Snail mRNA of the HK-2 cells compared with those of the negative control cells. The level of Snail mRNA was the highest in the HK-2 cells treated with 100 microg/L MCP-1.MCP-1 may induce the EMT of renal proximal tubular epithelial cells in vitro, and this effect may involve upregulation of Erk1/2 Map kinase signal pathways and Snail mRNA expression. P38MAPK or Rho kinase signal pathways can not be proven to be involved in MCP-1 induced EMT.

To examine the effect of Bone Morphogenetic protein-7 (BMP-7) on Monocyte chemoattractant protein-1 (MCP-1) induced epithelial-myofibroblast transition (EMT) in cultured renal proximal tubular cells (HK-2) and the relationship between TGF-beta1-smad 3 expressions and MCP-1 induced EMT.The cultured HK-2 cells were divided into six groups: a, negative control, b, treated with TGF-beta1 (5 ng/ml) as positive control, c, treated with MCP-1 (0.1, 1, 10, 50 ng/ml), d, treated with BMP-7 (0.1, 1, 10, 50 ng/ml), e. co-treated with MCP-1 (1 ng/ml) and MCP-1 neutralized antibody (1 ng/ml), f. co-treated with MCP-1 (1 ng/ml) and BMP-7 (50 ng/ml). alpha-Smooth Muscle Actin (alpha-SMA) mRNA expression of HK-2 cells was assessed with RT-PCR. Secretion of type I collagen was assessed with RT-PCR and ELISA, respectively. TGF-beta1 and Smad 3 expressions were assessed with Western blot.alpha-SMA mRNA expression significantly increased in HK-2 cells treated with MCP-1 (0.1, 1 ng/ml) compared with negative controls (5.97 +/- 0.35, 23.36 +/- 1.37 vs. 0.59 +/- 0.38, P0.01). alpha-SMA mRNA expression of HK-2 cells concomitantly treated with MCP-1 neutralized antibody or BMP-7 (50 ng/ml) and MCP-1 (1 ng/ml) significantly decreased than that in cells treated with MCP-1 (1 ng/ml) alone (1.93 +/- 0.34, 13.59 +/- 0.38 vs. 36.36 +/- 1.37, P0.01). Secretion of type I collagen of the cells treated with MCP-1 (0.1, 1 ng/ml) markedly increased compared with negative control (1751 +/- 34, 1876 +/- 45 vs. 1450 +/- 62; P0.01). The secretion of type I collagen of the supernatant were also significantly lower than that in cells treated with MCP-1 (1 ng/ml) alone (1462 +/- 56, 1596 +/- 34 vs. 1876 +/- 45, P0.05). The expression of TGF-beta1 and Smad 3 of HK-2 cells treated with MCP-1 (1 ng/ml) were markedly higher than that of negative controls, respectively (36.31 +/- 1.37 vs. 0.75 +/- 0.16, P0.01; 56.98 +/- 2.61 vs. 23.05 +/- 1.82, P0.01). The expressions of TGF-beta1 and Smad 3 in HK-2 cells treated concomitantly with MCP-1 neutralized antibody or BMP-7 (50 ng/ml) and MCP-1 (1 ng/ml) were markedly decreased than that treated with MCP-1 alone, respectively. (4.61 +/- 0.74, 23.74 +/- 2.14 vs. 36.31 +/- 1.37, P0.01; 19.63 +/- 1.65, 37.06 +/- 1.82 vs. 56.98 +/- 2.61, P0.01). The expressions of TGF-beta1 and Smad 3 in HK-2 cells treated concomitantly with MCP-1 neutralized antibody or BMP-7 (50 ng/ml) and MCP-1 (1 ng/ml) were markedly decreased than that treated with MCP-1 alone, respectively. (4.61 +/- 0.74, 23.74 +/- 2.14 vs. 36.31 +/- 1.37, P0.01; 19.63 +/- 1.65, 37.06 +/- 1.82 vs. 56.98 +/- 2.61, P0.01).The results documented that MCP-1 may induce EMT of HK-2 cells in vitro, and this effect is related to up-regulated expression of TGF-beta1 and Smad 3. BMP-7 may partially inhibit MCP-1-induced EMT and this effect is related to the downregulated expression of TGF-beta1 and Smad 3 of the cells. The results also suggest that MCP-1 induced EMT may involve the TGF-beta1-independent pathway of the cells.

To examine the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1, VEGFR2) in transdifferentiated human proximal tubular epithelial (HK-2) cell induced by transforming growth factor beta1 (TGFbeta1).The transdifferentiation of HK-2 cells was detected by evaluation of expression of alpha-SMA by cytoimmunochemistry and RT-PCR. The VEGF mRNA was evaluated with RT-PCR. The secreted VEGF in the culture media was measured with ELISA. The cellular VEGF, VEGFR1, and VEGFR2 were measured with Western blot.The immunostain of alpha-SMA were positive in HK-2 cell induced by TGFbeta1 at the concentration of 5 and 8 ng/ml for 72 h. The expression of alpha-SMA mRNA was induced by TGFbeta1 in concentration- and time-dependent manners. The expressions of mRNA and protein of VEGF were upregulated by TGFbeta1 at the concentration of 0.1 and 1 ng/ml for 72 h and at the concentration of 8 ng/ml for 12 h and 24 h when compared with the control. But expressions of mRNA and protein of VEGF were downregulated by TGFbeta1 at the concentration of 3, 5, and 8 ng/ml for 72 h and at the concentration of 8 ng/ml for 36, 48, and 72 h, respectively. Meanwhile, Protein levels of VEGFR1 and VEGFR2 were upregulated by TGFbeta1 in concentration- and time- dependent manners.Increased expression of VEGFR1 and VEGFR2 and two-phase change in VEGF expression occurred in the process of tubular epithelial transdifferentiation induced by TGFbeta1. Reduced expression of VEGF may contribute to tubular epithelial transdifferentiation in a vicious circle.

To observe the effect of bone morphogenetic protein-7 (BMP-7) on the transdifferentiation of cultured human tubular epithelial cell (HKC) induced by TGF-beta1 and to elucidate its possible mechanism.The cultured HKC cells were divided into 5 groups: serum-free group (negative control); single TGF-beta1 treated group (positive control); single BMP-7 treated group; combined TGF-beta1 and BMP-7 treated group; and BMP-7 pre-treated group. Expression of keratin of HKC cells was assessed by indirect enzyme immunohistochemistry (IEI), expression of alpha-smooth muscle actin (alpha-SMA) and E-cadherin by immunohistological method, percentage of alpha-SMA positive HKC cells by flow cytometry, and mRNA expression of alpha-SMA, TGF-beta1, and TGF-beta type II receptor by reverse transcription PCR.The expression of alpha-SMA and the percentage of alpha-SMA positive HKC cells markedly increased after having been treated by TGF-beta1 while the expression of E-cadherin and keratin decreased. In the group pre-treated with BMP-7 (50 ng/ml) and then added with TGF-beta1 (8 ng/ml), expression of alpha-SMA was significantly lower than in the positive control group, while expression of E-cadherin and keratin significantly higher than in the positive control group. Measurement of the percentage of alpha-SMA positive HKC found significant deference between the combined TGF-beta1 and BMP-7 treated group and the positive control group (9.7% vs 19.8%; 5.8% vs 19.8%; P0.05). Significant difference existed between the BMP-7 (50 ng/ml) pre-treated group and the positive control group (8.7% vs 19.8%, P0.05). mRNA expression of alpha-SMA was measured by RT-PCR and the results showed that it significantly decreased in the group treated or pre-treated with BMP-7 (50 ng/ml) (15% and 12% of the results in the positive control group, respectively). The mRNA expression levels of both TGF-beta1 and its type II receptor significantly decreased (28% and 19%; 47% and 36%, compared with the positive control group, respectively).Transdifferentiation of cultured renal epithelial cell induced by TGF-beta1 can be inhibittd by certain levels of BMP-7, cultured together with TGF-beta1 or pretreated. BMP-7 can prevent and inhibit the mRNA expression of TGF-beta1 and its type II receptor, which may be an important mechanism by which BMP-7 inhibit the transdifferentiation of renal tubular epithelial cell.

To examine the changes of expressions of orexin A, orexin receptor-1 (OX1R), prepro-orexin (Prepro-OX) mRNA, OX1R mRNA and ob-R of hypothalamus in rats with chronic renal failure (CRF).Sixty-two male Wister rats weighing 200-250 g were divided into three groups, including group 1 (normal, n = 5), group 2 (sham-operated, n = 25) and group 3 (CRF, n = 32). Hypothalamus orexin A was assayed by radioimmunoassay. Serum leptin was assayed by enzyme linked immunosorbent assay. The expression of Prepro-OX mRNA and OX1R mRNA of hypothalamus were measured by reverse transcription polymerase chain reaction, and expression of orexin A, OX1R and ob-R by immunohistochemistry. Automatic biochemical analyzer was used to measure the serum creatinine.Hypothalamus orexin A levels were negatively correlated (r = -0.63, P0.001) with serum leptin levels in the rats. The expression of hypothalamus Prepro-OX mRNA in CRF rats was significantly lower than that of sham-operation at week 12 (P0.01). Hypothalamus Prepro-OX mRNA levels were negatively correlated (r = -0.81, P0.001) with the levels of serum leptin and serum creatinine (r = -0.68, P0.05) in the rats at week 12. The expression of hypothalamus OX1R mRNA in CRF rats was lower than that of sham-operation at week 12 (P0.05). Specific immunoreactivity for orexin A was present in perikeryon of the hypothalamus neuron. Specific OX1R-like immunoreactivity was observed in some nerve fibres. Specific immunoreactivity for ob-R was present in membranes of the hypothalamus neuron. Hypothalamus neurons of orexin A-like specific immunoreactivity in CRF rats were significantly fewer than those in shamoperated rats at week 8. Hypothalamus neurons of OX1R-like specific immunoreactivity in CRF rats were similar to those in sham-operated rat at week 8. Hypothalamus neurons of ob-R-like specific immunoreactivity in CRF rats were significantly more than those in sham-operated rats at week 8.The lower hypothalamus orexin A levels may be induced by high serum leptin level in CRF rats. The lower expression of hypothalamus Prepro-OX mRNA in CRF rats may be one of the main causes inducing lower hypothalamus orexin A. The expression of OX1R in hypothalamus neurons is somewhat reduced and the expression of ob-R in hypothalamus neurons is somewhat raised in CRF rats. These remain to be studied further.

To explore whether there is phenotypic modulation of mesangial cells in streptozotocin (STZ) induced diabetic rats and study the effect of Tujian Mixture (TJM) on it.SD rats were divided into the normal control group (NC group, n = 8), the unilateral nephrectomized control group (QC group, n = 8), the STZ induced diabetes mellitus with unilateral nephrectomy model group (DM group, n = 8), the Valsartan treated group (VT group, n = 8) and the TJM treated group (ZY group, n = 9), rats in the latter two groups were modeled as in the DM group and treated with Valsartan (20 mg/kg.d) and TJM (20 g/kg.d) respectively for 12 weeks. The expression of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor-beta 1 (TGF-beta 1) in rats glomeruli were observed by immunohistochemistry assay, and the ratio of alpha-SMA and TGF-beta 1 positive area/total glomerule tuft area (SMA/GT and TGF/GT) were analyzed using computer-assisted image analysis software.In the NC and the QC groups, only trace of alpha-SMA positive staining was found. But there was prominant alpha-SMA positive staining in glomeruli of the DM group, with SMA/GT and TGF/GT increased significantly (P0.01), and marked increase of 24 hrs proteinuria excretion (P0.01). As compared with the DM group, the three indexes were all significantly lower in the VT and ZY groups (P0.01), and the lowering of proteinuria was more significant in the ZY group than that in the VT group (P0.01).The expression of alpha-SMA in glomeruli in STZ induced diabetic rats with unilateral nephrectomy is pronounced, indicating that phenotypic modulation of mesangial cells involvement in the pathogenesis of diabetic nephropathy. TJM and Valsartan can reduce 24 hrs proteinuria excretion, inhibit the phenotypic modulation of mesangial cells and the expression of TGF-beta 1 in glomeruli of diabetic rats, and the effect of TJM is more potent than that of Valsartan in lowering urinary protein excretion.

To explore the changes of orexin A and neuropeptide (NPY) in plasma and hypothalamus of rats with chronic renal failure (CRF).41 male Wister rats weighing 200 approximately 250 g were randomly divided into three groups: normal group, sham operation group, and CRF group (with the right kidney and 2/3 of the left kidney resected). A certain number of rats were decapitated 4, 8,and 12 weeks after respectively. Their hypothalami were removed and blood collected. Radioimmunoassay was used to measure the levels of orexin A and NPY in hypothalamus and plasma. Automatic biochemical analyzer was used to measure the serum creatinine.The serum creatinine level of CRF rats was both significantly higher than those of the sham operation rats at week 8 and week 12, respectively. The plasma orexin A level of CRF rats at week 12 was 264 pg/ml +/- 62 pg/ml, significantly higher than that of sham operation group (183 pg/ml +/- 56pg/ml, P = 0.039). The hypothalamus orexin A level of CRF rats were 10.5 fmol/mg +/- 2.7 fmol/mg wet weight at week 12, significantly lower than that of sham operation rats (17.4 fmol/mg +/- 3.9 fmol/mg wet weight, P = 0.023). The plasma NPY levels of CRF rats at week 8 and week 12 were significantly higher than those of the sham operation rats (7.1 pmol/ml +/- 1.7 pmol/ml vs 5.0 pmol/ml +/- 0.5 pmol/ml, P = 0.01; and 7.9 pmol/ml +/- 1.1 pmol/ml vs 4.8 pmol/ml +/- 1.1 pmol/ml, P = 0.0008). The hypothalamus NPY level of CRF rats at week 12 were 70 fmol/mg +/- 23 fmol/mg wet weight, significantly lower than that of the sham operation rats (113 fmol/mg +/- 31 fmol/mg wet weight, P = 0.03).Loss of renal function may diminish the excretion of orexin A and neuropeptide. The lowering of hypothalamus orexin A and neuropeptide Y levels may be one of the causes inducing anorexia in CRF.

To understand the clinical and pathological features of patients with Fanconi syndrome associated with renal function damage induced by Chinese herbs.Six cases with herb-induced renal failure associated with Fanconi syndrome were clinicopathologically analyzed.All six patients had kidney insufficiency after ingestion of Chinese herbs. All of them took the herbs containing aristolochia manshuriensis definitely. Four of the six presented with rapidly progressive acute renal failure and one with acute renal failure. All of them had similar clinical features, such as polydipsia, polyuria, anemia, glycosuria, aminoaciduria, increased urine beta(2) microglobin (beta(2)m) excretion and proximal tubular acidosis. Renal biopsies performed in 3 cases showed extensive hypocellular interstitial fibrosis, tubular atrophy and loss of tubules. Glomeruli were apparently intact.Nephropathy caused by Chinese herbs may be associated with Fanconi syndrome and renal failure.