1.Carotid ultrasound screening of ischemic cerebral apoplexy in high-risk group and exploration of behavioral influence factors
Ling QIN ; Fang TANG ; Chengqi ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(5):433-435
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.
3.Construction and Co-expression of Grass Carp Reovirus VP6 Protein and Enhanced Green Fluorescence Protein in the Insect Cells
Qin, FANG ; Eng Khuan SENG ; Wen, DAI ; Lan-lan, ZHANG
Virologica Sinica 2007;22(5):397-404
Grass carp reovirus (GCRV), a disaster agent to aquatic animals, belongs to Genus Aquareovirus of family Reoviridea. Sequence analysis revealed GCRV genome segment 8 (s8) was 1296 bp nucleotides in length encoding an inner capsid protein VP6 of about 43kDa. To obtain in vitro non-fusion expression of a GCRV VP6 protein containing a molecular of fluorescence reporter, the recombinant baculovirus, which contained the GCRVs8 and eGFP (enhanced green fluorescence protein)genes, was constructed by using the Bac-to-Bac insect expression system. In this study, the whole GCRVs8 and eGFP genes, amplified by PCR, were constructed into a pFastBacDual vector under polyhedron (PH) and p10 promoters, respectively. The constructed dual recombinant plasmid (pFbDGCRVs8/eGFP) was transformed into DH10Bac cells to obtain recombinant Bacmid (AcGCRVs8/eGFP) by transposition. Finally, the recombinant bacluovirus (vAcGCRVs8/eGFP) was obtained from transfected Sf9 insect cells. The green fluorescence that was expressed by transfected Sf9 cells was initially observed 3 days post transfection, and gradually enhanced and extended around 5days culture in P1(Passage1) stock. The stable high level expression of recombinant protein was observed in P2 and subsequent passage budding virus (BV) stock. Additionally, PCR amplification from P1 and amplified P2 BV stock further confirmed the validity of the dual-recombinant baculovirus. Our results provide a foundation for expression and assembly of the GCRV structural protein in vitro.
4.Functions of long noncoding RNAs and their roles in ocular diseases
Fang-Kun, ZHAO ; Yu, QIN ; Jing, LI ; Jin-Song, ZHANG
International Eye Science 2016;16(8):1469-1473
?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.
5.Association of non-alcoholic fatty liver with plasma homocysteine and the methylenetetrahydrofolate reductase gene polymorphism in patients of type 2 diabetes mellitus in Shanxi, China
Ling HU ; Qiaohui ZHANG ; Fang MIAO ; Jie QIN ; Fie LIU
Chinese Journal of General Practitioners 2009;08(6):385-388
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.
6.The anti-tumor effect of Cytochrome oxidase P450 3A4 combined with Cyclophosphamide in vitro
Wei ZHANG ; Jishi WANG ; Yuan YANG ; Qin FANG
Fudan University Journal of Medical Sciences 2009;36(4):384-388
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.
7.Diagnosis and treatment of blunt diaphragmatic rupture
Wenfeng ZHANG ; Qin FANG ; Yu LI ; Dongyi CHEN ; Shun XU
Chinese Journal of General Practitioners 2008;7(1):60-61
A retrospective analysis was carried out in 36 patients with blunt diaphragmatic rupture during March 1991 and March 2006. Twenty-two diagnoses were confirmed preoperatively, and the rest 14were confirmed perioperatively. Three patients underwent surgical treatment after no response to conservative therapy. Thoracotomy was performed in 21 patients, laparotomy in 9, thoracotomy plus laparotomy in 5 and combined thoraco-laparotomy in 1. Most diaphragmatic rupture may be caused by blunt collision and could lead to thoracoabdominal injury. In spite of high mortality rate, the condition is usually under diagnosed. Definite diagnosis and timely operation are important to increase survival rate and reduce mortality.
8.Expression and Identification of Inclusion Forming-related Domain of NS80 Nonstructural Protein of Grass Carp Reovirus
Chao FAN ; Lanlan ZHANG ; Chengfeng LEI ; Qin FANG
Virologica Sinica 2009;24(3):194-201
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.
9.High Level Expression of Grass Carp Reovirus VP7 Protein in Prokaryotic Cells
Lanlan ZHANG ; Jinyu SHEN ; Chengfeng LEI ; Xiaoming LI ; Qin FANG
Virologica Sinica 2008;23(1):51-56
Sequences analysis revealed Grass carp reovirus (GCRV) s10 was 909 nucleotides coding a 34 kDa protein denoted as VP7, which was determined to be a viral outer capsid protein (OCP). To obtain expressed OCP in vitro, a full length VP7 gene was produced by RT-PCR amplification, and the amplified fragment was cloned into T7 promoted prokaryotic expression vector pRSET. The recombinant plasmid,which was named as pR/GCRV-VP7,was then transformed into E.coli BL21 host cells. The data indicated that the expressed recombinant was in frame with the N-terminal fusion peptide. The over-expressed fusion protein was produced by inducing with IPTG, and its molecular weight was about 37kDa, which was consistent with its predicted size. In addition, the fusion protein was produced in the form of the inclusion body with their yield remaining steady at more than 60% of total bacterial protein. Moreover,the expressed protein was able to bind immunologically to anti-his-tag monoclonal antibody (mouse) and anti-GCRV serum (rabbit). This work provides a research basis for further structure and function studies of GCRV during entry into cells.
10.Effect of Heat Reinforcing Acupuncture Manipulation on Articular Synovium LDH, SDH and CCO in Rheumatoid Arthritis ;Rabbits
Xiaozheng DU ; Jinhai WANG ; Xiaoguang QIN ; Xiaoli FANG ; Xiaojiang ZHANG
Shanghai Journal of Acupuncture and Moxibustion 2016;35(10):1256-1261
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.