Objective To investigate the effects of co-transfection of the plasmid containing the wildtype (wt) INK4a/ARF gene on the chemosensitivity of human lung adenocarcinoma cell line A549. Methods Recombinant eukaryotic expression vectors pcDNA3-p16 INK4a and pcDNA3-p14 ARF were co-transfected into A549 cells by cationic liposome, in which the site of both genes was lost. By RT-PCR, immunocytochemistry and Western blotting analysis, G418 positive clone was obtained. The parental cell and negative control cell with plasmid pcDNA3-LacZ were used as controls. The 50% inhibition concentration (IC 50 ) of five commonly-used chemotherapy drugs and the apoptosis index (AI) of IC 50 were analysed respectively. Results Compared with the parental cells and negative control cells, the influence of the five drugs on IC50 was different. The values of IC 50 of doxorubicin and cisplatin decreased significantly but that of taxol, topotecan and vinorelbine remained unchanged. The apoptosis index of doxorubicin and cisplatin was much higher than that of other three drugs. Conclusion The chemosensitivity of a part of anti-cancer drugs could be changed by transfection of exogenous INK4a/ARF gene into lung adenocarcinoma cell A549.

Objective To investigate the knowledge awareness of medical staff in department of Obstetrics and Gynecology,on prevention of mother-to-child transmission of hepatitis B virus(HBV).Methods Medicatl staff,who attended national or local medical conferences of Obstetrics and Gynecology from July 29th to October 25th in 2011,were invited to complete a questionnaire containing questions on clinical implications of HBV serologic markers,relevant prevention measures and including intervention measures for both pregnant women and infants.Data were collected and analyzed by Chi-square test or Fisher's exact test.Results A total of 559 answers were analyzed.Of the 559 staff,85.0％(n=475) were doctors and 15.0％(n=84) were nurses.The percentages ofstaff from primary,secondary and tertiary hospitals were 13.2％(n=74),36.9％(n=206) and 49.9％ (n=279).Overall,90.0％ (503/559) participants agreed that positive hepatitis B surface antigen (HBsAg) was contagious,but 27.5％ (154/559) mistook that the presence of antibody against hepatitis B e antigen (anti-HBc) and/or antibody against hepatitis B core antigens (anti-HBc) with negative HBsAg was contagious.Totally,96.3％ (519/539) respondents knew that pregnant women should be screened for HBV infection,and those from secondary hospitals had more consciousness of HBV screening for pregnant women than those from tertiary hospitals [99.0％ (201/203) vs94.7％(250/264),x2=6.466,P=0.011].Although,95.3％ (511/536) realized that infants of mothers with positive HBsAg should receive hepatitis B immunoglobulin (HBIG) and vaccination,but 61.6％ (330/536) and 80.4％ (427/531) of the enrolled medical staff knew the exact regimen.With the available immunoprophylaxis,13.8％ (74/536) participants mistakenly believed that cesarean section may prevent HBV mother-to-child transmission,and only 13.0％ (69/532) correctly answered that neonates of women with positive HBsAg can be breast-fed.Additionally,54.4％ (290/533) participants mistakenly believed that HBsAg positive pregnant women needed HBIG injection during pregnancy to reduce HBV mother to-child transmission.Conclusions Medical staff in obstetric and gynecological department has essentially mastered the strategies of prevention of mother-to-child transmission of HBV,but much more details should be stressed.

Objective To explore the effect of amisulpride and risperidone in the treatment of patients with first-episode schizophrenia and its influence on social function.Methods 70 patients of schizophrenia conformed to the International classification of diseases tenth edition(ICD-10) were randomly divided into amisulpride group(observation group,35 cases) and risperidone group(control group,35 cases) by using the random number table method.The Positive and Negative Scale(PANSS) was used to evaluate the efficacy,the Scale of Social Function in Psychosis Inpatients(SSPI) was used to evaluate social function before and after 8 weeks of treatment.Results After 8 weeks treatment,the negative symptom factor score of the PANSS in the observation group was (15.04±3.55)points,which was improved significantly compared with (17.82±3.87)points in the control group,the difference was statistically significant(t=3.132,P<0.05).The scores of the field in movement and interaction,social activities and skills factor score and the total score of SSPI in the observation group were (15.49±3.54)points,(14.53±4.25)points,(39.25±8.27)points,respectively,which in the control group were (12.78±3.29)points,(10.01±3.78)points,(33.72±7.83)points,respectively,the differences between the two groups were statistically significant (t=3.317,4.701,2.873,all P<0.05).Conclusion Amisulpride is effective in improving the negative symptoms,social function in patients with schizophrenia,and the effect is better than risperidone.

Objective To understand the pattern of and influencing factors on leisure time physical exercises among adults in Zhejiang. Method Totally 17 437 residents aged 18 years and over selected by stratified cluster random sampling in Zhejiang in 2010 were investigated. Result Overall 21.82%(95%CI:17.12%-27.38%) of adults took part in leisure time physical exercises in 2010. The proportion was higher in urban (29.04%,95%CI:23.46%-35.33%) than in rural (18.81%,95%CI:12.89%-26.61%) (χ2=222.06, P<0.05) areas, and also higher in females (22.91%,95%CI:17.66%-29.15%) than in males (20.56%,95%CI:16.39%-25.48%) (χ2=13.94, P<0.05). Adults aged 18-24 and 55-74 years were more likely to take apart in physical exercises (χ2=266.73, P<0.05), and the lowest proportion was found among farmers (5.33%,95%CI:3.99%-7.11%) (χ2=2 078.40, P<0.05). These proportions both increased along with education level and family income increase (χ2=444.87, P<0.05;χ2=332.20, P<0.05). Overall 83.04%and 90.08%of physical exercisers took part in exercises at least 3 times per week and 30 minutes per time, only 30.67% of those reached moderate or vigorous intensity, and overall 5.38% (95%CI: 3.85%-7.48%) of adults took part in regular leisure time physical exercises in 2010. Multiple logistic regression showed that age, education level, occupation and chronic disease history have influence on regular physical exercises. Conclusion The leisure time physical exercises level was generally low among adults in Zhejiang. Young and middle-aged adults with less education and low income especially farmers should be put on emphasis.

Objective To explore the treatment method and clinical effect of congenital scoliosis caused by lumbosacral spine deformity in children.Methods From October 2000 to October 2015,a consecutive series of 21 congenital scoliosis due to lumbosacral spine deformity were treated in Department of Pediatric Orthopedics,the Third Affiliated Hospital of Zhengzhou University,including 12 male and 9 female,and the age was (63.1±47.3) months(3-144 months).The hemivertebrae,bone bridge,and the mixed type were found in 18 cases,2 cases,and 1 case,respectively.Hemivertebraes were on the left in 10 cases,on the right in 9 cases;involved L5 in 8 cases,S1 in 6 cases,multiple vertebrae malformation in 4 cases,wedged-shaped vertebrae involved L5 associated with contralateral bar in 1 case,unilateral unsegmented bar from L4 to S1 in 1 case,from L2 to L5 in 1 case.There were 4 cases combining with thoracic hemivertebrae,3 cases with ribs fusion and contralateral bar,16 cases(76%) with spinal cord malformations,3 cases(17%) with urinary system malformations,no congenital heart malformation.All the 21 patients were operated with one-stage posterior approach.Compared the scoliosis Cobb angle,trunk shift,operative time,blood loss volume,and complications among preoperation,post-operation and final follow-up.Results In this study,operative time was (278.9±83.1) min,blood loss volume was (355.3±249.0) mL,follow-up was (5.1±2.7) years.There were 18 cases operated with pedicle screw fixation,aged (72.9±44.1) months(16-144 months),operative time was (296.2±74.2) min,blood loss volume was(422.1±238.2) mL;the mean coronal Cobb angle of malformation area at preoperation,post operation and the final follow-up were(27.3±10.2)°,(11.0±5.9)°,(9.8±4.2)°,while the correction rate of post-operation and the final follow-up were 59.7%,64.1%.There were 3 cases operated by hemivenebra resection without pedicle screw fixation,aged 7.7 months(3-15 months),operative time was 168.3 min,blood loss volume was 103.3 mL.The mean coronal Cobb angle of malformation area at preoperation,post-operation and the final follow-up were 26.0°,13.6°,12.5°,while the correction rate of post-operation and the final follow-up were 47.7%,51.9%.The follow-up period was (5.1±2.7) years.The coronal Cobb angle of lumbosacral curve at preo-peration,post-operation and the final follow-up were (27.7±10.0)°,(11.3±5.4)°,(10.0±5.0)°,compared with preoperation,the differences of post-operation and the final follow-up were statistically significant (t=6.600,7.230,all P<0.05),with the correction rate of 59.2%,63.9%;the compensatory head side Cobb angle were(25.0±12.8)°,(11.9±8.2)°,(10.3±6.9)°,compared with preoperation,the differences of post-operation and the final follow-up were statistically significant (t=3.934,4.626,all P<0.05),with the correction rate of 52.4%,58.8%.Trunk shift was significantly improved.One case appeared fracture fixation rods breakage after 9 years,1 case appeared iliac screw breakage after 2 years but no complication such as infection,nerve injury.Conclusions The formation of obstacles and vertebral segmentation defects will lead to lumbosacral spine deformity,and scoliosis.Early one-stage posterior hemivertebra resection,breakage the bone bridge combined with pedicle screw fixation can obtain satisfactory clinical outcome.

Objective To investigate the effects of melatonin on the changes of superoxide dismutase ( SOD ) , reactive oxygen species (ROS),and malondialdehyde(MDA) in N2A cells under hypoxia conditions.Methods Randomly divided the primary cultured neuroblasto-ma cells of mouse into the control group ,hypoxia group ,and MT treatment group .The MT treatment group were given melatonin 5μg/mL for 24 h to set up the treatment model .Rsbiotech of the hypoxia group and MT treatment group were given gaseous mixture of 95%N2 and 5%CO2 ,The ischemia hypoxia model of N 2A cells was set up with cells in the oxidative stress state and cultured for 24 hours at low concentra-tions of serum .The content of SOD , ROS, and MDA was measured respectively by xanthine oxidase , fluorogenic quantitative detection and thiobarbituric acid chromatometry .Results The expression of SOD in hypoxia group and MT treatment group were significantly decreased compared with that in control group(P<0.05).The level of SOD in MT treatment group significantly recovered compared with hypoxia group with singnificant difference (P<0.05).The expression of MDA and ROS in hypoxia group and MT treatment group were significantly in-creased compared with that in control group (P<0.05).The level of MDA and ROS in MT treatment group significantly recovered compared with hypoxia group with singnificant difference (P<0.05).Conclsion Melatonin provides a protective effect on the secondary damage of nerve cells with hypoxia ischemia .The possible mechanism is melatonin could play the role of free radical scavenging and up -regulate the ex-pression level of antioxidants .

Objective To evaluate the utility of waist-to-height ratio ( WHtR ) in predicting metabolic syndrome ( MS) among non-overweight Chinese adults, and to investigate the difference by age and sex. Methods Based on a population-based cross-sectional survey on MS in Zhejiang province in 2010, data of 10 792 non-overweight (body mass index<24. 0 kg/m2 ) adults aged 18 years and older were analyzed by using the receiver operating characteristic(ROC)curve analysis and multiple logistic regression. Results The area under ROC curve (AUC) values of WHtR in MS prediction were 0. 765(95%CI 0. 740-0. 789) and 0. 786(95%CI 0. 765-0. 807), and odds ratios(OR) of WHtR for MS were 1. 23(95%CI 1. 20-1. 27) and 1. 22(95%CI 1. 20-1. 26) in non-overweight men and women, respectively. These two values were with significant difference among non-overweight adults with different agedandsexes(P<0.01). No-overweightmenhadsmallerAUCandORamong18-34agegroupwhilelargerAUCand OR among 35-49, 50-64, and 65-plus age groups than women. The AUC and OR showed slight difference among age groups in non-overweight men, while these two values decreased with age in non-overweight women. The optimal cutoff value was higher in non-overweight women (0. 509 vs 0. 498) than that in men. Non-overweight men aged 50-64 years old had higher cutoff value than those aged 18-34, 35-49, and 65-plus years, while among non-overweight women the cutoff values increased with age. Conclusion WHtR had preferable discriminatory power and association with MS among non-overweight Chinese adults. Further studies should be conducted to explore the difference by age and sex.

Obej ctive Knockout mice are widely used in the studies of joint diseases .This article investigated the effects of joint processing methods , collagenase types ,and collagenase digestion time on the number of primary fibroblast -like synoviocytes (FLSs) obtained from mice. Methods The hind legs of 6 of the 12 male mice were cut open from the hip joints , but not those of the other 6.FLSs were isolated using the type-Ⅳcollagenase digestion method and purified by differential digestion .Cell morphology was observed under the inverted microscope .The type, viability, and purity of the cells were determined by flow cytometry . Rse ults Significantly fewer FLSs were obtained from the mice with the hind legs cut open ( 19 133 ±115 ) than from those without (24 933 ± 503) (P<0.05).The numbers of FLSs collected from the cell suspension at 1, 2, 3, 4, 5,6 , and 7 hours after digestion were 700 ±300 , 600 ±100 , 15 200 ±900 , 5100 ±800 ,2700 ±300 , 900 ±200, and 300 ±100, respectively, the highest at 3 hours. There were statistically significant differences in the total number of FLSs obtained by type-Ⅳ and type-Ⅱ collagenase digestions (24900 ±500v s 18 100 ±400, P<0.05). Conclusion For in virt o culture of primary mouse FLSs, it is recommended that the hip joints be not cut open, and type-Ⅳcollagenase be used with cell sus-pension at 2-6hours after digestion .

Obej ctive Abnormal proliferation of T cells plays an important role in the development of autoimmune diseases. The article aimed to study the inhibitory effect of small molecule compound BD691 on T cell proliferation and its mechanism. Methods Human peripheral blood T-lymphocytes were isolated and purified by the immunomagnetic microbeads,then T cells were ac-tivated with anti-CD3/CD28 mAbs or alloantigen.The inhibitory effect of BD691 on activated T cell proliferation, the cytotoxic effect BD891 on resting T cells and the expression of activated T cells marker CD25 were measured by flow cytometry.Furthermore, ELISA was used to detect the secretion of cytokines associated with T cell differentiation. Results BD691 significantly inhibited the prolif-eration of T cells being stimulated by anti-CD3/CD28 mAb or alloantigen in a dose-dependent manner, and IC50 values are (8.5 ± 1.5)μmol/L and (7.2 ±1.3)μmol/L, respectively.However, BD691 had no obvious cytotoxic effects on resting T cells and periph-eral blood mononuclear cells, even at a high concentration ( up to 100μmol/L) .In T cells which were not activated by anti-CD3/CD28 mAb, the percentage of CD25+T cells is only 1.6%of the total cells, while the number increased to 68% after activating treatment.Mean-while, in T cells which were activated by 0, 3.3, 10, 30μmol/L BD691, no obvious change of CD25 expression were observed, while immunosuppressant FK506 (0.1μmol/L) significantly decreased the expression of CD25 +T cells (14.9%).In unactivated T cells, 95.6%cells were at G0/G1 phase, while after activation, the percentage of cells at G0/G1 phase reduced to 57.7%.In addition, BD691 inhibited the secretion of IFN-γ, IL-6 and IL-17 in activated T cells, but had no effects on the secretion of IL-2, IL-4 and IL-10. Co nclusion BD691 exerts no effects on T cell activation, but it inhibits T cell proliferation by inducing T cell cycling arrest at G0/G1 phase.Moreover, BD691 inhibits the secretion of key cytokines (such as IFN-γ, IL-6, IL-17) closely related to the differ-entiation of Th1 and Th17 cells.The results suggest that BD 691 is a potential lead compound to develop a new immunosuppressant for the inhibition of abnormal proliferation and differentiation of T cells.

Objective:To investigate any change in the effective connectivity between the bilateral anterior central gyruses after transcranial magnetic stimulation (TMS).Methods:Twenty-one healthy subjects were examined using resting state functional magnetic resonance imaging (rs-fMRI) before and after receiving continuous theta burst stimulation (cTBS). The brain atlas of the Institute of Automation of the Chinese Academy of Sciences was used for fine partitioning of the bilateral anterior central gyruses. Granger causality analysis was used to compare any changes in the effective connectivity between them.Results:After the cTBS inhibited the right M1 area, significant changes in effective connectivity among the sub-regions of the bilateral M1 area were observed. The effective connectivity of the right upper limb to the left upper limb and the left head to face were weakened, while that of the left upper limb to the right head, as well as of the face to the right upper limb was enhanced.Conclusion:For people whose right M1 area has been inhibited by cTBS, the effective connectivity changes in both upper limb functional areas of the M1 region reflect inter-hemispheric inhibition. Opposite changes were found in the trunk and upper limbs.