1.Establishment of human lung squamous carcinoma cell line CHLH-1
Hong-Cheng LIU ; Sheng-Dong HUANG ; De-Jun GONG ; Xiao-Hong LIU ; Yang YUAN ; Zhi-Yun XU ;
Academic Journal of Second Military Medical University 1999;0(12):-
Objective:To establish a human lung squamous carcinoma cell line and to study its biological characteristics. Methods:Lung squamous carcinoma specimens were freshly resected during operation;the tissues were incubated in vitro and the cell line was named CHLH-1.The biological characteristics of the cells were studied by light microscopy,electron microsco- py,chromosome analysis and transplantation experiment.Results:Cells from the specimens of the primary tumor,the CHLC- 1 cell line and the cells from transplanted tumor possessed the characteristics of malignant squamous epithelium under light and electron microscope.The cell growth curve,doubling time and mitotic index were also observed in vitro.Nuclear chromosome analysis revealed that the tumor was a subtriploid with a mode of 60-68 per cell.Tumor nodes were observed under the skin of nude mice by heterogenic transplantation.Conclusion:The characteristics of the established cell line suggest that it is a newly established human squamous carcinoma cell line.
2.Analysis on the rodents density monitoring data in Fangshan District of Beijing, 2016-2017
Hai-ying GONG ; Zhao-hui LIU ; Ying TONG ; Yong ZHANG ; Mei-de LIU ; Xiao-peng ZENG
Shanghai Journal of Preventive Medicine 2020;32(12):992-
Objective To understand species, population density, and seasonality of rodents in Fangshan District of Beijing and provide the information for prevention and control of rodents and rodent-borne diseases. Methods The rodents were captured by using the trap at-night method in Mukezhai, Xianfenggu, Liushilu, Gushanzhai, Wanjingxiangou.The rodents pieces were identified and the density was calculated. Results Totally 4 665 traps were placed and 268 rodents were captured during 2016-2017.Among the seven identified species, the proportion of
3.Isolation, expression analysis of a chilling induced cDNA from rice root with differential display: an evidence role for caffeine-sensitive calcium signal.
Kui-De YIN ; Xing-Mei ZHANG ; Shi-Qiang LIU ; Le-Gong LI
Chinese Journal of Biotechnology 2002;18(4):468-471
Chilling-sensitive rice varieties acquire chilling tolerance when their roots are exposed to water stress for short time. Caffeine-sensitive calcium signal was involved in this procedure. By using total RNA differential display, a chilling induced cDNA(ICT: induction of chilling treatment) was isolated from roots of chilling-sensitive rice variety. It was determined that it is a novel cDNA by homology searching. The transcript level of ict mRNA is up-regulated under chilling stress, it is decreased to low level when the samples were transferred to standard culture conditions. Pre-treated with mannitol for two hours is beneficial to inducing ICT level of expression. This chilling induction was inhibited by caffeine, suggesting that it may play a putative role in signal transduction of caffeine-sensitive calcium.
Cold Temperature
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DNA, Complementary
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genetics
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isolation & purification
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Gene Expression Profiling
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Gene Expression Regulation, Plant
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drug effects
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Mannitol
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pharmacology
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Oryza
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drug effects
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genetics
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Plant Roots
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drug effects
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genetics
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RNA, Messenger
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drug effects
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genetics
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metabolism
4.Detection of the mutation in HBV polymerase gene by RFLP PCR method in hepatitis B patients treated with lamivudine.
Zhuo LI ; Yan-bin GUO ; Wa HAO ; Zun-hui LIN ; Hai-ying JIN ; De-gong LIU
Chinese Journal of Experimental and Clinical Virology 2003;17(3):266-269
BACKGROUNDTo investigate the mutation of HBV polymerase gene in chronic hepatitis B patients treated with lamivudine.
METHODSThe restriction-fragment-length-polymorphism (RFLP) assay for HBV DNA sequence determination at the codon 528 and 552 in the HBV polymerase gene associated with lamivudine resistance in vitro. HBV DNA samples extracted from sera of 240 patients were subjected to PCR amplification with primer pairs F2/R2 (552), F3/R2 (528). Each PCR product was digested with Nde I or Nla III.
RESULTSSerum HBV DNA mutation was found in 51/240 patients (38/51M552V, 26/38L528M, 13/51M552I) after therapy for 52 weeks. DNA sequence analysis was performed on samples of 3 patients, and the results were consistent with those of RFLP assay.
CONCLUSIONThe RFLP assay was able to detect the mutation of HBV DNA at codon 552 and 528 which are the principal site of HBV DNA resistant to lamivudine. The specific PCR method for HBV DNA mutation is rapid, simple and specific.
Drug Resistance, Viral ; Gene Products, pol ; genetics ; Hepatitis B virus ; enzymology ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Reverse Transcriptase Inhibitors ; therapeutic use
5.Gene transfection efficiency and destination of recombinant adenovirus gene transfer through internal jugular vein injection in rats.
Yu-hang AI ; Li-na ZHANG ; Hua GONG ; Liu PENG ; Yang-de ZHANG
Journal of Central South University(Medical Sciences) 2005;30(6):653-656
OBJECTIVE:
To investigate the adenovirus-mediated LacZ gene expression and the destination in different organs of SD rats after the intravenous injection in rats.
METHODS:
Recombinant adenovirus vector containing LacZ was transferred to SD rats by injecting into the internal jugular vein. To identify the sites and periods of LacZ gene expression, X-gal staining was used to detect beta-gal level and period of LacZ gene expression of different organs in the transfected and non-transfected rats at different time intervals.
RESULTS:
On the 1st day after the injection, the lung, liver, kidney, and spleen expressed some beta-gal; on the 3rd day after the injection, the lung, liver, kidney, and spleen expressed beta-gal obviously; their peak levels were on the 7th day; the beta-gal level decreased on the 14th day; beta-gal expression disappeared in the most organs except the lungs on the 28th day. In all animals, the brain did not express any beta-gal.
CONCLUSION
The adenovirus-mediated exogenous gene transfer in the internal jugular vein may be an effective approach of gene therapy in some diseases in the lung, liver, and kidney.
Adenoviridae
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genetics
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Animals
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Female
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Gene Transfer Techniques
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Genetic Therapy
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Injections, Intravenous
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Jugular Veins
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Lac Operon
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genetics
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Lung
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metabolism
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Male
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Recombinant Proteins
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genetics
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Transfection
6.Estrogen receptor gene polymorphisms and HBV-induced liver cirrhosis.
Jian-ping XIE ; Xian-qiong GONG ; De-ming TAN ; Fei LIU ; Jian-liang ZHOU
Journal of Central South University(Medical Sciences) 2006;31(3):379-382
OBJECTIVE:
To investigate the relationship between estrogen receptor (ER) gene polymorphisms and HBV-induced cirrhosis.
METHODS:
Xba I and Pvu II polymorphisms of ER gene were analyzed in 98 patients with HBV-induced cirrhosis, 72 patients with chronic hepatitis B, and 84 healthy controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique.
RESULTS:
The frequencies of Pp genotype and P allele of ER gene in patients with HBV-induced cirrhosis were higher than those in patients with chronic hepatitis B and controls, while the frequencies of pp genotype and p allele of ER gene in patients with HBV-induced cirrhosis were lower than those in patients with chronic hepatitis B and controls (P < 0.05). The risk of HBV-induced cirrhosis occurrence in carriers with Pp and pp genotype was 2.23 folds than that in carriers with pp genotype. There was no significant difference in frequencies of allele and genotype in Xba I polymorphism among the HBV-induced cirrhosis group, the chronic hepatitis B group, and the control group (P > 0.05).
CONCLUSION
Pp genotype and P allele might be the susceptibility gene for HBV-induced cirrhosis while pp genotype and p allele might be the protective gene.
Adolescent
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Adult
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Aged
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Alleles
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Female
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Genotype
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Hepatitis B, Chronic
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complications
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genetics
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Humans
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Liver Cirrhosis
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etiology
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genetics
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Male
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Middle Aged
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Polymerase Chain Reaction
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Polymorphism, Genetic
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Polymorphism, Restriction Fragment Length
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Receptors, Estrogen
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genetics
7.Clinical outcomes of patients with hypertensive intracerebral hemorrhage in the thalamus received stereotactic combined with microsurgical treatment
Lü-Qian HUANG ; Ming GONG ; Zhong-An WANG ; De-Yong ZHANG ; Li-Wen YI ; Jun-Hui LIU
Chinese Journal of Neuromedicine 2012;11(8):815-818
Objective To explore the clinical outcomes of patients with hypertensive intracerebral hemorrhage in the thalamus received stereotactic combined with microsurgical treatment,and find the best treated method. Methods Forty-three patients with hypertensive intracerebral hemorrhage in the thalamus,admitted to our hospital from October 2009 to March 2011,were selected in our study; these patients were divided into stereotactic combined with microsurgical treatment group (observation group,n=22) and direct surgery group (control group,n=21).Preoperative hematoma,and hematoma at 1 week and 1 month after the treatments under CT were compared; clinical curative effect at 1 month after the treatments and activities of daily living (ADL) 3 months after the treatments were compared. Results The hematoma in the observation group 1 week and 1 month after the treatments was statistically different as compared with that in the control group (P<0.05).Clinical curative effect in the observation group (average rank=18.022) was better than that in the control group (26.166) with significant differences (P<0.05); the total effective rate in the observation group was 77.3% and that in the control group was 52.4% with significant differences (P<0.05).ADL scale indicated that the ability of the observation group (average rank=18.363) were better than that in the control group (25.809) with significant differences (P<0.05).The postoperative complication rate in the observation group (22.7%) was lower than that in the control group (42.8%) with significant differences (P<0.05). Conclusion Stereotactic combined with microsurgical treatment in patients with hypertensive intracerebral hemorrhage in the thalamus is safe and effective; this treatment can improve the postoperative living quality and enjoy few complications,which is worth in the clinical application.
8.Study on the relationship between Terra-MODIS image and the snail distribution in marshland of Jiangning county, Jiangsu province.
Bo ZHANG ; Zhi-ying ZHANG ; De-zhong XU ; Zhi-dong SUN ; Xiao-nong ZHOU ; Zi-li GONG ; Shi-jun LIU ; Cheng LIU ; Bin XU ; Yun ZHOU
Chinese Journal of Epidemiology 2003;24(4):257-260
OBJECTIVETo analyze the relationship between the normalized difference vegetation index (NDVI) and the snail distribution in marshland of Jiangning county in Jiangsu province, and to explore the utility of Terra-MODIS image map in the small scale snail habitats surveillance.
METHODSNDVI were extracted from MODIS image by vector chart of the snail distribution using ArcView 8.1 and ERDAS 8.5 software. The relationship between NDVI and the snail distribution were Investigated using Bivariate correlations and stepwise linear regression.
RESULTSThe snail density on marshland was positively correlated with the mean NDVI in the first ten-day of May and the maximum NDVI (N(20max)) in the last ten-day of May. Incidence of pixel with the live snail on marshland was positively correlated with the mean NDVI (N(2mean)) in the first ten-day of May. An equation Y(1) = 0.009 47 x N(20max) (R(2) = 0.73), Y(2) = 0.018 6 x N(2mean) (R(2) = 0.906) was established.
CONCLUSIONThis study showed that the Terra-MODIS satellite images reflecting the status of the vegetation on marshland in Jiangning county could be applied to the study to supervise the snail habitat. The results suggested that MODIS images could be used to survey the small scale snail habitats on marshland.
Animals ; Breeding ; China ; epidemiology ; Ecology ; Environmental Monitoring ; Epidemiological Monitoring ; Humans ; Satellite Communications ; Schistosomiasis japonica ; epidemiology ; transmission ; Seasons ; Snails ; growth & development ; parasitology ; physiology
9.Relationship between V617F mutation and 46/1 haplotype in JAK2 gene in patients with chronic myeloproliferative diseases and frequencies of 46/1 haplotype in different Chinese nationalities.
Zheng-Qin TIAN ; Ping ZHU ; Hong-Xing LIU ; Yan CHEN ; Fang WANG ; Yang ZHANG ; Wen TENG ; Yi-Wen GONG ; Jun-Yan XIA ; De-Cheng BAI ; Qian LIU ; Xue-Qiang WU
Journal of Experimental Hematology 2012;20(2):362-367
Somatic gene V617F mutation in JAK2 is a critical molecular and biological indicator to diagnosis of chronic myeloproliferative disease (MPD). This study was aimed to investigate the genetic background of V617F mutation in 46/1 gene haplotype in Chinese MPD patients, and the frequencies of 46/1 gene haplotype and V617F mutation in three nationalities of Chinese populations. Peripheral blood or bone marrow samples of 150 V617F mutation positive MPD patients, 123 V617F mutation negative MPD patients, 124 healthy Han individuals, 395 healthy Tibetan individuals and 315 healthy Yugu individuals were collected. The allele-specific multiplex PCR method was established, the presence or absence of V617F mutation, the presence or absence of 46/1 haplotype, and the relationship between V617F and 46/1 haplotype were easily identified by agarose gel image. The results showed that the V617F mutation located in the 46/1 haplotype of 88 cases (58.67) among 150 V617F-positive MPD cases. In 814 Chinese healthy individuals including Han, Tibetan, Yugu nationalities, the frequency of the 46/1 gene haplotype was 38.37 without difference in the frequency among different nationalities, and no V617F mutation was found in Chinese healthy populations, The frequency of the 46/1 gene haplotype was 43.09 in V617F mutation negative MPD patients and was 69.33 in V617F mutation positive MPD patients, the latter was obviously higher than former and than that in healthy Han individuals. In conclusion, a multiplex PCR method has been developed that is simple and useful to identify V617F mutation in JAK2 gene and its relationship to the 46/1 haplotype. In more than half of Chinese V617F-positive MPD patients, the V617F mutation locates in 46/1 haplotype in JAK2. The frequencies of 46/1 haplotype are statistically insignificant among Han, Tibetan and Yugu nationality populations.
Asian Continental Ancestry Group
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genetics
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Ethnic Groups
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genetics
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Female
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Haplotypes
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Humans
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Janus Kinase 2
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genetics
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Male
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Mutation
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Myeloproliferative Disorders
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genetics
10.The changes of extracellular matrix in adult degenerative nucleus pulposus cells with stiring microcarrier culture system in vitro.
Bin NING ; Hai-fei LIU ; Wei-ming GONG ; Kai ZHAO ; Hong-xia DU ; Yong LIU ; De-chun WANG ; You-gu HU
Chinese Journal of Surgery 2013;51(5):432-436
OBJECTIVETo evaluate the biological effect on the synthesis of the extracellular matrix (ECM) in the cultivation of adult degenerative nucleus pulposus cells using the stiring microcarrier system in vitro.
METHODSThirty-four specimens were collected after intervertebral fusion operations of the patients with intervertebral disc herniation diseases from September 2005 to May 2009. The specimens were then randomly allocated into 2 groups for in vitro cultivation: monolayer culture group and microcarrier culture group. On the exponential phase, SP-ABC immunohistochemical staining and Western blot quantitative analysis were conducted in the two groups to detect the collagen type I and II. Proteoglycan contents of two groups in different growth phases were detected with (35)S-sulfate incorporation assay.
RESULTThe expressions of collagen type I and II in microcarrier culture group were significantly higher than those in monolayer culture group: SP-ABC immunohistochemical staining (collagen type I: 32.5 ± 4.4 vs. 15.2 ± 1.2, t = 2.871, P < 0.01; collagen type II: 43.6 ± 4.1 vs. 23.1 ± 2.2, t = 2.375, P < 0.05); Western blot quantitative analysis (collagen type I: 0.62 ± 0.08 vs. 0.50 ± 0.06, t = 3.327, P < 0.01; collagen type II: 1.46 ± 0.08 vs. 0.86 ± 0.04, t = 2.453, P < 0.05). Nucleus pulposus cells cultivated in stiring microcarrier system showed significantly increased proteoglycan synthesis than monolayer culture group does on both exponential phase and stationary phase (exponential phase: 34 821 ± 312 vs. 21 046 ± 673, t = 2.134, P < 0.05; stationary phase: 45 134 ± 175 vs. 32 193 ± 713, t = 2.801, P < 0.01).
CONCLUSIONSThe expression of collagen type I, II and proteoglycan of adult degenerative nucleus pulposus cells are positive regulated by the stiring microcarrier system, which can be used in the mass amplification of the adult degenerative nucleus pulposus cells.
Adult ; Aged ; Cell Culture Techniques ; Collagen ; metabolism ; Extracellular Matrix ; metabolism ; Female ; Humans ; Intervertebral Disc ; cytology ; Male ; Middle Aged ; Proteoglycans ; metabolism ; Random Allocation ; Young Adult