Objective To establish the expression of exogenous tissue plasminogen activator (TPA) gene in human umbilical vein endotheliocytes to provide both theoretical basis and new method for gene therapy of ischemic heart disease and prevention of postoperative vessel re-stenosis. Methods Expression vector pcDNA3 1(+)TPA was constructed, and transfected into human umbilical vein endotheliocytes cultivated in vitro by lyposome. The exogenous TPA expression was observed. Results The expression vector pcDNA3 1(+)TPA could efficaciously express TPA in human umbilical vein endotheliocytes. The protein quantity of TPA in the transfected cells was 568 6ng/10 6cell/24h, when measured by enzyme-linked immunosorbent assay, while that in the non-transfected cells was 17 8ng/10 6 cell/24h. The exogenous TPA's activity in the transfected cells was 108 8IU/10 6cell/24h, when measured by chromogenic substrate assay, while that in the non-transfected cells was 5 6 IU/10 6cell/24h. Conclusion When pcDNA3 1(+)TPA was transfected into human umbilical vein endotheliocytes, exogenous TPA could be expressed efficaciously, which provides theoretical basis and new method for TPA gene therapy of ischemic heart disease.

To improve bond selectivity of recombinant β-glucuronidase in Escherichia coli (PGUS-E), based on the PGUS-E structure guidance, three key points R329, T369 and N467 were identified to be responsible for the bond selectivity of PGUS-E, and further saturation mutagenesis was conducted. Two positive mutants R329K and T369V were obtained by a combined selection technique of thin-layer chromatography and high performance liquid chromatography. Compared to PGUS-E, the bond selectivity of mutants R329K and T369V increased by 26.9% and 34.3%, respectively; whereas the biochemical properties such as pH and temperature profile were unchanged. Nevertheless, the activity was decreased compared to PGUS-E. These results further confirmed that sites R329 and T369 played important roles for the bond selectivity and activity. In summary, this study significantly increased the bond selectivity of PGUS-E by structure guided saturation mutagenesis, providing experimental support for elucidating the relationship between the structure and function of PGUS-E.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Escherichia coli ; metabolism ; Glucuronidase ; chemistry ; Industrial Microbiology ; Mutagenesis ; Recombinant Proteins ; chemistry ; Temperature

MicroRNAisatypeofsingle-strandednon-codingRNA,directlybondingtothecomplemen-tary target gene mRNA.That leads to the inhibition of mRNA translation of the target molecule in order to reduce expression of the target gene.MicroRNAs are not only involved in the regulation of inflammatory reac-tion,but also play important roles in the formation of neoplasm.The researches showing that the inflammatory reaction interacting with miRNAs results in the differential expressions of microRNAs in lung cancer have been widespreadly concerned.Along with the deepening of the research on the mechanisms of inflammatory reaction and differential expression of microRNAs,it will provide new strategies for the prevention,diagnosis and treat-ment of lung cancer.

China’s existing laws and regulations in telemedicine fall behind practice.As a result, telemedicine is confronted with such vague constraints as permission,legal relations and legal liability, validity of electronic medical records,telemedicine equipment standards,patients’right to know and their right to privacy,as well as disputes settlement.On such basis,the authors propose to improve the laws and regulations in this regard as soon as possible.

[Objective] To make a microscopic identification of medicinal material of Herba Rabdosiae Serrae (HRS) from different plant resources. [Methods] The dry leaves of the whole, the middle part and the upper-middle part from the plants of Isodon serra (Maxim.) Kudo, Isodon striatus (Benth.) Kudo, Isodon lophanthoides var. geradianus (Benth.) Hara, and Isodon lophanthoides Hara var. graciliflorus (Benth.) Hara were sliced by different methods. Structure of leaf epidermis, the transverse section of leaf and structure of leaf powder were observed under light microscope. [Results] There existed differences in epidermal cells, stomata distribution, nonglandular hair, glandular hair, small glandular hair and glandular scale of leaf epidermis from four kinds of medicinal plants of Herba Rabdosiae Serrae . And there also existed differences in epidermal cells, anticlinal wall and external-section wall of epidermal cells, nonglandular hair, glandular hair, glandular scale, stomata distribution, mesophyll, main vessels, and vascular bundle of leaf transverse section, as well as in epidermal cells, stomata distribution, glandular scale, nonglandular hair, glandular hair and ducts of leaf powder from four kinds of medicinal plants of Herba Rabdosiae Serrae. [Conclusion] Medicinal plants of different kinds of Herba Rabdosiae Serrae have different microscopic identification structure in leaf epidermis, leaf transverse section and leaf powder.

The paper introduces the mutual relationship among the innovation of four dimensions including new medical service concept,new patient interface,new service transmission system and new technology of telemedicine based on the illustration of four-dimensional model of service innovation to the telemedicine service,and makes an analysis by taking the telemedicine service innovation of remote medical center in Henan Province as an example,thus providing reference for the innovative development of telemedicine service.

Objective To compare the effect of conservative and operative treatment for cervical spinal cord injury without fracture and dislocation (CSCIWFD)and to detect mechanism of injury as well as its relationship to outcome.Methods A retrospective review was conducted on 688 patients with CSCIWFD treated from August 1994 to March 2013.There were 155 patients managed conservatively (conservation group) and 533 surgically (operation group).Neurological function improvement was compared between two groups to detect the correlation of patents' age and treatment methods with outcome.Results The patients were followed up for mean 17.9 months (range,3-36 months).Neurological function was estimated using Japanese Orthopedic Association (JOA) score:(1) the recovery rate of patients aged over 40 years in operation group was better than that in conservation group (P ＜0.05) ; (2) the recovery rate in patients aged under 39 years was unsatisfactory in both groups,with insignificant difference between the two groups (P ＞ 0.05).Conclusions Different age of patients with CSCIWFD has different injury mechanism,injury severity and outcome.Surgery provides better results than conservative treatment for patients aged over 40 years,but both results are poor for patients aged under 39 vears.

Objective To isolate enterovirus 71 from a death children,and analyze whether the neurovirulence was related to the variation of nucleotide and amino acid. Methods Enterovirus 71 was isolated from throat swabs which were colleted from Shandong Linyi People's Hospital. The full length genome was sequenced by amplification with RT-PCR and sequencing of 9 overlapped gene fragments covering full length of the genomes. The nucleotide and amino acid sequenced was aligned by BLAST, Bioedit and MEGA 4. Results A strain of enterovirus 71 was isolated and named as SDLY107. The full length was 7405 bp. The results of homology analysis of overall nucleotide sequence showed that strain Fuyang. Anhui. P. R. C/17.08/2 had highest homology (98.6%)with strain SDLY107, and the homology was 80.0% between strain SDLY107 with prototype strain BrCr/70,and 86. 5% between strain SDLY107 with nerve strain MS/87. Phylogenetic analysis showed that the phylogeny was close between SDLY107 with some isolated strains from Chinese Mainland, such as Beijing, Henan, Guangxi, Sbenzhen, Lanzhou, Fuyang, Chongqing and Zhejiang strains, which was clustered for C4 subtype. The results of amino acid sequence analysis showed that there were 2 mutations, E947D and K1873R, for strain SDLY107. Conclusion SDLY107 belonged to C4 subtype, amino acid mutations E947D and K1873R of which may be relevant to the pathogenicity of EV71.

Objective:To analyze gene expression profiling of left ventricular myocardium in patients with chronic congestive heart failure(CHF)caused by rheumatic heart disease(RHD)with the normal controls in order to identify CHF associated target genes. Methods:The gene expression profiles of left ventricular myocardium from patients with CHF by RHD and normal controls were obtained from six human whole-genomic oligonucleotide microarrays(Affymetrix HG U133 Plus 2.0 GeneChip). GeneSpring software was used to identify the differentially expressed genes in both groups,and bioinformatic analysis was applied to analyze the target genes associated with CHF. Real-time PCR was carried out to validate the expression of three target genes. Results:We identified 102 target genes associated with CHF which were classified into 7 gene clusters. Microarray results were further confirmed by real time PCR for three genes. ATF3 was markedly down-regulated,IGFBP2 and NPPB were notably up-regulated in the left ventricular myocardium samples from CHF patients. Conclusion:A lot of differentially expressed genes,obtained by using the whole-genomic expression profiling technology,might be a contributory factor for the initiation and progression of CHF and it helpful for the understanding of underlying pathophysiological implications. Further investigation on these genes would provide a strategy to identify genetic markers and molecular events associated with CHF caused by RHD.

BACKGROUND:Cell transplantation can repair damaged myocardial tissue.However,whether transplaned cells and host cells formed an effective electricity and mechanical couple or whether reconstruction of connexin (Cx) and arrhythmia formed,are still unclear.OBJECTIVE:It is hypothesized that arrhythmia can be treated by changing Cx levels and intervening abnormal GJ channel.Moreover,to explore the effects of bone marrow mesenchyma stem cells (MSCs) on the expression of Cx43 and Cx45 in rata with myocardial infarction.DESIGN,TIME AND SETTING:The randomized controlled animal study was performed at the Experimental Center,Guangxi Medical University from January 2008 to May 2009.MATERIALS:A total of 180 Wistar rats were randomly divided into four groups:normal control,sham operation,myocardial infarction,and MSCs,with 45 animals in each group.Each group was then divided into 3 subgroups (n=15) according to 4 weeks,8 weeks and 12 weeks post-transplantation.Additional 20 healthy,Wistar rats,aged 1 month,were selected to harvest MSCs.METHODS:The third passage of MSCs was induced by 5-aza like cardiomyocytes for 4 weeks,labeled with DAPI at 2 hours before transplantation.Models of acute myocardial infarction were established in all groups except sham operation group.At day 7 after model preparation,2×10~(10) /L MSCs were infused into the edge and center of myocardial infarcted region by multipoint injection.Rats in the myocardial infarction group were subjected to an equal volume of saline as wall as those in the normal control and sham operation groups.MAIN OUTCOME MEASURES:The mRNA expressions of Cx43,Cx45 were assayed by fluorescence quantitative PCR.RESULTS:The mRNA expression of Cx43 among each groups had no difference at weeks 4,8 and 12 after intervention in the normal areas.Compared to the normal areas,Cx43 mRNA reduced significantly at ischamic zone in the myocardial infarction group and MSCs group (P＜0.01),with notably increased of Cx45 mRNA expression (P＜0.01).Compared to myocardial infarction group,Cx43 mRNA expression was statistically higher in the MSCs group at the same points (P ＜ 0.01),and the Cx45 mRNA dramatically declined (P ＜ 0.01).CONCLUSION:Acute myocardial infarction reduces the mRNA expression of Cx43 and increases the Cx45 mRNA expression.The exogenous cells transplantation can upturn the mRNA expression of Cx43 in the border-zone of the infarcted area,and down-regulate the Cx45 mRNA expression in the border-zone of the infarcted area.