1.Detection and analysis of gene polymorphism in hepatitis B virus C region.
Yong-zheng JING ; Sheng-bo PU ; Guo-cai WU ; Jian YANG ; Xue-ying YU ; Zhen-hua GUO ; Guo-ying ZHANG ; Zhong-hua GAO
Chinese Journal of Experimental and Clinical Virology 2011;25(4):241-244
OBJECTIVETo explore the variations of gene C in hepatitis B viruses between hepatitis B patients and healthy carriers, and provide experimental evidences for analysis of virus gene mutations acting on the virus material science and response of the body to the virus.
METHODSThe virus DNA load in hepatitis B patients and healthy blood donors was investigated by real-time polymerase chain reaction (PCR). Gene sequence analysis was taken to detect gene polymorphism, and all the success samples were compaired with standard strain by DNAstar.
RESULTS(1)G Compared with standard strain, C region in all samples had mutations, there were 31 mutations in at least 2 samples (3 mutations in gene PreC and 28 mutations in gene C), including 9 missense mutations, 1 chain termination mutation and 21 synonymous mutation. Mutations nt 1827 c-->a and nt 2221 c-->t existed in all the samples, and most samples had 6 synonymous mutations. Four hepatitis B patients had mutation nt1896 g-->a, and another 4 patients had 2 mutations, namely, S87G and I97F (or 197L) in HBcAg CTL recognition episome. (2) The success ratio of amplification and sequencing of HBV DNA was closely associated with its copy numbers. In the present study, copy numbers of HBV DNA which were successfully amplified and sequenced were almost more than 40 193/ml.
CONCLUSIONSHBV genome were easily affected by nucleotide mutations, 2 residues had mutations in gene of C region, which is firstly reported, suggesting these mutations may be geographical restricted. Mutations in gene of C region may either change the structure and function of HBeAg and HBcAg, which may further induce the escape of immune clearance for HBV or influence the detection of HBsAg or HBeAg, which may creat new problems for the prevention, diagnosis and treatment of hepatitis B.
Female ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Mutation ; Polymorphism, Genetic
2.A Chinese girl with cystic fibrosis: a case report identified by sweat and genetic tests.
Yan CHENG ; Gang NING ; Bin SONG ; Ying-kun GUO ; Xue-sheng LI
Chinese Medical Journal 2012;125(4):719-719
Child
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Cystic Fibrosis
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diagnosis
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diagnostic imaging
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genetics
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Female
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Genetic Testing
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Humans
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Radiography
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Sweating
3.ITS sequences variation and phylogenetic analysis on 31 geographical populations of Notopterygium incisum.
Lu-cun YANG ; He-chun LIU ; Xue-li ZHOU ; Wen-hua XU ; Guo-ying ZHOU
China Journal of Chinese Materia Medica 2015;40(19):3748-3753
In this study, 31 Notopterygium incisum populations were analyzed using ITS sequences to investigate the genetic structure. The results showed that: the ITS region ranged in size from 634 to 635 bp and base composition was with high G + C content of 57.8%. Thirty-one polymorphic sites were detected from 402 sequences of 31 populations of N. incisum, and the proportion of polymorphic sites was 4.88%, in which parsimony informative sites were up to 12. And 31 haplotypes were identified based on these polymorphic sites. Molecular variance analysis (AMOVA) indicated that high genetic differentiation (57%) existed among population, and gene flow was low (N(m) = 0.38) among populations. Phylogenetic relationships of 31 haplotypes were analyzed using NJ method with N. forbesiias an out-group. Phylogenetic analysis showed that 31 haplotypes from different populations mixed together and did not form distinct geographically separated clades.
Apiaceae
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classification
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genetics
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Base Sequence
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China
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DNA, Intergenic
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genetics
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Gene Flow
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Genetic Variation
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Molecular Sequence Data
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Phylogeny
4.Separation, purification and primary reverse cholesterol transport study of Cordyceps militaris polysaccharide.
Shou-Dong GUO ; Ying-Jie CUI ; Ren-Zhong WANG ; Ren-Yuan WANG ; Wen-Xue WU ; Teng MA
China Journal of Chinese Materia Medica 2014;39(17):3316-3320
The authors designed to separate, purify and determine the monosaccharide composition of the polysaccharide from Cordyceps militaris, and study its effect on reverse cholesterol transport in vivo by isotope tracing assay. Polysaccharides were separate and purify by ion exchange column Q-sepharose Fast Flow and size exclusion column Sephacryl S200HR; the molecular weight and monosaccharide composition of the polysaccharides were determined by high performance gel permeation chromatography and high performance liquid chromatography coming with pre-column derivation, respectively. Finally, three purified polysaccharides CMBW1, CMBW2 and CMYW1 were obtained, their total carbohydrate contents were 87%, 89%, 95%, respectively; their protein contents were 6.5%, 1.3%, 2.8%, respectively; their molecular weights were 772.1, 20.9, 13.2 kDa, respectively; CMBW1 was composed of mannose, glucosamine, rhamnose, glucuronic acid, glucose, galactose and arabinose with a molar ratio of 7.25: 0.17: 1.29: 0.23: 6.30: 11.08: 0.79; CMBW2 was composed of mannose, glucosamine, galactose and arabinose with a molar ratio of 2.40: 0.16: 2.92: 0.24; CMYW1 was composed of mannose, glucosamine, glucuronic acid and glucose with a molar ratio of 0.59: 0.57: 0.45: 25.61. Polysaccharide at 50 mg x kg(-1) could significantly improve the transport of 3H- cholesterol to blood and excretion from feces. All of the three purified polysaccharides CMBW1, CMBW2 and CMYW1 were heteropolysaccharide; and they could improve reverse cholesterol transport in vivo, the underlying mechanisms are being studied.
Animals
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Biological Transport
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drug effects
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Cholesterol
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metabolism
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Chromatography, High Pressure Liquid
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instrumentation
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methods
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Cordyceps
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chemistry
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Mice
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Monosaccharides
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analysis
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isolation & purification
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Polysaccharides
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chemistry
;
isolation & purification
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pharmacology
;
Tritium
6.A case report of myelodysplastic/myeloproliferative disease unclassifiable with karyotype aberration of trisomy 8 and JAK2 mutation.
Kun LIU ; Chang-Xin YING ; Xue-Dong CHEN ; Xue-Yun ZHOU ; Kun-Yuan GUO
Journal of Experimental Hematology 2012;20(5):1139-1143
This study aimed to investigate the relationship between clinical features of myelodysplastic/myeloproliferative disease, unclassifiable (MDS/MPD-U), karyotype of chromosome and JAK2 mutation in 1 case. The clinical features, karyotype and JAK2 mutation of the patient with MDS/MPD-U were studied by means of bone marrow biopsy, karyotype analysis and ARMS-PCR technique. The results indicated that the typical micromegakaryocytes and thrombocytosis, karyotype aberration of trisomy 8 as well as JAK2 V617F mutation were found in this patient. It is concluded that the patient was diagnosed as MDS/MPD-U with trisomy 8 and JAK2 V617F mutation. The data of this patient will provide evidence for studying correlation of chromosome karyotype aberration with JAK2 V617F mutation and for evaluating prognosis of MDS/MPD-U.
Chromosomes, Human, Pair 8
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Female
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Humans
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Janus Kinase 2
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genetics
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Karyotyping
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Middle Aged
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Mutation
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Myelodysplastic-Myeloproliferative Diseases
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classification
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genetics
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Trisomy
7.Process of HIV-1 reverse transcription and its detection by using PCR.
Wen-Xue YAO ; Ying-Liang WU ; Ying GUO
Acta Pharmaceutica Sinica 2008;43(2):118-122
Human immunodeficiency virus (HIV) is a retrovirus, belongs to Lentiviridae family. As long as viral genetic material entering into host cytoplasm, double-strand DNAs synthesis occurs which is catalyzed by reverse transcriptase (RT) with viral plus-strand RNA as template. This reverse transcription is a key link of HIV-1 life cycle and an important target for anti-HIV drug development. The process of reverse transcription can be divided into several steps: formation of minus-strand strong-stop DNA; the first translocation; initiation of plus-strand DNA synthesis; and, the second translocation and the completion of both strands. These steps can be detected individually by using polymerase chain reaction (PCR) according to the amplified products on the region of R/U5, U3, U5/PBS and the sequence between LTR and Gag. In this review, we summarize the principle for detecting stages of HIV-1 reverse transcription by using PCR.
DNA Replication
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genetics
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DNA, Viral
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biosynthesis
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genetics
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HIV Reverse Transcriptase
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genetics
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metabolism
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HIV-1
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genetics
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metabolism
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Polymerase Chain Reaction
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methods
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RNA, Viral
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genetics
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Reverse Transcription
8.Comparative study on inner quality of various Chrysanthemum morifolium cultivated in Tongxiang city.
Tao WANG ; Qiao-Sheng GUO ; Xue-Gen SHEN ; Ying-Nuan LIANG ; Tao-Ying WANG ; Jian-Song ZHOU
China Journal of Chinese Materia Medica 2007;32(9):783-849
OBJECTIVETo study the quality and the chemical components of Chrysanthemum morifolium from Tongxiang city.
METHODChemical constituents of nine cultivars were compared in three types of index: chlorogenic acid, flavonoid and volatile oil.
RESULT AND CONCLUSIONThe content varied significantly. The content of chlorogenic acid in Jinjuerhao was 6.66%, the highest among the samples. Yizhongdabaiju showed the highest flavonoid and volatile oil with 9.49% and 3.30 mL x kg(-1) respectively.
China ; Chlorogenic Acid ; analysis ; Chrysanthemum ; chemistry ; classification ; growth & development ; Ecosystem ; Flavonoids ; analysis ; Flowers ; chemistry ; Oils, Volatile ; analysis ; Plant Preparations ; analysis ; chemistry ; standards ; Plants, Medicinal ; chemistry ; classification ; growth & development ; Quality Control ; Species Specificity
9.Changes and the clinical significance of muscle strength after different proportion tibial neurotomy.
Feng XUE ; Bao-guo JIANG ; Zhong-guo FU ; Dian-ying ZHANG
Chinese Journal of Surgery 2005;43(16):1095-1097
OBJECTIVETo study the changes of the muscle strength after the selective tibial neurotomy and the relationship between the changes and the quantities of neurotomy, and to discuss the clinical significances.
METHODSTwenty-four normal SD rats were divided into 4 groups with 6 in each. In group A, the left tibia nerve were cut off by 80%. 60% in group B, 40% in group C, 20% in group D, with the right as the control side. After 6 weeks measure the strength of the crural triceps and the weight of them.
RESULTSIn all the groups muscle weight and muscle strength decreased. 88.2% strength decreased on the average in group A, 54.2% in group B, 19.5% in group C, 4.7% in group D.
CONCLUSIONIt will not damage strength of the crural triceps to cut off below 40% tibial nerve in SD rats.
Animals ; Female ; Male ; Muscle Contraction ; physiology ; Muscle Denervation ; Muscle, Skeletal ; innervation ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Tibial Nerve ; surgery
10.Study on hepatitis C virus genotypes in Yantai district, Shandong province.
Guo-Ying ZHANG ; Sheng-Bo PU ; Xue-Ying YU ; Jian YANG ; Jian-Feng SHI ; Ai-Ling XU ; Zhen-Hua GUO ; Ming-Gang QIAO ; Jie ZHANG ; Yong-Zheng JING
Chinese Journal of Experimental and Clinical Virology 2012;26(4):259-262
OBJECTIVETo understand the hepatitis C virus (HCV) genotype distribution in Yantai district of Shandong province, and to explore whether the HCV genotypes was relevant to the injure of liver through the index of liver function.
METHODSUsing specific PCR primers to amplify the HCV RNA 5' UTR/NS5B,then PCR products were sequenced by genetic analyzer. The genotypes were identified by alignment to the GenBank reference sequences and construction the phylogenetic tree of 5' UTR.
RESULTSAmong 9 unpaid blood donors we detected two kinds of genotypes of 1b and 3a, respectively, 8 cases (88.9%) and 1 case (11.1%). Among 33 cases of hepatitis C patients we detected the 1b, 2a and 6a the three kinds of genotypes, respectively, 22 (66.7%), 10 (30.3%) and 1 (3.03%) cases. Subtype 1b is the advantage of popular genotype in HCV carriers from Yantai district, and the distribution of 1b was no significant difference in the different population (chi2 = 0.796, P = 0.373); The difference of indicator of liver damage in the different genotypes of subjects were significant (P < 0.05), the mean of ALT, AST of 2a-subtype carriers was significantly higher than the 1b-subtype population.
CONCLUSIONSThe genetic diversity of HCV in Shandong Yantai district was presented. The main genotypes were 1b-subtype, and 3a and 6a-subtype was detected firstly. The genotype of HCV were relevant to the liver damage indicators, 2a-subtype hepatitis C virus infection in the liver cells may play an important role in the disease process.
Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Hepatitis C ; epidemiology ; virology ; Humans ; Male ; Middle Aged ; Phylogeny ; Young Adult