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Objective: To investigate mechanisms behind the faster rehabilitation of limb fractures when associated with traumatic brain injury (TBI)., Methods: New Zealand rabbits were divided into TBI group and sham-operation group for four studies as follows: (1) blood and cerebrospinal fluid (CSF) were drawn on days 1, 3, and 7 to demonstrate changes in serum leptin, growth hormone (GH), insulin-like growth factor 1 (IGF-1), and CSF leptin; (2) bone defection was created by drilling in the tibial bone and either leptin or normal saline was injected into rabbit's cerebellomedullary cistern. X-ray was taken at 1 days, 2 weeks, and 5 weeks and evaluated by criteria to determine rate of bone healing; (3) FITC-labeled rabbit leptin was injected into TBI and sham-operation groups, and frozen sections of rabbit brain were observed to identify differences in central nervous system (CNS) leptin by fluorescence; (4) polymerase chain reaction (PCR) was used to evaluate the expression of leptin production by brain tissue., Results: Serum and CSF leptin, GH, and IGF-1 concentrations were found to be higher in the TBI group than the sham-operation group at days 1, 3, and 7 (P<0·05). CSF leptin of the TBI group was positively correlated with serum leptin on day 1 (P<0·05), and positively correlated with GH and IGF-1 on days 3 and 7 (P<0·05). X-ray criteria demonstrated that leptin administration caused significantly faster healing calluses at 3 and 5 weeks as compared to control animals (P<0·05). FITC-labeled leptin study demonstrated that TBI animals had stronger expression of leptin in the brain than sham-operated animals. However, PCR of brain tissue leptin showed no significant differences between TBI and sham-operated animals in the expression of leptin., Conclusions: Our study suggests that increased CSF leptin, likely from blood-brain barrier breakdown, combined with elevated serum GH and IGF-1 after TBI, leads to accelerated fracture healing.

Objective: The objective of this study is to evaluate the effect and mechanism of aging on iodinated-contrast-media-induced nephropathy in male rats., Methods: Twenty-four healthy male rats were initially divided into 12-month-old and 24-month-old age groups (adult and older age groups, respectively; n = 12/group); subsequently, each age group was randomly divided into saline control (NS) and contrast media (CM) groups (n = 6/group). CM (76% diatrizoate, 10 mL/kg b.w.) was given through the caudal vein. Urinary creatinine (Ucr) and serum creatinine (Scr) were detected by an automatic biochemical analyzer. The activities of renal malondialdehyde (MDA), superoxide dismutase (SOD), angiotensin-converting enzyme (ACE), angiotensin II (Ang II), and reduced form of nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) were determined by spectrophotometric assays with commercially available kits according to the manufacturers' protocols. Renal histological changes were observed by hematoxylin and eosin staining and scored semiquantitatively., Results: In diatrizoate-injected aged rats, Scr, the activities of ACE, Ang II, MDA, and NADPH oxidase in renal tissues were significantly increased (p < 0.01). The histologic scores were higher in the aged animals with CM treatment than those of control or adult rats (p < 0.01). There was an increasing trend but no significant statistical difference in renal ACE, Ang II, MDA, and NADPH oxidase or histologic scores in adult CM-injected rats compared with control animals (p > 0.05)., Conclusions: Older age is an aggravating factor of iodinated-contrast-media-induced nephropathy in male rats. Oxidative stress and the renin-angiotensin system (RAS) may play an important role in nephrotoxicity induced by iodinated contrast media, especially in aged male rats.

Background: There is rapidly accumulating evidence that use of oxidized hair dye causes various forms of nephrotoxic injury. However, the regulation and implication of the nephrotoxic injury resulting from p-aminophenol (PAP) and p-paraphenylenediamine (PPD), the main components of oxidized hair dye, remain unknown., Methods: To clarify the effect of PAP and PPD, we analyzed the proliferation, lactate dehydrogenase (LDH) levels, apoptosis and subsequent mRNA levels of caspase-3 in HK-2 cells stimulated with different concentrations of PAP or PPD. Cell proliferation was assessed by MTT assay. The production of LDH was determined by Hitachi 7170 biochemical analyzer. The apoptosis of cell was analyzed using flow cytometry and scanning electron microscopy. The mRNA levels of caspase-3 were quantitatively measured by real-time PCR., Results: The proliferation of HK-2 cells was significantly inhibited by PAP, PPD and each mixed with hydrogen peroxide (H2O2). The level of apoptosis of HK-2 cells, the mRNA levels of caspase-3 and LDH production by PAP or PPD stimulation was significantly higher than controls or after H2O2. A typical apoptotic morphological change was observed under electron microscopy in response to PAP or PPD., Conclusion: It appears that caspase-3 may play a key role in the nephrotoxic injury resulting from PAP, PPD or its oxidized form.

Objective: To elucidate the relationship of oxidative stress and specificity protein 1 (Sp1) in the process of epithelial-to-mesenchymal transdifferentiation (EMT) and also to investigate the molecular mechanism of protective effect of probucol on the pathogenesis of diabetic kidney disease (DKD)., Methods: Thirty male Sprague-Dawley (SD) rats were randomly divided into control group, diabetic group, and diabetic group under probucol therapy (n = 10 per group). The biochemical indicators including 24-h urinary total protein (24-h UTP) excretion, blood glucose (BG), lipids [triglycerides (TGs), total cholesterol (TC)], serum creatinine (Scr), creatinine clearance rate (Ccr), kidney tissue malondialdehyde (MDA) level, and glutathione peroxidase (GSH-Px) activity were assessed in all groups. The renal pathological changes were evaluated by hematoxylin and eosin (HE) and Masson staining. The protein expression of Sp1, α-smooth muscle actin (α-SMA), and E-cadherin was also measured and analyzed by immunohistochemistry and Western blotting., Results: Compared with the control group, the BG, TC, Scr, 24-h UTP, and MDA level of renal tissue increased significantly and the Ccr reduced in the rats of diabetic group (all p < 0.01). The pathological scores and the expression of Sp1 and α-SMA in renal tissue were up-regulated (p < 0.01) and the expression of E-cadherin was down-regulated significantly in the diabetic animals (p < 0.01). In the diabetic animals treated with probucol, the renal injuries were alleviated (p < 0.01)., Conclusions: Oxidative stress may play an important role in the EMT process of tubular epithelial cells. Probucol could ameliorate renal disease progression in this model of diabetic nephropathy, which might be due to an antioxidant action, down-regulation of Sp1 protein expression, and inhibition of renal tubular EMT.

Norcantharidin (NCTD) was shown in our previous studies to attenuate renal tubulointerstitial fibrosis in rat models with diabetic nephropathy (DN). The aim of this study was to determine the effects of NCTD on the expression of extracellular matrix (ECM) and TGF-β1 in HK-2 cells stimulated by high glucose and on calcineurin (CaN)/NFAT pathway. Whether or not the antifibrotic effect of NCTD on renal interstitium was dependent on its inhibition of CaN pathway was also investigated. Experimental concentrations of NCTD were verified by cytotoxic test and MTT assay. HK-2 cells were transfected with CaN small interference RNA (siRNA). The mRNA and protein expressions of FN, ColIV, TGF-β1, and CaN in HK-2 cells were detected by real-time PCR and western blot. The CaN/NFAT pathway was examined by indirect immunofluorescence and western blot. Our study revealed that NCTD concentrations over 5 mg/l had overt cytotoxicity on HK-2 cells. Meanwhile, both 2.5 and 5 mg/l NCTD inhibited HK-2 cell proliferation (P < 0.05). NCTD inhibited the upregulation of FN, ColIV, and TGF-β1 of HK-2 cells stimulated by high glucose (P < 0.05), and also significantly downregulated the expression of CaN mRNA and protein in HK-2 cells (P < 0.05). In addition, not only was the nuclear translocation of NFATc inhibited, but its protein level in the nucleus was also reduced. Following CaN siRNA transfection, CaN mRNA and protein expression were significantly decreased. In contrast, the protein levels of FN, ColIV, and TGF-β1 increased in HK-2 cells stimulated by high glucose (P < 0.05). However, NCTD treatment downregulated their expression. These results indicated that NCTD could decrease the expression of ECM and TGF-β1 in HK-2 cells stimulated by high glucose, downregulate CaN expression, and block the CaN/NFAT signaling pathway. However, the effect of NCTD on inhibition of the expression of ECM and TGF-β1 was not associated with its inhibition of the CaN/NFAT pathway.

The present study aims to observe the effects of NCTD (norcantharidin) on proliferation and FN (fibronectin) expression in human renal proximal tubular epithelial cell line (HK-2) induced by albumin in vitro. HK-2 cells were divided into control group, albumin group and different concentration of NCTD intervention groups. Proliferation of HK-2 cells was determined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide], FN protein in culture media of HK-2 cells was examined by ELISA, and FN mRNA was analysed by RT-PCR (reverse transcription-PCR). We chose less than 5.0 mg/l of NCTD as the experimental concentration for the cytotoxicity test. MTT score was higher in the albumin group than in the control group (P<0.05). As compared with that of the albumin group, MTT score and FN protein concentration decreased, FN mRNA significantly down-regulated in NCTD intervention groups respectively (P<0.05). Our study showed that NCTD could inhibit the albumin-induced cell proliferation and FN expression in HK-2 cells, which might further prove the anti-fibrotic role of NCTD in proteinuria-associated tubulointerstitial damage.

Objective: To investigate the effects of norcantharidin (NCTD) on tubulointerstitial fibrosis of diabetic nephropathy (DN) in streptozotocin-induced rat model., Methods: Sprague-Dawley rats were randomly divided into control group, model group, low-dose NCTD (0.05 mg/kg/day) group, and high-dose NCTD (0.1 mg/kg/day) group. The model group was induced by injection intraperitoneally with 30 mg/kg streptozotocin in 0.1 mol/L sodium citrate solution (pH 4.5), after high-calorie foods were given for 2 months. NCTD was administered daily after the DN rat model was built. Rats were sacrificed at the end of the third and the eighth week; renal fibrosis and the expression of FN, collagen IV, TGF-β1, and calcineurin (CaN) were detected by Masson and immunohistochemistry staining, respectively., Results: Tubulointerstitial fibrosis was observed in DN rats, this kind of pathological changes was ameliorated in NCTD treatment group (p < 0.05). The expressions of FN, collagen IV, and TGF-β1 protein increased in the tubulointerstitial field of DN rats compared with the rats in control group. NCTD treatment could dose-dependently decrease their expression and reverse the fibrotic degree (p < 0.05). Meanwhile, the expression of CaN was detected in tubular fields of normal kidney and increased in the tubulointerstitial field in DN rats. However, NCTD downregulated its expression in a dose-dependent manner (p < 0.05)., Conclusions: NCTD could downregulate FN, collagen IV, and TGF-β1 expression in tubulointerstitial fields and attenuate tubulointerstitial fibrosis in the early stage of DN rats. NCTD also alleviated the expression of CaN in tubules in DN. The relationship between the role of NCTD's anti-tubulointerstitial fibrosis and its inhibition to CaN expression remains to be further elucidated.

Background: The aim of this study was to investigate the correlation between the transforming growth factor (TGF)-beta1 gene -509C/T polymorphism and the susceptibility to primary nephrotic syndrome (PNS), and in particular to the severe degree of tubulointerstitial damage (TID) seen in Chinese., Methods: Ninety-eight PNS patients and 128 normal controls were studied. The extent of tubulointerstitial changes was evaluated and patients were divided into two groups according to the severe or mild degree of TID. The TGF-beta1gene -509C/T polymorphism was detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, and the serum level of TGF-beta1 was determined by enzyme-linked immunosorbent assay (ELISA)., Results: No statistical differences in genotype or allele frequency of the TGF-beta1 gene -509C/T were found between PNS and normal subjects. However, T allele and CT + T T genotype frequency were higher in the PNS with severe TID than the mild TID and controls. Additionally, the serum concentration of TGF-beta1 was significantly higher in the PNS with severe TID group than the other two groups and in the T T genotype individuals than the CC and CT genotype individuals. A logistic regression analysis demonstrated that TGF-beta1 gene -509C/T genotype was the risk factor of TID in PNS patients [OR (odd ratio) 2.34, confidence interval (CI) 0.98-3.46, p = 0.012]. CONCLUSION. TGF-beta1 gene -509C/T polymorphism was associated with severe TID. The higher value in serum concentration of TGF-beta1 was also associated with severe TID and the T T genotype/T allele. T allele gene might be the important risk factor for susceptibility.

Connective tissue growth factor (CTGF), a downstream mediator of transforming growth factor-beta1 (TGF-beta1) inducing fibrosis, has recently been implicated in peritoneal fibrosis. Extracellular matrix (ECM) accumulation and angiogenesis are characteristic changes in peritoneal fibrosis. In this study we investigated the effect of CTGF knockdown via interference RNA (RNAi) on ECM production and vascular endothelial growth factor (VEGF) expression induced by TGF-beta1 in human peritoneal mesothelial cells (HPMCs). Four CTGF short hairpin RNA (shRNA) expression constructs were generated using the pRetroSuper vector, and infectious retroviral particles were prepared to infect HPMCs. Expression levels of CTGF, fibronectin(FN), collagen 1 (col 1), laminin, and VEGF mRNA and protein were measured by semi-quantitative reverse transcription polymerase chain reaction and western blot assay. CTGF expression was increased after stimulation with TGF-beta1, but inhibited using each of the four independent CTGF shRNA constructs (P < 0.01). Moreover, expression of ECM proteins (FN, col 1, and laminin) and VEGF were upregulated after incubation with TGF-beta1, but elevated levels of ECM and VEGF induced by TGF-beta1 were significantly inhibited by RNAi (P < 0.01), but not by the empty retroviral vector (P > 0.05). From these results, we concluded that retrovirus-mediated CTGF shRNA can effectively inhibit ECM production and VEGF expression induced by TGF-beta1 in HPMCs. This study suggests that downregulation of CTGF may represent a potential therapeutic approach for peritoneal fibrosis through decreasing ECM accumulation and angiogenesis.

Objective: To observe the effect of Qingyi Decoction (QYD) in treating severe acute pancreatitis (SAP) and its impacts on blood levels of tumor necrosis factor-alpha (TNF-alpha), interleukin 6 and 8 (IL-6 and IL-8)., Methods: One hundred and ten patients of SAP were equally randomized into the treated group and the control group, they were treated with the same therapeutic program excepting that QYD was given only to the treated group. The post-treatment incidence of severe complication, mortality and operation transferring rate, as well as the changes of APACHE II scores and blood levels of TNF-alpha, IL-6 and IL-8 in patients of both groups were observed., Results: The incidences of the two severe complications, acute respiratory distress syndrome and intestinal paralysis, in the treated group was 3.6% and 5.4% respectively, while in the control group, 12.7% and 18.2%, showing statistical significance between groups (P < 0.05). The APACHE II score in the treated group decreased significantly on the 7th day, which was better than that in the control group (8.14 +/- 2.30 scores vs 3.35 +/- 2.20 scores, P < 0. 05). In addition, the efficacy in the treated group was also superior to that in the control group in terms of reducing mortality, operation transferring rate, and blood levels of TNF-alpha, IL-6 and IL-8 on the 7th and 9th day (P < 0. 05)., Conclusion: QYD could markedly improve the prognosis of SAP patients by way of lowering the blood levels of TNF-alpha, IL-6 and IL-8.

Background: Contrast-induced nephropathy is a serious complication of diagnostic and interventional procedures., Purpose: To evaluate the nephrotoxicity of high- and low-osmolar contrast media (HOCM, LOCM) on kidneys in Sprague-Dawley rats. Telmisartan was administered to confirm its protective role against nephrotoxicity induced by contrast media., Material and Methods: Sixty male rats were randomly divided into six groups (n=10/group). Glycerin was given to all rats except controls to induce renal injury. HOCM (diatrizoate) or LOCM (iohexol) (10 ml/kg b.w., 300 mg I/ml) was given through a caudal vein. Serum creatinine level was measured by an automatical biochemical analyzer. Caspase-3 activity and Angiotensin II (Ang II) level of renal tissue were detected by fluorometric method and radioimmunoassay, respectively. The renal injury was also assessed by hematoxylin and eosin and TdT-mediated deoxyuridine nick end-labeling staining., Results: In diatrizoate-injected rats, serum creatinine level was increased (P<0.001). There was no significant difference between iohexol animals and glycerol controls in the level of serum creatinine. The renal caspase-3 activity and Ang II levels in HOCM and LOCM groups were higher than those in glycerol control group (P<0.001). The percentage of apoptotic tubular cells and pathological scores were lower in the iohexol animals than that in the diatrizoate animals (P<0.001). In the groups pretreated with telmisartan, no increase in the levels of serum creatinine, renal Ang II, and caspase-3 activity was observed (P>0.05). The renal injuries induced by contrast media were alleviated., Conclusion: Both HOCM (diatrizoate) and LOCM (iohexol) could cause renal tubular cell apoptosis in the kidneys damaged by glycerin. LOCM was less toxic to rat kidneys than HOCM. Caspase-3 and Ang II might play a role in renal tubular cell apoptosis induced by contrast media. Telmisartan protected the renal tissue from nephrotoxicity induced by contrast media.

Norcantharidin (NCTD), the demethylated analog of cantharidin isolated from Mylabris, is an anticancer drug routinely used against various human cancers in China. The aims of this study are to learn if NCTD has a protective action against severe proteinuria and consequent interstitial inflammation and fibrosis, and if the inhibition of nuclear factor-kappaB (NF-kappaB) and connective tissue growth factor (CTGF) by NCTD might be involved. Male Sprague-Dawley rats with protein overload nephropathy induced by intraperitoneally injected bovine serum albumin were used as a model. The histopathological examination of kidney tissue in the 9th week by light microscopy and scanning electron microscopy revealed that inflammatory cells had extensively infiltrated into the tubulointerstitial areas with interstitial fibrosis. The administration of NCTD at 0.1 mg/kg/day to the bovine-serum-albumin-injected animal models effectively reduced the proteinuria, and prevented the proteinuria-induced interstitial inflammation and fibrosis. Expressions of the NF-kappaB p65 subunit and CTGF, detected by immunohistochemistry, Western blotting and reverse-transcription polymerase chain reaction, were upregulated in protein overload nephropathy and were attenuated by NCTD. Inhibition of the expressions of the NF-kappaB p65 subunit and CTGF was one beneficial effect of NCTD. These results suggest that in addition to the antiproteinuric action of NCTD, due to its anti-inflammatory and antifibrotic effects as shown in the present study, it may become a therapeutic agent for proteinuria and its associated chronic inflammatory and fibrotic nephropathy., ((c) 2008 S. Karger AG, Basel.)

Objective: To explore the protective effect of amlodipine on the cytotoxicity induced by contrast media (meglumine diatrizoate) in human kidney cells (HKC)., Methods: An HKC line was used. The experiment was divided into 4 groups: a model group (diatrizoate 111g/L), a prevention group (diatrizoate 111g/L+amlodipine 10(-5)mol/L), an amlodipine control group (amlodipine 10(-5)mol/L), and a culture medium control group (simple none blood serum DMEM-F12 medium). Cytotoxicity induced by meglumine diatrizoate was analysed by methyl thiazolyl tetrazolium (MTT) test, lactate dehydrogenase (LDH) assay, Hochest33258 fluorescence stained cytospins, and flow cytometric DNA analysis. The protein expression of Bax was determined by Western blot, and caspase-3 activity was examined by fluorometric method., Results: In the prevention group, the cell viability increased significantly (P<0.05), LDH levels decreased (P<0.05), and the apoptosis was lower than that of the model group (P<0.05) .Bax protein expression and caspase 3 activity decreased (P<0.05)., Conclusion: Amlodipine can inhibit the HKC apoptosis and protect the renal tubule cell from injury induced by meglumine diatrizoate.

Objective: To compare the nephrotoxicity of high- and low-osmolar contrast media (HOCM and LOCM), and to determine the protective role of fosinopril or telmisartan and its possible mechanism., Methods: Forty eight healthy SD rats were randomly divided into 6 groups: a normal control group, a glycerol control group, a low-osmolar contrast media (LOCM) group, a high-osmolar contrast media (HOCM) group, a fosinopril group, and a telmisartan group. Glycerine for inducing kidney damage was given to all rats except the normal control group. Twenty-four hours after the injection of glycerine, the mixed fosinopril suspension (10mg/kg) or telmisartan (5mg/kg) was poured into the stomach in the preventive group. Serum creatinine (SCr) and plasma angiotensin II (AngII) levels were detected by an automatical biochemical analyzer and radioimmunoassay; caspase-3 activity and claudin-1 expression of the renal tissue were detected by fluorometric method and immunohistochemical method. The renal injury was assessed by hematoxylin and eosin (HE) staining and terminal deoxynucleotide mediated nick and labeling (TUNEL) staining, respectively., Results: In diatrizoate-injected rats, SCr and AngII levels were increased (P<0.05). Expression of claudin-1 protein and caspase-3 activity in the renal tissue was upregulated. The histologic changes and percentage of apoptotic cells were milder in the LOCM rats than those in the HOCM rats. In the group pretreated with fosinopril or telmisartan, no increase in the levels of SCr and AngII was discovered. The expression of claudin-1 protein and caspase-3 activity was significantly lower than that in the HOCM group. The renal injuries induced by diatrizoate were alleviated., Conclusion: Both HOCM and LOCM could cause cellular apoptosis in the kidney.LOCM was less toxic to rat kidney than HOCM. Nephrotoxicity induced by HOCM might be related to caspase-3, claudin-1 and AngII. Fosinopril or telmisartan may protect the renal tissue from nephrotoxicity induced by diatrizoate.

Background: The peritoneum response to peritoneal dialysis can lead to fibrosis. The transforming growth factor beta1 (TGF-beta1) plays a key role in regulating tissue repair and remodelling after injury. Connective tissue growth factor (CTGF), a downstream mediator of TGF-beta1 inducing fibrosis, has been implicated in peritoneal fibrosis. Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis that can hasten peritoneal fibrosis. In this study, we investigated the effect of small interfering RNA (siRNA) of CTGF by pRETRO-SUPER (PRS) retrovirus vector on the expression of CTGF and VEGF in human peritoneal mesothelial cells., Methods: Retrovirus producing CTGF siRNA were constructed from the inverted oligonucleotides and transferred into packaging cell line PT67 with lipofectamine, and the virus supernatant was used to infect human peritoneal mesothelial cell (HPMC). The cells were divided into seven groups: low glucose DMEM, low glucose DMEM + TGF-beta1 5 ng/ml, low glucose DMEM + TGF-beta1 5 ng/ml + PRS-CTGF-siRNA(1-4) and low glucose DMEM + TGF-beta1 5 ng/ml + PRS. The expression of CTGF and VEGF were measured by semiquantitative RT-PCR and Western blot., Results: Low levels of CTGF and VEGF were detected in confluent HPMCs. Following stimulation with TGF-beta1, the levels of CTGF and VEGF were significantly upregulated (P < 0.01). Introduction of PRS-CTGF-siRNA(1-4) resulted in the significant reduction of CTGF mRNA and protein, and VEGF mRNA (P < 0.01), especially in groups PRS-CTGF-siRNA1 and PRS-CTGF-siRNA4. The introduction of PRS void vector did not have these effects (P > 0.05)., Conclusions: The expression of CTGF siRNA mediated by PRS retrovirus vector can effectively reduce the level of CTGF and VEGF induced by TGF-beta1 in cultured HPMCs. This study may provide potential therapeutic strategies to prevent the peritoneal fibrosis.

Background: Tubulointerstitial damage (TID) is an important mediator in the progression of chronic proteinuric nephropathies. Our aim in this study was to evaluate the relationship between several clinical predictors and TID in adult-onset primary nephrotic syndrome in China., Methods: One hundred ninety-five adult inpatients who were diagnosed with primary nephrotic syndrome based on clinical presentation and biopsy results were enrolled in this study from March 2003 to September 2005. The degree of TID was graded by a semiquantitative method including <2 score and >or=2 score., Results: In all patients, the rate of glomerulosclerosis was correlated with the severity of TID. Serum creatinine and uric acid (r = 0.183, p = 0.012 and r = 0.377, p = 0.00001, respectively) but not serum lipid or total 24-h urinary protein were related with TID. In 64 patients, urinary excretion of IgG (r = 0.443, p = 0.00001) but not of albumin, transferrin, retinal-binding protein, or alpha1-microglobulin were significantly associated with the extent of TID. Proteinuria selectivity index based upon IgG also correlated significantly with the extent of TID (p = 0.0001) (score 0-1 vs. score >or=2)., Conclusions: These results showed that serum creatinine and uric acid, the excretion of urinary IgG and proteinuria selectivity index based upon IgG, were highly correlated with the severity of TID in adult-onset primary nephrotic syndrome. These clinical parameters might be useful for predicting the development and progression of proteinuric nephropathy as independent risk factors.

Objective: To explore the effects of exogenous transforming growth factor-beta 1 (TGFbeta1) on peripheral nerve regeneration after the peripheral nerve injury and if TGFbeta1 regulates the expression of basic fibroblast growth factor (bFGF) in the anterior horn motoneurons of spinal cord during regeneration., Methods: Forty-eight rats were crushed on the right sciatic nerve and then randomly divided into 2 groups: TGFbeta1 group and NS group. In TGFbeta1 group, TGFbeta1 50 microL (0.1 microg/mL) was injected into the proximal nerve near to the crushed nerve and after the operation the injured leg was injected with equal TGFbeta1 whereas the NS was replaced in the NS group. The rats of each group survived for 3, 7, 14 and 21 days after the lesion. The bFGF expression in the anterior horn motoneurons of spinal cord was detected by immunohistochemistry (IHC). Semi-thin section and Fast Blue retrograde tracing were also performed with the rats surviving for 21 days to observe the regeneration of distal end in the injured right sciatic nerve., Results: The number of bFGF immunoreactive positive motoneurons in TGFbeta1 group was obviously higher than that of the NS group (P < 0.05). In the distal sciatic nerve of the rats treated with TGFbeta1, the number and diameter of regenerating myelinated axons and the thickness of myelinated sheath were more than those of the NS group (P < 0.05). The number of motoneurons in spinal cord and neurons in dorsol root ganglia (DRG) labelled with Fast Blue in the NS group was obviously lower than in the TGFbeta1 group (P < 0.01)., Conclusion: Exogenous TGFbeta1 plays an important role in promoting the peripheral nerve regeneration; TGFbeta1 up-regulates the bFGF expression in the anterior horn motoneurons of spinal cord during the peripheral nerve regeneration.

Objective: To determine the effect of transforming growth factor beta1 (TGFbeta1) antisense RNA on the synthesis and secretion of extracellular matrix in human peritoneal mesothelial cells (HPMC)., Methods: The recombinant human TGFbeta1 antisense eukaryotic expression vector was transfected into HPMC. Fibronectin (FN) and plasminogen activator inhibitor-1 (PAI-1) were detected by ELISA method. The expression of FN, collagen I (col I) and PAI-1 mRNA were detected by RT-PCR method., Results: The mRNA of FN, Col I,nd PAI-1 in HPMC were repressed by TGFbeta1 antisense RNA 24 hours after the transfection in comparison with the control group, which decreased 17%, 26%, and 9.6%, respectively. The protein of FN and PAI-1 were inhibited 48 hours after the transfection (P < 0.05)., Conclusion: Recombinant human TGFbeta1 antisense eukaryotic expression vector can inhibit the synthesis and secretion of extracellular matrix in HPMC which may be effective in gene therapy for peritoneal fibrosis.

Objective: To determine the influence of troglitanzone on the expression of mPGEs induced by high glucose or LPS in human glomeruli cultured in vitro., Methods: Human glomeruli were isolated and incubated by LPS or high glucose absence or presence with different concentrations of troglitazone. The concentration of mPGEs in the supernatant was measured by an enzyme-linked immunosorbent assay (ELISA)., Results: LPS (10 microg/ml) and 5% glucose could improve the concentration of mPGEs significantly in the supernatant by cultured human glomeruli, and troglitazone (10 - 20 nmol/L) could block its increase (P < 0.05)., Conclusion: High glucose and LPS can improve mPGEs expression in cultured human glomeruli, and troglitazone can block this effects. These data suggest that PPAR-gamma agonist troglitanzone may be relevant to the treatment of diabetic nephropathy and inflammation in glomerluli.

Objective: To study the effect of huangqi on peritoneal sclerosis and its possible mechanism., Methods: Isolated human peritoneal mesothelial cells (HPMC) was cultured. Cultured HPMC were treated with F12 without serum (control group), F12 with TGF-beta 1 (TGF-beta 1 group), F12 with TGF-beta 1 and huangqi 100 micrograms/ml (huangqi 1 group), F12 with TGF-beta 1 and huangqi 1 mg/ml (huangqi 2 group). Fibronectin(FN) and plasminogen activator inhibitor-1 (PAI-1) were detected by ELISA method. The expression of FN mRNA, PAI-1 mRNA and bFGF mRNA was detected by RT-PCR method., Results: 1. HPMC expressed FN, PAI-1, and bFGF. 2. Fn and PAI-1 significantly increased compared with control with 5 ng/ml TGF-beta 1 stimulated at 24 hours and 48 hours(P < 0.05); 3. The different concentration of huangqi decreased FN and PAI-1 expression compared with TGF-beta 1 group, especially 1 mg/ml huangqi at 24 hours(P < 0.05). 4. FN, PAI-1 and bFGF mRNA expression were higher in 12 hours after stimulation with TGF-beta 1 compared with control. FN, PAI-1 and bFGF mRNA expression were lower in 12 hours after stimulation with different concentration of huangqi (100 micrograms/ml and 1 mg/ml) than those in the TGF-beta 1 group., Conclusion: TGF-beta 1 promotes extracellular matrix synthesis and secretion of HPMC. Huangqi partly counteracts the synthesis of human peritoneal mesothelial cell's extracellular matrix by the stimulation of TGF-beta 1.

Objective: To establish a culture model of peritoneal mesothelial cells in vitro separated by infusing the digestive fluid in the rat abdominal cativity., Methods: Peritoneal mesothelial cells (PMC) were digested with 0.125% trypsin-0.01% EDTA Na2 in the abdominal cavity. We collected coelio-perfusate and cultured it. The isolated cells were identified by the phase contrast microscope, electron microscope and immunohistochemical analysis., Results: The confluent cells showed a uniform cobblestone-like appearance under the phase contrast microscope; numerous surface microvilli could be found under the electron microscope and the purity of the cells was over 95%. Immunohistochemical studies revealed a positive staining for cytokeratin and vimentin, but a negative staining for Factor VIII associated antigen and CD45., Conclusion: The model of rat peritoneal mesothelial cells has been established successfully and it will provide an experimental basis of studying the fibrosis in peritoneal dialysis.

Objective: To establish a reproducible model for culture of human peritoneal mesothelial cells (HPMCs), and observe the expressions of extracellular matrix (ECM) protein and interleukin-8 (IL-8) in HPMC., Methods: Mesothelial cells were isolated from human omenta by trypsin EDTA disaggregation. HPMCs were identified by morphology and streptomyces protein-peroxidase (SP) method. In the same way, expressions of fibronectin, collagen type III, V and transforming growth factor beta 1 (TGF beta 1) were measured. Expression of IL-8 mRNA and FN mRNA in the HPMCs was detected by reverse transcription (RT) and polymerase chain reaction (PCR) amplification., Results: Confluent HPMCs appeared multipolar under microscope; numerous surface microvilli and an abundant endoplasmic reticulum were observed. Cultured HPMCs coexpressed cytokeratin and vimentin, and synthesized fibronectin, TGF beta 1 and collagen types III (instead of type V). Expressions of IL-8 mRNA and FN mRNA were also observed in HPMCs., Conclusion: Establishment of cultured HPMCs model will provide the basis of the research in preventment of peritoneal fibrosis and roles of IL-8 in peritoneal dialysis.

The study focused on the relationship between protein loss and dialysis duration, results of PET and KT/Vurea in two groups of CAPD patients. 16 patients of one group were included in CAPD for only 30 days while 19 patients of the other group were included in CAPD for more than one year. The results showed that value of protein loss had no obvious significance between both groups. Compared with the patients with value of PET D/Pcr < 0.65, the patients with value of PET D/Pcr > or = 0.65 had no significant protein loss. Between the patients whose index of urea clearance > or = 2.0, and those index of uera clearance < 2.0 the protein loss had no difference. It is suggested that, in routine dialysis duration peritoneal transport of small solutes and dialysis adequency. The conclusion shows furtherly that transportion of large protein and small solutes through different ways.

Objective: To clarify the relationship between the renal tubular function and the efficacy of glucocorticoid, in adult patients with primary nephrotic syndrome., Methods: Pro- and post-therapy urinary RBP and NAG were determined double antibodies sandwich ELISA and color comparimetry with p-nitrate reductase, respectively, in eighty adult patients with primary nephritic syndrome, according to the concentrations of urinary protein, these patients were divided into three no remission group, partial remission group and fully remission group, fifty-one normal persons as control group. Renal tubular function parameters among groups were compared before and after therapy, and the results were analysed when those parameters were used to predict the sensibility of glucocorticoid therapy., Results: 1. There were no significant differences in urinary RBP, NAG and protein levels before treatment among patient groups (P > 0.05); 2. There were significant decreases in urinary RBP and NAG following glucocorticoid therapy in those response to glucocorticoid (P < 0.01); 3. There were significant differences in urinary protein, RBP and NAG among three groups of patients with primary nephrotic syndrome after treatment., Conclusion: Determination of urinary RBP may predict the sensibility of the therapy in the adult patients with primary nephrotic syndrome and its diagnostic efficiency is better than urinary NAG.

Purpose: To evaluate the nephrotoxicity of high- and low-osmolar contrast media (HOCM, LOCM) on kidneys in Sprague-Dawley rats. The protective role of amlodipine was studied., Material and Methods: Forty rats of both sexes were randomly divided into 5 groups (n=8/group) and glycerine for inducing renal failure was given to all rats except controls., Results: In diatrizoate-injected rats, blood urea nitrogen (BUN) and serum creatinine (SCr) were increased; levels of phospholipase A2 (PLA2), lipid peroxide (LPO) and calcium were also increased in renal tissues. There was no significant difference between LOCM (iohexol) animals and glycerol controls either in the renal levels of PLA2, LPO and calcium or in the levels of BUN and SCr. The histologic changes were milder in the LOCM animals than in the HOCM animals. In the group pretreated with amlodipine, no increase in the levels of BUN or SCr was discovered and the renal content of PLA2, LPO and calcium were significantly lower than in the HOCM group; the renal injuries induced by diatrizoate were alleviated., Conclusion: The HOCM, diatrizoate, was more toxic to rat kidneys than the LOCM iohexol; PLA2, LPO and calcium load played a role in producing renal function impairment induced by diatrizoate meglumine; amlodipine protected the renal tissue from nephrotoxicity induced by diatrizoate.

Objective: To investigate whether the small solutes transport and the efficiency of peritoneal dialysis can be improved through ventral movement., Methods: Eighteen continuous ambulatory peritoneal dialysis (CAPD) patients and 6 male New Zealand rabbits peritoneal dialysis models were observed. Through self and pre-post control, we compared the concentration ratio of the solutes in effluent dialyste(D) to the solutes in plasma(P), mass transfer area coefficient and the drained volume., Results: The animal experiments showed that the D/P value for BUN was different prominently when comparing the vibration and non-vibration groups in 45 min and 60 min separately(P < 0.05). On the contrary, no obvious difference existed between the two groups when comparing the D/P values for protein and drained volume. The clinical study showed: D/P value for BUN at the 2 and 4 hr and D/P value for creatinine at the 2 hr increased obviously either in the low or high frequency group when compared with the control group(P < 0.01, P < 0.01, P < 0.05, respectively). D/P for protein and drained volume had no difference compared with the control group(P > 0.05)., Conclusion: The ventral movement and vibration could efficiently increase the transport of small solutes such as BUN and creatinine while had no influence on the transport of large molecular solutes such as protein and the ultrafiltration volume. It suggested that increasing the movement of CAPD patients can improve the efficiency of peritoneal dialysis.

To explore the relationship between peritoneal transport of solutes and permeability of the peritoneum capillary as well as peritoneal stagnant fluid layer within dwell time, we observed the effects of nitroprusside and vibration on peritoneal transport of solutes in continuous ambulatory peritoneal dialysis(CAPD) patients. Twelve stable, routine CAPD patients were involved, who had no peritonitis for at least 4 weeks before the test. Standard peritoneal equilibrium tests(PETs) were performed, and mass transfer area coefficiency(MTAC) were calculated after adding nitroprusside to the dialysate or vibrating the patients's peritoneum. The concentrations of total protein, albumin and immunity globulin G in total drained dialysate were examined, and total drained volumes were recorded. Compared with the control, the MTACs value of BUN, Creatinine(Cr) increased significantly both in the nitroprusside group and vibration group(P < 0.05); the concentration of immunoglobin G in the total drained fluid was higher in the nitroprusside group than that in the control(P < 0.05); However, there was no significant difference in the total drained volume among the three groups. We conclude that nitroprusside and vibration can increase the peritoneal transport of small molecular solutes, and vibration has less influence on the loss of protein in CAPD. It suggests that moderated movement may improve the removal of the small molecules in CAPD patients.

To determine the relationship between the urinary endothelin (ET-1), nitric oxide (NO) levels and the clinical, pathologic types of primary glomerulonephritis (GN) patients, urinary levels of ET-1 and NO were detected in 27 patients with biopsy-proven primary GN and 12 normal controls by radioimmunoassay and by copper-plated and cadmium column reduction assay, respectively. The results showed that urinary ET-1 levels in the patients with primary GN were significantly higher than in normal controls (p < 0.01), while the urinary ET-1 levels in patients with moderate mesangial proliferation GN were significantly higher than those in patients with mild mesangial proliferation GN (p < 0.05). Urinary ET-1 levels in patients whose clinical feature was nephrotic syndrome were found to be higher than in patients whose clinical feature was nephritic syndrome. However, urinary NO levels were to the contrary (p < 0.05). The ratio of ET-1/NO in primary GN patients was significantly higher than that in normal controls, and it positively correlated with the 24-hour urinary excretion of protein. These results suggest that urinary ET-1 levels are related to the proliferation of mesangial cells. The imbalance between ET-1 and NO may be related to the pathogenesis of primary GN and the occurrence of proteinuria.

To determine whether there are early renal function parameters (RFP) which can be monitored to rapidly detect nephrotoxicity induced by contrast media (CM), we observed RFP in 16 patients with normal renal function before and after administration of CM. Forty-eight hours after diatrizoate meglumine administration, blood urea nitrogen (BUN) and serum creatinine (SCr) increased (p < 0.05). In all patients, acute tubular damage was revealed by early urinary RFP. Increases in levels of serum angiotensin-I-converting enzyme (ACE), beta(2)-microglobulin (beta(2)M) and urinary albumin (Alb) were associated with alterations in glomerular function. The changes in early RFP occurred earlier than those of BUN and SCr. The present study demonstrates that serum ACE, beta(2)M, urinary Alb, gamma-glutamyl-transpeptidase and N-acetyl-beta-D-glucosidase are sensitive parameters for the early assessment of subclinical nephrotoxicity induced by CM.