In this study, pEGFP-N1 was chosen as the reporter plasmid and transferred into Ctenopharyngodon idellus kidney (CIK) cells by electroporation, and the optimal electroporation conditions were determined by testing the transfection efficiency with different voltages, pulse times, plasmid amounts, and numbers of shocks. The results showed that the maximum electroporation efficiency was achieved under the following conditions in a 0.2 cm electroporation cuvette containing CIK cells (1.5 x 10(7)/mL, 200 microl): electric voltage 200 V, pulse time 45 ms, plasmid 30 microg, and one electric shock. The total genomic RNA of grass carp reovirus (GCRV) was extracted in this experiment and reversely transcribed into cDNA, which was used to amplify the gene segment of GCRV non-structural protein NS26 using designed specific primers. The PCR product was recombined into pEGFP-N1 vector. The fusion protein EGFP-NS26 was successfully and efficiently expressed in the CIK cells by electroporation, which was confirmed by both fluorescent imaging and Western blot analysis. This experiment laid a foundation for further functional studies of the non-structural protein NS26 of GCRV.
Animals ; Cell Line ; Cyprinidae ; Electroporation ; Fish Diseases ; virology ; Gene Expression ; Kidney ; virology ; Reoviridae ; genetics ; physiology ; Reoviridae Infections ; veterinary ; virology ; Viral Nonstructural Proteins ; genetics ; metabolism

OBJECTIVE:To compare home-made mu'an-eye-gel(acyclovir plus honey) with commercial aciclovir(ACV)-eye-gel in releasing drug characteristics in vitro.METHODS:The in vitro drug release test was conducted by the third method of dissolution determination stated in Chinese Pharmacopeia together with bag filler method.The cumulative drug-releasing percentage and the acyclovir amount in mu'an-eye-gel versus ACV-eye-gel were determined by UV spec-trophotometry,and the accumulative releasing drug percentages of the two preparations were computed and their drug release behaviors w ere compared.RESULTS:The in vitro releasing behaviors of mu'an-eye-gel followed the Weibull kinetic equa-tion,however the vitro releasing behavior of commercial ACV-eye-gel followed the zero order kinetic equation,and the T80%and Q8 h had statistical significances between(mu'an-eye-gel:T80%=3.156?0.013(h),Q8 h=93.28?0.010(%);ACV-eye-gel:T80%=10.16?0.009(h),Q8 h=67.85?0.025(%)) 2 kinds of preparation.CONCLUSION:Mu'an-eye-gel is superior to the commercial ACV ophthalmic gel in both releasing velocity and accumulative drug release percentage.

The paper is about the systematic studies of biological characteristics of 15 stains rhizobia isolated purified from Acacia confusa grew in Guangxi karst environment.The results showed that there were typical characteristics of rhizobia.there were negative reaction about use of 3-ketolactose and beef extract peptone nutrient agar medium,and positive reaction about use of starch and citrate medium,and produce acid in reaction of BTB and litmus milk medium,(NH4)2HPO4 was used as nitrogen sources and both four monosaccharides and three disaccharides could be utilized as carbon sources in 15 strains rhizobia isolated Acacia confuse.Among the 15 strains for the tests,11 strains could deoxidize the nitrate of medium into nitrite,14 strains could grow well on NaCl solution concentration 3.0 %～4.0 %,14 strains could grow at 39℃,13 strains may grow on highest pH12 and 4 strains on lowest pH4 cultrue medium.15 strains can grow in 10% and 11 strains in 10%～30% of CaCO3 solution concentration.

Objective To study the feasibility of using heat and moisture exchangers (HME)as an alternative to heated humidifiers (HH) in patients undergoing mechanical ventilation. Methods 266 pa-tients with mechanical ventilation admitted to our ICU over the recent 3 years were allocated to the experi-mental group (humidification with a heat and moisture exchanger) and the control group (with heated hu-midifier), and the effect of humidification, the reserved time of artificial airway, the time on mechanical yen-tilation, the time of stay in ICU, the ineidenee of ventilator-associated pneumonia (VAP) and the mor-tality rate were comparatively studied and analyzed. Results Significant differences were found between the experimental and the control group in effect of humidification, insufficiency of humidification or excessive hu-midification, airway spasm and time on mechanical ventilation and time of stay in ICU. The incidence of VAP in the control group was significantly higher than that in the experimental group. There were no significant dif-ference between the two groups in the reserved time of artificial airway and the mortality rate. There were no accident of humidification occurred in the experimental group while there were one case complicated with air-way burn and 11 eases complicated with choking with water in the control group. Conclusions We conclude that HH can be replaced by HME on mechanical ventilation while disease evolution and effect of humidification should be monitored closely and keep HME unobstructed.

Objective To explore the setting of logos on tobacco control information at outlets for retails and restaurants in 12 selected cities of China.Methods For all the shops for retail of tobacco,alcohol,food and restaurants under survey in 333 blocks of 12 cities (Beijing,Tianjin,Shanghai,Qingdao,Hangzhou,Shaoxing,Suzhou,Nantong,Zhenjiang,Chengdu,Xining and Harbin),setting and contents of logos on tobacco control information,inside and outside them were examined.Results 45 700 objectives were included in the study.Among all types of retail shops,the identification rate of tobacco control information at the entrance and inside were 3.6％ and 4.4％,with an overall identification rate as 7.0％.The overall rate at the entrance of all the restaurants was 4.6％ which was larger than the ones at the retail shops.Our result showed that there were differences between cities and types of establishments and higher rates seen in the larger ones.Of all the places that having had placement of information on tobacco control,only 18.5％ of them had put them both inside and outside.Slogans or images on "No Smoking" were the main forms of information but less than 10％ of them would show signs as ‘exclusive non-smoking'.Conclusion Data from our survey showed that the identification rate of tobacco control information was at a low level in 12 cities,and differences were seen between cities,size of establishment,that called for improvement of the existing tobacco control policies in China.

Objective:To investigate the expression of Fas、FasL and the apoptosis of liver cancer cell line HepG2 transfected with LIGHT and IFN-? gene mediated by Cationic liposome.Methods:HepG2 cells were divided into two groups(the solo transfection of LIGHT gene and the combined transfection of LIGHT and IFN-? genes) and the control groups(no transfection).HepG2 cells were cellected at 12h,24h and 48h after transfection.The apoptosis of HepG2 cells and the expression of Fas and FasL of the HepG2 cells were investigated with flow cytometry.Results:After transfection,the apoptosis of HepG2 cells increased,and the apoptosis of combined transfection group was higher than the solo transfection of LIGHT(P

Objective To investigate the effect of PDCA cycle method on mother-infant bedside nursing.Methods A total of 200 mothers and infants were randomly divided into the observation group and the control group, each with 100 cases.The observation group applied PDCA cycle method for continuous quality management of every aspect of mother-infant bedside nursing , while the control group received conventional nursing.Mothers’ satisfaction, correctness of health education knowledge and grasp of neonatal nursing skills were compared between two groups .Results Ninety-two mothers in the observation group and 80 mothers in the control group were satisfied with health education , with statistically significant difference (Z=-2.391 1, P<0.05).Satisfaction rate on other items was also higher in the observation group than in the control group , but without any statistically significant difference (P>0.05).Mothers’ correct rate of the advantage of breast-feeding, breast nursing methods and postpartum nutrition activities was respectively 95%,97%,96% in the observation group , higher than 86%, 88%, 87% in the control group , and the differences were statistically significant (χ2 =4.710,5.837,5.207,2.445, respectively; P <0.05).Mothers’ grasp rate of neonatal shower, newborn massage, neonatal umbilical nursing was respectively 90%,92%,90% in the observation group, higher than 80%,82%,85%in the control group , with statistically significant differences (χ2 =8.37, 8.59,7.02, respectively;P<0.01).Conclusions PDCA cycle method for continuous quality management of every aspect of mother-infant bedside nursing can improve mothers ’ satisfaction, as well as their health education knowledge and neonatal nursing skills .

OBJECTIVETo investigate the association between haplotypes of S447X and Hind III polymorphisms of lipoprotein lipase (LPL) gene and serum lipids in a population-based twin cohort study in China.

METHODSTwin subjects were collected based on the twin registry system of China. All twins were investigated by a standard questionnaire and physical examinations. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect the genotypes of S447X and Hind III polymorphisms. Linkage disequilibrium test and haplotypes were estimated between two polymorphisms.

RESULTSNine hundred and eighty-seven pairs of twins were eligible for analysis. The two polymorphisms of LPL gene were significantly linkage disequilibrium. In female twins, the H- allele of Hind III polymorphism was significantly related to lower levels of triglycerides(TG) and lower risk of high TG dislipidemia, but those associations disappeared after adjusting the polymorphism of S447X. The H- X haplotype of those two polymorphisms was significantly related to lower TG and TG/HDL (decreasing 12.9% and 14.9% respectively), as well as significantly to lower risk of high TG dislipidemia (OR = 0.40).

CONCLUSIONThe haplotypes of S447X and Hind III polymorphisms were significantly related to the favorable effect of lipids,but this effect was mostly determined by the polymorphism of S447X, while the effect of Hind III polymorphism was indirectly influenced by the linkage disequilibrium with S447X polymorphism.

Adult ; Female ; Haplotypes ; genetics ; Humans ; Linkage Disequilibrium ; genetics ; Lipids ; blood ; Lipoprotein Lipase ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length ; genetics ; Twins ; blood ; genetics

Objective To study the information support of pregnant women undergoing assisted reproductive technology (ART) and their information resources.Methods 105 pregnant women undergoing assisted reproductive technology at Reproductive Center in our hospital were investigated with information support questionnaire.Results The score of information support for pregnant women undergoing assisted reproductive technology was (96.73±11.34), ranging in the middle. 68.57% of their information came from the medical staff.Conclusions Nurses should provide a wide and multi-channel information support for pregnant women undergoing assisted reproductive technology in conformity with their education level.

BACKGROUND:The angiogenesis may be related to the proliferation of neural stem ceils,but there is still no unified view.OBJECTIVE:To observe the influence of angiogenesis on neural stem cell proliferation in the subventricular zone of rats after focal cerebral ischemia/reperfusion.METHODS:Male Sprague-Dawley rats were randomly divided into normal group,sham group,vascular endothelial growth factor (VEGF)+cerebral ischemia/reperfusion group,normal saline (NS)+cerebral ischemia/reperfusion group.The injection was done via the lateral cerebral ventricle.Then,each group was subdivided into four groups (1,2,7,14 days after ischemia/reperfusion).Focal cerebral ischemia/reperfusion models were made by the thread method.After modeling,the corresponding intervention was given in each group.The expression changes of Nestin and vWF mRNA in the subventricular zone were detected in all groups by immunohistochemical staining and real-time PCR,respectively.RESULTS AND CONCLUSION:There was a certain increase in vWF and Nestin positive expression in the subventricular zone after cerebral ischemia/reperfusion.At 7 days after ischemia,the expression of vWF mRNA and Nestin reached the peak,indicating the proliferation of neural stem cells in the subventricular zone after cerebral ischemia/reperfusion is associated with the time of angiogenesis.In addition,the expression of vWF mRNA and Nestin was significantly higher in the VEGF+cerebral ischemia/reperfusion group than the other two groups,indicating angiogenesis could promote the proliferation of neural stem ceils in the subventricular zone of rats after cerebral ischemia/reperfusion.