BACKGROUND:Human antigen R (HuR) is a ubiquitously expressed RNA-binding protein that modulates gene expression at the post-transcriptional level. While cytoplasmic HuR expression was identified as a marker in epithelial-mesenchymal transition (EMT) process of several types of cancer, its role in diabetic nephropathy (DN) remains unclear.METHODS:Renal biopsies from Type 2 diabetic patients and STZ-induced DN rats were stained for HuR and EMT markers. Redistribution of HuR was detected by immunostaining and western blot in high glucose stimulated cells. RNAi was used to supress HuR expression. The binding affinity for EMT-related genes was evaluated by immunoprecipitation.RESULTS:Cytoplasmic HuR expression was elevated in human and rat DN specimens along with EMT changes compared to normal controls. HuR shuttling between nucleus and cytoplasm facilitated epithelial to mesenchymal transition in renal epithelial cells. The suppression of HuR partially inhibited EMT of high glucose stimulated HK-2 cells. Furthermore, HuR bound to 3'-UTRs of critical cytokines or transcription factors mRNA involved in EMT process.CONCLUSION:Acquired phenotypic traits of EMT were partially through the enhanced HuR-binding proteins and its post-transcriptional regulation role in DN.

AIM:The study was aimed to analyze whether ACE rs267604983 polymorphism was related to the onset of diabetic nephropathy (DN).METHODS:80 DN patients and 78 healthy controls were enrolled in the study. The differences of age, sex, body mass index (BMI), systolic blood pressure (SBP) and diastolic blood pressure (DBP) between two groups were analyzed. The genotyping of ACE rs267604983 was conducted by the technology of PCR-HRM. Odds ratio (ORs) with 95% CI were used to evaluate the relationship of ACE rs267604983 with DN susceptibility.RESULTS:The AA genotype of ACE rs267604983 was remarkably associated with the risk for DN (OR = 2.90, 95% CI = 1.12-7.51). In addition, for the A allele carriers, the risk for DN increased 1.87 fold (OR = 1.87, 95% CI = 1.16-3.01). The subgroup analysis showed that the AA genotype was found higher in normal albuminuria group than other groups (P = 0.006), while AG genotype was higher in macro albuminuria group (P = 0.036).CONCLUSION:ACE rs267604983 polymorphism is associated with the risk of DN. AA genotype and A allele may increase the risk for DN. Furthermore, AA and AG genotypes may have effects on the subgroups of DN.

目的:探讨肾炎康复片辅助标准激素治疗气阴两虚型肾病综合征（NS）的临床疗效以及对血小板数量（PLT）和血浆垂体腺苷酸环化酶激活肽（PACAP）的影响。方法:152例NS患者参照随机按分层随机法分为治疗组80例和对照组72例;两组患者均采取常规支持治疗和对症处理;对照组采用泼尼松,首始剂量l mg·kg-1·d-1,最大剂量不超过60 mg·d-1,连续口服8~12周,再以每2周减少原使用量的10%内服,最后以10 mg·d-1的剂量维持治疗,根据具体情况维持6~12个月,同时给予对症治疗。治疗组在对照组治疗的基础上加服肾炎康复片,5片/次,3次/d,两组均连续治疗12个月。检测治疗前后两组患者的24 h尿蛋白定量,血清白蛋白（ALB）,总胆固醇（TC）和甘油三脂（TG）水平;监测两组肝功能,记录治疗过程中不良反应发生率;比较两组治疗前后中医（TCM）单项症状评分;检测两组治疗前后PLT和血浆PACAP水平。结果:治疗组临床总有效率为97.5%,对照组为79.17%,治疗组明显优于对照组（P＜0.01）;治疗组治疗后24 h尿蛋白定量,TG和TC水平明显低于对照组,血清ALB水平显著高于对照组（P＜0.01）;治疗后治疗组患者的向心性肥胖和总体不良反应发生率明显少于对照组（P＜0.05）;治疗组治疗后TCM各单项指标评分均明显低于对照组,比较差异均有统计学意义（P＜0.01）;治疗组治疗后PLT数量明显低于对照组,血浆PACAP水平显著高于对照组,比较差异均有统计学意义（P＜0.01）。结论:肾炎康复片辅助标准激素治疗气阴两虚型NS能提高临床疗效,减少不良反应发生,降低患者血小板数量和升高血浆PACAP水平。

It has been well documented that survivin has multiple functions including cytoprotection, inhibition of cell death, and cell cycle regulation, particularly at the mitotic stage of the cell cycle, all of which favor cancer survival. Its expression in normal tissue is developmentally regulated, and any type of deregulation in survivin expression favors cancer survival. Gastric cancer is one of the most common malignancies and the second most common cause of cancer-related mortality worldwide. The molecular mechanisms involved in the transformation and progression of gastric cancer remain unclear. In the present study, we investigated the effect of lentiviral vector-mediated survivin shRNA delivery in gastric cancer cell lines. Lentiviral-mediated survivin shRNA was used to knock down survivin expression in gastric cancer cell lines SGC-7901, MGC-803 and MKN-28. The Τranswell chemotaxis and the CCK-8 assays were used to assess the migration and proliferation of the tumor cells, respectively. TUNEL assay was used to detect apoptosis. Quantitative real-time PCR and western blot analysis were used to quantify mRNA and protein levels, respectively. Our results demonstrated that lentiviral-mediated RNAi markedly suppressed the survivin expression in all three gastric cancer cell lines. Significant decrease in survivin mRNA and protein expression were detected in the gastric cancer cell lines stably transfected with the lentiviral survivin shRNA vector, and knockdown of survivin also significantly inhibited the proliferation and migration in the gastric cancer cells and tumorigenicity in a xenograft animal model. Our results indicated that aberrant high cytoplasmic survivin expression in gastric cancer cells is associated with increased proliferation index and tumor growth. In conclusion, our results suggest that lentiviral-mediated gene therapy has the potential to be developed into a novel therapeutic strategy for the treatment of gastric cancer.

Abnormal proliferation of human mesangial cells was the earliest pathological character in chronic kidney disease and linked to the accumulation of extracellular matrix and glomerular sclerosis. Multifunctional Angiotensin (AngII) had been emerged as a key player in initiation and progression of fibrogenic processes in kidney. In mesangial cells, treatment with the proliferation stimulus AngII triggered the escalated cyclinD1 expression, where its association with HuR increased dramatically. In our study, it was demonstrated that both in vivo and in vitro HuR redistribution in dysregulated mesangial cell proliferation accompanied by an abundant cyclinD1 expression following the AngII treatment. ActinomycinD experiments revealed that AngII stabilized cyclinD1 mRNA in human mesangial cells via HuR. Furthermore, employing the RIP-Chip assay yielded cyclinD1 mRNA with a higher affinity to HuR in mesangial cells induced by AngII compared with the normal ones in vitro study. Analysis of a cyclinD1 mRNA directly implicated HuR in regulating cyclinD1 production: cyclinD1 translation increased in HuR-shuttling cells induced by AngII and declined in cells in which HuR levels were lowered by RNA interference. We proposed that the release of HuR-bound mRNAs via an AngII–cyclinD1–HuR regulatory axis was implicated in the evolution of proliferative kidney diseases, providing us a novel therapeutic strategy to treat glomerular disease.

The aim of the present study was to clarify the association between lipid metabolism and the atherosclerosis in early-stage chronic renal failure at the molecular level and to explore the efficacy of decorin on chronic renal failure. Sprague Dawley rats receiving 5/6 nephrectomy and Sham surgery were divided into control and experimental groups. Sprague Dawley rats receiving 5/6 nephrectomy were divided into control and experimental groups, and the experimental group was further subdivided into rats receiving treatment with fibroblasts (FBs) transfected either with empty vector and with a decorin (DCN) gene. The dynamic levels of triglyceride (TG), total cholesterol (T-Ch) and total phospholipid (T-PL) were detected on the 10th, 30th and 60th days. The body weight, blood lipid levels, renal function and renal tissue were observed after four weeks, and transforming growth factor-βl and protein expression was detected by immunohistochemistry. In total, 4 weeks after treatment, the DCN expression in the renal tissue of rats treated with DCN-transfected FBs was significantly increased compared to that in the control rats. The results showed that the levels of the three lipids in the aortic arches were slightly elevated on the 10th day compared with those in the control group, and the TG level was significantly increased on the 30th day. The levels of T-Ch, TG and T-PL in the aortic arches were significantly elevated on the 60th day. The TG and T-Ch levels in the plasma and aortic tissues of Sprague Dawley rats receiving 5/6 nephrectomy without any treatment and after receiving treatment with FBs transfected with empty vector were significantly increased compared with those in the control group. The increased T-Ch and decreased T-PL levels in the erythrocyte membrane increased the rigidity of the erythrocyte and decreased erythrocyte deformability. In conclusion, highly expressed DCN mitigated renal fibrosis and thus delayed renal failure as well as mitigating the abnormal lipid metabolism of the chronic renal failure.

Distal esophageal adenocarcinoma is a highly aggressive neoplasm. Despite advances in diagnosis and therapy, the prognosis is still poor. Stathmin (STMN-1) is a ubiquitously expressed microtubule destabilizing phosphoprotein. It promotes the disassembly of microtubules and prevents assembly. STMN-1 can cause uncontrolled cell proliferation when mutated and not functioning properly. Recently, found to be overexpressed in many types of human cancers. However, its clinical significance remains elusive in distal esophageal adenocarcinoma. Here, we reported for the first time that STMN-1 is highly overexpressed in adenocarcinomas of the distal esophagus and strongly associated with lymph node metastasis.

Background and Aim: We have reported previously that RNA interference targeting stathmin1 (STMN1) gene in human gastric cancer cells inhibits proliferation in vitro and tumor growth in vivo. Based on these observations, in the present study, the possibility that local injection of lentivirus-delivered stathmin shRNA would induce regression of the established human gastric cancer xenograft in animal model was investigated.Methods: BALB/c nude mice were inoculated subcutaneously into the right armpit with human gastric cancer cells SGC-7901(2 x 106 cells in 200 mu L phosphate-buffered saline) to develop a xenograft model of human gastric cancer. When tumor reached suitable size, mice were randomly divided into two groups. STMN1 shRNA group (n = 6) were given local injection of lentivirus-delivered STMN1 shRNA, and the non-silencing shRNA group (n = 6) were administered with local injection of lentivirus-delivered non-silencing shRNA. Quantitative reverse transcription-polymerase chain reaction and Western blot were used to verify the knockdown of the gene expression in dissected tumor at mRNA and protein level, respectively.Results: Experimental therapy on the nude mice model bearing subcutaneous tumor of SGC-7901 cells showed that local administration of STMN1 shRNA effectively regressed the pre-established tumors. Stathmin shRNA-treated tumors were significantly regressed as compared with that of the tumor injected with non-silencing shRNA (P < 0.05). Tumor weight was significantly decreased in STMN1-treated group as compared with non-silencing shRNA group (P < 0.05). Quantitative reverse transcription-polymerase chain reaction and Western blot showed downregulation of STMN1 gene expression in STMN1 shRNA group as compared with non-silencing shRNA group (P < 0.05).Conclusion: These findings highlight the potential use of local injection of lentivirus-delivered shRNA for the treatment of early localized human gastric carcinoma.

BACKGROUND:Prognosis of esophageal squamous cell carcinoma (ESCC) is stage-specific; however, some patients with the same stage have different survival outcomes. Clinically, it is significant to explore the biological marker to predict patient's outcome. We investigated the association between the stathmin1 gene (STMN-1) expression and the prognosis of patients who underwent Ivor-Lewis esophagectomy.METHODS:A total of 162 patients who suffered from midthoracic stage IIA ESCC and completely resected with Ivor-Lewis esophagectomy were studied for STMN-1 expression by qRT-PCR in fresh-frozen tissue and validated by immunohistochemistry in matched formalin fixed-paraffin embedded tissue samples. STMN-1 level was evaluated as a prognostic factor in ESCC. SPSS 21.0 software was used to analyze the relationship between STMN-1 expression and clinicopathological characteristics and survival probability.RESULTS:The overall 3- and 5-year survival was 72.20 and 42.00 % respectively. Ninety-four patients (58.02 %) experienced disease recurrence with a disease-free interval of 21.50 ± 1.20 months. qRT-PCR result showed that STMN-1 mRNA level in patients who were alive at the end of follow-up was lower compared with patients who died during the follow-up period (p < 0.05). Immunohistochemical results showed that 94 patients had STMN-1 protein overexpression (58.02 %), patient with STMN-1 overexpression had worse survival compared with patients who had low STMN-1 expression (p = 0.00). Cox regression analysis revealed that STMN-1 protein expression and T classification are independent prognostic factors.CONCLUSIONS:Even localized ESCC are potential to relapse with poor prognosis. This study demonstrates that STMN-1 level is an independent prognostic factor after Ivor-Lewis esophagectomy. In addition, assessment of STMN-1 level could improve stratification of stage IIA ESCC patients.

AIMS:High cardiovascular mortality in patients with end-stage renal disease is closely associated with arterial medial calcification (AMC) caused by hyperphosphatemia, the mechanism of which associated hormones (FGF-23, klotho) and osteochondrogenic events is unclear. We examined the effect of Lanthanum carbonate on AMC via regulating the abnormalities in phosphorus metabolism of uremic rats.MAIN METHODS:45 healthy SD rats were randomly divided into 3 groups: Normal group (n=15), CRF group (n=15), CRF diet supplemented with 2% La (n=15). AMC in great arteries were evaluated by VonKossa. Osteochondrogenic specific genes were analyzed by Immunohistochemistry and qRT-PCR. Serum FGF-23 and klotho levels were detected by ELISA kit.KEY FINDINGS:Serum phosphate was markedly increased in CRF group (6.94 ± 0.97 mmol/L) and 2%La group (5.12 ± 0.84 mmol/L) at week 4, while the latter became hypophosphatemic (2.92 ± 0.73 mmol/L vs CRF group, p<0.01) at week 10. Inhibitory effect of 2%La on development of AMC was reflected by downregulated Runx2, Osterix, BSP, Osteocalcin and collagenII and a reduction of FGF-23 at week 4(vs CRF group, p<0.01) but not week 10.SIGNIFICANCE:Beneficial effects of Lanthanum carbonate on progression of AMC in CRF could be mainly due to the decreased phosphate retention and FGF-23 in early stage and likewise a reduction of bone-associated proteins via osteochondrogenic pathway. Lanthanum carbonate has no effect on soluble klotho and serum FGF-23 in late stage of CRF.

Arterial medial calcification (AMC) is frequent prevalence in patients with end stage renal disease. Evidence about hyperphosphatemia induced anabolic crosstalk between osteoblast and osteoclast in AMC of uremia is rare. Lanthanum carbonate as an orally administered phosphate-binding agent to reduce phosphate load and ameliorate AMC, but direct evidence is missing.

Objective To establish the model of acute renal rejection in rats after kidney transplantation and investigate the pathomechanism of transplanted tissue.Methods Seventy-eight normal male SD rats were divided into normal group(group A,n=15),pseudo-surgery group(group B,n=15),non-acute graft rejection group(group C,n=16,SD rats as donor),and acute graft rejection group(group D,n=16,Wistar rats as donor).The renal function of these four groups was evaluated at the seventh day after transplantation operation.Grafts were harvested at the seventh day and were observed the pathological changes under the light microscope.Acute rejection semiquantitative scores were analyzed according to the standard of Banff 97.Results The serum levels of urea nitrogen,creatinine and acute rejection semiquantitative scores were significantly higher in group C and group D than those in group A(P0.05),and were higher in group D than those in group C(P0.01).Conclusion Wistar-SD rats kidney transplantation can be used for acute rejection model after kidney transplantation.

Context: Multiple organ dysfunction syndrome (MODS) is a major cause of death in critically patients. It has been hypothesized that inactivation or removal of pro-inflammatory molecules may prevent or reverse MODS. Objective: The purpose of this paper was to investigate the efficacy of continuous veno-venous hemodiafiltration (CVVHDF) as treatment for MODS in an established animal model. Materials and methods: Male Beagle dogs (n = 18) were used to establish the model and were randomly assigned to a CVVHDF, sham, or control group. The serum levels of ALT, AST, Cr, BUN, PaO2, and PaCO2 were measured as functional makers of major organs. Apoptosis, DLA-DR expression, and cytokine levels of peripheral monocytes were determined. Results: The MODS model was successfully established. After CVVHDF treatment, the WBC and neutrophil counts were lower and the monocyte count and percentage were greater, but these were unchanged in the sham and control groups. Apoptosis of CD14+ monocytes was significantly lower in the CVVHDF group than in the sham and control groups. The fraction of DLA-DR+ monocytes and IL-1β secretion was significantly greater in the sham and control groups than in the CVVHDF group. Moreover, IL-4 secretion increased significantly in the CVVHDF group but not in the control group. Discussion and Conclusion: Our study of an experimental model of MODS indicated that MODS leads to significant disruption of physiological and immune functions. CVVHDF treatment alleviated some of these symptoms due to the improvement of monocyte function, reduction of monocyte apoptosis, and increase of anti-inflammatory cytokines.

Acute graft rejection is one of the most common and serious post complications in renal transplantation, noninvasive diagnosis of acute graft rejection is essential for reducing risk of surgery and timely treatment. In this study, a non-targeted metabonomics approach based on ultra performance liquid chromatography (UPLC) coupled with quadrupole time-of-flight mass spectrometry (MS) is used to investigate the effect of acute graft rejection in rat renal transplantation on metabolism. To collect more metabolite information both hydrophilic interaction chromatography and reversed-phase liquid chromatography were used. Using the partial least squares-discriminant analysis, we found that the change of metabonome in a sham-operated group and a non-graft rejection group had a similar trend, while that of the acute graft rejection group was clearly different. Several discriminating metabolites of the acute graft rejection were identified, including creatinine, phosphatidyl-cholines, lyso-phosphatidylcholines, carnitine C16:0, free fatty acids and indoxyl sulfate etc. These discriminating metabolites suggested that acute graft rejection in renal transplantation can lead to the accumulation of creatinine in the body, and also the abnormal metabolism of phospholipids. These findings are useful to understand the mechanisms of the rejection, it also means that a UPLC-MS metabonomic approach is a suitable tool to investigate the metabolic abnormality in the acute graft rejection in renal transplantation.

Objective To study the effect of continuous venovenous haemodiafiltration (CVVHDF) on the clearances of various solutes in multiple organ dysfunction syndrome (MODS)dogs.Methods Dogs were subjected to hemorrhagic shock plus resuscitation and endotoxemia to set up MODS model,then they were randomly divided into 2 groups: MODS group (M group)and MODS+CVVHDF group (M+C group).A PRISMA pre-dilution system with AN-69 filter was applied.Solute clearance of nitric oxide(NO),urea nitrogen(UN),creatinine (Cr),interleukin-6 (IL-6),interleukin-10 (IL-10),tumor necrosis factor-α (TNF-α) and endotoxin (LP) was determined with same dialysate and ultrafiltrate volume during CVVHDF.The clearance of various solutes (Kd) was calculated by the concentration of the cleared solutes in effluents (E) based on the formula Kd=(ExQE)/P.Results Clearances of NO,UN,Cr were greater than those of IL-6,IL-10,TNF-α,LP during the treatment of CVVHDF.Conclusion CVVHDF could effectively reduce the levels of various solutes in the development of MODS by convection and absorption.

外周血单核细胞对多器官功能障碍综合征(MODS)的炎性反应和免疫反应均起重要作用.单核细胞表面白细胞抗原DR(HLA-DR)表达和机体免疫功能密切相关.本研究通过观察连续性静脉-静脉血液透析滤过(CVVHDF)治疗MODS犬CD14+单核细胞HLA-DR的动态变化,以期阐明CVVHDF治疗MODS对机体免疫系统的影响及其疗效机制.