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 investigate the quantitative expression of mRNA levels of human NOV and WT1 genes in renal cell carcinoma ( RCC). Methods Using quantitative real-time RT-PCR analysis, we quantified NOV and WT1 mRNA levels in the samples from 57 patients (40 men and 17 women; mean age,65 years) with RCC. Of them,39 cases had T1 stage tumor, 13 had T2 stage and 5 had T3a stage by UICC pathologic staging;and 14 cases had G1 tumor,23 had G2 and 20 had G3 by Fuhrman grading. The expression levels of NOV and WT1 genes,and their association were analyzed. Results The median level of NOV mRNA expression was significantly lower in RCC ( 1. 17) than in normal kidney tissue (4. 32, P

Objective To investigate the effects of nephroblastoma overexpressed (NOV) on proliferation,adhesion,migration and invasion of remal cell curcinomai (RCC) cells. Methods We constructed a NOV expression plasmid and transfected the plasmid into RCC cell line 786-O and analyzed the effects of NOV expression on proliferation,adhesion,migration and invasion of RCC cells by growth curve assay,WST-1 assay,cell adhesion assay,matrigel invasion assay and transwell migration assay. Results The stable NOV transfected 786-O cells (786-O-NOV) showed decreased growth rate,at 48 h and 72 h,the proliferation activities of 786-O-NOV cells were inhibited by 29.14％ and 32.46％ the proliferation activities of empty vector cells were inhibited by 9.25％ and - 8.16％,respectively,compared to 786-O cells (P ＜0.05); while the 786-O cells transfected with empty vector (786-O-mock) had no difference with 786-Ocells.Adhesion assay indicated significantly increased adhesion of 786-O-NOV cells to fibronectin (0.26 ±0.03) and laminin (0.28 ±0.04),compared to 786-O cells (0.15 ±0.01,0.12±0.10) and 786-O-mock cells (0.14 ±0.02,0.13 ± 0.08).Invasion assay displayed that the numbers of cells penetrated through matrigel membrane were significantly higher in 786-O-NOV cells (240.25 ± 23.12) compared to 786-O cells ( 56.16 ± 6.25 ) and 786-O-mock cells ( 50.28 ± 7.13 ).Migration assay displayed that the numbers of cells passed through polycarbonate filters were significantly higher in 786-O-NOV cells (267.25 ± 20.94) compared to 786-O cells ( 66.10 ± 5.68 ) and 786-O-mock cells ( 56.28 ± 4.11 ).Conclusion NOV exhibits anti-proliferative effects on RCC cells; however,it promotes adhesion,migration and invasion of RCC cells.

Objective To study the relationship between reactive oxygen species (ROS) as well as total antioxidant capacity ( TAC ) within follicle fluid and body mass index ( BMI ) in patients with polycystic ovary syndrome (PCOS).Methods All patients enrolled in this study were infertile women receiving in vitro fertilization-embryo transfer (IVF-ET) treatment.55 PCOS patients were divided into over-weight group ( n =23 ) and non-over-weight group ( n =32 ).Another 55 age-matched non-PCOS women were also divided into control group ( n =30) and overweight group ( n =25 ).Plasma sex hormone,triglycerides,and total cholesterol were determined.On oocyte retrieval day after ovarian stimulation,ROS and TAC in follicular fluid were assayed.Results Subjects in over-weight and PCOS over-weight groups had higher triglycerides than those in control and PCOS non-over-weight groups [ ( 1.9 ±1.1,1.7 ± 0.9,1.0 ± 0.5,1.2 ± 0.7 ) mmol/L,respectively,all P＜0.05],so did total cholesterol [ ( 4.8 ± 1.2,5.2 ± 1.1,4.0 ± 0.6) mmol/L,respectively,all P＜0.05].In PCOS over-weight group,ROS and ROS/TAC within follicular fluid were ( 35.4 ± 6.7 ) RLU/S and 39.8 ± 22.0,both were higher than those in the other 3 groups ( all P＜0.05).TAC [ (0.8 ± 0.5 ) Mm] was lower in PCOS over-weight group than that among the other 3 groups( all P＜0.05 ).ROS/TAC was higher in PCOS non-over-weight group than that of control group ( 26.5 ± 14.5 vs 14.2 ± 12.5,P＜0.05 ).Univariate analysis showed that both ROS and ROS/TAC within follicular fluid in PCOS patients were positively correlated with BMI ( r =0.34 and r =0.32,both P＜0.05 ).Conclusion Abnormal oxidative stress exists in follicular fluid of PCOS patients,and the oxidative stress parameters show positive correlation with BMI.

ObjectiveTo evaluate the efficacy of radical transurethral bladder tumor resection plus chemotherapy for the treatment of muscle-invasive bladder cancer. Methods Thirty-two patients,who were diagnosed muscle-invasive bladder cancer by preoperative CT and cystoscopy and not tolerating or rejecting radical cystectomy were treated by transurethral resection of bladder tumor (TURBt).The maximum diameter of tumor ranged from 1 - 5 cm,3 cm on average.After conventional intravenous chemotherapy ( docetaxel 75 mg/m2 + oxaliplatin 130 mg/m2),and given intravenous therapy (HCPT 20 mg + 20 ml saline).Regular cystoscopy was used to monitor tumor recurrence.The examination was performed quarterly in the first 2 years post operation,twice a year since the third year.ResultsThe tumors of 32 cases were resected completely.Operative time were 15 -70 min,the blood loss was 10 -150 ml,without serious complications during the operation.Pathological report showed 32 cases of transitional cell carcinoma.Clinical stages were T2a 20 cases,T2b 12 cases.Pathological grade were G2 13 cases,G3 19 cases.There was no bone marrow suppression,anemia or other severe complications was seen in 32 cases that received chemotherapy.3 of which manifested as low fever,mild nausea,and headache,respectively,having a rest 2 to 3 days the symptoms disappeared.32 patients were followed up for 3 - 60 months,a mean of 28 months.After 1 year the recurrence rate was 9.4％ (3/32),after 2 years was 12.5％ (4/32).The TNM stage of these recurrence cases were 4 cases with T2a and 3 cases with T2b.12 patients died,5 patients died of bladder cancer metastasis.Other 20 patients were survival with no recurrence.ConclusionRadical TURBt plus chemotherapy could be a treatment for the selected patients with muscle invasive bladder cancer.

d best embryo transferred.

Objective To investigate the feasibility of adrenal sparing surgery for selected renal cell carcinoma (RCC), since the necessity of routine adrenalectomy during radical nephrectomy for RCC has been challenged in recent years.Methods 178 patients underwent perifascial nephrectomy in a 16-year period. Of these patients, 96 had ipsilateral adrenal gland preserved during nephrectomy and 82 underwent concomitant adrenalectomy. 75 excised adrenal specimens were examined pathologically. Disease specific survival rates were assessed according to the pathological stage of the tumors.Results Of the 75 patients, 53 presented a macroscopically normal adrenal gland without patho- histological changes. The other 22 patients were suspected to have adrenal metastasis intraoperatively, while only 5 of them were confirmed to have adrenal involvement by histopathology. Two patients in the adrenal gland preservation group developed ipsilateral adrenal recurrence and synchronous or metachronous contralateral adrenal metastasis during follow-up, although both were documented to have a normal-appearing adrenal gland intraoperatively. Five patients with adrenal metastasis and 2 patients with adrenal recurrence had large renal tumors. The survival difference among subgroups of patients undergoing adrenalectomy or with adrenal gland left in situ was not statistically significant.Conclusion Adrenal sparing surgery could be done for patients with small renal tumors along with macroscopically normal ipsilateral adrenal glands.

ObjectiveTo investigate the single nucleotide polymorphisms (SNPS) in the coding regions of the human ABCA2 gene and to determine the association of some of these SNPs with gallstone disease in a Chinese population. MethodsThe exons and part of the introns of the ABCA2 gene were sequenced using a fluorescent labeling automatic method in 24 patients with gallstone disease to identify and characterize the SNPs in a Chinese population. For SNPs in the exons, case-control studies were performed on patients and controls. ResultsTwelve SNPs were found within a 16911 bp region of the ABCA2 gene. Among them, two were in the exons, ten in the introns and five were novel SNPs. There was no significant difference in the SNPs genotype between the patients and the controis. ConclusionsThere is an important ethnic difference in the SNPs distribution of the human ABCA2 gene. The distribution of SNPs in the coding regions of the human ABCA2 gene is not significantly different between the patients and the controls.

Objective To survey birth defects of neonates conceived by using various types of in vitro fertilization and embryo transfer (IVF-ET) between 1998 and 2007 in Shanghai. Methods From 1998 to 2007, 8507 neonates from 6551 pregnancies conceived through assistant reproductive technology (ART) from 7 reproductive medicine center in Shanghai were enrolled in this retrospective study, including Shanghai Ji-Ai Genetics and IVF Institute, Shanghai Jiaotong University School of Medicine affiliated Renji Hospital,Buijin Hospital, China Welfare Institute International Maternal and Infant Health Hospital, Shanghai First Maternity and Infant Health Hospital, Shanghai the Ninth People's Hospital and the Second Military Medical University affiliated Changhal Hospital. The clinical data about the type and incidence of birth defect were analyzed. Meanwhile, the factors associated with birth defect were investigated, such as various ART,gender, mother age, numbers of gestational sac, the source and quality of sperm. Results The birth defect rate was 1.08% (92/8507) and the most remarkable malformation occurred in circulation system [34% (31/92)]. The rates of major congenital malformations were 1.21% (34/2799) in fresh conventional IVF-ET, 1.07% (20/1871) in IVF-frozen-thawed embryo transfer(IVF-FET), 1.04% (23/2212) in fresh intracyto plasmic sperm injection (ICSI)-ET and 0.92% (15/1625) in ICSI-FET, which did not show statistical difference (P>0.05). There was also no significantly different incidence of malformations between 1.12% (49/4371) in male neonates and 1.02% (42/4136) in female neonates (P >0.05).However, the occurrence of congenital malformation is positively related with maternal age, the rates were 0.84% (41/4884) in mother's age less than 30 years and 1.77% (16/902) in mother' age more than 35 years, which reached statistical difference (P < 0.05). It also showed remarkable different incidence among 0.53% (25/4679) in singletons, 1.59% (57/3576) in twins and 3.97% (10/252) in triplets (P< 0.05). No remarkable difference of malformation rate among sperm sources used in fertilization were observed, including 1.09% (81/7419)in ejaculated sperm, 1.21% (7/579)in percutaneous epididymal aspiration (PESA) and 0. 79% (4/509) in donor sperm (P>0.05). Conclusions The overall incidence of birth defect in neonates from ART is similar to those conceived naturally and is not associated with different types of IVF-ET, embryo cryopreservation, sperm sources. However, maternal age and multiple pregnancies coder the higher possibility of birth defect.

Objective To investigate the effects of hyperbaric oxygen combined with function training on motor function in children with cerebral palsy. Methods From December 2012 to December 2013 during the treatment,72 cases chil-dren with cerebral palsy were selected as the object of study confirmed in our hospital, two groups of children were given intensive training of motor function (Bobath method), Hyperbaric oxygen therapy, 1 time a day, continuous treatment for 10 d, intermittent 20 d was 1 course of treatment, continuous treatment after the 3 curative effect observation. And the observation group was combined with hyperbaric oxygen treatment, the change of GMF score before and after treatment were compared between the two groups after treatment. Results The total effective rate of observation group patients after the treatment was 94.4%, significantly higher than the control group (66.7%), the difference was significant (χ2=11.238, P<0.05) between two groups. Before treatment, GMF scores of two groups were not different significantly (P>0.05). After treatment, GMF scores of two groups decreased significantly (t=2.421, 2.235, P<0.05), and the GMF scores of observation group after treatment were sig nificantly lower than the control group, the difference was signifi-cant (t=2.387, P<0.05). Conclusion Training of hyperbaric oxygen combined with function training on motor function of relatively simple and the effect of the children is better than intensive training, two methods combined can obviously improved the movement function of children with cerebral palsy so as to further improve the effect of treatment of chil-dren with cerebral palsy, is worthy of extensive promotion and application.