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.

[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.

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.

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.

Antithrombin III (AT III) is the most important anti-clotting substance. Recombinant human antithrombin III (rhAT III) expressed in transgenic goat milk attracts more and more attention. Develop an effective purification route for rhAT III is vital to its industrial production. An efficient purification method was developed for the rapid purification of rhAT III by isoelectric precipitation and heparin affinity chromatography. First, casein was effectively removed by isoelectric precipitation. rhAT III was further purified by heparin affinity chromatography. In the process of heparin affinity chromatography, the effects of pH and temperature on the stability of rhAT III were studied, and the effects of operating conditions, elution gradient, flow rate and sample loaded, on the purification efficiency were also studied. Under the optimized conditions, the protein recovery of rhAT III was about 90% with purity over 99%, while its activity recovery was about 50%. Such a purification process is very simple and effective, and it would provide a valuable reference for the further scaling-up of industrial production.
Animals ; Animals, Genetically Modified ; Antithrombin III ; biosynthesis ; Chromatography, Affinity ; Female ; Goats ; Heparin ; Humans ; Mammary Glands, Animal ; metabolism ; Milk ; chemistry ; Recombinant Proteins ; biosynthesis

Objective To investigate whether resting state-fMRI (RS-fMRI) based on local consistency (ReHo), am?plitude low-frequency fluctuate (ALFF) and fALFF can add meaningful information on preoperative localization of epilepto?genic zone in patients with malformations of cortical development (MCD). Methods Ten epilepsy patients with MCD were studied with RS-fMRI using a 3.0 T scanner. The resting state data were preprocessed and analyzed using SPM8 and REST to generate the activation map. Abnormal ReHo, ALFF and fALFF related blood oxygen level dependent (BOLD) signal changes were compared to video EEG (VEEG),PET,MRI findings and the final result of a comprehensive evaluation-de?fined epileptogenic zone. For operated patients, postoperative resection and histology were compared to BOLD responses. Re?sults The results of spike localization of RS-fMRI were consistent with VEEG, PET, MRI findings and final comprehensive evaluation-defined epileptogenic zone in 6, 8, 6, 7 of 10 investigations. Six operated patients (including two negative results of MRI examination) revealed local abnormal changes but not visible on structural MRI, which was confirmed cortical malfor?mations by pathology after operation (2 heterotopia and 4 cortical dysplasia). Conclusion RS-fMRI may help to delineate the epileptic focus in epilepsy patients with MCD.

PurposeTo determine the feasibility and accuracy of dual source CT (DSCT) in assessing myocardial delayed-enhancement and left ventricular wall thickness of hypertrophic cardiomyopathy (HCM) in comparison with cardiac magnetic resonance (CMR).Materials and MethodsEighty patients with HCM confirmed by clinical diagnosis were enrolled in the study. DSCT images and CMR images were acquired at the arterial and lag phases. According to 17-segment model provided by American Heart Association, the left ventricular wall thickness and location of delayed-enhancement were verified, and the correlation of these two methods were analyzed in terms of the diagnosis of myocardial delayed enhancement (MDE).Results1360 myocardial segments for 80 patients were assessed. The left ventricular wall thickness determined by DSCT was significantly correlated with MR results (r=0.88,P<0.01). DSCT and MDE showed substantial agreement on per-patient (n=74,Kappa=0.751,P<0.05) and per-segment (n=1238, Kappa=0.746,P<0.01) levels. For dense myocardial delayed enhancement, CT findings were significantly correlated with those of CMR (r=0.89, P<0.01), but CT scan slightly underestimated the lesion scope of fibrosis. Bland-Altman analysis showed that CT and MRI were different in measuring the lesion volume of myocardial delayed enhancement (mean standard deviation was 2.71%).ConclusionThe cardiac CT examination provides comprehensive information in coronary artery and myocardial assessment, and MDE-DSCT is also effective in the diagnosis of myocardial fibrosis in HCM since it can be used in assessing myocardial fibrosis.