Ustekinumab (UST) is a human IgG1 monoclonal antibody that targets to the share p40 subunit of interleukin-12(IL-12) and IL-23. Evidence has shown that UST therapy is well tolerated and effective in inducing clinical response in refractory CD(Crohn's disease) and dose escalation is effective in recapturing response in over half of the patients. However, no predictive factor has been reported to be helpful for UST treatment in clinical practice. Additionally, there were few reports about therapeutic drug monitoring (TDM) of UST administration in China due to its late launch time in Chinese market and lack of experience in clinical use. Herein, we establish and validate the first UST-trough concentrations (TCs) -related nomogram in China for predicting endoscopic remission in refractory CD to facilitate clinical decision making.

Inflammatory bowel disease (IBD), characterized primarily by gastrointestinal inflammation, predominantly manifests as Crohn’s disease (CD) and ulcerative colitis (UC). It is acknowledged that Inflammation plays a significant role in cancer development and patients with IBD have an increased risk of various cancers. The progression from inflammation to carcinogenesis in IBD is a result of the interplay between immune cells, gut microbiota, and carcinogenic signaling pathways in epithelial cells. Long-term chronic inflammation can lead to the accumulation of mutations in epithelial cells and the abnormal activation of carcinogenic signaling pathways. Furthermore, Immune cells play a pivotal role in both the acute and chronic phases of IBD, contributing to the transformation from inflammation to tumorigenesis. And patients with IBD frequently exhibit dysbiosis of the intestinal microbiome. Disruption of the gut microbiota and subsequent immune dysregulation are central to the pathogenesis of both IBD and colitis associated colorectal cancer (CAC). The proactive management of inflammation combined with regular endoscopic and tumor screenings represents the most direct and effective strategy to prevent the IBD-associated cancer.

Background Long noncoding RNAs (lncRNAs) have been implicated playing important roles in human urologic cancers. Up to date, quite a few lncRNAs have been implicated as promising biomarkers for tumor early detection and prognosis monitoring. Methods In the present study, microarray analysis was initially performed to screen the differentially expressed lncRNAs between bladder cancer tissues and paired adjacent non-cancerous tissues (n=3).Subsequent qRT-PCR validation was conducted using tissue samples from 95 patients with bladder cancer. Results Results showed that the expression level of lncRNA-n336928 (noncode database ID: n336928) was significantly higher in bladder cancer tissues compared to that in adjacent noncancerous tissues (P<0.001). Chi-square test showed that expression of lncRNA-n336928 was positively correlated with bladder tumor stage and histological grade (P<0.001). Kaplan-Meier survival analysis revealed that patients with bladder cancer with high expression of lncRNA-n336928 had shorter overall survival time compared to patients with low expression of lncRNA-n336928. Multivariate analysis indicated that lncRNA-n336928 was an independent prognostic factor for overall survival for bladder cancer patients. Conclusions our study shows that high expression of lncRNA-n336928 is associated with the progression of bladder cancer, and that lncRNA-n336928 might serve as a biomarker for prognosis of bladder cancer

Objective To constructe the rat-specific EZH2 overexpression plasmid and the EZH2 catalytic subunit SET domain deletion plasmid,as well as assess their levels of expression in 293T cells.Methods The RNA extracted from rat cardiac tissue was reverse-transcribed into cDNA,the cDNA of catalytic subunit SET domain deleted EZH2 was amplified by overlap PCR,then the PCR amplification was achieved using the EZH2 gene as template.Products through PCR amplification and vector were cut with double enzyme,and then ligated the target fragments,purified by gel extraction,using T4 ligase.The ligated products were transferred into competent cells.After plate coating,monoclonal picking,shaking cultivation and sequencing procedures,the plasmids were extracted and transfected into 293T cells using liposome-mediated method.Results Western Blot and RT-PCR assays showed recombinant plasmids DNA can successfully overexpress EZH2 gene in 293T cells.Conclusions Two plasmids were successfully constructed.This study laid the foundation for the further study on the relationship between EZH2 and myocardial hypertrophy,and the function of EZH2 catalytic subunit.

The aim of the current study was to investigate the biological effect on T24 cells and human umbilical vein endothelial cells (HUVECs) of transfection with brain-specific angiogenesis inhibitor-1 (BAI-1). The recombinant plasmid pReceiver-M61-BAI-1 was transfected into human superficial bladder tumor cells (T24) and HUVECs, in parallel with the vector control. mRNA and protein expression levels of BAI-1 were then detected by quantitative polymerase chain reaction (qPCR) and western blotting, respectively. Cell apoptosis of T24 cells and HUVECs prior and subsequent to transfection with BAI-1 was analyzed by flow cytometric analysis. Proliferation of T24 cells and HUVECs prior and subsequent to transfection of BAI-1 was assessed by the MTT method. T24 cells and HUVECs transfected with pReceiver-M61-BA1-1 were classed as the experimental group; T24 cells and HUVECs transfected with p-Receiver-M61 were the control group. qPCR and western blotting methods confirmed that there was positive expression of BAI-1 in T24 cells and HUVECs transfected with pReceiver-M61-BAI-1, however BAI-1 was not expressed in T24 cells and HUVECs transfected with pReceiver-M61. The results of the MTT assay demonstrated that absorbance was markedly reduced in HUVECs at 12, 48 and 72 h subsequent to transfection with pReceiver-M61-BAI-1 when compared with that of the control group and in T24 cells transfected with p-Receiver-M61-BAI-1. Furthermore, flow cytometry results also indicated that the apoptotic rate of HUVECs transfected with p-Receiver-M61-BAI-1 was significantly increased compared with that of the control group and T24 cells transfected with p-Receiver-M61-BAI-1. BAI-1 was observed to markedly inhibit the proliferation of vascular endothelial cells in vitro, however, no direct inhibition by BAI-1 was observed in T24 cells. In conclusion, BAI-1 is suggested to be a potential novel therapautic target for the inhibition of tumor neovascularization.

In mouse testes, Musashi-1 (Msi-1) was predominantly expressed in the cytoplasm and nuclei of Sertoli cells. Here we demonstrate that knockdown of Msi-1 in Sertoli cells altered the levels and distribution of blood-testis barrier (BTB)-associated proteins. Moreover, Msi-1 knockdown in vivo disrupted BTB functional structure and spermatogenesis. In addition, we report a novel role of Msi-1 in regulating Sertoli cells survival following heat-induced injury. Endogenous Msi-1 protein in heat-treated Sertoli cells was recruited to stress granules. The formation of stress granules was considerably disrupted, and apoptosis was significantly up-regulated in Msi-1-knockdown Sertoli cells after heat treatment. p-ERK1/2 acted downstream of stress granule formation, and inhibition of p-ERK1/2 signaling triggered Sertoli cell apoptosis upon heat stress. In conclusion, we demonstrate that Msi-1 is critical for constructing a functional BTB structure and maintaining spermatogenesis. We also note a role for Msi-1 in regulating Sertoli cell fate following heat-induced injury, likely through the induction of stress granule formation and subsequent activation of p-ERK1/2 signaling.

Objectives The endothelial lipase gene (LIPG) is one of the important genes in the metabolism of high-density lipoprotein cholesterol (HDL-C) and may be involved in the pathogenesis of coronary artery disease (CAD). Materials and methods To investigate the relationship between the common single nucleotide polymorphisms (SNPs) 584C/T (rs2000813) and −384A/C (rs3813082) in the LIPG gene and CAD, allele and genotype frequencies of the two SNPs were analysed in 287 Chinese patients with CAD and 367 controls by the high-resolution melting curve (HRM) method. Results For 584C/T, no significant difference in polymorphic distribution was observed between patients and controls. However, the frequencies of allele C (20.2% vs 15%, p=0.013, OR=1.437, 95% CI 1.078 to 1.915) at −384A/C were significantly increased in patients compared with controls. Haplotype analysis also showed that haplotype CT (12.37% vs 8.72%, p=0.035, OR=1.478, 95% CI 1.034 to 2.112) was significantly higher in patients compared with controls. Conclusions These results suggested that the SNP −384A/C in the LIPG gene may be associated with risk for CAD and the LIPG gene may play a role in CAD in the Han Chinese.

Bacillus Calmette-Guérin (BCG) reduces the recurrence and progression of non-muscle invasive bladder cancer. The present study aimed to investigate the impact of a recombinant hIFN-α2b-secreting BCG (rBCG) on the mouse bladder MB49 cell line and an orthotopic mouse model of bladder cancer. MB49 cells were cultivated in the presence or absence of rBCG, BCG or BCG+hIFN-α2b. Cellular morphology and viability were assessed by microscopy and CCK-8 assay, respectively. Apoptosis was assessed by acridine orange, Hoechst 33258 staining and flow cytometry. MHC-I expression was assessed by flow cytometry. MB49 cells were transplanted into the bladders of C57BL/6 mice administered BCG, rBCG or BCG+hIFN-α2b. Local tissue Fas expression and T cell subsets were assessed by immunohistochemistry. Peripheral blood TNF-α and IL-12 levels were measured by ELISA, and circulating T lymphocyte subsets by flow cytometry. BCG, rBCG and BCG+hIFN-α2b increased the distortion and death of MB49 cells, yet rBCG reduced the proliferation and enhanced apoptosis most substantially. Apoptosis was increased after a 24-h co-culture with rBCG or BCG+hIFN-α2b. Mice administered rBCG survived longer than mice administered BCG (p<0.001), yet this result was not significantly different from mice administered BCG+hIFN-α2b. The average bladder weight was reduced by administration of rBCG (p<0.001). Fas expression and peripheral blood mTNF-α and mIL-12, cell counts of polymorphonuclear leukocytes, monocytes, T lymphocytes and CD4+/CD8+ ratios were significantly increased by all BCG treatments (p≤0.05), yet monocyte and T lymphocyte counts were higher in mice administered rBCG than in mice treated with BCG or BCG+hIFN-α2b (p=0.000). These results indicate that in an orthotopic murine bladder cancer model rBCG possesses superior antitumor activity to BCG+hIFN-α2b.

Objective: Human murine double minute 2 protein (MDM2) is mainly a negative regulator of p53 tumor suppressor pathway. We aimed to investigate the association between MDM2 SNP309 polymorphism and bladder cancer risk. Methods: A total of 535 bladder cancer patients and 649 health controls were recruited for our study. MDM2 SNP309 T>G polymorphism was genotyped by polymerase chain reaction-ligase detection reaction method. Logistic regression was used to analyze the relationship between the genotype and susceptibility of bladder cancer. Kaplan-Meier estimates and log-rank test were obtained to analyze the association between the genotype and risk of recrudesce in nonmuscle-invasive bladder cancer patients. A multivariable Cox proportional hazards model was fitted to identify independent prognostic factors. To further investigate the association, we conducted a meta-analysis including six studies. Results: The frequency of the MDM2 SNP309 T>G polymorphism showed no significant difference between cases and controls (all P>0.05). In the stratification analysis, the results showed that G allele carriers were prone to have a significant decrease in risk of low-grade bladder cancer (adjusted odds ratio: 0.613, 95% confidence interval: 0.427-0.881), and G variant was associated with a significantly reduced risk of recurrence in nonmuscle-invasive bladder cancer patients with or without chemotherapy (P<0.05). The results of the meta-analysis showed that G allele and GG genotype of MDM2 SNP309 polymorphism were significantly associated with increased risk of bladder cancer in Caucasians (both P<0.05), and no association was observed in total populations and Asians (P>0.05). Conclusion: MDM2 SNP309 T>G polymorphism has no influence on bladder cancer risk in Asians, but this single nucleotide polymorphism may be associated with genetic susceptibility of bladder cancer among Caucasians.

Genome-wide association studies (GWAS) have identified a number of genetic variants associated with risk of bladder cancer in populations of European descent. Here, we assessed association of two of these variants, rs11892031 (2q37.1 region) and rs401681 (5p15.33 region) in a Chinese case-control study, which included 367 bladder cancer cases and 420 controls. We found that the AC genotype of rs11892031 was associated with remarkably decreased risk of bladder cancer (adjusted odds ratio (OR), 0.27; 95% confidence interval (CI), 0.09–0.81; p = 0.019), compared with the AA genotype of rs11892031; and that CT/CC genotypes of rs401681 were associated with significantly increased risk of bladder cancer (adjusted OR, 1.79; 95% CI, 1.10–2.91; p = 0.02), compared with the TT genotype of rs401681. We further conducted stratification analysis to examine the correlation between single nucleotide polymorphism (SNP) rs11892031/rs401681 and tumor grade/stage. Results showed that heterogeneity in ORs of tumor categories was not significant for either rs11892031 or rs401681 (p > 0.05), indicating that the two SNPs seemingly do not associate with tumor grade and stage of bladder cancer in our study population. The present study suggests that the SNPs rs11892031 and rs401681 are associated with bladder cancer risk in a Chinese population. Future analyses will be conducted with more participants recruited in a case-control study.

CYP2C19 genetic polymorphisms influence clopidogrel response and clinical outcomes of cardiovascular disease. However, data on their relationship in stroke patients are scarce. We aimed to investigate the influence of CYP2C19 polymorphisms on platelet reactivity and clinical outcomes in ischemic stroke patients treated with clopidogrel. A total of 211 patients were enrolled. All patients were given clopidogrel treatment and underwent CYP2C19 genotyping and platelet function testing by flow cytometry including adenosine diphosphate-induced platelet aggregation (ADP-PAg) and platelet activation markers (PAC-1, CD62P and CD63). The modified Rankin Scale (mRS) was used and ischemic events were evaluated. A total of 129 (61.1%) of the 211 enrolled patients were carriers of CYP2C19 loss-of-function (LOF) alleles (⁎2, ⁎3). After clopidogrel therapy for 7 days, the levels of ADP-PAg, PAC-1, CD62P and CD63 were higher in carriers than noncarriers. CYP2C19 carriage was associated with more frequent high residual platelet reactivity. CYP2C19 polymorphisms alone could explain 12.9%, 4.3%, 8.9% and 5.5% of the inter-individual variability of ADP-PAg, PAC-1, CD62P and CD63 after clopidogrel treatment, respectively. At 6-month follow-up, 38 (19%) patients were scored poor prognosis and 15 (7.6%) ischemic events were observed. Carriers had poorer prognosis than noncarriers (P=0.025). No significant association of CYP2C19 carriage with ischemic events was found. Multiple regression analysis showed that CYP2C19 carriage was an independent predictor of poor prognosis (odds ratio, 3.01; 95% confidence interval, 1.23–7.38; P=0.016). In conclusion, carriage of the CYP2C19 LOF allele has significant influence on clopidogrel response and prognosis in patients with ischemic stroke.

Cytochrome P450 (CYP) 2C19 is a very important drug metabolizing enzyme. Although the single nucleotide polymorphisms (SNPs) of CYP2C19 G681A and G636A have been suggested that they may increase the incidence of cardiovascular events, the relationship between SNPs in CYP2C19 and cerebral ischemic stroke (CIS) are unclear. The aim of this study was to investigate the correlation between the distribution of G681A and G636A polymorphisms in CYP2C19 gene and the risk of CIS in Chinese.

Nuclear factor-kappa B (NF-κB), a cell survival signal, is involved in carcinogenesis. Polymorphism of NF-κB1 is associated with cancer by several studies. This study aims to perform a comprehensive meta-analysis of studies and determine the association between the NF-κB1-94ins/del ATTG promoter polymorphism and cancer. Twenty-five case–control studies (7,281 cases and 10,039 controls) were included. We used odds ratios (ORs) to assess the strength of the association, and 95 % confidence intervals (CIs) to identify precision of the estimate. Overall, NF-κB1-94ins/del ATTG promoter polymorphism was significantly associated with decreased susceptibility to cancer in overall population under homozygote (for DD vs. WW: OR = 0.74, 95 % CI = 0.58–0.96), recessive (for DD vs. WD+WW: OR = 0.82, 95 % CI = 0.69–0.99), dominant (for DD+WD vs. WW: OR = 0.84, 95 % CI = 0.71–1.00), and allele (for D vs. W: OR = 0.88, 95 % CI = 0.78–0.98) model. Subgroup analysis for ethnicity found that NF-κB1-94ins/del ATTG promoter polymorphism was significantly associated with decreased susceptibility to cancer in Asians (for DD vs. WW: OR = 0.54, 95 % CI = 0.40–0.74; for WD vs. WW: OR = 0.75, 95 % CI = 0.69–0.81; for DD vs. WD+WW: OR = 0.70, 95 % CI = 0.55–0.90; for DD+WD vs. WW; OR = 0.66, 95 % CI = 0.56–0.78; for D vs. W: OR = 0.75, 95 % CI = 0.65–0.86), but the association was not found in Caucasians. The findings suggest that NF-κB1-94ins/delATTG promoter polymorphism is significantly associated with decreased susceptibility to cancer in overall and Asian population.

New treatment strategies are required for renal cell carcinoma (RCC) due to its relative insensitivity to conventional radio- and chemotherapies. The promising strategy of tumor inhibition using human telomerase reverse transcriptase (hTERT)-controlled herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) in the hTERT promoter-driven HSV-TK/GCV suicide gene system was investigated. Tumor volume, weight, relative proliferation rate, and cell-apoptosis levels were examined in mice injected with adenovirus (Ad)-hTERT-HSV-TK and GCV. Increased cell death occurred following treatment with Ads carrying hTERT-HSV-TK/GCV or cytomegalovirus promoter-controlled (CMV)-HSV-TK/GCV for human RCC 786-0 and fibroblast MRC-5 cells. In mice, Ad-hTERT-HSV-TK/GCV more specifically inhibited tumor and RCC xenograft growth than Ad-CMV-HSV-TK/GCV (P < 0.05). Furthermore, Ad-hTERT-HSV-TK/GCV did not significantly damage normal fibroblasts or organ systems (heart, lung, liver, brain, kidney, and spleen). Thus, Ad-hTERT-HSV-TK/GCV is an effective RCC inhibitor in human cells in vitro and in vivo mouse models, indicating potential usefulness in RCC-targeted gene therapy.

Objective:This study aims to compare the difference in the expression and localization of brain-specific angiogenesis inhibitor 1 (bai-1) between human normal and clear cell carcinoma of kidney tissues at different clinical stages. this study also aims to explore the mechanism of bai-1-mediated inhibition of tumor endothelial cell proliferation. Methods:A total of 133 human normal and cancerous kidney tissues were obtained. the tissue localized more than 4 cm away from the cancerous kidney tissue was identified as the normal control kidney tissue. immunohistochemical staining was conducted to detect the expression of bai-1, vegf, mvd, and p53. the expression was detected with the respective antibodies for quantitative statistical analysis. fresh renal cell carcinoma tissue samples were obtained from 27 patients, and 15 tissue samples were obtained 4 cm away from the cancer. the expression of bai-1 in the renal cell carcinoma tissue was examined through western blot analysis. Results:The bai-1 expression level in the normal kidney tissues is much higher compared with that in the kidney cancer tissues, which ranged from low and moderate to high differentiated stages. statisti-cal analysis results revealed a correlation between the expression of bai-1 and vegf, mvd, and p53 proteins. Conclusion:The expression level of bai-1 in kidney cancer tissues is very low, even difficult to detect, compared with the increased expression of vegf and cd34. this finding suggests that bai-1 can inhibit tumor angiogenesis. the level of p53 is positively correlated with the expression of bai-1 ac-cording to the data derived with the spearman software, suggesting the existence of crosstalk between these two molecules.

目的:探讨膀胱尿路上皮癌(膀胱癌)石蜡包埋组织中提取MicroRNA的可行性,分析miR-17-5p在膀胱癌石蜡包埋组织中的表达情况与临床病理特征及术后复发的关联性.方法:荧光定量RT-PCR法检测50例新鲜膀胱癌组织和对应的石蜡包埋组织中miR-17-5p基因表达的相关性,分析不同年度间181例石蜡包埋膀胱癌组织中miR-17-5p基因的表达,并与患者术后复发进行相关性分析.结果:石蜡包埋膀胱癌组织与新鲜冷冻组织中miR-17-5p的表达密切相关(r=0.809,P＜ 0.001);不同年度间石蜡组织的miR-17-5p表达稳定,无统计学差异.miR-17-5p表达量与肿瘤临床特征明显相关,低表达患者其术后复发率明显增高(P＜0.05).结论:石蜡包埋膀胱癌组织与新鲜冷冻组织中miR-17-5p的表达有一致性,用石蜡组织提取MicroRNA是可行的,膀胱癌组织中miR-17-5p作用与其表达量相关,低表达是术后复发的独立因素.

Objective: To detect chromosomal aneuploidy in the urine cells of patients with urothelial carcinoma and to evaluate the results of fluorescence in situ hybridization (FISH) as a non-invasive diagnostic method for detecting urothelial carcinoma. Methods: Fresh urine samples from 100 patients suspected of having urothelial carcinoma and 20 healthy subjects (controls) were collected, and urine cytology and FISH were conducted. Abnormalities on chromosome 3, 7, 17 and 9p21 (p160) were found by FISH. Through the comparison and statistical analysis of the experimental results and pathology, the sensitivity and specificity of the two methods were established, as well as a threshold for the difference between high and low-grade carcinoma. Results: Urine samples of patients with urothelial carcinoma have many more chromosomal abnormalities than samples from the control group. The sensitivity and specificity of urine cytology were 71% and 80%, respectively. The sensitivity and specificity of FISH were 88% and 80%, respectively (P< 0.01). The sensitivity of FISH was higher than urine cytology. The receiver operating characteristic (ROC) curve was drawn using the sensitivity and specificity data. The areas under the curve were 0.758 for urine cytology and 0.842 for FISH. Conclusion: FISH is a valuable and non-invasive detecting method for patients suspected of having urothelial carcinoma. The overall sensitivity of FISH is higher than that of urine cytology, but the specificity of the two methods is similar.