Grass carp reovirus (GCRV), a disaster agent to aquatic animals, belongs to Genus Aquareovirus of family Reoviridea. Sequence analysis revealed GCRV genome segment 8 (s8) was 1296 bp nucleotides in length encoding an inner capsid protein VP6 of about 43kDa. To obtain in vitro non-fusion expression of a GCRV VP6 protein containing a molecular of fluorescence reporter, the recombinant baculovirus, which contained the GCRVs8 and eGFP (enhanced green fluorescence protein)genes, was constructed by using the Bac-to-Bac insect expression system. In this study, the whole GCRVs8 and eGFP genes, amplified by PCR, were constructed into a pFastBacDual vector under polyhedron (PH) and p10 promoters, respectively. The constructed dual recombinant plasmid (pFbDGCRVs8/eGFP) was transformed into DH10Bac cells to obtain recombinant Bacmid (AcGCRVs8/eGFP) by transposition. Finally, the recombinant bacluovirus (vAcGCRVs8/eGFP) was obtained from transfected Sf9 insect cells. The green fluorescence that was expressed by transfected Sf9 cells was initially observed 3 days post transfection, and gradually enhanced and extended around 5days culture in P1(Passage1) stock. The stable high level expression of recombinant protein was observed in P2 and subsequent passage budding virus (BV) stock. Additionally, PCR amplification from P1 and amplified P2 BV stock further confirmed the validity of the dual-recombinant baculovirus. Our results provide a foundation for expression and assembly of the GCRV structural protein in vitro.

Antineoplastic Agents ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology

Objective To investigate the vaginal flora characteristics,clinical symptoms and signs of lactating women 6-8 weeks after delivery and the influence factors.Methods Two hundred and twenty-three puerpera 6-8 weeks after delivery who went to outpatient department of Peking University First Hospital for postpartum review (study group) and 200 normal non-pregnant women (control group) from April to May,2010 were enrolled into this study.All in study group were evaluated by a questionnaire regarding their symptoms,sexual behavior,breast-feeding situation and delivery mode.Vaginal secretions from upper 1/3 of vagina were collected and evaluated by vaginal microbial evaluation system.Bacterial vaginosis (BV) was diagnosed if Nugent score ≥ 7.Enumeration data between groups was compared by t test,and quantitative data by Chi-square test.Results The density of vaginal flora in study group was different from that of control group (x2 =96.302,P＜0.001) ; so did the flora diversity of two groups (x2 =69.871,P＜0.001).There was no difference between mean age of study group [(30.5 ± 3.9) years] and control group [(30.9 ± 2.4)years],P＞0.05.The cesarean section rate was 52.7％ (89/169),13 women (7.7％) had began their sexual behavior before interview,116 women (70.3％) took breast-feeding.Dominant bacteria in the vagina of lactating women were Gram-positive cocci (69/175,39.4％) and Gram-positive bacillus (54/175, 30.9％), which was different from control group (Gram-positive bacillus,180/189,95.2％),P＜0.001.The vaginal pH of study group was 5.0±0.5,which was higher than that (＜4.5) of control group.In study group,incidence of lower hydrogen peroxide level of vagina was 84.3 ％ (177/210) ; positive rate of sialidase was 6.7 ％ (14/210) ; positive rate of leukocyte esterase was 91.0％ (191/210).A large number of postpartum women had an abnormal Nugent score,most of them showed a state of intermediate BV (132/195,67.7％) or BV (8/195,4.1％),while a few patients had symptoms of BV (23.5％,33/140).Sexual behavior had no significant effect on postpartum vaginal flora.The proportion of abnormal vaginal flora in breast feeding mothers (74.1 ％,86/116) was significantly higher than that (57.1％,28/49) in artificial feeding mothers,P=0.034.Delivery mode also affected postpartum vaginal flora.Women underwent cesarean section were more susceptible to abnormal vaginal flora (77.5％,69/89) than those underwent vaginal delivery (57.5％,46/80),x2 =7.726,P=0.005.Conclusions Vaginal flora of postpartum women 6-8 weeks after delivery had significant difference from that of nonpregnant women.The incidence of asymptomatic abnormal vaginal flora was high.Breastfeeding and delivery mode are influence factors of abnormal vaginal flora.

Objective To investigate the suppression effect of RNA interference(RNAi) on the expression of epidermal growth factor receptor(EGFR) of ovarian cancer SKOV3 cell line. Methods SKOV3 cells were transfected with synthetic EGFR sequence-specific dsRNA by lipofectamin 2000.The expression level of EGFR mRNA and protein were detected by RT-PCR and Western blotting,respectively.The proliferation of transfected SKOV3 cells was detected by WST-1 cell proliferation assay.The abilities of adhesion,migration and invasion were detected by cell adhesion assay,cell migration assay and Matrigel invasion assay,respectively. Results The suppression rates on EGFR mRNA and protein by sequence-specific dsRNA-EGFR were 73.65% and 58.94%,respectively.The cell proliferation was inhibited by 22.54%,35.76% and 31.07%,respectively at 24,48 and 72 h after transfection.The adhesion of sequence-specific dsRNA transfected SKOV3 cells for 30 min and 90 min were reduced by 12.11% and 18.66%,respectively.The migration and invasion of SKOV3 cells were decreased by 25.47% and 22.08%,respectively. Conclusion The down-regulation of EGFR expression of ovarian cancer by sequence-specific dsRNA can lead to the suppression of proliferation,adhesion,migration and invasion of ovarian cancer cells.

Objective To explore the clinical effects of atomization inhalation pulmicort respules in treatment with pediatric acute infectious laryngitis. Methods 104 cases with pediatric acute infectious laryngitis admitted in Xiaoshan Hospital of Zhejiang Province from January 2011 to July 2013 were radomly divided into experimental group(n=52)and control group(n=52). Patients in experimental group were given atomization inhalation pulmicort respules,and patients in control group were received dexamethasone by intravenous injection. The symptom improvement,disappearing times of symptoms and therapeutic effect in two groups were observed and compared. Results Compared with control group,the improvement scores were higher,and disappearing time of symptoms were shorter in experiment group(P<0.05). The therapeutic effect in experimental group was up to 100%, while control group was 90.74%,and the difference between them was statistically significant(P<0.05). Conclusion Compared with dexamethasone, atomization inhalation pulmicort respules has advantages in treatment with pediatric acute infectious laryngitis,with better efficacy,shorter treatment time and fewer side effects.

OBJECTIVETo investigate the exerted force in different phases of the female menstrual cycle, as well as the changes in estrogen (E2), osteocalcin (OCN), receptor activator of nuclear factor-kappa B ligand (RANKL), and osteoprotegerin (OPG) levels in the gingival crevicular fluid (GCF) during orthodontic tooth movement, to provide a theoretical basis for the selection of the best opportunity for efficient tooth movement.

METHODSTwelve women (aged 18 years to 28 years) with extracted first premolars had been selected. Six women in the group were randomly selected as the menstrual period group, whereas the remaining six were assigned to the ovulation period group. Right canines were retracted with 1.5 N NiTi close coil spring. GCF samples were collected prior to the force exertion experiments at 0 (T0), 15 (T1), 30 (T2), and 45 d (T3). The levels of E2, OCN, OPG and RANKL in GCF were measured by chemiluminescence and enzymelinked immunosorbent assay.

RESULTSThe E2 and OCN levels were significantly higher in the ovulation period group than in the menstrual period group (P < 0.05). No significant differences were found in RANKL and OPG levels between the two groups (P > 0.05). Finally, no significant difference was found in RANKL/OPG ratio between the two groups (P > 0.05).

CONCLUSIONExerted force on teeth during the menstrual period may promote rapid tooth movement.

Objective To investigate the relationship between cognitive function and emotional status in functional constipation patients.Methods Thirty-five functional constipation patients (the functional constipation group) and 24 healthy controls (the HCs) were enrolled in an event-related potential (ERP),behavior and electroeneephalogram study while performing an oddball auditory task.Response time,accuracy,latency and P300 amplitude were compared between the two groups.The Hamilton depression rating scale (HAMD-17),Hamilton anxiety scale (HAMA),symptom check list-90 (SCL-90) and Eysenck personality questionnaire (EPQ) were administered prior to the oddball task.Results The average HAMD-17,HAM A and SCL-90 (the somatization,obsessive-compulsive disorder,depression,anxiety and psychosis dimensions) scores of the functional constipation group differed significantly from those of the HCs.The intro-extraversion and neurosis dimensions of the EPQ were also significantly different.There was no significant difference in response time or latency between the two groups.The ERP results showed that the P300 amplitudes at the F3 and F7 sites were significantly different between the functional constipation group and the HCs.But there was no significant difference in P300 latencies at F3 or F7 between the two groups.Conclusions The findings suggest that patients with functional constipation are more susceptible to depression and anxiety,as well as somatization,obsessive-compulsive tendencies and other neurotic personality characteristics.Patients may employ psychological defense mechanisms to avoid the depression and anxiety.The ERP results imply there may be the implicit cognitive dysfunction with emotion in patients with functional constipation.

BACKGRUND:The high prevalence of hepatitis B virus(HBV) is related with the sexual characteristics in drug abusers.How about the relationship between prevalence of HBV/hepatitis C virus(HCV) co infection and the sexual characteristics in drug abusers? OBJECTIVE:To investigate the relationship between sexual characteristics and HBV/HCV co infection in drug abusers. DESIGN:A cross sectional survey taking the drug abuser as the subjects. SETTING:A department of clinical laboratory of a municipal hospital,an epidemiological department of the college of public health of a university,a professional drug abuser treatment center. PARTICIPANTS:Totally 452 male and 320 female drug abusers were randomly selected from the Changsha Xinkaipu Drug Abusers Treatment Center and Zhuzhou Baimalong Female Drug Abusers Treatment Center as the subjects from 1st to 31st January 2001. INTERVENTIONS:A self designed questionnaire was used to investigate the sexual characteristics of drug abusers.HBV surface antigen and HCV antibody were detected in the serums of the participants with ELISA. MAIN OUTCOME MEASURES:The relationship between HBV/HCV co infection and the sexual behavior, numbers of sexual partners,and the usage of condom in drug abusers. RESULTS:The total prevalence of HBV/HCV co infection in drug abusers was 21.24% ; the prevalence of HBV/HCV co infection in male drug abusers was 29.6% ),which was significantly higher than that in females(9.4% )(χ 2=46.016,P< 0.01).There was no significant difference in the prevalence of HBV/HCV co infection between male and female drug abuser whether they had or had not sexual behaviors(χ 2=3.775,P >0.05).The prevalence of HBV/HCV co infection was in an ascending tendency with the increase of sexual partner(χ 2=9.846,P< 0.05).The using frequency of condom was significantly higher in females than in males(U=14.80,P< 0.01). CONCLUSION:There is no correlation between sexual experience and prevalence of HBV/HCV co infection;male drug abusers have higher prevalence of co infection than females;the number of sexual partners is correlated with the prevalence of HBV/HCV co infection,the more sexual partners,the higher prevalence is.The protective effect of condom is greater in female drug abusers than in males.

OBJECTIVE:To discuss the accuracy of detecting tacrolimus(FK506) concentration in whole blood by ELISA and its application in organ transplantation.METHODS:The ELISA was used to detect FK506 valley point concentrations in 5 liver transplanted recipients at different postoperative periods .Based on the results combined with clinical findings, the dosages were adjusted.The accuracy of the method and guidance action in FK506 clinical application were described.RESULTS: In 131 detections of 350 samples, the results of quality control samples were all in the control range .The detecting RSDs of high and low concentration quality control samples were 9.09% and 11.90%, respectively.The control range of FK506 concentrations in whole blood were similar to that reported in literature .CONCLUSION :ELISA is a sensitive and specific method for routine FK506 concentration monitoring in whole blood,which is suitable for application in hospital.

Objective To discuss the effect of two different warming methods on the circulation of free flaps during operation and seek for an optimal warming method which is benificial for the circulation of free flaps, and thus provide clinical evidences for intraoperative care. Methods A total of 96 patients undergoing free flap transplantation were divided into routine warming group and diversification warming group randomly (n=48 each). In routine warming group, temperature in the operating room was set at routine temperature and flaps were douched by routine heated saline. In diversification warming group, the temperature in the operation room, fluids used for washing wounds and flaps were warmed, fluids for intravenous application were pre-warmed by a fluid warmer, in addition, the operation table where patients were lying on was covered with warmer blanket. The influence of two different warming methods on the circulation of free flaps was observed. Results The scores of flap elasticity and capillary refill time in routine warming group were 2.02 ± 0.79 and 2.04 ± 0.81, respectively, vs. 2.50 ± 0.61 and 2.48 ± 0.6 in diversification warming group, showing diversification warming group was statistically better than routine warming group (Z=1.949, 3.872, P<0.05). In addition, the flap survival rate in routine warming group was 81.2%(39/48) vs. 95.8%(46/48) in diversification warming group, showing significantly better results in diversification warming group (χ2=4.02, P < 0.05). Conclusions The diversification warm keeping method can effectively promote the circulation and survival of flaps, improve clinical prognosis, accelerate postoperative rehabilitation, therefore is worthy of being recommanded its clinical application.