Efficacy and safety of Abelmoschus manihot for primary glomerular disease: a prospective, multicenter randomized controlled clinical trial.

BACKGROUND: Abelmoschus manihot, a single medicament of traditional Chinese medicine, has been widely used to treat kidney disease. This is the first randomized controlled clinical trial to assess its efficacy and safety in patients with primary glomerular disease. STUDY DESIGN: Prospective, open-label, multicenter, randomized, controlled, clinical trial. SETTING & PARTICIPANTS: From May 2010 to October 2011, a total of 417 patients with biopsy-proven primary glomerular disease from 26 hospitals participated in the study. INTERVENTIONS: A manihot in the form of a huangkui capsule, 2.5 g, 3 times per day; losartan potassium, 50mg/d; or combined treatment, a huangkui capsule at 2.5 g 3 times per day, was combined with losartan potassium, 50mg/d. The duration of intervention was 24 weeks. OUTCOMES & MEASUREMENTS: The primary outcome was change in 24-hour proteinuria from baseline after treatment. Change in estimated glomerular filtration rate (eGFR) from baseline after treatment was a secondary outcome. The 24-hour proteinuria was measured every 4 weeks and eGFR was measured at 0, 4, 12, and 24 weeks. RESULTS: Mean baseline urine protein excretion was 1,045, 1,084, and 1,073 mg/d in the A manihot, losartan, and combined groups, respectively, and mean eGFR was 108, 106, and 106 mL/min/1.73 m2, respectively. After 24 weeks of treatment, mean changes in proteinuria were protein excretion of -508, -376, and -545 mg/d, respectively (P=0.003 for A manihot vs losartan and P<0.001 for the combined treatment vs losartan). Mean eGFR did not change significantly. The incidence of adverse reactions was not different among the 3 groups (P>0.05), and there were no severe adverse events in any group. LIMITATIONS: Results cannot be generalized to those with nephrotic syndrome or reduced eGFR. CONCLUSIONS: A manihot is a promising therapy for patients with primary kidney disease (chronic kidney disease stages 1-2) with moderate proteinuria.

Lanthanum carbonate for the treatment of hyperphosphatemia in CKD 5D: multicenter, double blind, randomized, controlled trial in mainland China.

BACKGROUND: Serum phosphorus control is critical for chronic kidney disease (CKD) 5D patients. Currently, clinical profile for an oral phosphorus binder in the mainland Chinese population is not available. OBJECTIVE: To establish the efficacy, safety, and tolerability of lanthanum carbonate in CKD 5D patients. DESIGN: Multicenter, randomized, double blind, placebo-controlled study. A central randomization center used computer generated tables to allocate treatments. SETTING: Twelve tertiary teaching hospitals and medical university affiliated hospitals in mainland China. PARTICIPANTS: Overall, 258 hemodialysis or continuous ambulatory peritoneal dialysis (CAPD) adult patients were enrolled. INTERVENTION: After a 0-3-week washout period and a 4-week lanthanum carbonate dose-titration period, 230 patients were randomized 1:1 to receive lanthanum carbonate (1500 mg-3000 mg) or placebo for a further 4-week maintenance phase. MAIN OUTCOME MEASURES: Efficacy and safety of lanthanum carbonate to achieve and maintain target serum phosphorus concentrations were assessed. RESULTS: In the titration phase, serum phosphorus concentrations of all patients decreased significantly. About three-fifths achieved target levels without significantly disturbing serum calcium levels. At the end of the maintenance period, the mean difference in serum phosphorus was significantly different between the lanthanum carbonate and placebo-treated groups (0.63±0.62 mmol/L vs. 0.15±0.52 mmol/L, P < 0.001). The drug-related adverse effects were mild and mostly gastrointestinal in nature. CONCLUSION: Lanthanum carbonate is an efficacious and well-tolerated oral phosphate binder with a mild AE profile in hemodialysis and CAPD patients. This agent may provide an alternative for the treatment of hyperphosphatemia in CKD 5D patients in mainland China. TRIAL REGISTRATION: No. ChiCTR-TRC-10000817.

[Treatment of chronic primary glomerulopathy patients of Shen deficiency and dampness heat syndrome by yishen qingli granule combined low-dose Tripterygium wilfordii multiglycoside tablet: a clinical efficacy observation].

OBJECTIVE: To evaluate the clinical efficacy and safety of treatment of chronic primary glomerulopathy (CPG) patients of Shen deficiency and dampness heat syndrome (SDDHS) by Yishen Qingli Granule (YQG) combined with low-dose Tripterygium Wilfordii multiglycoside Tablet (TWT). METHODS: Totally 231 CPG patients of SDDHS were enrolled in this study (including 60 patients from First Affiliated Hospital of Nanjing University of Chinese Medicine, 58 from First Affiliated Hospital of Nanjing Medical University, 46 from Xinqiao Hospital of Third Military Medical University, 35 from First Affiliated Hospital of Guangzhou University of Chinese Medicine, 14 from First Affiliated Hospital of Soochow University, and 18 from Wuxi Affiliated Hospital of Nanjing University of Chinese Medicine). They were randomly assigned to the control group (116 cases) and the trial group (115 cases) according to block group method. There were 217 cases in the safety analysis set (109 cases in the trial group vs 108 cases in the control group), and 203 cases in the full analysis set (99 cases in the trial group vs 104 cases in the control group). All patients received basic treatment such as ACEI/ARB. Furthermore, YQG (consisting of raw astragalus 10 g, prepared Polygonum Multiflorum 10 g, Pyrrosia 10 g, 1.5 g each package, containing 10 g of crude drugs) was additionally given to patients in the trial group, each package, twice daily. The TWT (10 mg) was given, twice a day. The TWT dose was adjusted according to 24 h urinary total protein (UTP). The placebos of YQG and TWT were administered to those in the control group. The treatment course consisted of 24 weeks and the follow-up visit lasted for 24 weeks. The biochemical indices were observed before and after treatment including 24 h UTP, urine red cell count (U(RBC)), renal functions (BUN, SCr), blood routine test (WBC), and liver functions (SGPT, SGOT). Reverse reactions such as gastrointestinal discomfort, skin rash, and irregular menstruation were also observed. RESULTS: Compared with the control group, the total effective rate was better in the trial group (82.83% vs 61.54%, P < 0.01). Results of stratified comparison of UTP showed better efficacy in the trial group (0.8-3.0 g/24 h, P < 0.01). The UTP decline occurred in the trial group after 8 weeks of treatment, with stable action, showing statistical difference when compared with the control group (P < 0.01). In the trial group, U(RBC) level decreased after treatment but changed more significantly. But there was no statistical difference in the changes when compared with the control group (P > 0.05). After treatment, there were no statistical difference in safety indicators such as WBC, SGPT, and SGOT between the two groups after treatment (P > 0.05). CONCLUSION: On the basis of basic treatment such as ACEI/ARB, application of YQG combined with low-dose TWT had better effect in controlling proteinuria of CPG patients, and could help stabilizing their conditions with less adverse reactions.

Uraemic serum induces dysfunction of vascular endothelial cells: role of ubiquitin-proteasome pathway.

The ubiquitin-proteasome pathway (UPP) has been indicated to contribute to dysfunction of endothelial cells (ECs). Nevertheless, the relationship between UPP and vascular complications of uraemia remains unknown. We aimed to determine whether the UPP is activated in vascular ECs when cultured with uraemic serum, and to examine the role of the UPP on dysfunction of ECs in uraemia. Rabbit aortic endothelial cells (RAECs) were cultured with normal serum or different concentrations of uraemic serum. The expression of the ubiquitin-activating enzyme (E1), an indicator of the UPP, was detected by real-time RT-PCR and Western blot; proteasome activity was determined by fluorescence spectrophotometry; and nuclear factor-κB (NF-κB) activity and expression, as well as tumour necrosis factor-α (TNF-α) expression, were also detected. We found that the expression of E1 and the activities of three kinds of proteasomes were increased significantly in RAECs after incubation with uraemic serum. Proliferation of RAECs was increased significantly by incubation with 3-15% uraemic serum but decreased markedly when incubated with uraemic serum above 15% (increased apoptosis). Incubation of RAECs with uraemic serum induced increased NF-B DNA-binding activity and nuclear translocation of NF-κB, decreased nitric oxide production and increased expression of TNF-α, which is the final effector of inflammatory activation of cells. All of these responses in RAECs were suppressed by the specific proteasome inhibitor, MG132. The inhibition of inflammatory responses by MG132 was further supported by a parallel experiment with pyrrolidine dithiocarbamate, a specific inhibitor of κNF-B. These findings suggest that the UPP was activated in RAECs by administration of uraemic serum, and played a pivotal role in the dysfunction of vascular ECs, such as inflammatory activation.

Effects of 4-phenylbutyric acid on the process and development of diabetic nephropathy induced in rats by streptozotocin: regulation of endoplasmic reticulum stress-oxidative activation.

Oxidative stress may contribute to the pathogenesis of diabetic nephropathy (DN), although the precise regulatory mechanism is still unclear. Recent reports have shown that chemical molecular chaperone 4-phenylbutyric acid (4-PBA) can suppress oxidative stress by attenuating endoplasmic reticulum (ER) stress. We therefore hypothesized that 4-PBA could provide renoprotection through the suppression of oxidative stress in DN rats. Male Sprague-Dawley (SD) rats were randomly divided into three groups: a normal control (NC) group, a streptozotocin (STZ)-induced DN model group, and a DN plus 4-PBA (1g/kg) treatment group. At the end of 4, 8, and 12 weeks, hydroxyproline content, NADPH oxidase activity and the expression of phosphorylation of inositol-requiring enzyme-1alpha (p-IRE1alpha), p47phox, nitrotyrosine (NT) and NF-E2-related factor 2 (Nrf2) in the kidneys of all rats were determined; malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity in serum and urine were also detected; renal nuclear factor kappaB (NF-kappaB) activity in all of the rats was examined at the end of 12 weeks. Compared with the NC group, the DN rats showed a significant increase in hydroxyproline content, NADPH oxidase activity, NF-kappaB activity, the expression of p-IRE1alpha, p47phox, NT and Nrf2 in renal tissue; markedly, MDA levels were higher and SOD activity was lower in serum and urine of DN rats than in NC rats for the indicated time. These alterations were inhibited by the administration of 4-PBA. These findings first demonstrated that treatment with 4-PBA significantly inhibits the process and development of diabetic nephropathy in rats through the regulation of ER stress-oxidative activation.

Conditioned medium from renal tubular epithelial cells stimulated by hypoxia influences rat bone marrow-derived endothelial progenitor cells.

BACKGROUND AND AIMS: It has been well documented that endothelial progenitor cells (EPCs) participate in neovascularization in adults and that rarefaction of peritubular capillaries (PTCs) is closely associated with progressive kidney disease. Therefore, we investigated whether EPCs were influenced by conditioned medium (CM) of renal tubular epithelial cells (RTECs) stimulated by hypoxia, to provide evidence for EPCs transplantation in vivo in the future. METHODS: Mononuclear cells of rat bone marrow were isolated by density gradient centrifugation and were cultured according to previously described techniques. RTECs were cultured primarily with routine tissue block adhering wall method. In addition, CM was harvested from RTECs cultivated for 48 h in 5% O(2). EPCs proliferation and migration were evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and transwell. The protein and mRNA expression of stromal cell-derived factor (SDF-1), vascular endothelial growth factor (VEGF), angiogenin 1 (Ang-1), and C-X-C chemokine receptor 4 (CXCR4) was separately assessed by Western blot, enzyme-linked immunosorbent assay (ELISA), and reverse transcriptase-polymerase chain reaction (RT-PCR) methods. RESULTS: We showed that hypoxia increased the expression of SDF-1 and VEGF in RTEC. In addition, hypoxic CM improved proliferation, migration, and expression of VEGF, Ang-1, and CXCR4 of EPCs. CONCLUSIONS: These results suggest that hypoxic CM improves neovascularization of EPCs and may also be considered as therapeutic agents to supply the potent origin of reconstituion of PTCs of progressive kidney disease.

[Potential effect of uremic medium on cell proliferation and apoptosis and the activation of nuclear factor-kappa B pathway of aortic endothelial cells in rabbit culture].

OBJECTIVE: To investigate the potential effect of uremic medium on cell proliferation and apoptosis of aortic endothelial cell (AEC), two key processes in the development of atherosclerosis, in rabbit culture. And to understand the effects of uremic medium on the activation of nuclear factor-kappa B (NF-κB) pathway and cytokines expression of AEC. METHODS: Rabbit AEC were cultured with growth media supplemented with pooled sera from normal rabbits or those with chronic renal failure. The 80% confluent AEC were incubated for 24 h with media supplemented with pools of control or uremic sera. Cell proliferation was assessed by a MTT assay and cell cycle detected by flow cytometry. Hoechst33342 assay and flow cytometry were used to investigate the apoptotic effect of uremic medium in AEC. The expression of mRNA and protein levels for NF-κB, IκBα were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. NF-κB P65 nuclear translocation was analyzed by immunofluorescence. The activity of NF-κB was measured by electrophoretic mobility shift assay (EMSA). Concentrations of TNF-α and IL-6 in culture supernatants were evaluated by ELISA, and the expression of protein for TNF-α in cell lysates by Western blot. RESULTS: Uremic medium induced proliferation in the lower concentration range of 3%-10% while promoted apoptosis in the higher concentrations (> 10%). Uremic serum increased NF-κB mRNA (0.35 ± 0.05 vs 0.26 ± 0.02, P < 0.01) and protein (1.67 ± 0.15 vs 0.41 ± 0.05, P < 0.01) expression, decreased IκBα mRNA (0.13 ± 0.03 vs 0.24 ± 0.04, P < 0.01) and protein (0.29 ± 0.06 vs 0.65 ± 0.08, P < 0.01) expression. Uremic serum enabled NF-κB p65 nuclear translocation and increased NF-κB DNA binding activity. An increased secretion of cytokines IL-6 and TNF-α. in AEC was observed after a treatment of 10% uremic sera in a time dependent manner. The expression of TNF-α in AEC exposed to 10% uremic sera also increased significantly (0.37 ± 0.04 vs 0.14 ± 0.03, P < 0.01). CONCLUSION: Uremic medium induces the activation of AEC. A lower level of uremic medium accelerates the proliferation of AEC while a higher level induces the apoptosis of AEC. The increased proliferation may be related to a higher NF-κB activity and the expression of inflammation cytokines. Although the enhanced atherosclerosis can not be explained on the basis of an apoptotic process, the proliferative status can contribute to intimal proliferation, an earlier step in the development of atherosclerosis.

[Protective effects of metanephric mesenchymal cells on acute renal tubular damage: experiment with rats].

OBJECTIVE: To study the protective effects of metanephric mesenchymal cells (MMCs) on acute renal tubular damage and explore its possible mechanism. METHODS: MMCs were isolated and cultured from 13-day-old embryonic rats and labeled with 5-bromodeoxyuridine. Seventy-two male SD rats were randomly divided into 3 equal groups: MMC group, receiving MMC injection instantaneously when ischemia/reperfusion (I/R) renal injury was induced, I/R group, undergoing I/R to establish acute renal tubular damage models, and sham operation group. Six rats from each group were killed at different time points: 24 h, 48 h, 72 h, and 96 h later. Blood sample was collected from the vena cava inferior, to examine the serum creatinine (SCr) and blood urea nitrogen (BUN). Specimens of kidney underwent microscopy. Apoptosis was conformed by TUNEL assay. Immunohistochemistry was used to detect the protein expression of Bcl-2 and Bax. The distribution of MMCs labeled with 5-bromodeoxyuridine in kidney was observed by immunofluorescence technique. RESULTS: The SCr and BUN levels in different time points of the MMC group were both significantly lower than those of the I/R group (both P <0.05), HE staining showed that pathological damage of the MMC group was less than that of the I/R group (P <0.05). TUNEL results investigated that the number of apoptosis renal tubular epithelial cells of the MMC group was (13.4 +/- 3.2/HPF), significantly less than that of the I/R group [(25.4 +/- 5.2/HPF)]. In comparison with the I/R group, there were more Bcl-2 positive cells and fewer Bax positive cells in the MMC group. BrdU-labeled MMCs began to occur in the renal tissue (60 +/- 6/HP) In the 72 h subgroup of MMC group, and number of BrdU-labeled MMCs, the 96 h subgroup was (143 +/- 8/HP), significantly higher than that of the 72 h subgroup (P<0.05). CONCLUSION: MMCs have the ability to protect renal function in acute renal tubular damage in rats, migrate and repopulate in the I/R injured renal tubules, and inhibits renal tubular epithelial cell apoptosis. The mechanism may be involved in regulating the expression of Bcl-2 and Bax.

[A study of the clinical classification of 3638 cases of mushroom poisoning].

OBJECTIVE: To study the clinical classification and characteristics as well as prognosis of mushroom poisoning. METHODS: We retrospectively analyzed 191 papers published domestically for 3466 cases of mushroom poisoning from 1995 to 2004 and studied the Xinqiao Hospital data of 172 cases of mushroom poisoning treated from 1980 through 2004. We made a retrospective investigation and clinical classification of all the 3638 cases of mushroom poisoning. RESULTS: Among the 3638 patients, clinical manifestations as gastroenteritis were found in 571, all of them were cured. The most common symptoms were those of acute renal failure being found in 1450 cases; 1414 were cases (97.5%) and 36 died (2.5%). Symptoms of toxic hepatitis were found in 1010 cases, 841 were cases (83.3%) and 169 died (16.7%). Psychoneurological disorder was manifested in 214 cases; 197 were cases (92.1%) and 17 died (7.9%). Erythrolysis was found in 73 cases; 71 were cases (97.3%) and 2 died (2.7%). The therapeutic effect and prognosis of mushroom poisoning with different clinical manifestation varied very significantly (P < 0.001). Of the 320 cases clinically unclassified, multiple organ dysfunction syndrome was found in 222 cases; 98 were cases (44.1%) and 124 died (55.9%). Definite classification could not be made in 98 cases; 90 were cases (91.8%) and 8 died (8.2%). CONCLUSIONS: The clinical classification of mushroom poisoning was usually of four types as described in the textbooks or special articles, but it should be of five types according to the analysis of the data of the present study, i.e. gastroenteritis type, acute renal failure type, toxic hepatitis type, psychoneurological disorder type and erythrolysis type. It is not clear whether there is a mixed type or not. Further investigation is needed in this respect.

Protein-bound P-cresol inhibits human umbilical vein endothelial cell proliferation by inducing cell cycle arrest at G0/G1.

P-cresol is a typical protein-bound uremic toxin, which is retained in patients with renal failure. It is not known whether protein-bound P-cresol exhibits the toxicity in humans. This study aims to investigate the endothelial toxicity of protein-bound P-cresol. Cultured human umbilical vein endothelial cells (HUVEC) were treated with unbound or human serum albumin-bound (HSA, 4 g/dL), P-cresol (0, 20, 40, 80 µg/mL) for 24, 48, 72 h, respectively. Cell viability was determined by using cell counting kit-8 (CCK-8) assay. Cell apoptosis and cell cycle were assessed by using flow cytometry. The expression of cell cycle proteins in HUVEC were analyzed by using western blot and double immunofluorescent labeling assay. The results indicated that the viability of HUVEC was dose- and time-dependently inhibited by the protein-bound P-cresol (77.56% inhibition at 72 h, P<0.05) and unbound P-cresol (80.65% inhibition at 72 h, P<0.05). Most HUVECs were arrested at G0/G1 phase by both protein-bound P-cresol (79.63% inhibition at 72 h, P<0.05) and unbound P-cresol (81.27% at 72 h, P<0.05). Both protein-bound and unbound P-cresol enhanced the expression of p21Cip1 (0.62 and 0.60, both P<0.05) and suppressed the expression of cyclin D1 (0.49 and 0.53, both P<0.05) in a dose-dependent manner. In conclusion, unbound and protein-bound P-cresol inhibit the HUVEC proliferation by inducing cell cycle arrest at G0/G1 phase in a dose- and time-dependent manner, which associates with the up-regulation of p21Cip1 and down-regulation of cyclin D1.

Clinical analysis of 156 cases of multiple organ failure caused by fish bile.

OBJECTIVE: To analyze the clinical features and prognosis of patients suffering from fish bile poisoning. METHODS: A total of 156 multiple organ failure (MOF) patients caused by fish bile poisoning were hospitalized in our department over the past 28 years. The patients' symptoms, examination results, treatment and outcomes were collected and analyzed. RESULTS: All patients' first symptom was gastrointestinal discomfort, including unbearable nausea and intractable vomiting. The symptoms that followed were oliguria or anuria, edema, headache, fatigue, jaundice, palpitation, alimentary tract hemorrhage, gross hematuria, dyspnea, shock, tachycardia, bradycardia, arrhythmia, coma, and cardiac arrest. The symptom severity and cohort were different among different patients. Twenty-one cases received gastroscopy, which exhibited diffuse gastric mucosal bleeding. Twelve patients received renal biopsy, which exhibited focal necrosis of tubular epithelial cells. One patient received a liver biopsy, which exhibited extensive hepatocyte necrosis. All patients received blood purification therapy. Of the four patients who died, 4 out of 5 organs had failed. The general mortality rate was 2.6%. CONCLUSIONS: Compared with the MOF caused by trauma and sepsis, the fish bile poisoning MOF has a much lower morality rate. However, patients with higher age, more underlying diseases, and more organ failure tended to have a worse prognosis.

Costimulatory molecule VSIG4 exclusively expressed on macrophages alleviates renal tubulointerstitial injury in VSIG4 KO mice.

BACKGROUND: Activation and infiltration of T cells and macrophages are key features of renal tubulointerstitial injury. The costimulatory molecule V-set and immunoglobulin domain-containing protein-4 (VSIG4), which is exclusively expressed on macrophages, is capable of inhibiting the T cell response. However, it is unclear whether VSIG4 is involved in renal tubulointerstitial injury. This study was designed to investigate the role of VSIG4 in renal tubulointerstitial injury and the related T cell infiltration. METHODS: The unilateral ureteric obstruction (UUO) model of renal inflammation and tubulointerstitial fibrosis was established in VSIG4 transgenic knock-out C57BL/6 mice (VSIG4(-/-)) and wild-type C57BL/6 mice (VSIG4(+/+)). Comparative analysis of renal biological indices were assessed by quantitative real-time PCR and immunofluorescence staining. RESULTS: Both the VSIG4(-/-) and VSIG4(+/+) mice showed UUO-related temporal changes in renal expression of CD3, CD4 and CD8 T cell markers, with the protein levels being significantly lower in the VSIG4(+/+) UUO mice. Moreover, at each time point examined the UUO VSIG4(+/+) mice showed significantly lower renal mRNA levels of the cytokines interleukin (IL)-2, interferon- and tumor necrosis factor-, but significantly higher IL-10, than the UUO VSIG4(-/-) mice. CONCLUSIONS: The macrophage-expressed VSIG4 may act to alleviate renal tubulointerstitial injury via inhibition of T cell infiltration and secretion of inflammation related factors.

Effect of chronic renal failure medium on the ubiquitin­proteasome pathway of arterial muscle cells.

The ubiquitin­proteasome pathway (UPP) is involved in the occurrence and development of atherosclerosis through inhibitor of κB (IκB) degradation which activates nuclear factor-κB (NF­κB). However, the correlation between UPP and vascular complications of uramia remains unknown. The aim of the present study was to determine whether the UPP is activated in aortic smooth muscle cells (ASMCs) when cultured with uremic serum and to examine the role of the UPP on the dysfunction of ASMCs in uremia. ASMCs were cultured with pooled normal sera or chronic renal failure sera. The mRNA expression levels for ubiquitin (Ub) and Ub-activating enzyme (E1) were analyzed using reverse transcription PCR and levels of the ubiquitinated proteins E1 and IκBα were measured using western blot analysis. The enzymatic activities of three 20S proteasomes were examined using specific fluorogenic peptide substrates. Compared with normal serum, chronic renal serum increased E1 mRNA and protein expression of rabbit ASMCs (both P<0.01). In addition, the mRNA expression of Ub also increased and the expression of IκBα was observed to decrease significantly (both P<0.01). Ubiquitinated proteins in the normal and chronic renal failure groups were not found to be significantly different, but the activity of proteasomes increased significantly (P<0.01). Chronic renal failure medium induced the activation of the UPP in ASMCs.

Optimized project of traditional Chinese medicine in treating chronic kidney disease stage 3: a multicenter double-blinded randomized controlled trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Stage 3 is the key phase of chronic kidney disease. Traditional Chinese medicine (TCM) has been used for the treatment of chronic kidney disease. But a large sample trial is desirable. MATERIALS AND METHODS: A total of 578 Chinese patients with primary glomerulonephritis in CKD stage 3 were randomly assigned to three groups: patients received TCM (TCM group), benazepril (Ben group), TCM combined with benazepril (TCM+Ben group). Patients were followed up for 24 weeks. The primary endpoint was the time to the composite of 50% increased of serum creatinine, end stage renal disease or death. RESULTS: eGFR in the TCM and the TCM+Ben group were improved (week 24 vs. baseline, P<0.05) while eGFR in the Ben group was decreased (week 24 vs. baseline, P>0.05). 24h urinary protein excretion (UP) and urinary albumin/creatinine (UAlb/Cr) were decreased in the TCM+Ben (week 24 vs. baseline, P<0.05) and the Ben group (week 24 vs. baseline, P>0.05). UP and UAlb/Cr were increased in the TCM group to week 12, then were stable (week 24 vs. baseline, P<0.05). The hemoglobin in the TCM group was also improved (week 24 vs. baseline, P<0.05). The accumulative survival rate in the TCM+Ben group was higher than that in the TCM group and the Ben group (P=0.044). Side effects in the TCM group were the lowest in these groups (P<0.05). The patients with dry cough in the TCM+Ben group and the Ben group were increased as compared with the TCM group (P<0.05). Hyperkalemia happened less frequently in the TCM group as compared with the other two groups (P=0.052). CONCLUSIONS: For the patients with CKD stage 3, TCM can improve eGFR and hemoglobin with lower side effects. Benazepril significantly decreased the proteinuria. Chinese medicine integrated with benazepril can ameliorate renal function and decrease proteinuria synergistically.

Attenuation of diabetic nephropathy in diabetes rats induced by streptozotocin by regulating the endoplasmic reticulum stress inflammatory response.

The endoplasmic reticulum (ER) is capable of sensing metabolic and stress parameters and integrating intra- and extracellular signals to support a coordinated cell response. In the present study, we verified the hypothesis that 4-phenylbutyric acid (4-PBA), a chemical chaperone, prevented the progression of diabetic nephropathy (DN). Male Sprague-Dawley rats were randomly divided into 3 groups: a normal control group, a DN group, and a DN model plus 4-PBA treatment group (PBA). The DN model was induced by injection of streptozotocin with uninephrectomy. The dosage of 4-PBA treatment was gavaged at a dose of 1 g/kg body weight each day for 12 weeks. The expression of the ER stress indicators significantly increased in the kidney of DN rats within the indicated period. Moreover, the expression of phosphorylated c-JUN NH(2)-terminal kinase, the monocyte chemoattractant protein-1, and the final fibrotic effector all elevated markedly in the kidney of DN rats. Urinary protein excretion rate and the concentration of urinary monocyte chemoattractant protein-1 were higher than those in the normal control group. Treatment with 4-PBA can suppress the expression of the glucose-regulated protein 78 and the phosphorylation of the PKR-like ER kinase, both of which are ER stress indicators; renoinflammatory signal; and the expression of inflammatory cytokines and fibrosis factors. It also can inhibit the increase in urinary protein excretion rate and urinary monocyte chemoattractant protein-1. In conclusion, 4-PBA exerts a marked renoprotective effect possibly due to modulating ER stress and related inflammatory cascade.

Prevention of diabetic nephropathy in rats through enhanced renal antioxidative capacity by inhibition of the proteasome.

AIMS: Oxidative stress may play an important role in the pathogenesis of diabetic nephropathy (DN). Recent studies have shown that the ubiquitin-proteasome pathway (UPP) and oxidative stress have interaction. We aimed to investigate whether inhibiting the proteasome has a preventive effect on DN through suppression of renal oxidative stress. MAIN METHODS: Male Sprague-Dawley rats were randomly divided into three groups: a normal control (NC) group, a streptozotocin-induced DN model group, and a DN plus MG132 (10 µg/kg) treatment group. KEY FINDINGS: Increased 24-h urinary protein excretion rate (UPER) and renal pathological changes were all improved after MG132 administration. Furthermore, enhanced renal 26S proteasome activity and concentration in DN rats were effectively reduced after MG132 administration. Increased p47phox and nitrotyrosine (NT) expressions in kidneys of DN rats were decreased after MG132 treatment. Renal mRNA and protein expressions of NF-E2 related factor 2 (Nrf2) were up-regulated by MG132 in comparison to DN alone. Decreased renal mRNA expression of superoxide dismutase 1 (SOD1), catalase (CAT) and glutathione peroxidase (GPx) in DN rats was heightened after MG132 intervention. Depressed activities of renal SOD, CAT and GPx in DN rats were also improved by MG132 treatment. Increased renal nuclear factor κB (NF-κB) activity was inhibited after MG132 administration in DN rats at the end of 12 weeks. SIGNIFICANCE: Our present data suggest that inhibition of the proteasome by low-dose MG132 has a preventive effect on DN development and progression in rats through the up-regulation of antioxidant genes.

[Effects of costimulatory molecule VSIG4 exclusively expression in macrophage on renal interstitial fibrosis].

AIM: To investigate the relationship between costimulatory molecule VSIG4 expression in macrophage and mice renal interstitial fibrosis. METHODS: A total of 40 healthy male C57B6 mice (VSIG4(-/-); and VSIG4(+/+);) were divided into two groups, VSIG4(-/-); tubulointerstitial Nephritis (n=20) and VSIG4(+/+); tubulointerstitial Nephritis (n=20), mice received left urethral obstruction operation(UUO) then were killed at 7 d before operation, 3 d, 7 d and 14 d post operation for both groups.Blood SCr and BUN were detected. Renal interstitial fibrosis was measured by HE stain. The expression of TGF-beta1, MMP-2, in UUO mice(VSIG4(-/-); and VSIG4(+/+);) were detected by immunohistochemistry. RESULTS: SCr and BUN levels in the two groups have no significantly different. TGF-beta1 was significantly increased in the VSIG4(-/-); UUO mice group in comparison with the other group, while MMP-2 was reduced. CONCLUSION: The interstitial inflammatory cell infiltration and tubular lesion of VSIG4(-/-); UUO mice were increased, with the expression of TGF-beta1 increased and MMP-2 reduced. VSIG4 may play a role in inhibiting interstitial fibrosis.

[Hypoxia responsive element regulated herpes simplex virus-thymidine kinase system enhances killing effect of gancyclovir on Ewing's sarcoma cell line under hypoxic condition].

OBJECTIVE: To find out a possible approach to improve the effectiveness of radiotherapy and chemotherapy for Ewing's sarcoma by constructing a eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia and to evaluate the effects of this HRE regulated HSV-TK system on killing effect of gancyclovir (GCV) on Ewing's sarcoma cell line SK-ES under hypoxic condition. METHODS: The HRE was synthesized according to the literature and cloned into the enhancer site of pIRES(2)-EGFP vector to obtain the pHRE recombinant plasmid. The HSV-TK was amplified by PCR and cloned into the multiple clone site of pIRES(2)-EGFP and pHRE to obtain pTK and pHRE-TK recombinant plasmid. The human Ewing's sarcoma cell line SK-ES was transfected by pTK or pHRE-TK recombinant plasmid with liposome and then was exposed to normoxic (21% oxygen) or hypoxic (3% oxygen) condition. The expression of enhanced green fluorescent protein (EGFP) was monitored by fluorescent microscopy. The sensitivity of human Ewing's sarcoma cell line SK-ES transfected with pTK or pHRE-TK recombinant plasmid to the anti-tumour drug GCV was determined with the method of tetrazolium (MTT) after treating with GCV for five days. RESULTS: (1) The result of sequencing showed that the recombinant plasmid pHRE contained HRE, and that the recombinant plasmid pTK and pHRE-TK contained HSV-TK gene in the sense direction. (2) Comparison of fluorescent optical density (FOD) showed that (1) the EGFP FOD value of pHRE and pHRE-TK group cells exposed to hypoxia was significantly higher than those exposed to normoxia (P < 0.01); (2) when the cells were exposed to hypoxia, the EGFP FOD value of pHRE and pHRE-TK group cells was significantly higher than that of pTK and empty vector group (P < 0.01); (3) there was no significant difference among the four groups of cells when they were exposed to normoxia (P > 0.05). (3) Comparison of the sensitivity of four groups of cells to GCV showed that (1) the cells in pHRE-TK and pTK groups were much more sensitive to GCV than the cells in pHRE group under hypoxia condition (P < 0.01), the higher the GCV concentration, the greater the difference; (2) the cells of pHRE-TK group were more sensitive to GCV than those in pTK group under hypoxic condition (P < 0.01), but was almost equally sensitive under normoxic condition (P > 0.05); (3) the pHRE-TK group cells had higher sensitivity to GCV under hypoxia than normoxia (P < 0.01) while the pTK group cells had almost the same sensitivity to GCV under hypoxia and normoxia (P > 0.05). CONCLUSION: (1) The eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia was constructed successfully. (2) HRE could up-regulate expression of EGFP by SK-ES cells under hypoxia condition. (3) HRE could enhance the killing effect of HSV-TK/GCV system on human Ewing's sarcoma cell line SK-ES under hypoxic condition.

Clinical comparisons of 1,498 chronic renal failure patients with and without tuberculosis.

OBJECTIVE: To investigate the clinical characteristics of chronic renal failure (CRF) patients suffering from tuberculosis. METHODS: Clinical materials from CRF inpatients in our department suffering from tuberculosis from 1997 to 2003 were investigated and compared with similar data from tuberculosis patients without CRF in Chongqing City (China) during the same period. RESULTS: Of the 1,498 tested CRF inpatients, 71 suffered from tuberculosis, more than half of which suffered from atypical type V tuberculosis (lesion spot uncertain). The incidence of tuberculosis in the CRF inpatients (4740/100,000) was significantly higher than that in Chongqing City (151/100,000; P<.001). We found an inverse association between renal function and tuberculosis; the worse the renal function, the higher the tuberculosis incidence. CRF patients suffering from tuberculosis had fewer positive antituberculosis-antibody and purified protein-derivative tests (12.7% and 15.5%, respectively) than did tuberculosis patients without CRF (72.1% and 58.1%, respectively; P<.05). In CRF patients with tuberculosis, the two-thirds dosage antituberculosis drug treatment was effective. Twelve cases had good outcome after a 1-year course of antituberculosis treatment and then kidney transplantation. In contrast, 4 patients who received less than half a year of antituberculosis treatment died after kidney transplantation, in association with relapsed tuberculosis. CONCLUSION: CRF patients are susceptible to tuberculosis, especially to atypical tuberculosis outside the lung. Thus, antituberculosis treatment might be necessary in CRF patients found to be suffering from chronic fevers of unknown origin. Furthermore, it appears to be safe for patients with CRF plus tuberculosis to receive kidney transplantation after 1 year of antituberculosis treatment.

[Sequence-dependent cleavage of HBV DNA by combining with triple helix-forming oligodeoxyribonucleotides modified with manganese porphyrin in vitro].

OBJECTIVE: To observe the ability of triple helix-forming oligonucleotides (TFO) modified with manganese porphyrin to combine with and cleave HBV DNA fractions. METHODS: The ends of TFO were modified with manganese porphyrin and acridine; At 37 degrees C and pH 7.4 condition in vitro, TFO modified with manganese porphyrin and acridine were bound with 32P labeled HBV DNA fragments, the affinity and specificity binding to target sequence were tested by electrophoretic mobility shift and DNase 1 footprinting assays, the ability to cleave HBV DNA fractions was observed with cleavage experiments. RESULTS: TFO modified with manganese porphyrin and acridine could bind to target sequence in a sequence-dependent manner with Kd values of 3.5 x 10(-7) mol/L and a relative affinity of 0.008. In the presence of KHSO5, TFO modified with manganese porphyrin and acridine could cleave target sequence in the region forming triple DNA. CONCLUSION: In the presence of KHSO5, TFO modified with manganese porphyrin and acridine could cleave target HBV-DNA in sequence-dependent manner.