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1.
Clinical features of chronic hepatitis B patients with YMDD mutation after lamivudine therapy 总被引：5，自引：0，他引：5
INTRODUCTION Chronic hepatitis B(CHB)remains a major public health problem,affecting more than350mil-lion people worldwide.Cirrhosis,liver failure,or hepatocellular carcinoma will develop in approxi-mately15to40percent of infected patients(Ganem and Prince,2004).Lamivudine,an oral nucleoside analogue,inhibits HBV replication(de Clercq,2001;2004;Lai et al.,1997;1998;Marcellin et al.,2004).It can markedly reduce serum HBV DNA levels and normalize alanine aminotransferase(ALT)levels … 相似文献
2.
Detection of PCV2 DNA by SYBR Green I-based quantitative PCR 总被引：5，自引：0，他引：5
We developed an assay for the detection and quantitation of porcine circovirus type 2 (PCV2) with the SYBR Green I-based real-time PCR. The real-time PCR provides a broad dynamic range, detecting from 103 to 1011 copies of DNA per reaction. No cross-reactions were found in specimens containing PCV1. Because of the high sensitivity and specificity of the assay with a relatively rapid and simple procedure, real-time PCR can be used as a routine assay for the clinical diagnosis of PCV2 infection. In this study we applied real-time PCR assay to 80 clinical samples, collected from 40 pigs with postweaning multisystemic wasting syndrome (PMWS) and 40 healthy pigs in comparison with conventional PCR assay. In 56 of 80 samples, PCV2 DNA was de-tected by conventional PCR assay. All samples positive for PCV2 DNA in conventional PCR assay were also positive in real-time assay, and 12 of 24 samples that tested negative for PCV2 DNA in the conventional assay were tested positive in real-time PCR assay. Real-time PCR assay increased the number of samples in which PCV2 was detected by 15%. It is, therefore, considered to be a useful tool for the detection of PCV2. 相似文献
3.
Objective: To identify the mutations of iduronate-2-sulfatase (IDS) gene, to reveal its mutation features, and to establish a basis for genetic counseling and prenatal gene diagnosis of Hunter syndrome. Methods: Urine glycosaminoglycans (GAGs) assay, PCR and DNA sequencing were performed to detect mutation of IDS gene of the patient and his parents. Results: The result showed that the patient was: DS( ), HS( ), KS(-), CS(-), and that both of his parents were negative. A frame-shift deletion mutation (1062 del 16) was identified in exon 7 of the patient's IDS gene. His parents' genotypes were normal. Conclusion:The patient's mutation was not inherited by his parents but a novel one. The mutation probably altered the primary structure and tertiary structure of IDS enzyme protein remarkably and lowered the activity of IDS enzyme greatly. Therefore it is supposed to be the direct cause of the disorder. 相似文献
4.
Feng Chen Yong-hui Dang Chun-xia Yan Yan-ling Liu Ya-jun Deng David J. R. Fulton Teng Chen 《Journal of Zhejiang University. Science. B》2009,10(10):711-720
The purpose of this study was to investigate mitochondrial DNA (mtDNA) hypervariable segment-I (HVS-I) C-stretch variations and explore the significance of these variations in forensic and population genetics studies. The C-stretch sequence variation was studied in 919 unrelated individuals from 8 Chinese ethnic groups using both direct and clone sequencing ap-proaches. Thirty eight C-stretch haplotypes were identified, and some novel and population specific haplotypes were also detected. The C-stretch genetic diversity (GD) values were relatively high, and probability (P) values were low. Additionally, C-stretch length heteroplasmy was observed in approximately 9% of individuals studied. There was a significant correlation (r=-0.961, P<0.01) between the expansion of the cytosine sequence length in the C-stretch of HVS-I and a reduction in the number of up-stream adenines. These results indicate that the C-stretch could be a useful genetic maker in forensic identification of Chinese populations. The results from the Fst and dA genetic distance matrix, neighbor-joining tree, and principal component map also suggest that C-stretch could be used as a reliable genetic marker in population genetics. 相似文献
5.
WU Xiao-mo§ FU Jing-geng§ GE Wang-zhong ZHU Jiang-yan WANG Jun-yong ZHANG Wei QIAN Wei HUO Ke-ke 《Journal of Zhejiang University. Science. B》2007,8(2):81-87
INTRODUCTION Hepatocellular carcinoma (HCC) is the sixth most common cancer in new cases per year (626 000) and the third most common cancer resulting in death (598 000) (Parkin et al., 2005). China is the country most seriously suffering from HCC, accounting for 55% of total cases and deaths worldwide per year. The clinical behavior of HCC is heterogeneous and difficult to predict, and in patients undergoing resec-tion, recurrence rates can be as high as 50% in two years and life ex… 相似文献
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7.
Feng-wei Song Bin-bin Chen Zhao-hui Sun Li-ping Wu Su-juan Zhao Qi Miao Xia-jing Tang 《Journal of Zhejiang University. Science. B》2013,14(6):479-486
Objective
To screen mutations in FERM domain-containing protein 7 (FRMD7) gene in two Chinese families with X-linked idiopathic congenital nystagmus (XLICN).Methods
Common ophthalmic data and peripheral blood of two Chinese XLICN families (families A and B) were collected after informed consent. Genomic DNA was prepared from the peripheral blood of members of the two families and from 100 normal controls. Mutations in the FRMD7 gene were determined by directly sequencing polymerase chain reaction (PCR) products.Results
We identified a novel mutation c.980_983delATTA compound with c.986C>A mutation in the 11th exon of FRMD7 in family B, and a previously reported splicing mutation c.782G>C (p.R261G) in family A. The mutations were detected in patients and female carriers, while they were absent in other relatives or in the 100 normal controls.Conclusions
Our results expand the spectrum of FRMD7 mutations in association with XLICN, and further confirm that the mutations of FRMD7 are the underlying molecular mechanism for XLICN. 相似文献8.
Xu-fei Tan Feng Chen Shan-shan Wu Yu Shi Dong-cheng Liu Zhi Chen 《Journal of Zhejiang University. Science. B》2010,11(3):221-226
Objective: To find new protein biomarkers for the detection and evaluation of liver injury and to analyze the relationship between such proteins and disease progression in concanavalin A (Con A)-induced hepatitis. Methods: Twenty-five mice were randomly divided into five groups: an untreated group, a control group injected with phosphate buffered saline (PBS), and groups with Con A-induced hepatitis evaluated at 1, 3 and 6 h. Two-dimensional gel elec-trophoresis (2-DE) and mass spectrometry (MS) were used to identify differences in protein expression among groups. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to verify the results. Results: In mice with Con A-induced hepatitis, expression levels of four proteins were increased: RIKEN, fructose bisphosphatase 1 (fbp1), ketohexokinase (khk), and Chain A of class pi glutathione S-transferase. Changes in fbp1 and khk were confirmed by qRT-PCR. Conclusion: Levels of two proteins, fbp1 and khk, are cleady up-regulated in mice with Con A-induced hepatitis. 相似文献
9.
In order to develop a model for screening the agonists of human 132-adrenoceptor from Chinese medicinal herbs extracts, we used a cell-based functional assay based on a common G protein-coupled receptor (GPCR) regulation mechanism and destabilized enhanced green fluorescent protein (d2EGFP) reporter gene technique. The positive cell clone was confirmed by real-time polymerase chain reaction (PCR) and imaging analysis. To assess the value of this model, we screened over 2000 high performance liquid chromatography (HPLC)-fractionated samples from the ethanol extracts of Chinese medicinal herbs. Six fractions (isolated from Panax japonicus, Veratrum nigrum, Phellodendron amurense, Fructus Aurantii Imrnaturus, Chaenomeles speciosa, and Dictamnus dasycarpus) showed significant effects on active reporter gene expression, three of which (isolated from Phellodendron amurense, Fructus Aurantii lmmaturus, and Chaenomeles speciosa) were selected for further concentration re-sponse analysis and the half maximal effective concentration (ECI/2 max) values were 4.2, 2.7, and 4.8 μg/ml, respectively.Therefore, this reporter gene assay was suitable for screening β2-adrenoceptor agonists. The results suggest that the six herbal extracts are the possible agonists of β2-adrenoceptor. 相似文献
10.
INTRODUCTION Platelet activating factor (PAF), a kind of pho-spholipids compound, synthesized and secretedmainly by neutrophilic granulocyte, platelet, is ahighly potent chemical mediator in inflammationand allergic reactions and induces microvascularleakage in several tissues (Chung, 1997). PAFs arefew in vivo, have short half life (only about 30seconds) and rapidly convert to lyso-PAF that hasno bio-activity and is both the metabolite and pre-cursor of PAF (Rao, 1998). Lyso-P… 相似文献
11.
Objective: To establish a new assay for platelet-activating factor (PAF), to compare it with bio-assay; and to discuss its significance in some elderly people diseases such as cerebral infarction and coronary heart disease. Methods: To measure PAF levels in 100 controls, 23 elderly patients with cerebral infarction and 65 cases with coronary heart disease by reversed phase high-performance liquid chromatographic technique (rHPLC). Results: rHPLC is more convenient, sensitive, specific, and less confusing, compared with bio-assay. The level of plasma PAF in patients with cerebral infarction was higher than that in the controls (P<0.01), and in patients with coronary heart disease. Conclusion: Detection of PAF with rHPLC is more reliable and more accurate. The new assay has important significance in PAF research. 相似文献
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万德年 《黄冈职业技术学院学报》2004,6(1):60-63
SARS是由新型冠状病毒感染引起的一种以发热、咳嗽等为主要症状 ,传染性极强、病死率高的急性呼吸道传染病 ,病人能迅速发展成急性呼吸窘迫综合症而死亡。本文对该病的病原学、流行病学、发生发展过程、临床诊断及综合防制等进行了回顾和综述。 相似文献
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目的：探讨荧光定量PCR检测HBV DNA与HBVm的关系。方法；采用荧光定量PCR法和酶联免疫吸附试验（ELISA）对159份HBVm8种不同阳性模式及67份HBVm全阴模式血清进行HBV DNA检测。结果；HBsAg、HBcAb、HBeAe阳性者HBV DNA阳性率为89．83％；HBsAg、HBcAb、HBeAb阳性者HBV DNA阳性率为28．81％；HBsAb、HBcAb、HBeAb阳性者HBV DNA阳性率为16．66％；HBVm全阴性者HBV DNA阳性率为17．91％。结论：荧光定量PCR法检测HBV DNA是乙型肝炎病毒复制最直接可信的指标．可反映HBV真实感染和复制状态．特别是低复制状态，明显优于ELISA法检测HBVm，具有重要临床意义。 相似文献
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目的研究脑梗塞患者血浆脂蛋白(a)水平.方法以酶联免疫吸附法(ELISA)测定44例脑梗塞患者与38名健康人血浆脂蛋白(a)浓度.结果脑梗塞患者的脂蛋白(a)浓度显著高于正常健康人.结论血浆高脂蛋白(a)水平是脑梗塞的重要危险因素. 相似文献
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目的：通过比较国标法、real—timePcR法和LAMP方法,得到适合基层实验室快速检测单核细胞增生李斯特菌的方法。方法：分别用国标法、real-timePcR法和环介导的等温扩增（100p—mediated isothermal amplification,LAMP）法检测李斯特菌,并进行方法比较。结果：三种方法对单增李斯特菌的检测结果一致．国标法整个检测过程需5～7d;real—timePCR法实验条件要求高,成本高,检测需1．5～2．5d;LAMP法实验条件低,成本低,检测时限与real—timePCR相同。结论：LAMP~检测单增李斯特菌灵敏度高、特异性强、快速高效和低成本,适合基层实验室应急检测和现场监测使用。 相似文献
16.
Feng Gao Xin-yang Hu Xiao-jie Xie Qi-yuan Xu Ya-ping Wang Xian-bao Liu Mei-xiang Xiang Yong Sun Jian-an Wang 《Journal of Zhejiang University. Science. B》2010,11(8):608-617
Mesenchymal stem cell(MSC)transplantation has shown a therapeutic potential to repair the ischemic and infracted myocardium,but the effects are limited by the apoptosis and loss of donor cells in host cardiac microenvironment.The aim of this study is to explore the cytoprotection of heat shock protein 90(Hsp90)against hypoxia and serum deprivation-induced apoptosis and the possible mechanisms in rat MSCs.Cell viability was determined by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Apoptosis was assessed by Hoechst 33258nuclear staining and flow cytometric analysis with annexin V/PI staining.The gene expression of Toll-like receptor-4(TLR-4)and V-erb-b2 erythroblastic leukemia viral oncogene homolog 2(ErbB2)was detected by real-time polymerase chain reaction(PCR).The protein levels of cleaved caspase-3,Bcl-2,Bcl-xL,Bax,total-ERK,phospho-ERK,totaI-Akt,phospho-Akt,and Hsp90 were detected by Western blot.The production of nitric oxide was measured by spectrophotometric assay.Hsp90 improves MSC viability and protects MSCs against apoptosis induced by serum deprivation and hypoxia.The protective role of Hsp90 not only elevates Bcl-2/Bax and Bcl-xL/Bax expression and attenuates cleaved caspase-3 expression via down-regulating membrane TLR-4 and ErbB2 receptors and then activating their downstream PI3K/Akt and ERK1/2 pathways,but also enhances the paracrine effect of MSCs.These findings demonstrated a novel and effective treatment strategy against MSC apoptosis in cell transplantation. 相似文献
17.
Chuan ZUO Xi-sheng XIE Hong-yu QIU Yao DENG Da ZHU Jun-ming FAN 《Journal of Zhejiang University. Science. B》2009,10(5)
Astragalus mongholicus (AM) derived from the dry root of Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao is a widely used traditional Chinese medicine. The present study investigated the potential role of AM on renal fibrosis on a rat model of unilateral ureteral obstruction (UUO). We divided 48 Sprague-Dawley rats randomly into 4 groups: sham-operated group (Sham), untreated UUO group, AM-treated (10 g/(kg.d)) UUO group, and losartan-treated (20 mg/(kg.d)) UUO group as positive control. Haematoxylin & eosin (HE) and Masson staining were used to study the dynamic histological changes of the kidneys 7 and 14 d after operation. The expressions of fibronectin (FN), type I collagen (coII), hepatocyte growth factor (HGF), transforming growth factor-pi (TGF-βl), and a-smooth muscle actin (a-SMA) were analyzed by real-time polymerase chain reaction (PCR), immunohistochemistry staining, and Western blot. Results show that, similar to losartan, AM alleviated the renal damage and decreased the deposition of FN and colI from UUO by reducing the expressions of TGF-pi and a-SMA (P<0.05), whereas HGF increased greatly with AM treatment (P<0.05). Our findings reveal that AM could retard the progression of renal fibrosis. The renoprotective effect of AM might be related to inhibition of myofibroblast activation, inducing of HGF and reducing of TGF-β1 expression. 相似文献
18.
目的 :探讨HSP70和HSP2 7在食管癌组织中的表达情况。方法 :采用SP免疫组织化学染色检测 5 9例食管癌组织中HSP70和HSP2 7。结果 :HSP70和HSP2 7的总阳性率分别为 6 4.4 1% (38/5 9)和 5 8.6 2 % (34/5 8) ,HSP70和HSP2 7的共同阳性率为 30 .0 0 % (9/30 )。结论 :在食管癌中HSP70和HSP2 7普遍表达。 相似文献
19.
The Catabacteriaceae is a new bacterial family with a unique member: Catabacter hongkongensis is a strictly anaerobic, non-sporulating, Gram-positive coccobacillus that is phylogenetically related to some clostridial clusters. Little is known of its epidemiology and environmental distribution, but the inclusion of its 16S rRNA gene sequence in GenBank has allowed it to be detected qualitatively. As a first approach for prospective surveys, a real-time polymerase chain reaction (PCR) procedure to identify C. hongkongensis has been developed. The presence of Catabacteriaceae in 29 water bodies subjected to possible human or animal impact has been investigated. Four of them were positive. The results confirm that highly polluted water can contain C. hongkongensis. 相似文献
20.
Bao-zhong Zhang Xin Zhang Xiao-ping An Duo-liang Ran Yu-sen Zhou Jun Lu Yi-gang Tong 《Journal of Zhejiang University. Science. B》2009,10(6):479-482
Site-directed mutagenesis (SDM) has been a very important method to probe the function-structure relationship of proteins. In this study, we introduced an easy-to-use, polymerase chain reaction (PCR)-based SDM method for double-stranded plasmid DNA, with a designed restriction site to ensure simple and efficient mutant screening. The DNA sequence to be mutated was first translated into amino acid sequence and then the amino acid sequence was reversely translated into DNA sequence with degenerate codons, resulting in a large number of sequences with silent mutations, which contained various restriction endonuclease (RE) sites. Certain mutated sequence with an appropriate RE site was selected as the target DNA sequence for designing a pair of mutation primers to amplify the full-length plasmid via inverse PCR. The amplified product was 5'-phosphorylated, circularized, and transformed into an Escherichia coli host. The transformants were screened by digesting with the designed RE. This protocol uses only one pair of primers and only one PCR is conducted, without the need for hybridization with hazardous isotope for mutant screening or subcloning step. 相似文献