A systematic analysis of the typeⅢ secretion genes of Aeromonas hydrophila strain AH-1 by constructing ahyR and ahyI mutant revealed that they are under quorum-sensing control. This observation was supported by the down-regulation of the TTSS genes in the presence of lacZ-TTSS gene promoter and the corresponding advanced secretion of AexT in ahyI mutant.

Virginiamycin acetyltransferase D (VatD) plays a vital rule in streptogramins resistance by chemically inactivating streptogramin A. Therefore, it is desirable to discover novel small molecular weight inhibitors of VatD via state-of-the-art virtual screening techniques. This "cocktail" strategy by combining VatD inhibitor with streptogramins may provide new therapeutic opportunity for resistant bacteria infections. Structure-based virtual screening method (molecular docking) was applied to rank and score a chemical database containing 300 000 commercially available compounds against the VatD substrate binding site. Twenty six out of the 200 top scored compounds from the docking calculation were selected and submitted to the VatD enzymatic inhibition assay. The plasmid pRSET B/vatD was constructed and transformed into E.coli (trxB) host cells for over-expression, and VatD enzyme was purified and validated by showing acetyltransferase activity to Virginiamycin M1. Three out of these 26 tested compounds showed enzymatic inhibition on VatD with IC50 168.6, 91.0 and 55.2 μmol·L-1, separately. Other compounds could not be dissolved in the system and/or had little effect on the enzyme (IC50>200 μmol·L-1). To our knowledge, it is first time that small molecular weight organic compounds were identified as VatD inhibitors. It is expected that the VatD inhibitors identified at present study could serve as lead compounds for the further development of the novel therapeutic agents to overcome streptogramins resistance.

Objective　To study the effects of estradiol-17β (17β-E2) on the synthesis of GnRH and GH in submandibular gland of rat. Method　Immunohistochemical ABC method was used to observe the localization of GnRH and GH in submandibular gland of rat.Results　　The synthsis of GnRH was promoted and the synthesis of GH was inhibited by 17β-E2 in rat's submandibular gland.　Conclusion　The 17β-E2 may play an important regulative role in the synthesis of GnRH and GH.

Objective To investigate the effect of optimized radiological examination strategy on iatrogenic radiation exposure in severe trauma patients so as to provide scientific basis for standardized application of radiological examination.Methods A controlled, three-stage intervention study from April 2010 to November 2011 was carried out.From April 2010 to July 2010, a pre-intervention study was conducted and enrolled 60 patients [43 males, 17 females;age (50 ± 14)years, age range 23-78 years].From August 2010 to March 2011, optimized strategies of radiological examination were implemented, including improving clinicians' knowledge to the standardization of radiological examination and iatrogenic radiation injury and limiting frequency of CT scans through the electronic medical record.From April 2011 to November 2011, post-intervention study was conducted and enrolled 100 patients (81 males, 19 females;age (47 ± 14) years, age range 18-79 years].During this period, major trauma patients were analyzed with respect to the clinical information, radiation examination frequency, ionizing radiation dose and influencing factors.Radiation examination frequency and radiation dose were compared before and after the intervention.Results Radiological examinations were mainly X-ray and CT before the implication of optimized strategies.Of the 60 patients, median frequency of X-rays and CT scan was 6.0(3.0-11.0) and 10.0(8.0-13.8).Median frequency of CT scan was positively correlated with the injury severity score (ISS) and ICU length of stay (r =0.369 and 0.523, P ＜ 0.05).Of the 100 patients, median frequency of CT scan was significantly reduced after the optimization of radiological examination (8.0 vs.10.0, P ＜ 0.05).Total frequency of radiological examination was significantly reduced as well (13.6 vs.17.8, P ＜0.01).There was no significant difference in the treatment success rate before and after the optimization of radiological examination (85.0％ vs.88.3％, P ＞ 0.05).When the frequency of head and chest CT scan was limited, the frequency of radiological examination, radiation exposure and radiological examination expenses were greatly reduced.Conclusions Too much X-ray,CT or other radiological examinations are noted in major trauma patients during the treatment period.Improved understanding of radiation-induced injury, optimizing radiological examination and controlling the repeated radiological examinations of the same site contribute to reducing iatrogenic radiology exposure without affecting the outcome.

Objective To estimate the clinical value of thinprep cytologic test (TCT) in screening cervical lesion. Methods 4234 TCT samples were interpreted according to the Besthesda System (TBS), 272positive cases (ASC-US or above) were taken colposcopic examination and biopsy. Results The coincidence of the results between TCT and biopsy in low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL), squamous cell carcinoma (SCC), adenocarcinoma (AC) were 85.71%(12/14), 100 % (20/20), 75 % (3/4) and 100 % (2/2), respectively. The positive rates of over ASC-H by TCT and of over CIN Ⅰ by biopsy were 23.16 % (63/272) and 24.26 % (66/272), respectively. There is no difference between two positive rates (x2 = 0.868, P = 0.581). Conclusion TCT and histopathological diagnosis had a high coincidence, a combination of both can greatly enhance HSIL and cervical cancer and reduce the incidence of missed diagnosis. TCT would be a rapid and convenient method for screening cervical cancer.

Objectlve To assess the clinical effects and safety of intravitreal injection of conbercept and macular photocoagulation for non-ischemic macular edema secondary to branch retinal vein occlusion (BRVO).Methods A total of 50 patients (50 eyes) with non-ischemic macular edema following BRVO were retrospectively analyzed.Patients were divided into 2 groups:ICI group (26 eyes) received intravitreal injection of conbercept,laser group (24 eyes) received standard-of-care grid laser for macular edema.The best-corrected visual acuity (BCVA) and central macular thickness (CMT) were observed before and 1 week,1 month,2 months and 3 months after treatment.Then,the changes in pre-treatment and post-treatment were compared,and the related complications were recorded.Results The difference of BCVA before treatment was not significant between two groups (P ＞ 0.05).BCVA at 1 week,1 month,2 months and 3 months after treatment in two groups were all improved,the differences were statistically significant compared with before treatment (all P ＜ 0.05),and the differences were statistically significant between two groups after treatment (all P ＜0.05).At 3 months after treatment,BCVA of 18 patients (69.23％) in ICI group and 8 patients (33.33％) in ICI group improved 2 lines.The difference of CMT before treatment was not significant between two groups (P ＞ 0.05),CMT at 1 week,1 month,2 months and 3 months after treatment in two groups were all decreased,the differences were statistically significant compared with before treatment (all P ＜ 0.05),and the differences were statistically significant between two groups after treatment (all P ＜0.05).13 eyes received repeat intravitreal injection in ICI group,including 2 eyes at 1 month,7 eyes at 2 months with CMT ＞250 μm,and 3 eyes at 3 months.No severe side effect related with drug and intravitreal injection occurred in the two groups.Conclusion Mean BCVA change and CMT change are significantly greater in the intravitreal injection of conbercept than the standard-of-care grid laser group for the macuiar edema secondary to non-ischemic BRVO.

Objective To systematically evaluate the efficacy,safety and recent curative effects of delta-shaped anastomosis in totally laparoscopic distal gastrectomy (ICBI) for gastric carcinoma.Methods Literatures in English and Chinese comparing ECBI and ICBI published up to November 2015 were searched from international and national online databases.Meta-analysis was conducted by Review Manager 5.3 soft ware.Results There are eleven studies involved,with 2020 gastric cancer patients,including,1 169 ECBI cases and 851 ICBI cases.Meta-analysis showed that there was no significant statistical differences between ECBI group and ICBI group in operative time,resection margin length,overall postoperative complications and anastomosis-related complications.(all P ＞ 0.05).When compared to ECBI,the estimated blood loss was significantly less in ICBI,and ICBI with more retrieved lymph nodes,faster recovery of gastrointestinal function,less use of painkiller and shorter postoperative hospital stay (all P ＜ 0.05).Conclusion ICBI is safe and feasible in treatment of gastric cancer,and has a good short-term effect.

Objective To explore a novel and highly specific small-molecule TNFβinhibitor by using computer-aid?ed virtual screening and cell-based assays in vitro. Methods Computer-aided drug design and virtual screening were used to design and identify chemical compounds that targeted TNFβbased on the crystal structure of the TNFβ-TNFR1 com?plex. The effect of the small-molecule compound against TNFβ-induced cytotoxicity of L929 cells was detected by MTT as?say, and the efficacy of the compound to inhibit TNFβ-induced apoptosis of L929 cells was determined by flow cytometry as?say. The impact of the compound on L929 cell cycle was examined by Propidium Iodide (PI) staining and flow cytometry, and the influence of the compound on TNFβ-triggered signal pathway was analyzed by Western blot assay and Ultra VIEW VOX 3D Live Cell Imaging System. Results No.35 compound (named as C35 thereafter) could effectively inhibit TNFβ-induced cell death in a dose dependent manner, and the half-maximum inhibition concentration (IC50) was 8.19μmol/L. Furthermore, C35 had lower cytotoxicity and minimal effect on L929 proliferation. Here we further revealed that C35 could affect TNFβ-induced apoptotic pathway by blocking the activation of Caspase 3, and markedly reduce L929 cell apoptosis induced by TNFβ. Conclusion A novel TNFβsmall-molecule inhibitor was identified by combining computer-aided virtual screening with functional assays, and which could block TNFβ-triggered apoptotic pathway and efficiently inhibit the cell death in?duced by TNFβ.

Objective To investigate the effect of glucocorticoid and bisphosphonate on Hedgehog signaling pathway in human bone mesenchymal stem cells (hBMSCs) and bone tissue.Methods ① Bone biopsy test:forty cases of systemic lupus erythematosus (SLE) patients were divided into 2 groups:20 cases in newly diagnosis group,20 cases in the GCs group (the dosage of glucocorticoids was higher than 1 mg·kg-1·d-1).Patients in the GCs group was randomly divided into 2 groups:10 cases in the control group were without anti-osteoporosis treatment,10 cases in treatment group received alendronate 70 mg once a week.All of the patients had bone marrow puncture after24 weeks,and the value of average optical density of Gli1 was tested with immunohistochemistry.② Cell culture:hBMSCs cultured in normal medium were intervened with rh-SHHN and methylprednisolone (10-3 mol/L,10-5 mol/L) after successfully identified.Quantitative polymerase chain reaction (PCR) was used to detectthe mRNA expression of Gli1.The final calcium nodules was detected by Alizarin red staining.hBMSCs cultured in osteogenesis medium were intervened with bisphosphonate and methylprednisolone (10-3 mol/L) after successfully identified.Quantitative PCR was used to detect the expression of Gli1 mRNA.Alizarin red staining was used to detect the final calcium nodules.Comparisons between the two groups were carried out using t-test while the difference analysis of multi-groups were tested by factorial analysis.Results The average optical density of Gli1 in the GCs group (47±7) was less than the newly diagnosed group (61±12) (t=4.442,P＜0.01),and it was less in the control group (51±6) than in the treatment group (42±6) (t=3.701,P=0.002).When normally cultured,high and moderate concentration of methylprednisolone suppressed the mRNA (0.38±0.04; 0.68±0.24) (F=8.748,P＜0.01) expression of Gli1 which was initially stimulated by rh-SHHN (2.01 ±0.38).And the final calcium nodules in groups which contained methylprednisolone were much less than rh-SHHN only group.When hBMSCs were cultured in osteogenesis medium,compared with the methylprednisolone group (0.024±0.011),the expression of Gli1mRNA(0.034-0.006) (t=7.62,P＜0.01) and the final calcium nodules were significantly improved by bisphosphonate.Conclusion High and moderate doses of glucocorticoids inhibit hBMSC osteogenesis by inhibiting Gli1.Low concentration of alendronate can not only stimulate hBMSC osteogenesis differentiation but also can remit the inhibition effects of GCs through the way of Hedgehog.

Objective To summarize the diagnostic efficacy of ADC value for differentiation of benign and malignant lymph nodes on diffusion MRI with Meta-analysis. Methods Published papers on differentiation of benign and malignant lymph nodes with ADC value were searched and reviewed.Quality evaluation was performed for the eligible papers before data extraction.Test for heterogeneity was performed first,then appropriate model was selected to calculate the weighted mean difference,sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odds ratio,pretest and posttest probability.The potential of ADC value for differentiation of benign and malignant lymph nodes was assessed qualitatively and quantitatively.Results Fifteen papers including 735 cases and 1963 lymph nodes were selected.According to Meta-regression analysis,subgroup analysis and robust analysis,two studies with benign lymph nodes in patients with benign lesion and one study using chemical shift saturation technique were excluded because of their impact on the robustness of the pooled results. The weighted mean difference (WMD) between malignant and benign lymph nodes was -0.355 × 10-3mm2/s[95％ confidence interval (CI):-0.423 ×10-3- -0.288 × 10-3 mm2/s].Although the cutoff of ADC value for differentiation in each study was different,the diagnostic efficacy was stable,the pooled sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odds ratio and area under summarized receiver operator's curve were 0.87 (95％ CI:0.79-0.92),0.87 (95％ CI:0.82-0.90),6.5 (95％ CI:4.7-9.2),0.15 (95％ CI:0.09-0.25 ),43 ( 95％ CI:21-87 ),0.93 ( 95 ％ CI:0.90-0.95 ).The posttest malignancy probability of benign lymph node indicated by ADC was 6％,while that of malignant lymph node was 72％.Conclusion The ADC value can be used to differentiate benign and malignant lymph nodes with good sensitivity and specificity noninvasively.