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.

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.

Following an analysis of the problems and challenges of hospitals regulation in China and a comparative study of situations at home and abroad,this paper presented six policy proposals for the reform of public hospitals regulatory system in China.The policy suggestions included definite regulatory objectives,defined relationship between public hospitals and government,strengthening both economic regulation and social regulation, establishing multivariate regulatory system, introducing flexible regulatory methods and full utilization of the third-party impact,which met the requirements of public hospital in healthcare reform in China and propelled the reform forward.

Objective To explore the effect of Yangxueqingnao granule in treatment of cerebral small vessel disease (SVD)patients with mild cognitive dysfunction (MCI).Methods 52 patients with SVD related MCI were selected.They were treated with Yangxueqingnao granule,each 4.0 g,3 times a day,3 months for a course of treat-ment.Montreal cognitive scale (MoCA),simple mental state scale (MMSE)and event related potential (P300)were detected before and after treatment,and the clinical effect was observed.Results After a course of treatment by Yan-gxueqingnao granule,the cognitive function and memory of 52 patients were improved to some extent.In addition to the attention of an MoCA meter to a foreign project[Visual space:before taking the medicine (2.92 ±0.26)points,after taking the medicine (3.25 ±0.66)points;Named:before taking the medicine (2.26 ±0.70)points,after taking the medicine(2.92 ±0.49)points;Language:before taking the medicine (1.47 ±0.70)points,after taking the medicine (2.17 ±0.98)points;Abstraction:before taking the medicine (0.45 ±0.38)points,after taking the medicine (0.68 ±0.63)points;Delayed recall:before taking the medicine (1.67 ±0.74)points,after taking the medicine (2.52 ±1.50)points;Directional:before taking the medicine (4.73 ±0.35 )points,after taking the medicine (5.52 ±0.57)points ]and total score[Before taking the medicine (18.75 ±0.66)points,after taking the medicine (19.12 ±1.45)points],the differences were statistically significant (the total score:t =7.56,P =0.000;Visual space:t =5.86,P =0.002;Named:t =5.42,P =0.000;Be careful:t =1.23,P =0.121;Language:t =4.52,P =0.000;Abstraction:t =2.65,P =0.001;Delayed memory:t =7.96,P =0.000).While the total score of MMSE scale [before taking the medicine (25.36 ±1.89)points,after taking the medicine (28.53 ±2.91 )points],memory [before taking the medicine (2.64 ±0.42)points,after taking the medicine (2.75 ±0.53)points]and recall [before taking the medicine (1.52 ±0.48)points,after taking the medicine (1.98 ±0.78)points ],the differences were statistically significant(the total score:t =2.78,Memory:t =1.95,Recall:t =3.43,all P <0.05).P300 before and after treatment[before taking the medicine P300 latency of (389 ±21)ms,after taking the medicine P300 latency of (341 ±18)ms)],the difference was significant (t =12.514,P <0.001).Conclusion Yangxueqingnao granule had therapeutic effect in patients with SVD -MCI.

Objective To evaluate the effects of transtheoretical model and motivational interviewing on medication?compliance in patients with percutaneous coronary intervention (PCI) post-operation. Methods One hundred patients with PCI post-operation were randomized into the observation group (n = 50) and the control group (n = 50). The medication compliance, 6-min walk test and major adverse cardiovascular events (MACE) were compared between two groups. Results At 1 month, 3 months and 6 months post-intervention, the medication compliance in the observation group was significantly increased (P < 0.05, respectively). The 6-min walk test in the observation group was significantly increased compared with the control group. The MACE rate in the observation group and the control group was 10.0% and 36.0%, respectively, with significant difference (P< 0.05). Conclusion The transtheoretical model and motivational interviewing can effectively increase the medication compliance, improve the postoperative rehabilitation, and decrease the MACE rate.

OBJECTIVE:To provide reference for drug management in clinical trial process in hospital. METHODS:To summarize the common problems of current drug control in clinical trial process and analyze their harmfulness. Some countermeasures were put forward to resolve those problems. RESULTS & CONCLUSIONS:At present uniform management mode hasn’t been established for drug control in clinical trial institutions. Realizations on the importance of drug control are different from one another. There are several factors which affect clinical trials even result in undesirable consequence. The institution should formulate scientific,effective and practical management standards and operation process to improve the quality of drug control and ensure safety, scientificity and reliability of clinical trials.