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Expression of Yes-associated protein in non-small cell lung cancer and its relationship with clinical pathological factors 被引量:18
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作者 SU Li-li MA Wei-xia YUAN Jian-feng SHAO Yang XIAO Wei JIANG Shu-juan 《Chinese Medical Journal》 SCIE CAS CSCD 2012年第22期4003-4008,共6页
Background Yes-associated protein (YAP) plays an important role in signal transduction and gene transcription regulation in normal cells, with elevated and over-expressed YAP levels observed in various malignant tum... Background Yes-associated protein (YAP) plays an important role in signal transduction and gene transcription regulation in normal cells, with elevated and over-expressed YAP levels observed in various malignant tumors. The aim of this study was to investigate the expression of YAP in non-small cell lung cancer (NSCLC), and to study the possible relationship of YAP expression with the occurrence and development of NSCLC. Methods YAP expression was assessed in 40 cases of NSCLC tumor tissues by immunohistochemistry, and their protein and mRNA levels were evaluated through Western blotting and reverse transcription-polymerase chain reaction (PCR), respectively. Normal lung tissues obtained from the same patient were used as control. Statistical analysis was performed to correlate the YAP expression to clinical pathological factors, such as tumor type, stage and grade. Results YAP-positive expression was found in 28 (70%) of the 40 cases of NSCLC, which included 10 cases of squamous cell carcinoma (25%), 17 cases of adenocarcinoma (42.5%) and 1 case of squamous adenocarcinoma (2.5%). In the 28 YAP-positive cases, 19 cases showed lymph node metastasis and were classified in TNM stage II + III (47.5%); the other nine cases showed no lymph node metastasis (22.5%) and were classified in the TNM stage I. There was no relationship between YAP expression and patients' age, gender or tumor histological grades. However, YAP showed significant over expression in late period of T stage (P=0.012), TNM stage (P=0.039), and lymph node metastasis (P=-0.013), respectively. Notably, YAP-positive expression was significantly higher in adenocarcinoma than that in squamous cell carcinoma (P=0.041). Conclusions Over-expression of YAP was associated with NSCLC, especially lung adenocarcinoma. The high YAP expression in late period of tumor stage and lymph node metastasis may indicate that YAP expression could be an early marker for NSCLC tumorigenesis. 展开更多
关键词 non-small cell lung cancer Yes-associatedprotein IMMUNOHISTOCHEMISTRY Western blotting reverse transcription-polymerase chain reaction
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Role of microRNAs in gastric cancer 被引量:15
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作者 Hideyuki Ishiguro Masahiro Kimura Hiromitsu Takeyama 《World Journal of Gastroenterology》 SCIE CAS 2014年第19期5694-5699,共6页
Although gastric cancer(GC)is one of the leading causes of cancer-related death,major therapeutic advances have not been made,and patients with GC still face poor outcomes.The prognosis of GC also remains poor because... Although gastric cancer(GC)is one of the leading causes of cancer-related death,major therapeutic advances have not been made,and patients with GC still face poor outcomes.The prognosis of GC also remains poor because the molecular mechanisms of GC progression are incompletely understood.MicroRNAs(miRNAs)are noncoding RNAs that are associated with gastric carcinogenesis.Studies investigating the regulation of gene expression by miRNAs have made considerable progress in recent years,and abnormalities in miRNA expression have been shown to be associated with the occurrence and progression of GC.miRNAs contribute to gastric carcinogenesis by altering the expression of oncogenes and tumor suppressors,affecting cell proliferation,apoptosis,motility,and invasion.Moreover,a number of miRNAs have been shown to be associated with tumor type,tumor stage,and patient survival and therefore may be developed as novel diagnostic or prognostic markers.In this review,we discuss the involvement of miRNAs in GC and the mechanisms through which they regulate gene expression and biological functions.Then,we review recent research on the involvement of miRNAs in GC prognosis,their potential use in chemotherapy,and their effects on Helicobacter pylori infections in GC.A greater understanding of the roles of miRNAs in gastric carcinogenesis could provide insights into the mechanisms of tumor development and could help to identify novel therapeutic targets. 展开更多
关键词 MICRORNA Gastric cancer Reverse transcription-polymerase chain reaction CHEMOSENSITIVITY Helicobacter pylori Circulating MicroRNA
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Increased CD163 expression is associated with acute-on-chronic hepatitis B liver failure 被引量:13
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作者 Hong Ye Li-Yuan Wang +1 位作者 Jing Zhao Kai Wang 《World Journal of Gastroenterology》 SCIE CAS 2013年第18期2818-2825,共8页
AIM: To assess CD163 expression in plasma and peripheral blood and analyze its association with disease in acute-on-chronic hepatitis B liver failure (ACHBLF) patients. METHODS: A retrospective study was conducted fro... AIM: To assess CD163 expression in plasma and peripheral blood and analyze its association with disease in acute-on-chronic hepatitis B liver failure (ACHBLF) patients. METHODS: A retrospective study was conducted from January 1, 2011 to January 1, 2012. Forty patients with ACHBLF (mean age 44.48 ± 12.28 years, range 18-69 years), 40 patients with chronic hepatitis B (CHB) (mean age 39.45 ± 12.22 years, range 21-57 years) and 20 ageand sex-matched healthy controls (mean age 38.35 ± 11.97 years, range 28-60 years) were included in this study. Flow cytometry was used to analyze the frequency of CD163+ peripheral blood mononuclear cells (PBMCs) and surface protein expression of CD163. Real-time transcription-polymerase chain re-action was performed to assess relative CD163 mRNA levels in PBMCs. Plasma soluble CD163 (sCD163) levels were measured by enzyme-linked immunosorbent assay. Clinical variables were also recorded. Comparisons between groups were analyzed by Kruskal-Wallis H test and Mann-Whitney U test. Statistical analyses were performed using SPSS 15.0 software and a P value < 0.05 was considered statistically significant. RESULTS: Flow cytometry showed that the population of CD163+ PBMCs was significantly greater in ACHBLF patients than in CHB patients and healthy controls (47.9645% ± 17.1542%, 32.0975% ± 11.0215% vs 17.9460% ± 6.3618%, P < 0.0001). However, there were no significant differences in mean fluorescence intensity of CD163+ PBMCs within the three groups (27.4975 ± 11.3731, 25.8140 ± 10.0649 vs 20.5050 ± 6.2437, P = 0.0514). CD163 mRNA expression in ACHBLF patients was significantly increased compared with CHB patients and healthy controls (1.41 × 10 -2 ± 2.18 × 10 -2 , 5.10 × 10 -3 ± 3.61 × 10 -3 vs 37.0 × 10 -4 ± 3.55 × 10 -4 , P = 0.02). Plasma sCD163 levels in patients with ACHBLF were significantly increased compared with CHB patients and healthy controls (4706.2175 ± 1681.1096 ng/mL, 1089.7160 ± 736.8395 ng/mL vs 435.9562 ± 440.8329 ng/mL, P < 0.0001). In ACHBLF patient 展开更多
关键词 Acute-on-chronic hepatitis B LIVER failure Model for END-STAGE LIVER disease CD163 Soluble CD163 Real-time transcription-polymerase chain reaction
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Monoclonal antibody-based serological methods for maize chlorotic mottle virus detection in China 被引量:11
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作者 Jian-xiang WU Qiang WANG +3 位作者 Huan LIU Ya-juan QIAN Yan XIE Xue-ping ZHOU 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2013年第7期555-562,共8页
Maize chlorotic mottle virus (MCMV) infects maize plants and causes significant losses in corn production worldwide. In this study, purified MCMV particles were used as the immunogen to produce monoclonal antibodies (... Maize chlorotic mottle virus (MCMV) infects maize plants and causes significant losses in corn production worldwide. In this study, purified MCMV particles were used as the immunogen to produce monoclonal antibodies (MAbs) and polyclonal antibodies (PAbs). Four murine MAbs (4B8, 8C11, 6F4, and 9G1) against MCMV were obtained through the hybridoma technology. The triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA), dot-immunobinding assay (DIBA), and immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) using the MAb 4B8 were then developed for sensitive, specific, and rapid detection of MCMV in fields. MCMV could be detected in infected leaf crude extracts at dilutions of 1:327 680, 1:64000, and 1:3276800 (w/v, g/ml) by TAS-ELISA, DIBA, and IC-RT-PCR, respectively. One hundred and sixty-one maize field samples showing virus-like symptoms and sixty-nine symptomless maize field samples from ten different provinces of China were collected and screened for the presence of MCMV using the established serological methods. A phylogenetic tree was constructed based on the full length CP genes and Chinese MCMV isolates formed one branch with Thailand isolates. The detection results demonstrated that MCMV is one of most prevalent viruses infecting maize in the Yunnan and Sichuan provinces of China. 展开更多
关键词 Maize chlorotic mottle virus (MCMV) Immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) Triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) Monoclonal antibody (MAb) Dot-immunobinding assay (DIBA)
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Correlation between MMP-2 and NF-κ B expression of intracranial aneurysm 被引量:10
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作者 Wei-Tao Cheng Ning Wang 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2013年第7期570-573,共4页
Objective:To investigate the correlation between expressions of MMP-2 and NF-κB in the intracranial aneurysm wall,and explore their role in the mechanism of the occurrence,growth and rupture of intracranial aneurysms... Objective:To investigate the correlation between expressions of MMP-2 and NF-κB in the intracranial aneurysm wall,and explore their role in the mechanism of the occurrence,growth and rupture of intracranial aneurysms.Methods:RT-PCR was used to detect the expression of MMP-2 and NF-κB mRNA of 30 cases of intracranial aneurysm tissue and 10 cases of normal intracranial arterial tissue:Immunohistochemical method was used to detect the expression of MMP-2 and NF-κB protein.Results:the semi-quantitative analysis of MMP-2 and NF-κB in aneurysms tissues and normal tissues were statistically significant different from each other(P【0 05).Immunohistochemical staining results showed NF-κB was expressed in different layers.The expression of them were positive in intimal and medial,and the expression sites were located in the nucleus.MMP-2 were expressed in different layers of the aneurysm wall,and the expressions were positive in media and extima.The MMP-2 and NF-κB-positive expression of aneurysm wall were significantly higher than in normal cerebral arteries(P【0.05).MMP-2 and NF-κB mRNA expression showed positive correlation in the aneurysm wall tissue(r = 0.689,P = 0.005). Conclusions:The expressions of MMP-2 and NF-κB in the intracranial aneurysm wall tissue were significantly higher than in the normal intracranial arterial tissues.They have a synergistic effect on the formation of intracranial aneurysms. 展开更多
关键词 INTRACRANIAL ANEURYSMS Nuclear factor-κB Matrix METALLOPROTEINASE-2 Reverse transcription-polymerase Chain reaction IMMUNOHISTOCHEMISTRY
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Overexpression of polo-like kinase1 predicts a poor prognosis in hepatocellular carcinoma patients 被引量:11
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作者 Zi-Li He He Zheng Hui Lin Xiong-Ying Miao De-Wu Zhong 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第33期4177-4182,共6页
AIM: To elucidate the role of overexpressed polo-like kinasel (PLK1)in hepatocellular carcinoma (HCC). METHODS: We prospectively collected clinicopathological, immunohistochemical and semi-quantitative reverse t... AIM: To elucidate the role of overexpressed polo-like kinasel (PLK1)in hepatocellular carcinoma (HCC). METHODS: We prospectively collected clinicopathological, immunohistochemical and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) data from 135 HCC patients undergoing successful hepatectomy. The correlations between PLK1 mRNA expression and clinicopathologic variables were analyzed by Mann-Whitney U test. Prognostic factors were identified by univariate and multivariate analyses. RESULTS: Immunohistochemical results showed overexpression of PLK1 was mainly found in tumor tissues compared with tumor-free tissue. A similar mRNA result was obtained by semi-quantitative RT-PCR. A total of 111 samples were positive for PLK1 mRNA expression. The positive expression was correlated with venous invasion, tumor nodules and Edmondson grade. Furthermore, 1, 3, 5-year survival rates in the positive expression group were significantly lower than the negative control group. Multivariate analysis showed that positive PLK1 expression was an independent risk factor for HCC. CONCLUSION: PLK1 could be a potential biomarker for diagnosis and therapy for HCC. 展开更多
关键词 Hepatocellular carcinoma IMMUNOHISTOCHEMISTRY Reverse transcription-polymerase chain reaction Survival analysis Polo-like kinase 1
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Expression of programmed death 1 and its ligands in the liver of autoimmune hepatitis C57BL/6 mice 被引量:7
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作者 CAO Jin LIU Feng-Xia YU Meng-xue 《Chinese Medical Journal》 SCIE CAS CSCD 2009年第16期1941-1946,共6页
Background Autoimmune hepatitis (AIH) is a chronic inflammatory liver disease with unknown etiology. Programmed death 1 (PD-1) and its ligands (PD-L1 and PD-L2), BT-H1/PD-L1 and B7-DC/PD-L2, are new CD28-B7 fami... Background Autoimmune hepatitis (AIH) is a chronic inflammatory liver disease with unknown etiology. Programmed death 1 (PD-1) and its ligands (PD-L1 and PD-L2), BT-H1/PD-L1 and B7-DC/PD-L2, are new CD28-B7 family members that are involved in the regulation of immune responses. Previous observation suggests that PD-1 system plays an inhibitory role in regulating peripheral blood T cells, B cells and myeloid cells, thus their abnormality may be related to autoimmune diseases. This study aimed to explore the role of PD-1/PD-L1, L2 system in the pathogenesis of AIH. Methods The mice model of experimental autoimmune hepatitis (EAH) was established in C57BL/6 mice and the expression levels of PD-1 and PD-L1, L2 in the murine liver and the cytokines, including interferon (IFN)-γ, tumor necrosis factor (TNF)-α and interleukin (IL)-4 in the spleen were detected using reverse transcription-polymerase chain reaction (RT-PCR), and the results were compared with those of normal controls. Results The expression levels of PD-1, PD-L1, PD-L2 mRNA were higher in EAH compared with normal controls (P 〈0.05), the PD-L2/PD-1 ratio was relatively lower in EAH (EAH -0.08±0.35, normal controls 0.52±0.07, P=0.009). In the EAH, the expression of the three cytokines were all upregulated compared with normal controls. PD-L1 had a positive correlation with the expression of IFN-γ (r =0.289, P 〈0.05), while PD-L2 showed a positive correlation with both expressions of IL-4 (r=0.378, P 〈0.01) and IFN-γ (r =0.261, P 〈0.05). While TNF-α showed no correlation with PD-L1 (r=0.044, P=0.736) or PD-L2 (r=0.127, P=0.335). Conclusions The expression of PD-1/PD-L1, L2 is upregulated in EAH and regulated by IFN-γ and IL-4. PD-1 system may play an important role in the pathogenesis of AIH. 展开更多
关键词 autoimmune hepatitis programmed death 1 programmed death 1 ligands reverse transcription-polymerase chain reaction CYTOKINE
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Diagnosis of nervous necrosis virus in orange-spotted grouper,Epinephelus coioides, by a rapid and convenient RT-PCR method 被引量:6
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作者 MU Yinnan LIN Kebing +1 位作者 CHEN Xinhua AO Jingqun 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2013年第10期88-92,共5页
Viral nervous necrosis (VNN) causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is i... Viral nervous necrosis (VNN) causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is important to block its outbreak. In this study, a reverse transcription-polymerase chain reaction (RT-PCR) was developed for the rapid, convenient, and sensitive detection of the VNN pathogen, nervous necro- sis virus (NNV), in the grouper. The whole process was completed within 3.5 h from the RNA extraction to PCR product visualization. The detection limit of this method was 200 copies of NNV RNA standard, which corresponded to 200 copies of virus particles. This RT-PCR method was specific to the NNV detection with no cross-reactivity to other fish viral disease pathogens, such as infectious pancreatic necrosis virus (IPNV), infectious hematopoietic necrosis virus (IHNV), spring viraemia of carp virus (SVCV), epizootic haematopoietic necrosis virus (EHNV), and large yellow croaker iridovirus (LYC1V). With this method, the orange-spotted grouper (Epinephelus coioides) fry from hatcheries with or without incidence of the VN- N epidemic in Fujian Province were detected. The results showed that all or 93% of the fry from the two hatcheries with incidence of the epidemic were diagnosed as positive, while 40% or 25% of fry from the t- wo hatcheries without the VNN epidemic were also detected as NNV positive, indicating that this RT-PCR method can be used for rapid, sensitive detection of NNV infection and applied in the VNN epidemic alert. 展开更多
关键词 viral nervous necrosis nervous necrosis virus orange-spotted grouper (Epinephelus coioides) reverse transcription-polymerase chain reaction DETECTION
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Detection of circulating hepatocellular carcinoma cells in peripheral venous blood by reverse transcription-polymerase chain reaction 被引量:5
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作者 Yang Liu Meng-Chao Wu +1 位作者 Guang-Xiang Qian Bai-He Zhang From the Institute of East Hepatobiliary Surgery, Second Military Medical University, Shanghai 200438, China 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2002年第1期72-76,共5页
Objective: To detect circulating hepatocellular carcino-ma by demonstrating hepatocellular carcinoma cells orhepatocyte-associated mRNA in the nuclear cell com-ponent of peripheral blood (PBL).Methods: Peripheral bloo... Objective: To detect circulating hepatocellular carcino-ma by demonstrating hepatocellular carcinoma cells orhepatocyte-associated mRNA in the nuclear cell com-ponent of peripheral blood (PBL).Methods: Peripheral blood (5 ml) samples were ob-tained from 93 patients with hepatocellular carcinoma(HCC) and from 33 control subjects (9 with liver cir-rhosis after hepatitis B,14 with chronic hepatitis B,10with normal liver function). To identify HCC cells inperipheral blood, liver-specific human alpha-fetopro-tein (AFP) mRNA was amplified from total RNA ex-tracted from whole blood by reverse transcription-polymerase chain reaction.Results: AFPmRNA was detected in 50 blood samplesfrom the HCC patients (50/93, 53.8%). In contrast,there were no clinical control patients whose samplesshowed detectable AFPmRNA in PBL. The presence ofAFPmRNA in blood seemed to be correlated with thestage (by TNM classification) of HCC, the serum AFPvalue, and the presence of intrahepatic metastasis,portal vein thrombosis, tumor diameter and/or distantmetastasis. In addition, AFPmRNA was detected in theblood of 21 patients with metastasis at extrahepaticorgans (100%) in contrast to 29 (40.3%)of 72 pa-tients without metastasis.Conclusion: The presence of AFPmRNA in peripheralblood may be an indicator of malignant hepatocytes,which might predict hematogenous spreading metasta-sis of tumor cells in patients with HCC. 展开更多
关键词 liver neoplasms ALPHA-FETOPROTEIN MRNA reverse transcription-polymerase chain reaction
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Expression and functional characterization of platelet-derived growth factor receptor-like gene 被引量:5
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作者 Guo, Feng-Jie Zhang, Wei-Jia +6 位作者 Li, Ya-Lin Liu, Yan Li, Yue-Hui Huang, Jian Wang, Jia-Jia Xie, Ping-Li Li, Guan-Cheng 《World Journal of Gastroenterology》 SCIE CAS CSCD 2010年第12期1465-1472,共8页
AIM:To investigate the role of platelet-derived growth factor receptor-like gene(PDGFRL)in the anti-cancer therapy for colorectal cancers(CRC).METHODS:PDGFRL mRNA and protein levels were measured by reverse transcript... AIM:To investigate the role of platelet-derived growth factor receptor-like gene(PDGFRL)in the anti-cancer therapy for colorectal cancers(CRC).METHODS:PDGFRL mRNA and protein levels were measured by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry in CRC and colorectal normal tissues.PDGFRL prokaryotic expression vector was carried out in Escherichia coli(E.coli),and purified by immobilized metal affinity chromatography.The effect of PDGFRL protein on CRC HCT-116 cells was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT),clone counting,cell cycle,and wound healing assay.RESULTS:Both RT-PCR and immunohistochemistry showed that the expression of PDGFRL in colorectal normal tissues was higher than in cancer tissues.Recombinant pET22b-PDGFRL prokaryotic expression vector was successfully expressed in E.coli,and the target protein was expressed in the form of inclusion bodies.After purification and refolding,recombinant human PDGFRL(rhPDGFRL)could efficiently inhibit the proliferation and invasion of CRC HCT-116 cells detected by MTT,clone counting and wound healing assay.Moreover,rhPDGFRL arrested HCT-116 cell cycling at the G0/G1 phase.CONCLUSION:PDGFRL is a potential gene for application in the anti-cancer therapy for CRC. 展开更多
关键词 Platelet-derived growth factor receptorlike Colorectal cancer Prokaryotic expression Reverse transcription-polymerase chain reaction IMMUNOHISTOCHEMISTRY
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Methods for the extraction and RNA profiling of exosomes 被引量:5
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作者 Emily Zeringer Mu Li +6 位作者 Tim Barta Jeoffrey Schageman Ketil Winther Pedersen Axl Neurauter Susan Magdaleno Robert Setterquist Alexander V Vlassov 《World Journal of Methodology》 2013年第1期11-18,共8页
AIM:To develop protocols for isolation of exosomes and characterization of their RNA content.METHODS:Exosomes were extracted from He La cell culture media and human blood serum using the Total exosome isolation(from c... AIM:To develop protocols for isolation of exosomes and characterization of their RNA content.METHODS:Exosomes were extracted from He La cell culture media and human blood serum using the Total exosome isolation(from cell culture media)reagent,and Total exosome isolation(from serum)reagent respectively.Identity and purity of the exosomes was confirmed by Nanosight?analysis,electron microscopy,and Western blots for CD63 marker.Exosomal RNA cargo was recovered with the Total exosome RNA and protein isolation kit.Finally,RNA was profiled using Bioanalyzer and quantitative reverse transcriptionpolymerase chain reaction(q RT-PCR)methodology.RESULTS:Here we describe a novel approach for robust and scalable isolation of exosomes from cell culture media and serum,with subsequent isolation and analysis of RNA residing within these vesicles.The isolation procedure is completed in a fraction of the time,compared to the current standard protocols utilizing ultracentrifugation,and allows to recover fully intact exosomes in higher yields.Exosomes were found tocontain a very diverse RNA cargo,primarily short sequences 20-200 nt(such as mi RNA and fragments of m RNA),however longer RNA species were detected as well,including full-length 18S and 28S r RNA.CONCLUSION:We have successfully developed a set of reagents and a workflow allowing fast and efficient extraction of exosomes,followed by isolation of RNA and its analysis by q RT-PCR and other techniques. 展开更多
关键词 EXOSOMES MICROVESICLES Cell culture media SERUM RNA Quantitative reverse transcription-polymerase chain reaction SEQUENCING
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Preliminary research on myosin light chain kinase in rabbit liver 被引量:6
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作者 Bin Ren~1 Hua-Qing Zhu~2 Zhao-Feng Luo~1 Qing Zhou~2 Yuan Wang~2 Yu-Zhen Wang~1 1 Department of Biochemistry and Molecular Biology,University of Science and Technology of China,Hefei 230027,Anhui Province,China2 Laboratory of Molecular Biology,Anhui Medical University,Hefei 230032,Anhui Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第6期868-871,共4页
AIM: To study preliminarily the properties of myosin light chain kinase (MLCK) in rabbit liver. METHODS: The expression of MLCK was detected by reverse transcription-polymerase chain reaction(RT-PCR); the MLCK was obt... AIM: To study preliminarily the properties of myosin light chain kinase (MLCK) in rabbit liver. METHODS: The expression of MLCK was detected by reverse transcription-polymerase chain reaction(RT-PCR); the MLCK was obtained from rabbit liver, and its activity was analyzed by gamma-(32)P incorporation technique to detect the phosphorylation of myosin light chain. RESULTS: MLCK was expressed in rabbit liver, and the activity of the enzyme was similar to rabbit smooth muscle MLCK, and calmodulin-dependent. When the concentration was 0.65 mg x L(-1), the activity was at the highest level. CONCLUSION: MLCK expressed in rabbit liver may catalyze the phosphorylation of myosin light chain, which may play important roles in the regulation of hepatic cell functions. 展开更多
关键词 ANIMALS HEPATOCYTES Liver Myosin Light Chains Myosin-Light-Chain Kinase PHOSPHORYLATION RABBITS Research Support Non-U.S. Gov't
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Expression of SSX-1 and NY-ESO-1 mRNA in tumor tissues and its corresponding peripheral blood expression in patients with hepatocellular carcinoma 被引量:3
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作者 LU Yun WU Li-qun +2 位作者 LU Zheng-hua WANG Xin-jian YANG Jing-yang 《Chinese Medical Journal》 SCIE CAS CSCD 2007年第12期1042-1046,共5页
Background Cancer-testis antigen (CTA) is a family of the most noticeable tumor antigens which could be potential tumor markers for cancer diagnosis. In this research we aimed to investigate the expression of SSX-1 ... Background Cancer-testis antigen (CTA) is a family of the most noticeable tumor antigens which could be potential tumor markers for cancer diagnosis. In this research we aimed to investigate the expression of SSX-1 and NY-ESO-1 mRNA, two members of the CTA family, in tissue and peripheral blood of patients with hepatocellular carcinoma (HCC) to assess their feasibility for the immunotherapy and diagnosis of HCC and the association of their expression levels with diverse clinical indicators. Methods Thirty-six north Chinese patients with HCC and 30 normal controls were enrolled in this study. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of SSX-1 and NY-ESO-1 mRNA in tumor tissues and corresponding levels in peripheral blood of patients. Results The positive rates of SSX-1 and NY-ESO-1 mRNA expression were 61.1% (22/36) and 11.1% (4/36), respectively, in cancer tissues; 38.9% (14/36) and 5.6% (2/36), respectively, in the corresponding peripheral blood samples. No positive expression of either SSX-1 or NY-ESO-1 mRNA was detected in the samples of cancer-adjacent tissues, cirrhotic tissues, normal liver tissue or the peripheral blood of control patients. No significant relationship was found between the expression of these two genes and clinical indicators such as age, gender, tumor size, extent of differentiation, serum a-fetoprotein (AFP) level or infection with hepatitis B virus (P〉0.05). The short term recurrence rate was 46.2% (6/13) in patients whose peripheral blood expressed SSX-1 mRNA, while the recurrence rate in patients with negative SSX-1 mRNA was 28.6% (4/14). Conclusions SSX-1 and NY-ESO-1 antigens might be new potentially promising targets for antigen-specific immunotherapy for HCC. High specific expression of SSX-1 and NY-ESO-1 mRNA suggested that we could apply them as tumor markers. The short term recurrence rate was significantly higher in patients whose peripheral blood expressed SSX-1 mRNA, suggesting tha 展开更多
关键词 cancer-testis antigen hepatocellular carcinoma reverse transcription-polymerase chain reaction SSX-1 NY-ESO-1
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Expression of interleukin 6 in brain and colon of rats with TNBS-induced colitis 被引量:3
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作者 Wang, Kai Yuan, Chuan-Ping +7 位作者 Wang, Wei Yang, Zhan-Qing Cui, Wei Mu, Lian-Zhi Yue, Zhan-Peng Yin, Xiu-Ling Hu, Zhong-Ming Liu, Ju-Xiong 《World Journal of Gastroenterology》 SCIE CAS CSCD 2010年第18期2252-2259,共8页
AIM:To characterise expression of interleukin 6(IL-6),a potent proinflammatory cytokine,in the occurrence and development of inflammatory bowel disease(IBD) and investigate its effect on neuroimmunomodulation and immu... AIM:To characterise expression of interleukin 6(IL-6),a potent proinflammatory cytokine,in the occurrence and development of inflammatory bowel disease(IBD) and investigate its effect on neuroimmunomodulation and immune homeostasis regulation.METHODS:In this study,rats with colitis induced by trinitrobenzene sulfonic acid(TNBS) were sacrificed on days 3,7,14,21 and 28 after induction.In the controls,the TNBS was just replaced by equivalent amount of phosphate buffered solution(PBS,0.01 mol/L).IL-6 mRNA expression in brain and colon tissues in each phase was evaluated by real-time reverse transcriptionpolymerase chain reaction,and cellular localisation and protein level of IL-6 was determined by immunohistochemistry.RESULTS:At day 7,mRNA expression of IL-6 was significantly higher in the colon and brain of IBD rats than that of the controls.The protein level was also significantly higher in colon,hypothalamus and cerebral cortex of IBD rats compared with the controls.So there are similar temporal trends in IL-6 mRNA expression and protein levels in all positions with a persistent increase to a peak at day 7,followed by a decline and gradual return to normal levels.CONCLUSION:These results revealed that changes in IL-6 expression in brain and colon tissues occur in different phases of IBD.Therefore,we propose that the nerve centre regulates and controls the occurrence and development of IBD via IL-6. 展开更多
关键词 Inflammatory bowel disease Interleukin 6 NEUROIMMUNOMODULATION Real-time transcription-polymerase chain reaction Immunohistochemistry
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Significance of combined detection of LunX mRNA and tumor markers in diagnosis of lung carcinoma 被引量:3
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作者 Hui Yu Xiuying Huang +4 位作者 Zhihua Zhu Yi Hu Wei Ou Lanjun Zhang Ningning Zhou 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2014年第1期89-94,共6页
Objective:To evaluate the significance of combined detection of LunX mRNA,carcinoembryonic antigen (CEA),neuron-specific enolase (NSE),and cytokeratin 21-1 fragment (CYFRA21-1) in clinical diagnosis of lung car... Objective:To evaluate the significance of combined detection of LunX mRNA,carcinoembryonic antigen (CEA),neuron-specific enolase (NSE),and cytokeratin 21-1 fragment (CYFRA21-1) in clinical diagnosis of lung carcinoma.Methods:Based on the quantitative RT-PCR and chemiluminescence immunoassay,the expression levels of LunX mRNA,CEA,NSE,and CYFRA21-1 in 113 patients with lung carcinoma (case group) and 30 healthy participants (control group) were detected.Meantime,the sensitivity,specificity,and accuracy of the combination detection were also explored.Results:The positive rates of LunX mRNA in peripheral blood and CEA,NSE,and CYFRA21-1 in serum were significandy higher in case group than those in control group (x2=17.295,16.825,19.148,and 17.450; P<0.05).There was no statistical significance when positive rate of LunX mRNA was evaluated among different pathological types (x2=0.047,P>0.05).The positive rate of LunX mRNA in stage Ⅰ + Ⅱ,Ⅲ,and Ⅳ had a significandy increasing tendency (x2=10.565,32.462,P<0.05).The positive rate of CYFRA21-1 was highest in squamous carcinoma (78.5%),the positive rate of NSE was highest in small cell carcinoma (86.7%),and the positive rate of CEA wag highest in lung adenocarcinoma (80.4%).The sensitivity and accuracy of the combination detection were 91.1% and 88.1%,respectively.Conclusions:The combined detection of LunX mRNA and tumor markers (TMs) including CEA,NSE,and CYFRA21-1 in peripheral blood is helpful to increase the diagnostic accuracy of lmg cancer.Also,it can inform the pathological typing of lung carcinoma. 展开更多
关键词 Lung cancer reverse transcription-polymerase chain reaction LunX mRNA carcinoembryonic antigen (CEA) neuron-specific enolase (NSE) cytokeratin 21-1 fragment (CYFRA21-1)
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Quality of life of COVID-19 recovered patients:a 1-year follow-up study from Bangladesh
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作者 Mohammad Delwer Hossain Hawlader Md Utba Rashid +27 位作者 Md Abdullah Saeed Khan Mowshomi Mannan Liza Sharmin Akter Mohammad Ali Hossain Tajrin Rahman Sabrina Yesmin Barsha Alberi Afifa Shifat Mosharop Hossian Tahmina Zerin Mishu Soumik Kha Sagar Ridwana Maher Manna Nawshin Ahmed Sree Shib Shankar Devnath Debu Irin Chowdhury Samanta Sabed Mashrur Ahmed Sabrina Afroz Borsha Faraz Al Zafar Sabiha Hyder Abdullah Enam Habiba Babul Naima Nur Miah Md.Akiful Haque Shopnil Roy K.M.Tanvir Hassan Mohammad Lutfor Rahman Mohammad Hayatun Nabi Koustuv Dalal 《Infectious Diseases of Poverty》 SCIE CAS CSCD 2023年第4期101-102,共2页
BackgroundThe COVID-19 pandemic posed a danger to global public health because of the unprecedented physical,mental,social,and environmental impact affecting quality of life(QoL).The study aimed to find the changes in... BackgroundThe COVID-19 pandemic posed a danger to global public health because of the unprecedented physical,mental,social,and environmental impact affecting quality of life(QoL).The study aimed to find the changes in QoL among COVID-19 recovered individuals and explore the determinants of change more than 1 year after recovery in low-resource settings.MethodsCOVID-19 patients from all eight divisions of Bangladesh who were confirmed positive by reverse transcription-polymerase chain reaction from June 2020 to November 2020 and who subsequently recovered were followed up twice,once immediately after recovery and again 1 year after the first follow-up.The follow-up study was conducted from November 2021 to January 2022 among 2438 individuals using the World Health Organization Quality of Life Brief Version(WHOQOL-BREF).After excluding 48 deaths,95 were rejected to participate,618 were inaccessible,and there were 45 cases of incomplete data.Descriptive statistics,paired-sample analyses,generalized estimating equation(GEE)analysis,and multivariable logistic regression analyses were performed to test the mean difference in participants’QoL scores between the two interviews.ResultsMost participants(n=1710,70.1%)were male,and one-fourth(24.4%)were older than 46.The average physical domain score decreased significantly from baseline to follow-up,and the average scores in psychological,social,and environmental domains increased significantly at follow-up(P<0.05).By the GEE equation approach,after adjusting for other factors,we found that older age groups(P<0.001),being female(P<0.001),having hospital admission during COVID-19 illness(P<0.001),and having three or more chronic diseases(P<0.001),were significantly associated with lower physical and psychological QoL scores.Higher age and female sex[adjusted odd ratio(aOR)=1.3,95%confidence interval(CI)1.0–1.6]were associated with reduced social domain scores on multivariable logistic regression analysis.Urban or semi-urban people were 49%less likely(aOR=0.5,95%CI 0.4– 展开更多
关键词 Quality of life Health-related quality of life Reverse transcription-polymerase chain reaction COVID-19 BANGLADESH
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Msi1基因沉默对人肝癌HepG2细胞的影响 被引量:4
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作者 王燕 章建国 +2 位作者 张曙 靳钦 卞婷婷 《交通医学》 2014年第2期115-118,共4页
目的:探讨siRNA沉默Msi1基因表达后,人肝癌HepG2细胞的增殖与凋亡的情况。方法:设计合成针对Msi1 mRNA序列的siRNA多条序列,分别转染人肝癌HepG2细胞。使用RT–PCR方法检测各组细胞Msi1基因表达情况;Western blot法检测survivin蛋白、ca... 目的:探讨siRNA沉默Msi1基因表达后,人肝癌HepG2细胞的增殖与凋亡的情况。方法:设计合成针对Msi1 mRNA序列的siRNA多条序列,分别转染人肝癌HepG2细胞。使用RT–PCR方法检测各组细胞Msi1基因表达情况;Western blot法检测survivin蛋白、caspase3蛋白表达情况;MTT法、平皿克隆实验检测HepG2细胞增殖情况;流式细胞术检测各组HepG2细胞凋亡情况。结果:(1)RT-PCR结果:转染后24 h后,序列a,b,c转染效率分别为65%,64%及62%。其抑制率分别是Msi1 siRNAa(68%)、Msi1 siRNAb(56%)、Msi1 siRNAc(35%)。(2)流式细胞术检测各组细胞凋亡:空白组、阴性组、脂质体组、Msi1 siRNAa组凋亡率分别为(4.14±0.26)%、(4.51±0.78)%、(4.19±1.21)%,(16.8±0.26)%,Msi1 siRNAa组与空白对照组比较差异有统计学意义(P<0.05)。(3)Msi1 siRNAa最能有效抑制肝癌HepG2细胞中Msi1表达,与空白对照组比较,Msi1 siRNAa组HepG2细胞生长、增殖速度明显减缓(P<0.05);survivin蛋白表达水平显著下调(P<0.05),而caspase3蛋白表达水平上调(P<0.05);Msi1 siRNAa组凋亡率增高(P<0.05)。结论:沉默Msi1可抑制人肝癌HepG2细胞的增殖,诱导其凋亡。 展开更多
关键词 原发性肝癌 Msi1基因 凋亡 增殖 HepG2细胞 逆转录聚合酶链反应 Western BLOT检测 流式细胞术 Msi1 Reverse transcription-polymerase Chain Reaction (RT-PCR) Flow Cytometry(FCM)
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Insulin-like growth factor receptor-1 overexpression is associated with poor response of rectal cancers to radiotherapy 被引量:4
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作者 Xiao-Yu Wu Zhen-Feng Wu +7 位作者 Qin-Hong Cao Che Chen Zhi-Wei Chen Zhe Xu Wei-Su Li Fu-Kun Liu Xue-Quan Yao Gang Li 《World Journal of Gastroenterology》 SCIE CAS 2014年第43期16268-16274,共7页
AIM: To explore the potential correlation between insulin-like growth factor receptor-1 (IGF-1R) expression and rectal cancer radiosensitivity.
关键词 Insulin-like growth factor-1 receptor Rectal carcinoma Preoperative radiotherapy IMMUNOHISTOCHEMISTRY Reverse transcription-polymerase chain reaction
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Combination of Cytogenetic Analysis and Molecular Screening in Patients with de novo Acute Myeloid Leukemia 被引量:2
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作者 耿哲 张恒 +5 位作者 王迪 肖毅 王娜 李春蕊 黄亮 周剑峰 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2012年第4期501-510,共10页
Nowadays the role of genetic findings in determining the diagnosis,therapy and prognosis of acute myeloid leukemia(AML) has become more valuable.To improve and validate the detection of clonal chromosomal aberrations ... Nowadays the role of genetic findings in determining the diagnosis,therapy and prognosis of acute myeloid leukemia(AML) has become more valuable.To improve and validate the detection of clonal chromosomal aberrations in leukemia,we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction(RT-PCR) and fluorescence in situ hybridization(FISH),and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML.Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients,and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH.The PCR products in some suspected cases were tested by two-directional sequencing.The results showed that except unqualified samples,fusion genes were detected by multiplex RT-PCR in 211 of 474 patients(44.51%),including AML1-ETO,CBFβ-MYH11,PML-RARα,PLZF-RARα,NPM-RARα,MLL rearrangements,BCR-ABL,DEK-CAN,SET-CAN,TEL-PDGFR,TLS-ERG,AML1-MDS1(EVI-1).In 373 patients,who took both multiplex RT-PCR and karyotype analysis,the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373(42.89%) and 179/373(47.98%) respectively,and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373(57.90%).The PCR results from 11 cases 'normal' in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing.It is concluded that karyotype studies remain the cornerstone for genetic testing;conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML,especially for the cryptic or submicroscopic aberrations.Once a genetic marker has been identified by combined analysis,it could be used to monitor residual disease during/after chemotherapy,by quantitative RT-PCR and/or FISH. 展开更多
关键词 acute myeloid leukemia chromosome aberration KARYOTYPING multiplex reverse transcription-polymerase chain reaction fluorescence in situ hybridization
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In advanced gastric cancer:Prognosis and treatment of patients with positive peritoneal cytology 被引量:2
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作者 Francesco Frattini Stefano Rausei +3 位作者 Corrado Chiappa Francesca Rovera Luigi Boni Gianlorenzo Dionigi 《World Journal of Gastrointestinal Surgery》 SCIE CAS 2013年第5期135-137,共3页
Positive peritoneal cytology in gastric cancer is classified as M1 disease by the 7thEdition of American Joint Committee on Cancer staging system.With the introduction of laparoscopy and peritoneal washing cytology in... Positive peritoneal cytology in gastric cancer is classified as M1 disease by the 7thEdition of American Joint Committee on Cancer staging system.With the introduction of laparoscopy and peritoneal washing cytology in the staging of gastric cancer a new category of patients has been identified.These are patients with no macroscopic peritoneal metastases but with peritoneal cytology positive(P0C1).Prognosis and treatment of such patientsrepresent a controversial issue.We evaluate the state of the art of staging system in gastric cancer and discusss tandardisation in staging and treatment procedures.There is still a lack of uniformity in the use of laparoscopy with peritoneal cytology in clinical decision making and in the surgical treatment for gastric cancer.Survival of this patient subset remains poor.Multimodal therapies and new therapeutic strategies are required to improve the survival of these patients. 展开更多
关键词 Advanced gastric cancer PERITONEAL WASHING CYTOLOGY STAGING LAPAROSCOPY Reverse transcription-polymerase chain reaction
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