1.Baseline investigation of medical aid following Wenchuan Eathquake
Xunchui CHEN ; Peng KUN ; Jian LI ; Chunli YI ; Lan YAO
Chinese Journal of Hospital Administration 2010;26(3):194-198
Objective This paper aims to make clear the post-disaster medical needs of residents in the disaster areas and changes among the poor population, for the purpose of policy recommendations on post-disaster medical assistance. Methods Such methods as in-home questionnaire survey and literature study were used to study the income, prevalence rate and mental health of 4 380 households in 10 counties (cities and districts) in the disaster areas. Results In the hardest hit areas, 80. 4% and 52. 8% of residents in rural and urban areas are dependent on government relief respectively;the proportion of poor population resulting from the disaster rose from 5% before the disaster to over 75% afterwards in the hard-hit areas, while such a population rose to 15% in the hardest-hit areas and about 10% in general disaster areas;It is estimated that the new medical aid funds in Sichuan Province will reach 350 million yuan, two-fold that of the budget for 2009;In the hardest hit areas, the two-week prevalence rate per thousand people is twice that found in the third survey of health services in 2003, characterized of acute respiratory illness and rheumatoid;In the hardest-hit areas, nearly 70% of the residents are exposed to high mental health risks. Conclusion Recommendations: Strengthening the raising and use of medical aid fund;dynamic management for the population in need of post-disaster medical aid;Developing appropriate medical aid packages to fit post-disaster health needs and postdisaster disease characteristics;and consolidating the connection between medical aid and primary medical insurance system.
2.Construction of Lmdd-LMP2A, an attenuated Listeria vaccine strain expressing the Epstein-Barr vi-rus latent membrane protein 2A (EBV-LMP2A) and evaluation of its anti-tumor effects against na-sopharyngeal carcinoma
Wei ZHAO ; Xiaoqin CHEN ; Xin WAN ; Ci CHENG ; Zhe LIN ; Dongju FENG ; Kun YAO ; Yun CHEN
Chinese Journal of Microbiology and Immunology 2015;(3):207-212
Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.
3.Relationship between hormone therapy in women with ovarian malignancy and prognosis
Li LI ; Zhongmian PAN ; Xinqiu CHEN ; Kun GAO ; Wei ZHANG ; Yuan LUO ; Zhongqiang YAO ; Xinqiang LIANG ; Wei HUANG ; Desheng YAO
Chinese Journal of Obstetrics and Gynecology 2008;43(11):843-848
Objective To explore the relationship between hormone therapy (HT) in women withovarian malignancy and prognosis. Methods HT was used in 31 patients with ovarian cancer after surgery,and 44 eases with ovarian eaneer served as controL The expression of estrogen receptor (ER)α, ERβ andprogesterone receptor (PR) was detected by immunohistoehemieal staining respectively. The level of serumealeitonin and transforming growth factor α (TGFα) was detected by radio-immune and enzyme-linkedimmunosorbent assay pre- or post-surgery, as well as half a year to one year later post-surgery respectively inthese eases. The survival curve of Kaplan-Meier and log-rank test as well as scale risk of Cox model wereused to analyze the relationship between HT and prognosis of ovarian cancer. Results ( 1 ) The results oflog-rank test showed that there was no difference in survival curve of patients with or without HT [ (1108±52), (1086±43) d; P=0.940] ; the results of scale risk of Cox model also showed that HT was not anindependent prognosis factor for patients with HT. (2) There was no relationship with HT and theaccumulated survival in patients with either positive or negative expression of ERa, ERβ and PR in tissue;as well as between HT and the level of serum TGFα pre-, post-surgery, or half a year to one year aftersurgery. (3) The level of serum caleitonin in patients without HT post-surgery half a year to one year laterwas higher than that pre-surgery [ (141±13), (95±11) μg/; P<0.05], but there was no significantdifference between patients with HT half a year to one year later past-surgery and pre-surgery [ (90±18)μg/L, (93±14) μ/L; P>0.05]. (4) There was a significant difference in body and emotion function between HT and without HT groups [(1.84±1.50), (1.45±0.82); (12.69±10.20), (12.90±11.61); P<0.05], as well as in sex quality and autonomic nerve maladjustment and in the special listmade [(1.05±0.74), (1.77±1.08); (10.10±3.21), (13.09±4.30); P<0.05]. ConclusionsThere is no adverse influence on prognosis in using of HT for patients with ovarian cancer after surgery. HTfor patients with ovarian cancer post-surgery can help keep a stable level of scmm calcitonin as well asimprove the quality of life.
4.The aberrant effective connectivity of the limbic system to happy face in patients with depression
Xiaoxue LIU ; Zhijian YAO ; Lingling HUA ; Jianhuai CHEN ; Rui YAN ; Kun BI ; Qing LU
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(11):993-995
Objective To investigate the effective connectivity of limbic circuit in patients with major depressive disorder when they recognized dynamic positive face expressions,aiming to discuss the possible mechanism of emotion processing in depressed patients.Methods Eighteen depressive patients and eighteen well-matched healthy control volunteers participated in the experiment.All subjects were asked to recognize the emotion face during the magnetoencephalograph (MEG) scanning.The regions of interested (ROI) brain areas included the orbital frontal cortex (OFC),the anterior cingulated cortex (ACC),the amygdala (AMYG),the hippocampus and the insula.The MEG data were preprocessed by the SPM8 software and further analyzed by the Granger casual model (GCM).The non-parameter permutation was used to compare the value of effective connectivity between the healthy controls and the depressed patients.Results Compared with healthy controls,the effective connectivity from the ACC to the AMYG (P=0.0052),from the OFC to the AMYG(P=0.0046),from the Hippocampus to the ACC (P=0.0016),and from the ACC to the Hippocampus (P=0.0042)was significantly reduced in depressed patients.Conclusion The depressed patients display decreased interaction of the limbic circuit during the happy facial emotion processing,indicating that the depressed patients are unable to deal with the positive stimuli,and to certain extent,explaining the abnormal neuropathophysiological mechanism of positive stimuli in MDD.
5.Assessment of ventricular function in children with single ventricle after cavopuimonary connection using velocity vector imaging
Lijun WU ; Yuqi ZHANG ; Kun SUN ; Lijun CHEN ; Yanjun ZHAO ; Liping YAO
Chinese Journal of Ultrasonography 2012;21(1):6-9
ObjectiveTo evaluate the feasibility and accuracy of velocity vector imaging (VVI) in assessing ventricular function in children with single ventricle after cavopulmonary connection.Methods Thirty children with single ventricle after cavopulmonary connection were enrolled in this study,30 agematched normal children were served as control group.The systolic peak velocity,displacement,strain and strain rate in this two groups measured in 6 segments by VVI were compared.dp/dt of single ventricle was estimated by atrioventricular regurgitation using simplified Bernoulli equation.Results Strain and strain rate were significantly lower in all 6 segments in children with single ventricle after cavopulmonary connection compared with values in normal children( P <0.05,respectively),strain rate of the basal segment at the rudimentary chamber correlated best with dp/dt (r =0.72,P <0.01).Conclusions Segmental ventricular dysfunction was observed in children with single ventricle after cavopulmonary connection,and could be assessed accurately using velocity vector imaging.
6.Exploration of setting up comprehensive medical biochemistry experiment
Bin LI ; Yongchun CHEN ; Kun LIU ; Yuan GAO ; Yun SHI ; Min YAO ; Lingling JIANG
Chinese Journal of Medical Education Research 2012;(11):1117-1119
Comprehensive biochemistry experiment,which is interlocked and has difficulties in a certain degree,requires considerable knowledge,multiple techniques and long time.In order to ensure the smooth progress of the experiment,biochemistry and molecular biology department of Hebei medical university has taken following measures in teaching preparation and teaching implementation:building a specialized laboratory;performing collective lesson preparation and pre-experiments;technical teaching;teaching with multimedia equipments;students submitting experimental preparatory reports before class and then completing the experiments in groups.These measures achieved the intended purpose of setting up a comprehensive experiment.
7.Human Herpesvirus 7 Glycoprotein B (gB) , gH, gL, gO Can Mediate Cell Fusion
Jian XU ; Kun YAO ; Jie DOU ; Jian QIN ; Wenrong XU ; Yun CHEN ; Quanzhang YIN ; Feng ZHOU
Progress in Biochemistry and Biophysics 2007;34(11):1202-1209
Human herpesvirus 7 (HHV-7) infection is dependent on the functions of structural glycoproteins at multiple stages of the viral life cycle. These proteins mediate the initial attachment and fusion events that occur between the viral envelope and a host cell membrane, as well as cell to cell spread of the virus. To characterize the HHV-7 glycoproteins that can mediate cell fusion, a cell-based fusion assay was used. 293T cells expressing the HHV-7 glycoproteins of interest along with a luciferase reporter gene under the control of the T7 promoter were cocultivated with SupT1 cells transfected with T7 RNA polymerase. HHV-7 glycoproteins gB, gH, gL and gO can mediate the fusion of 293T cells with SupT1 cells, and the fusion can be inhibited by anti-CD4 mAbs. Thus, the coexpression of HHV-7 gB, gO, gH and gL is sufficient and necessary for HHV-7 induced membrane fusion, and one of these glycoproteins or protein complex formed by these glycoproteins might be the ligand(s) of CD4 molecule.
8.DNA barcoding the medicinal plants of the genus Paris.
Yingjie ZHU ; Shilin CHEN ; Hui YAO ; Rui TAN ; Jingyuan SONG ; Kun LUO ; Jing LU
Acta Pharmaceutica Sinica 2010;45(3):376-82
DNA barcoding is a technique in which species identification and discovery are performed by using short and standard fragments of DNA sequences. In this study, eleven species of Paris, including seven varieties, were sampled. Five chloroplast sequences, psbA-trnH, rpoB, rpoC1, rbcL, matK, and one nuclear marker, the second internal transcribed spacer (ITS2) of ribosomal DNA, were amplified and sequenced. The PCR amplification and sequencing efficiency, intra- and inter-specific divergence and barcoding gap were used to evaluate different loci, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The ITS2 sequences in the studied samples of Paris were amplified and sequenced successfully using primers designed by our group, while matK showed low level in the amplification and psbA-trnH was difficult for sequencing because of over 800 bp and poly (A) structure. Analysis of the intra- and inter-specific divergence and barcoding gap showed ITS2 was superior to other loci. The ITS2 showed a much higher percentage of success (100%) in identification than other five loci, none of which indicated more than 50% except matK (52.9%). The 2-locus combination of rbcL+matK didn't improve ability of authentication. In addition, the rate of successful identification with ITS2 kept 100% when the samples were expanded to 67 samples of 29 species. In conclusion, ITS2 can be used to correctly identify medicinal plants of Paris, and it will be a potential DNA barcode for identifying medicinal plants of other taxa.
9.Construction and identification of recombinant retroviral vector and stable cell line expressing latent membrane protein 2A of Epstein-Barr virus
Yun CHEN ; Feng ZHOU ; Beicheng SUN ; Genyan LIU ; Bing WANG ; Kun YAO
Journal of Cellular and Molecular Immunology 2009;25(11):1013-10,151,019
AIM: To clone EBV-LMP2A gene, construct and identify the recombinant retroviral vector and stable cell strains expressing EBV LMP2A. METHODS: The full-length EBV LMP2A gene was generated by RT-PCR amplification from B95.8 cells which contain complement nucleotide sequence of EBV LMP2A gene. The gene was ligated to T-vector and sequenced to construct retroviral vector consisting with LMP2A. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pGEZ-LMP2A and two auxiliary viral vectors pHIT456 and pHIT60 by lipofectAMINE2000. Viral titration was performed according to the instructions of the manufacturer. To establish L929 cell line stable expressing LMP2A, L929 cells were infected with recombinant retrovirus three times and selected by Zeocine. The Zeocine-resistant clones (L929/LMP2A) were screened for LMP2A expression by RT-PCR and Western blot. RESULTS: The recombinant retrovirus vector carrying LMP2A gene was constructed successfully. Transfection yield a titer of 5×10~8 infectious particles/L. The infected L929 cells were selected by Zeocine. Results of RT-PCR and Western blot indicated that L929 transgenetic cells could stably express EBV-LMP2A. CONCLUSION: The L929 cell line stably expressing LMP2A provides suitability for extraction of the LMP2A protein and preparations of the vaccine for the therapy of EBV-associated diseases.
10.Constructing a p53-fused dual luciferase reporter and verifying its function
Jing TAN ; Xianzhen JIANG ; Leye HE ; Zhiqiang JIANG ; Yuxin TANG ; Kun YAO ; Binghai CHEN ; Juan XUE
Journal of Central South University(Medical Sciences) 2013;38(11):1135-1140
Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.