Objective To study the transfection efficiency, protein expression, and effect of adenovirus-mediated transfection on microvascular endothelial cells transfected by angiopoietin-related protein 2(Ad. ARP2)gene. Methods Mice coronary microvascular endothelial cells(CMECs) were isolated, cultured and transferred by Ad-ARP2. The transfection efficiency and cellular toxicity of adenovirus vector to CMECs were detected by immunofluorescence staining. Expression of Ad. ARP2 in CMECs and the secreted materials in culture medium were measured by Western blot and ELISA and compared among groups of Ad. ARP2, Ad. null, and PBS control. Vascular endothelial cells were incubated with conditional medium containing secreted ARP2, and effects on cells sprouting were observed in matrigel. Results Adenovirus-transfected CMECs showed a very high efficiency. When multiplicity of infection (MOD was 200, the transfection efficiency was 93. 5% ,and no harmful effect on CMECs growth was found. When CMECs were transfected with Ad. ARP2, there was a high ARP2 expression, which was significantly different from that with Ad. Null or PBS. The conditional medium containing ARP2 had an excellent ability to stimulate sprout of CMECs which phenomenon could not be seen in the control groups. Conclusions Adenovirus vector can be transferred into CMECs efficiently and safely. Ad. ARP2 gene transfection allows a high transient expression, and the expression products can stimulate the sprout of microvascular endothelial cells in vitro very well.

·Age-related macular degeneration (AMD) and retinitis pigmentosa (RP) are common outer retinal degenerative problems, which are also the predominant causes of most blinding retinal diseases. Retinal prosthesis is a promising solution for such photoreceptor degeneration diseases.Most of current concepts for a retinal prosthesis are based on neuronal electrical stimulation. In the past twenty years, retinal prosthesis has been developed in two different directions: epiretinal prosthesis and subretinal prosthesis. Each prosthesis technique has its advantages and disadvantages. For epiretinal prosthesis, it is easier to be implanted and has the advantage of keeping most of the electronics in the vitreous cavity, off the retinal surface, which greatly helps in dissipating the heat generated by the implant device. In this paper, a brief overview of retinal prostheses concepts is introduced. After that, several important aspects of epiretinal electrical stimulation will be discussed. Moreover, some practical epiretinal prosthesis devices developed by researchers in United States, Germany and Japan in the past have been reviewed. We hope that the devices will be used widely in the near future.

Objective:To choose the best vector for the expression of CS3 fimbriae. Methods: The CS3 operon was cloned into different plasmid vectors such as pUC19 and pTrc99A. The expression of CS3 was monitored by whole-cell ELISA and SDS-PAGE analysis. The assembly of CS3 fimbriae was detected with electron microscopy. Results:The expression level of CS3 fimbriae using plasmid pUC19 as carrying vector was the highest, and the insertion orientation of CS3 gene into the plasmid has a little effect on its expression level. The expression of CS3 fimbriae was confirmed by SDS-PAGE analysis and electron microscopy.Conclusions:The promotor of CS3 itself played the key role in the expression of CS3 fimbriae and the copy number of plasmid was the main factor to affect the expression level.

Objective To find the risk and hidden danger of blood bacterial contamination existence at present and to investigate the method for carrying out the bacterial contamination related quality monitoring.Methods According to the requirements of the blood collection and supplying related laws and regulations standards,the blood bacterial contamination situation was monitored by supervising the personnel during the blood collection and supplying,key equipments,key materials,environment monitoring and sterility test of blood products.Results The total eligible rate of quality monitoring indicators of blood bacterial contamination reached 99.8％,the eligible rate of blood products bacterial contamination monitoring was 100 ％.Conclusion The hygiene quality of blood collection process and blood products all are in good condition.

To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

ObjectiveTo discuss the expression of VEGF and COX-2 in breast cancer and their correlation with Clinical relationships.MethodsWith Elivision immunohistochemistry,the expression of VEGF and COX-2 was evaluated in 60 samples with primary breast cancer,and it was determined that the correlation of their expression and the clinical features including age,tumor size,clinical TNM staging,pathological grading,armpit lymph node metastasis and prognosis. ResultsThe expression of VEGF had close correlation with tumor size, the clinical TNM staging, pathological grading, and armpit lymph node metastasis (P ＜0.05),but had no correlation with age.The expression of COX-2 had close correlation with the clinical TNM staging pathological grading, and armpit lymph node metastasis (P ＜0.05), but had no correlation with age and tumor size. The expression of VEGF had positive correlation with that of COX-2 (r =0.2615, P ＜0.05);In the death group, the expression of VEGF and COX-2 was significantly higher than the survival group (P ＜0.05).ConclusionThe up-regulated expression of VEGF and COX-2 closely correlated with tumor take place, evolution, metastasis and soake of breast carcinoma. The expression of COX-2 had positive correlation with that of VEGF.Combined detection would be helpful in screening the patients of breast cancer with high risks of recurrence and metastasis, judge the prognosis, so it is good for further treatment.

OBJECTIVETo understand if the Neuraminidase (N1) of Influenza A virus at the surface of yeast-displaying system, eukaryotic expression system and the infected cells could be used for anti-NA Abs screening, their activities and bindings to five candidate Abs were assayed.

METHODSThe surface NA expression was obtained by transfecting by recombinant NA constructors with specific tag-labels or live virus infection. The functional activity was measured by the fluorescent assay. Their bindings to the Abs were detected by flow cytometry.

RESULTSThe surface NAs presenting on the yeast-displaying system and eukaryotic expression system exhibited functional NA activities as the NA at the surface of virus-infected cells which showed affinities to Ab1, 4, and 5. The same bindings to Abl and 5 were found in the surface NA expressed by eukaryotic expression system while minor binding was observed in the yeast displayed-NA.

CONCLUSIONThe epitopes of yeast-displayed NA may be different from the NAs present at eukaryotic expression system and the infected cells which more likely suitable for the screening of anti-NA Abs.

Antibodies ; immunology ; Antigens, Surface ; genetics ; immunology ; Cell Line ; HEK293 Cells ; Humans ; Influenza A virus ; genetics ; immunology ; Neuraminidase ; genetics ; immunology ; Protein Binding ; Recombinant Proteins ; genetics ; immunology

Adolescent ; Adult ; Behavior, Addictive ; epidemiology ; Child ; Female ; Humans ; Inpatients ; statistics & numerical data ; Internet ; Male ; Young Adult

Objective To observe the effects of comprehensive measures of changing grain,selenium supplementation,off-site education and resettlement on prevention of children's Kashin-Beck disease in Aba state.Methods Fifty eight villages in Aba Kashin-Beck disease areas were chosen as intervention points in Aba state Sichuan province from 2007 to 2011.Based on the implementation of prevention and control measures,the villages were divided into off-site education + changing grain + selenium supplementation group and resettlement + off-site education + changing grain + selenium supplementation group,Geletuo town of Seda county,Ganzi state was selected as a control point,and right-hand anteroposterior X-ray examination(including the wrist) was carried out on children aged 6-13 from 2007 to 2011 annually.Clinical and X-ray diagnosis of Kashin-Beck disease was made in accordance with the Diagnostic Criteria of Kaschin-Beck Disease(GB 16003-1995).The effects of prevention and control measures were evaluated by comparing the child X-ray detection rate before and after the implementation of the measures.Results The average X-ray positive detectable rate of children in the intervention points was 2.07％(66/3181),2.72％ (69/2540),1.16％ (35/3017),0.56％ (19/3397) and 0.56％ (24/4273),respectively from 2007 to 2011,with a downward trend (x2trend =66.74,P ＜ 0.01).There was a downward trend in the average X-ray positive detectable rate of children in off-site education + changing grain + selenium supplementation group [1.60％(29/1809),2.63％ (39/1484),1.29％ (25/1941),0.64％ (15/2332),0.42％ (10/2379)] and resettlement + off-site education + changing grain + selenium supplementation group [2.70％ (37/1372),2.84％ (30/1056),0.93％(10/1076),0.38％ (4/1065),0.74％(14/1894)] (x2trend=30.97,35.19,all P ＜ 0.01).The average X-ray positive detectable rate of children in the intervention group was 0 from 2007 to 2010,and was 1.61％ (1/62) in 2011.The difference of X-ray positive detectable rate was not statistically significant in the control group in the 5 years from 2007 to 2011.The difference of children's X-ray positive detectable rate was not statistically significant between control group and intervention group.Conclusions The effect of taking changing grain,selenium supplementation,off-site education and resettlement comprehensive measures to prevent children's Kashin-Beck disease is not significant in those places where the state of Kaschin-Beck disease is not active.

Objective To study the effect of Shuangteng Bitong Cataplasm on TLR4, MyD88 and NF-κB and inflammatory cytokines in rats with collagen-induced arthritis; To discuss relevant mechanism of action. Methods A CIA model was made by subcutaneous injection of collagen emulsion from rat tail. Rats were randomly divided into normal group,model group,Shuangteng Bitong Cataplasm high-dose and low-dose groups.The contents of TNF-α, IL-1β and IL-6 in plasma were evaluated by ELISA, and the expression levels of TLR4, MyD88 and NF-κB in synovial tissues were evaluated by RT-PCR and Western blot. Results Compared with the model group, the contents of TNF-α, IL-1β and IL-6 in serum decreased remarkably in the Shuangteng Bitong Cataplasm low-dose and high-dose group, and the expression levels of TLR4, MyD88 and NF-κB in synovial tissues decreased remarkably. Conclusion Shuangteng Bitong Cataplasm can inhibit inflammatory reaction and regulate the expressions of relevant factors in synovial tissues to realize the anti-flammatory effects.