1.Cloning,Sequence Analysis and Expression in E.coli of the EP0 Gene of Pseudorabies Virus Ea Strain
Liu-Rong, FANG ; Huan-chun, CHEN ; Shao-bo, XIAO ; Xiang-Ru, MA ; Ge-fei, WANG
Virologica Sinica 2001;16(2):183-187
The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea,and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.
2.Research advances in atopic dermatitis.
Chun-Ping SHEN ; Huan XING ; Lin MA
Acta Academiae Medicinae Sinicae 2009;31(1):103-106
Atopic dermatitis (AD) is a chronic, recurrent, and inflammatory skin disease. It usually occurs in children with a clinical manifestation of severe itching. In recent years the incidence of AD is slowly increasing, which severely impacts the physical and mental health of children. This article summarizes the epidemiology, influencing factors, and management of this diseases.
Dermatitis, Atopic
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epidemiology
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etiology
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therapy
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Humans
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Prognosis
4.Protective effect of water soluble CoQ10 on rotenone-induced apoptosis in PC12 cells
Yu-Min JIANG ; Hai-Ning LI ; Shao-Qing LIN ; Yan-Yan CHEN ; Jing AN ; Chun-Huan MA ; Nan-Nan HUAN ; Jiang CHENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2018;39(4):514-518
Objective To investigate the protective effect and the underlying mechanism of water soluble coenzyme Q10 (CoQ10)against rotenone induced injury on PC12 cells model.Methods PC12 cells were cultured with rotenone,water-soluble CoQ1 0 was added to the culture media 3 hours prior to the rotenone incubation.We determined cell viability by CCK8;reactive oxygen species (ROS)was detected by spectrophotometer;and Bcl-2, Bax,active Caspase-3,Caspase-9 and apoptosis-inducing factor (AIF)were measured by Western blotting after 24-hour rotenone incubation.Results After the treatment by rotenone,cell viability decreased significantly (P<0.01)and ROS level increased (P<0.01).CoQ10 could improve PC12 cell viability (P<0.01)and reduce the level of ROS (P<0.01).Western blotting experiments showed that CoQ10 could reduce rotenone-induced Caspase-9 (P<0.05),active Caspase-3 (P<0.05)and Bax (P<0.01)expressions,increase the expression of Bcl-2 (P<0.01),and prevent nuclear translocation of AIF (P<0.05).Conclusion CoQ10 has a protective effect on rotenone-induced apoptosis in PC12 cells,the mechanism of which may be through scavenging ROS in cells;decreasing caspase-9 ,active caspase-3 and Bax expressions;and increasing the expression of Bcl-2 ;and preventing AIF nuclear translocation.
5.Deletion of spiramycin 3-O-acyltransferase gene from Streptomyces spiramyceticus F21 resulting in the production of spiramycin I as major component.
Lin-Zhuan WU ; Chun-Yan MA ; Yi-Guang WANG ; Jian-Lu DAI ; Jing-Yan LI ; Huan-Zhang XIA
Chinese Journal of Biotechnology 2007;23(4):612-617
Spiramycin (SP) belongs to the 16-member macrolide antibiotics. It contains three components,namely SP I, SP II and SP III, which differ structurally in the acylation moieties on the C3 of the lactone. The SP I component contains a hydroxyl group at C3. SP II, and SP III are formed by further acetylation or propionylation of the C3 of SP I, by the same 3-O-acyltransferase (3-O-AT) . The study focused on simplifying spiramycin components. Theoretically, disruption/deletion of the 3-O-AT gene will reduce/stop the acylation of SP I to SP II and SP III. In this study, degenerated primers were designed according to the conserved regions of 3-O-acyltransferase, MdmB and AcyA in the medicamycin and carbomycin producers of S. mycarofaciens and S. thermotolerans, respectively, and an 878bp DNA fragment was amplified from the spiramycin-producer of S. spiramyceticus F21. Blast analysis of the 878bp DNA fragment suggested that it encoded the 3-O-acyltransferase (3-0-AT, sspA) gene for spiramycin biosynthesis. The flanking regions of this 878bp DNA fragment were then amplified by single-oligonucleotide-nested PCR, and a total of 4.3 kb DNA was obtained (3457nt among the 4.3kb fragment was sequenced, and deposited in GenBank DQ642742),covering the whole putative 3-O-acyltransferase gene, sspA. The sspA was then deleted from the S. spiramyceticus F21 genome by double cross-over homologous recombination, mediated by temperature-sensitive plasmid pKC1139. A comparison was done of the components of spiramycins produced by the sspA-deleted mutant strain with that of the parent strain by HPLC analysis, which showed that sspA-deleted mutant produced SP I (72%), SP II (18%), and SP III (9.6%), whereas parent strain produced SP I (7.8%), SP II (67%), and SP III (25%), respectively, demonstrating the role of ssp A in the acylation of SP I into SP II and SP III. The ssp A-deleted mutant strain obtained in this study may be used for the production of SP I, or may serve as a good starter for the construction of spiramycin derivatives.
Acyltransferases
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genetics
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Aminoglycosides
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biosynthesis
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Gene Deletion
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Genes, Bacterial
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genetics
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Genetic Engineering
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methods
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Streptomyces
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enzymology
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genetics
6.Factors associated with attitudes toward tobacco control policy in public places among adults in three counties of China
Cui-Zhu MEI ; Shao-Jun MA ; Xue-Fang XU ; Jun-Fang WANG ; Chun-Ping WANG ; Ai-Ping CHEN ; Gong-Huan YANG
Chinese Journal of Epidemiology 2009;30(6):549-553
Objective To investigate the attitudes of urban and rural community members toward total banning on smoking in public places and to explore the factors associated with these attitudes, in three counties/cities in China. Methods A cross-sectional study was conducted in three counties/cities in 2004, including Xin' an county of Henan province, Anyi county of Jiangxi province, and Mianzhu city of Sichuan province. A total of 5642 residents at age of 18-69 years old were interviewed face-to-face with a uniform questionnaire by locally-trained interviewers, through a random three-stage stratified sampling in each county. Factors were assumed to be associated with attitudes, using chi-square test in univariate analysis and non-conditional logistic regression model in multivariate analysis. Results The prevalence of current smoking among respondents was 44.3%. 80.7% (1379/1709) of the current smokers reported smoking often or sometimes in public places. Only 9.6% (479/4983) of the respondents reported that their indoor workplaces had totally banned on smoking. 43.5% of the respondents supported a total smoking ban strategy in pubic venues. The results of multivariate logistic regression model showed that eight factors were significantly associated with support for the total smoking ban in public places included region, residency, age, gender, education, smoking status, awareness about passive smoking hazards, and hearing of any message on tobacco control through media differences of rates regarding the factors as: residents in urban to rural areas (OR=1.29), elderly to youngsters (30-49 vs. 18-29, OR=1.46; 50-69 vs. 18-29, OR=1.71), female to male(OR=1.27), high-educated to less-educated ones, quitters to current smokers (OR=1.90), nonsmokers to current smokers (OR=2.01). Those who know messages on health hazards of passive smoking (OR=2.26), or heard of message on tobacco control through media (OR= 1.43). Conclusion Results from our study revealed that a thorough smoke-flee policy in public places should be developed and implemented in these three counties/cities.
7.Music intervention on mechanically ventilated patients:a randomized control trial
Lin HAN ; Ji-Ping LI ; Yue-Huan ZHONG ; Wei-Guang MA ; Xiao-Chun YANG
Chinese Journal of Modern Nursing 2010;16(14):1613-1616
Objective To examine the effects of music intervention on the physiological variables and the anxiety level of mechanically ventilated patients. Methods A total of 150 patients with mechanical ventilation were randomly divided into the music group, the headphone group and the control group. The patients were investigated by using the C-STAI scale before and after the study period. The indicators including heart rate, respiratory rate,saturation of oxygen and blood pressure were recorded before and after the study period. Results Significant differences were found in state anxiety before and after the test in music group and headphone group (P < 0. 01 ).The difference ( post-pre ) of state anxiety within three groups revealed significant differences ( P < 0. 01 ).Significant differences were found on HR, RR, SBP and DBP before and after the test in music group (P <0. 01 ).Conclusions Music and headphone can reduce anxiety in mechanically ventilated patients and the effects of the former are better. Music can steady the heart rate, saturation of oxygen and blood pressure for patients with mechanical ventilation.
8.Clinical study on the indications of 33 acupoints.
Yuan-Yuan ZHENG ; Zeng-Bin MA ; Liang-Xiao MA ; Nan-Nan GUO ; Chun LI ; Yan-Ping WANG ; Kai CHENG ; Huan YANG ; Wan-Ning LIU ; Lü-Wei JIN ; Jiang ZHU
Chinese Acupuncture & Moxibustion 2008;28(6):417-422
Multi-central randomized controlled method was used to scientifically verify indications of 33 acupoints and provide definite clinical basis for the indications of single acupoint. Of the 52 studies, 40 studies showed that the therapeutic effect in acupuncture observation groups were better than the control group; 11 studies showed similar therapeutic effect of the two groups, and 1 study showed the acupuncture observation group was worse than the control group. Therefore, results indicate that in a certain observation cycle, acupuncture at single acupoint have different effects on diseases.
Acupuncture Points
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Humans
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Randomized Controlled Trials as Topic
9.The current status of passive smoking in Chinese families and associated factors.
Chun-Ping WANG ; Xue-Fang XU ; Shao-Jun MA ; Cui-Zhu MEI ; Jun-Fang WANG ; Ai-Ping CHEN ; Gong-Huan YANG
Chinese Journal of Preventive Medicine 2008;42(3):186-191
OBJECTIVETo understand the prevalence of passive smoking in Chinese families and discuss its associated factors, as to providing scientific evidence for establishing tobacco control measures in China.
METHODCross-sectional survey: from June to September, 2004, we randomly selected six counties in three different provinces ( Mianzhu and Xichong of Sichuan Province; Anyi and Hukou of Jiangxi Province; Xinan and Yanshi of Henan Province) and performed face-to-face questionnaire survey on citizens between 18 and 69 years old. All the data were double independently input by professional data entry company to ensure data accuracy. The prevalence of home passive smoking exposure in families with different demographic characteristics was described by using prevalence, and the possible correlated factors of home passive smoking exposure as independent variables, multiple factors were analyzed using Logistic Stepwise Regression Analysis method.
RESULTSThe analysis on 8142 nonsmokers revealed that the rate of passive smoking was 28.42%, with 27.38% of male and 28.93% of female suffering from passive smoking. All 87.19% of the smokers would smoke in front of their families. As many as 42.14% of the nonsmokers would offer cigarettes to their guests, while about 46.82% of the nonsmokers would suggest smokers to smoke outdoor. Home restriction on tobacco was extremely rare and only 6.33% of all the families completely forbade smoking. Multivariate logistic regression analysis of non-conditions revealed that, there was a lower level of involuntary tobacco smoke exposure in female, older age group, lower education level, divorced, or widowed families. There was no difference in involuntary tobacco smoke exposure between town dwellers and county dwellers, but such difference did exist in different districts.
CONCLUSIONThe three provinces under investigation should have severe involuntary tobacco smoking exposure. Gender, age, literacy level, occupation and region should be all factors that influence the status of involuntary tobacco smoking exposure in different families. There is a high percentile that smokers would smoke in front of their families and kids and a relative low pressure against smoking from nonsmokers. Cigarette offering is very prevalence. The knowledge and attitude about passive smoking should be separated from the situation of passive smoking exposure.
Adolescent ; Adult ; Aged ; China ; Factor Analysis, Statistical ; Family ; Female ; Humans ; Male ; Middle Aged ; Sampling Studies ; Surveys and Questionnaires ; Tobacco Smoke Pollution ; statistics & numerical data ; Young Adult
10.The clinical and experimental analyses of a multiple myeloma patient with derivative der(Y;1).
Tong WANG ; Jing-ying QIU ; Xiao-lan MA ; Xiao-peng JIA ; Yan-ping WANG ; Hui-jun YU ; Huan LI ; Chun-rong TONG
Chinese Journal of Medical Genetics 2010;27(2):214-216
OBJECTIVETo investigate the clinical significance of a rare chromosome abnormality der(Y)t(Y;1) in a patient with multiple myeloma (MM).
METHODSThe chromosome spread was prepared after 24 h culture of bone marrow. G-banding technique was used to analyze the karyotype. Fluorescence in situ hybridization (FISH) was performed to ascertain the origin of abnormal chromosome detected by conventional karyotypic analysis. Flow cytometry was used to detect the expression of the CD38/CD138/ZAP70. Immunoelectrophore was applied to identify the type of immunoglobulin.
RESULTSA complex pattern of chromosome rearrangement was observed: 92,XXYY[3]/49,X,der(Y)t(Y;1)(q12;q21),t(11;14)(q13;q32),+18,+20,+21[47]/49,X,idem,del(13q22),ace[1]/98,XX,der(Y)t(Y;1) x 2,+18,+18,+20,+20,+21,+21[10]/46,XY[19]. The result was confirmed by metaphase-FISH. The type of immunoglobulin was IgD with the level of 6.24g/L. The CD38/CD138 was positive but ZAP70 was negative.
CONCLUSIONStructural abnormality of chromosome Y is rare in blood malignancy. Most of them were described in myelodysplastic syndrome or myeloproliferative disorders. It is the first report of der(Y)t(Y;1) abnormality in multiple myeloma.
Chromosome Aberrations ; Chromosomes, Human, Y ; genetics ; Haplotypes ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; therapy ; Treatment Outcome