ObjectiveTo investigate the effect of arsenic trioxide (ATO) on anti-dsDNA antibody and the expression of DNA methyltransferase 1-mRNA and CD11a-mRNA in lupus MRL/lpr mice.Methods MRL/lpr mice were divided into three groups:the ATO group,the sodium chloride(NS) group,and the cyclophosphamide(CTX) group.The control group consisted of 20 syngeneic normal C57/BL mice,which were subdivided into the ATO group and the NS group.After two-month treatment,all mice were sacrificed.Blood routine test was conducted by SYSMEX KX21.The anti-dsDNA antibody in the serum were detected by ELISA.The expression of DNMT1 and CD11a was determined by RT-PCR.ANOVA and paired t test were used for statistical analysis.Results① The serum level of anti-dsDNA antibody(0.89±0.07) and the gray scale value of CD11a-mRNA(0.43±0.25) in the ATO group were much lower than those in the NS group of MRL/lpr mice(1.77±0.28,P＜0.01; 0.99±0.31,P＜0.05),while the gray scale value of DNMTI-mRNA (0.32±0.30) was significantly higher than that in the NS group(0.16±0.26,P＜0.05 ).② The serum levels of anti-dsDNA antibody was low in both the ATO group and the CTX group (0.90±0.07,0.66±0.22),and it was higher in the ATO group than that in the CTX group (P＜0.05).The gray scale value of DNMT1 mRNA in the ATO group (0.32±0.30) was higher than that in the CTX group (0.16±0.18,P＜0.05) in MRL/lpr mice,and the gray scale value of CD11a mRNA in the ATO group(0.43±0.25) was lower than that in the CTX group (0.86±0.31,P＜0.05) in MRL/lpr mice.③ There was no difference in above parameters between the ATO-group and NS group in C57/BL mice(P＞0.05).ConclusionArsenic trioxide can reduce the serum level of auto-antibody and reverse low methylation.But it has no effect on normal mice.

SUMMARY National Institutes of Health (NIH) released the biomedical research project NIH Roadmap Initiatives, including 3 themes, new pathways to discovery,research teams of the future,and re-engineering the clinical research enterprise. The purpose of the project is to catalyze to transform our new scientific knowledge into tangible benefits for people. Now,Mostly of the Project have begin to carry into practice.

Objective To investigate the reason of persisting positive rapid plasma reagin (RPR) in serofast syphilis patients, and to provide reference for clinical treatment and prognosis.Methods A total of 33 serofast patients and 23 healthy controls were enrolled in this study.The percentages and absolute counts of CD3+, CD4+, CD8+ T lymphocytes and natural killer (NK) cells were detected by flow cytometry.The comparison of two groups was analyzed by independent sample t test, and the correlation between change of lymphocyte subgroups and RPR titer in serofast syphilis patients was analyzed by bivariate linear correlation method.Results Compared with healthy controls, the percentages of CD3+, CD8+ T lymphocytes in serofast syphilis group were both increased significantly (75.75±5.76)% vs (68.37±5.80)%, (t=4.69, P<0.01);(27.34±7.02)% vs (24.33±1.95)%, (t=2.34, P=0.025), while both the percentage and absolute count of NK cells were significantly decreased (7.32±4.48)% vs (14.87±6.26)%, (t=5.269, P<0.01);(136.2±83.4)/μL vs (298.8±166.9)/μL, (t=4.311, P<0.01).RPR titer of the patients was negatively correlated with both percentage and absolute count of CD4+ T lymphocytes (r=-0.476 and-0.515, respectively, both P<0.01), and it was positively correlated of CD8+ lymphocytes (r=0.588 and 0.305, P<0.01 and P=0.804).Conclusion The imbalance of immune response of lymphocyte subsets observed in serofast syphilis may explain the RPR titers change.

To explore the diagnostic values of miRNA141 and miRNA375 for prostate cancer.The expressions of miRNA141 and miRNA375 in serum samples from 30 prostate cancer (PCA) patients,40 benign prostatic hyperplasia patients and 50 normal controls were detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).The expression of miRNA141 in tissue samples from prostate cancer was 13.4±7.2 folds of that from benign prostatic hyperplasia while the expression of miRNA375 in tissue samples from prostate cancer was 28.4 ± 10.3 folds of that from benign prostatic hyperplasia.The differential expression of miRNA141 and miRNA375 in sera of prostate cancer suggested that miRNA141 and miRNA375 might be potential tumor markers in clinical practice.

Objective To investigate the common bacteria and change in antimicrobial resistance in an intensive care unit (ICU)in the past 5 years,and provide evidence for rational use of antimicrobial agents.Methods Bacteria isolated from ICU patients in a tertiary first-class hospital from 2009 to 2013 were collected,identified,and per-formed antimicrobial susceptibility testing.Results A total of 1 196 bacteria isolates were isolated in 2009-2013, the top five species were Acinetobacter baumannii (A.baumannii,29.60%),Pseudomonas aeruginosa (P . aeruginosa,14.38%),Staphylococcus aureus (S .aureus ,12.21 %),Escherichia coli (E.coli,12.21 %),and Klebsiella pneumoniae (K .pneumoniae ,11 .37%).Resistance rates of S .aureus to oxacillin,gentamycin, clindamycim,ciprofloxacin,and rifampicin showed a decreasing tendency from 2009 to 2013(all P <0.05),and no strain was detected to be resistant to vancomycin during 5 years.Resistance rates of P .aeruginosa to ceftazidime, cefepime,aztreonam,gentamycin,amikacin,tobramycin,and piperacillin/tazobactam decreased gradually (all P <0.05),imipenem resistance rates were 32.26% -46.43% in 2009 -2012 and 16.00% in 2013;A.baumannii maintained a low level resistance to amikacin,tobramycin,and compound sulfamethoxazole(all P <0.05),resistance rates to imipenem were all >80% in 2009-2011 and 10.53% in 2013,A.baumannii had high resistance rates to most antimicrobial agents(resistance rates were >80%)during 5 years.Resistance rates of K .pneumoniae and E. coli to piperacillin/tazobactam,cefazolin,cefepime,amikacin,and aztreonam had a decreased tendency in 2009 -2013(all P <0.05).Conclusion The common bacteria causing infection in the ICU of this hospital showed a down-ward trend,which may be related to the introduction of national policies and management of hospital,continuous management of antimicrobial agents is suggested,antimicrobial agents should be used rationally to prevent the in-crease of bacterial resistance.

Objective To explore the role of receptor for advanced glycation end products (RAGE) in HMGBl-mediated CD4+ T cells differentiation to Th1/Th2.Methods CD4+ T lymphocytes isolated from the spleens of male BALB/C mice by magnetic beads were suspended in RPMI-1640 with 10％ FCS in 2× 106cell/well on 96-well cell culture plates in vitro.The cells were randomly divided 4 groups according to concentration of HMGB1 treatment:control group,10 ng/mL group,100 ng/mL group,1 000 ng/mL group after stimulation with ConA in 3 μg/mL for 12 hours.IL-4 and IFN-γ levels in culture supernatants were quantitated by ELISA kits after HMGB1 stimulation for 12,24,and 48 h.According to the results,cultured cells were exposed to HMGB1 in 100 ng/mL for 24 h in the following experiments.The cells were randomly divided into 4 groups:control group,A group,B group,C group,and each group were cultured with ConA in 3 μg/mL for 24 h.The cells of control group and other three groups were stimulated with PBS or 100 μg/L HMGB1 for 24 h.The cells of B,C groups were incubated with 1/200 diluted 5 μg/L anti-RAGE Abs (anti-bodies) or PBS for 2 h before HMGB1 stimulation.The cell suspension was obtained to detect the levels of IL-4 and IFN-γ by EILSA and the protein levels and mRNA expressions of RAGE,CATA-3 were detected by western-blot and real-time fluorescent quantitative PCR,respectively.ResuRs Compared with control group,CD4+ T cells incubated with increasing concentrations of HMGB1 (100,1 000 ng/mL) for 24 h resulted in a decrease in IFN-γ/IL-4 ratio (P＜0.05).When CD4+ T cells were exposed to 100 ng/mL HMGB1 for 12 h,IFN-γ/IL-4 ratio was markedly increased.However,CD4+ T cells treated with 100 ng/mL HMGB1 for 24,48 h,IFN-γ/IL-4 ratio was markedly inhibited (P＜0.05).Compared with control group,protein levels and mRNA expressions of RAGE and GATA3 of cells in A group were significantly increased (P＜0.05),and IFN-γ/IL-4 ratio of cell suspension in A group and B group was significantly down-regulated (P＜0.05).Compared with A group,IFN-γ/IL-4 ratio of cell suspension in C group was increased (P＜0.05),and expression of GATA3 mRNA was down-regulated (P＜0.05).Compared with A group,protein level of RAGE of cells in C group was significantly down-regulated (P＜0.05),but protein level of RAGE of cells in C group was still increased compared with control group (P＜0.05).Conclusion Th1/Th2 differentiation induced by HMGB1 on CD4+ T lymphocytes in vitro was at least partly mediated by over-activating RAGE/GATA3 pathway.

Objective:This study aims to construct a prognostic model of bladder cancer (BLCA) based on lncRNA.Methods:BLCA lncRNA expression data and clinical information were downloaded from TCGA. Univariate Cox regression was used to evaluate the correlation between the expression level of each lncRNA and overall survival (OS), and the lncRNAs with a corrected P-value<0.01 were selected as candidate predictors. In the training queue, the prediction model is constructed by methods such as least absolute shrinkage and selection operator, and multi-factor stepwise Cox regression, and verified in the verification queue at the same time.. Evaluation the area under the curve of time-dependent receiver operating characteristic (tROC) and Harrel C index. According to the median risk score of the prediction model, patients were divided into high-risk group and low-risk group and the differences in clinicopathological characteristics between the two groups were compared by t-test or chi-square test. Results:Establish a BLCA prognostic model based on 13 lncRNAs, of which LINC01465, ARHGAP5-AS1, ZFHX4-AS1, MAFG-AS1 are prognostic risk factors (β regression coefficients are 0.32, 0.16, 0.06, 0.20, respectively, all>0), and the rest are protection factors (β regression coefficients are all<0); the prediction model of the overall survival in the first year, the third year, and the fifth year in the complete cohort has an area under the tROC curve of 0.79, 0.82, and 0.80 respectively, and the Harrell C index is 0.74. Its predictive ability is better than the previously published BLCA prognostic model based on lncRNA. Adjusting for confounding factors including age and tumor stage found that the risk score of this model was an independent poor prognostic factor for overall survival in BLCA patients (hazard ratio 4.05; P<0.001). Comparison of clinicopathological characteristics of patients in the high-risk and low-risk groups showed that in the high-risk group, there were more old patints (70.0 vs. 66.1, P<0.001), more non-papillary patients (74.2% vs. 61.2, P=0.005), more high-stage patients (37.6% vs. 28.0%, P<0.001 for stage Ⅳ patients), and more high-grade tumors (98.0% vs. 92.0%, P=0.005). Conclusion:In this study, a prognostic model of bladder cancer based on 13 lncRNAs was constructed. This model has good predictive ability and can provide value for clinical decision-making and patient consultation.

The link of hedgehog (Hh) signaling activation to human cancer and synthesis of a variety of Hh signaling inhibitors raise great expectation that inhibiting Hh signaling may be effective in human cancer treatment. Cyclopamine (Cyc), an alkaloid from the Veratrum plant, is a specific natural product inhibitor of the Hh pathway that acts by targeting smoothened (SMO) protein. However, its poor solubility, acid sensitivity, and weak potency relative to other Hh antagonists prevent the clinical development of Cyc as a therapeutic agent. Here, we report properties of cyclopamine tartrate salt (CycT) and its activities in Hh signaling-mediated cancer in vitro and in vivo. Unlike Cyc, CycT is water soluble (5-10 mg/mL). The median lethal dose (LD50) of CycT was 62.5 mg/kg body weight compared to 43.5 mg/kg for Cyc, and the plasma half-life (T1/2) of CycT was not significantly different from that of Cyc. We showed that CycT had a higher inhibitory activity for Hh signaling-dependent motor neuron differentiation than did Cyc (IC50 = 50 nmol/L for CycT vs. 300 nmol/L for Cyc). We also tested the antitumor effectiveness of these Hh inhibitors using two mouse models of basal cell carcinomas (K14cre:Ptch1(neo/neo) and K14cre:SmoM2(YFP)). After topical application of CycT or Cyc daily for 21 days, we found that all CycT-treated mice had tumor shrinkage and decreased expression of Hh target genes. Taken together, we found that CycT is an effective inhibitor of Hh signaling-mediated carcinogenesis.
Animals ; Carcinoma, Basal Cell ; pathology ; Cell Differentiation ; drug effects ; Embryonic Stem Cells ; cytology ; Hedgehog Proteins ; antagonists & inhibitors ; metabolism ; Mice ; Motor Neurons ; cytology ; Plants, Medicinal ; chemistry ; Receptors, G-Protein-Coupled ; antagonists & inhibitors ; metabolism ; Signal Transduction ; drug effects ; Skin Neoplasms ; pathology ; Smoothened Receptor ; Solubility ; Tartrates ; blood ; pharmacology ; Tumor Burden ; drug effects ; Veratrum ; chemistry ; Veratrum Alkaloids ; blood ; isolation & purification ; pharmacology