Objective To investigate carotid atherosclerosis and its relevant risk factors of ischemic cerebral apoplexy in high-risk population and to explore new modes of behavioral influencing factors based on health management platform.Methods Using the carotid ultrasound screening of personnel undergoing physical examination,the differences of the related risk factors were studied among 1 152 cases of carotid atherosclerosis with differ ent gender and different age and different disease periods.Results High-risk population between 40-49 years wasmainly focused on mild lesions,and the numbers of serious lesions increased significantly among people older than 70;male patients numbered 51 were significantly much more than the females numbered 21 (P＜0.01)during mild lesion period,there was no statistical significance between male numbered 36 and female numbered 29 during serious lesion period (P＞0.05).Setting mild and serious period respectively as the dependent variables,and hypertension,overweight,dyslipidemia,diabetes mellitus,smnoking,physical activity and atrial fibrillation as the independent variables to conduct logistic regression analysis,it showed that mild lesions were related to overweight,hypertension and dyslipidemia.The risk factors of serious lesion period included hypertension,overweight,diabetes mellitus,dyslipidemia,atrial fibrillation,smoking and lack of physical activity.The female detection rates of hypertension and dyslipidemia in female patients,which were 83.8％ and 44.1％ respectively,were higher than those in male patients,which were 67.6％ and 31.0％ respectively.The detection rates of smoking and arterial fibrillation in female patients,which were 0.7％ and 0.3％ respectively,were lower than those in male patients,which were 20.7％ and 1.0％ respectively.Conclusions Early detection of carotid atherosclerosis lesions and its relevant risk factors as soon as possible,and early intervention influencing factors to mild lesions to prevent the atherosclerosis from getting worse.A new model of zero level prevention can be developed to control of ischemic stroke based on health management platform.

Objective To clone CYP3A4 gene and to construct CYP3A4 recombinant mammalian expression vectors which are transferred into K562 cells for expression,and detect the anti-tumor effect of CYP3A4 combined with Cyclophosphamide (CPA) in vitro.Methods Mammalian expression vector containing CYP3A4 gene cloned from human hepatocytes by RT-PCR were constructed and transferred into K562 cells via liposome. The expression of CYP3A4 was detected by RT-PCR and Western blot respectively. MTT detected the anti-tumor effect of CYP3A4 recombinant mammalian expression vectors combined with CPA.Results We cloned CYP3A4 gene and constructed the recombinant mammalian expression vectors pcDNA3, 1/myc-His A (+)-CYP3A4 successfully. Both RT-PCR and Western blot showed significantly higher mRNA and protein expression of CYP3A4 in gene-transfected group than in empty vector-transfected controls and in empty cells controls. The IC_(50) values were 1. 090 16 mmol/L in gene-transfected group, which were significantly lower than in other two groups and could be reduced and increased by the revulsant and inhibitor respectively.Conclusions CYP3A4 recombinant mammalian expression vectors had anti-tumor effect cooperating with CPA, and the effect could be increased and reduced by the revulsant and inhibitor of CYP3A4 respectively.

Grass carp reovirus (GCRV), a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies, also called viral factories. Sequences analysis revealed the nonstructural protein NS80, encoded by GCRV segment 4, has a high similarity with uNS in MRV(Mammalian orthoreoviruses), which may be associated with viral factory formation. To understand the function of the uNS80 protein in virus replication, the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region (335.742) was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28oC. In addition, serum specific rabbit antibody was obtained by using super purified recombinant NS80(335.742) protein as antigen. Moreover, the expressed protein was able to bind to anti-his-tag monoclonal antibody (mouse) and NS80(335-742) specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.

To investigate the relationship between plasma level of homocysteine(Hcy) and the methylenetetrahydrofolate reductase ( MTHFR ) gene polyroorphism with non-alcoholic fatty liver in patients with type 2 diabetes mellitus (T2DM). Methods In a case-control study, plasma levels of Hcy, folic acid (FA), vitamin B12 (VitB12), glycosylated hemoglobin Alc (HbAlc), fasting blood glucose (FBG), total cholesterol and triglyceride were measured in 159 T2DM patients with and without non-alcoholic fatty liver ( NAFL), as well as 52 normal controls. Mutation of the C677T of MTHFR gene was determined by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) for all of them. Results Patients of T2DM both without NAFL (96 case) and with NAFL had higher prevalence of hyperhomocysteinemia (Hhcy) (49% and 21%, respectively ) than normal controls did (4 cases, 8% ) (P<0.05), while patients of T2DM with NAFL had higher prevalence of Hhcy than those without it did (P <0. 05). Plasma level of Hey positively correlated to genotype frequency of the MTHFR gene, plasma 0levels of HbAlc and FBG in patients of T2DM, with coefficients of correlation of 0.248, 0.423 and 0.242, respectively (P < 0.05). Results of multiple logistic regression analysis showed that course of the disease, body mass index, plasma levels of FBG and Hcy all were independent risk factors for non-alcoholic fatty liver in patients with T2DM. Conclusions Hhey was an independent risk for non-alcoholic fatty liver and plasma level of Hey was influenced by frequency of the TT genotype of the MTHFR gene, plasma levels of FA and VitB12, as well as metabolic disturbance in patients with T2DM.

?Advances in genome-wide analysis have revealed that up to 90%of the human genome is transcribed.However, only approximately 1% of RNA transcripts encode proteins, and the remaining transcripts are noncoding RNAs.Noncoding RNAs can be roughly divided into small noncoding RNAs (<200nt ) and long noncoding RNAs ( LncRNAs, >200nt ). Small noncoding RNAs include microRNAs, transfer RNAs and small nucleolar RNAs, whereas the long noncoding RNAs comprise ribosomal RNA, natural antisense transcripts, etc. Although the biosynthesis and biological activities of microRNAs are well studied through bioinformatics and active biological molecules analysis, the understanding of LncRNAs on these aspects is still limited.LncRNAs play multiple roles in regulating gene transcription and translation, and epigenetics.Aberrant LncRNAs expression can occur in various pathological processes and significantly related to the pathogenesis or poor prognosis of ophthalmological diseases. In this review, we will focus on the characteristics and regulatory functions of LncRNAs that are commonly associated with ophthalmological diseases.

Objective To establish an easier and better method for primary culture of rabbit fibroblast-like synoviocytes(FLS) by modified tissue cell culture.Method Healthy rabbit joint synovial layers were cut into small pieces and siphoned off water attached on the tissue with sterile filter paper and then cultured.The growth status and morphology were observed.The growth curve of the 3rd passage cells was measured by CCK-8 assay, and the expression of vimentin was detected by immunocytochemistry.Results The FLS cultured in vitro exhibited a spindle-shaped appearance and could rapidly expand.The cell growth curve was typical of S type, and the cells highly expressed vimentin.Conclusions Primary culture of rabbit FLS by the improved explant culture method is successfully established.The method is simple and highly efficient.It provideds a new method for isolation of FLS.

BACKGROUND:Currently, fibroblast-like synoviocytes from the synovial tissue of patients with rheumatoid arthritis or adjuvant arthritis rats are isolated and cultured mainly using enzyme digestion method, but tissue explant adherent method is rarely reported in primary culture. OBJECTIVE:To establish a culture method of fibroblast-like synoviocytes from adjuvant arthritis rabbits and to investigate cell biological characteristics. METHODS:Rabbit models of adjuvant arthritis were prepared by injected with adjuvant at multiple sites and times, the knee synovial tissues were obtained, and fibroblast-like synoviocytes were separated and cultured by the tissue explant adherent method. Furthermore, the cell morphology was observed. The cell proliferation was measured using cell counting kit-8 assay, and the expression of Vimentin was detected by immunofluorescence.RESULTS AND CONCLUSION:The primary cultured rabbit synoviocytes were long spindle fiber-like and coincided with the fibroblast-like synoviocytes. Moreover, cells were positive for Vimentin. In conclusion, fibroblast-like synoviocytes from rabbits with adjuvant arthritis are successfully isolated and cultured by the tissue explant method.

Moyamoya disease (MMD) is a chronic and progressive cerebrovascular disease which is characterized by the bilateral internal carotid artery ends and (or) stenosis or occlusion of anterior cerebral artery and middle cerebral artery initial segments,compensatory proliferation of small blood vessels in the skull base and formation of abnormal vascular network.Its etiology and pathogenesis remains unclear.The present studies speculate that MMD may be a polygenic disease,inflammation,immune response,abnormal cytokine secretion,endothelial progenitor cell change and nitric oxide level change are associated with the occurrence and development of MMD.This article reviews the advances in research on the genetics and pathophysiological mechanism of MMD.

Objective To investigate the regulating effect of heat reinforcing acupuncture manipulation on body energy metabolism enzymes in rheumatoid arthritis (RA) and preliminarily explain the mechanism of heat-producing action of heat reinforcing acupuncture manipulation. Method Forty chinchilla rabbits were randomized into normal, model, equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups. A model of cold syndrome-type RA was made by ovalbumin induction and exposure to low temperature in the other four groups not including the normal group. From two days after successful model making, the normal and model groups were grabbed and fastened (bound) by the same way as for the acupuncture groups, 30 min once daily. The equal reinforcement and reduction group received even reinforcing-reducing method;the twirling reinforcement group, twirling reinforcement method;the heat reinforcing group, heat reinforcing acupuncture manipulation. The needle was manipulated for 1 min and retained for 30 min once daily, for a total of seven days. The RA rabbit knee joint circumference was measured and the inflammation score was recorded according to synovial histopathological sections before and after treatment. After the completion of intervention, the rabbits were sacrificed and the articular synovium was rapidly separated for frozen sections. Articular synovium lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and cytochrome oxidase (CCO) activities were measured by histochemical staining. Result After acupuncture intervention, the RA rabbit knee joint circumference was shortened in all the equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups, but the shortening effect on the RA rabbit knee joint circumference was better in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05);the inflammation score recorded according to the RA rabbit synovial histopathological sections was also decreased more in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05). Synovial LDH integral optical density, total positive area and area percentage were significantly higher in the model group than in the normal group (P<0.05), and significantly lower in the equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups than in the model group(P<0.05) and also in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05). Rabbit synovial SDH and CCO activities were significantly higher in the model group than in the normal group (P<0.05). Rabbit synovial SDH and CCO integral optical density, total positive area and area percentage were significantly higher in the equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups than in the model group (P<0.05) and also in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05). Conclusion Heat reinforcing acupuncture manipulation has a definite therapeutic effect on RA and can increase SDH and CCO activities to enhance aerobic metabolism and produce more local energy in a rabbit model of RA. That may be the mechanism by which heat reinforcing acupuncture manipulation produces heat.

Objective:To isolate rabbit fibroblast-like synoviocytes(FLSs) by suspended explant culture method. Methods:The healthy rabbit joint synovial layers were obtained. The cells were cultured by suspended explant culture method and compared with the explants adherent culture method. The growth status and morphology were observed. The growth curve of the 3rd passage cells was measured by CCK-8 assay,and the expression of vimentin was tested by immunocytochemistry. Results: The FLS obtained by the two methods exhibited a spindle-shaped appearance and could rapidly expand. The cell growth curve was typical of S type, and the cells highly expressed vimentin. Conclusion:Primary culture of rabbit fibroblast-like synoviocytes by suspended explant culture method were successfully established. The method was simple and highly efficient. It provided a new method for the isolation of FLS in vitro.