AIM: To investigate the potential involvement of leptin in carcinogenesis of hepatocellular carcinoma (HCC) and to elucidate the etiology, carcinogenesis and progress of HCC.METHODS: Expressions of Ob gene product, le...AIM: To investigate the potential involvement of leptin in carcinogenesis of hepatocellular carcinoma (HCC) and to elucidate the etiology, carcinogenesis and progress of HCC.METHODS: Expressions of Ob gene product, leptin and its receptor, Ob-R were investigated in 36 cases of HCC spedmens and corresponding adjacent non-tumorous liver tissues with immunohistochemical staining. The effect of leptin on proliferation of Chang liver cell line and liver cancer cell line SMMC-7721 was studied with cell proliferation assay (MTT).RESULTS: Leptin expression was detected in 36 cases of adjacent non-tumorous liver tissues (36/36,100%) with moderate (++) to strong (+++) intensity; and in 72.22%(26/36) of HCC with weaker (+) intensity (P<0.05). Thirty of 36 (83.33%) cases of adjacent non-tumorous liver tissues were positive for Ob-R, with moderate (++) to strong (+++) intensity. In HCC, 11/36 (30.56%) cases were positive, with weak (+) intensity (P<0.05). In cell proliferation assay, leptin inhibited the proliferation of Chang liver cells. The cell survival rate was 10-13% lower than that of the untreated cells (P>0.05). Leptin had little effect on the proliferation of liver cancer cells (/)>0.05).CONCLUSION: High level expression and decreased or absent expression of leptin and its receptor in adjacent non-tumorous liver cells and HCC cells, inhibitory effect of leptin on the proliferation of normal Chang liver cells and no effect of leptin on proliferation of liver cancer cells,may provide new insights into the carcinogenesis and progression of human HCC. It could be assumed that leptin acting as an inhibitor and/or promoter, is involved in the process of carcinogenesis and progress of human HCC.展开更多
AIM: To study the effect of cholecystokinin-octapeptide (CCK-8) on lipopolysaccharide (LPS) -induced pulmonary artery smooth muscle cell (PASMCs) injury and the role of heine oxygenase-1 (HO-1), and to explore the reg...AIM: To study the effect of cholecystokinin-octapeptide (CCK-8) on lipopolysaccharide (LPS) -induced pulmonary artery smooth muscle cell (PASMCs) injury and the role of heine oxygenase-1 (HO-1), and to explore the regulation mechanism of c-Jun N-terminal kinase (JNK) and activator protein-1 (AP-1) signal transduction pathway in inducing HO-1 expression further. METHODS: Cultured PASMCs were randomly divided into 4 or 6 groups: normal culture group, LPS (10 mg/L), CCK-8 (10^-6 mol/L) plus LPS (10 mg/L) group, CCK-8 (10^-6 mol/L) group, zinc protoporphyrin 9 (ZnPPIX) (10^-6 mol/L) plus LPS (10 mg/L) group, CCK-8 (10^-6 mol/L) plus ZnPPIX and LPS (10 mg/L) group. Seven hours after LPS administration, ulterstructrual changes and content of malondialdehyde (MDA) of PASMCs in each group were investigated by electron microscopy and biochemical assay respectively. HO-1 mRNA and protein of PASMCs in the former4 groups were examined by reverse transcriptase polymerase chain reaction (RT-PCR) and immunocytochemistry staining. Changes of c-fos expression and activation of JNK of PASMCs in the former 4 groups were detected with immunocytochemistry staining and Western blot 30 min after LPS administration. RESULTS: The injuries of PASMCs and the increases of MDA content induced by LPS were alleviated and significantly reduced by CCK-8 (P<0.05). The specific HO-1 inhibitor-ZnPPIX could worsen LPS-induced injuries and weaken the protective effect of CCK-8. The expressions of c-fos, p-JNK protein and HO-1 mRNA and protein were all slightly increased in LPS group, and significantly enhanced by CCK-8 further (P<0.05). CONCLUSION: HO-1 may be a key factor in CCK-8 attenuated injuries of PASMCs induced by LPS, and HO-1 expression may be related to the activation of JNK and activator protein (AP-1).展开更多
AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtracti...AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtractive hybridization (SSH) technique and to clone genes associated with its transactivation activity, and to pave the way for elucidating the pathogenesis of hepatitis B virus infection.METHODS: pcDNA3.1(-)-complete S containing full-length HBV S gene was constructed by insertion of HBV complete S gene into BarmH-I/Kpn I sites. HepG2 cells were cotransfected with pcDNA3.1(-)-complete S and pSV-lacZ.After 48 h, cells were collected and detected for the expression of β-galactosidase (β-gal). Suppression subtractive hybridization and bioinformatics techniques were used.The mRNA of HepG2 cells transfected with pcDNA3.1(-)-complete S and pcDNA3.1(-) empty vector was isolated,and detected for the expression of complete S protein by reverse transcription polymerase chain reaction (RT-PCR)method, and cDNA was synthesized. After digestion with restriction enzyme RcaI, cDNA fragments were obtained.Tester cDNA was then divided into two groups and ligated to the specific adaptors 1 and 2, respectively. After tester cDNA had been hybridized with driver cDNA twice and underwent nested PCR twice, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out within E. coli strain DH5α. The cDNA was sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR) amplification.RESULTS: The complete S mRNA could be detected by RT-PCR in HepG2 cells transfected with the pcDNA3.1(-)-complete S. The activity of β-gal in HepG2 cells transfected with the pcDNA3.1(-)-complete S was 6.9 times higher than that of control plasmid. The subtractive library of genes transactivated by HBV complete S protein was constructed successfully. The amplified library contains 86 positive clones. Colony PCR showed that 86 clones contained DNA inserts of 200-1 000 bp, respectively.Sequence analysis was performed in 35 clones randomly,and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 33 coding sequences were obtained. These cDNA sequences might be target genes transactivated by complete S protein of HBV. Moreover, two unknown genes were discovered, full length coding sequences were obtained by bioinformatics techniques,one of them was named complete S transactivated protein 1 (CSTP1) and registered in GenBank (AY553877).CONCLUSION: The complete S gene of HBV has a transactivating effect on SV40 early promoter. A subtractive cDNA library of genes transactivated by HBV complete S protein using SSH technique has been constructed successfully. The obtained sequences may be target genes transactivated by HBV complete S protein among which some genes coding proteins are involved in cell cycle regulation, metabolism, immunity, signal transduction, cell apoptosis and formation mechanism of hepatic carcinoma.展开更多
AIM: To investigate the correlation of enhancement features of hepatocellular carcinoma (HCC) revealed by single-level dynamic spiral CT scanning (DSCT) with tumor microvessel density (NVD), and to determine the valid...AIM: To investigate the correlation of enhancement features of hepatocellular carcinoma (HCC) revealed by single-level dynamic spiral CT scanning (DSCT) with tumor microvessel density (NVD), and to determine the validity of DSCT in assessing in vivo tumor angiogenic activity of HCC. METHODS: Twenty six HCC patients were diagnosed histopathologically. DSCT was performed for all patients according to standard scanning protocol. Time-density curves were generated, relevant curve parameters were measured,and gross enhancement morphology was analyzed. Operation was performed to remove HCC lesions i to 2 weeks following CT scan. Histopathological slides were carefully prepared for the standard F8RA immunohistochemical staining and tumor microvessel counting. Enhancement imaging features of HCC lesions were correlatively studied with tumor MVD and its intra-tumor distribution characteristics. RESULTS: On DSCT images of HCC lesions, three patterns of time-density curve and three types of gross enhancement morphology were recognized. Histomorphologically, the distribution of positively stained tumor endothelial cells within tumor was categorized into 3 types. Curve parameters such as peak enhancement value and contrast enhancement ratio were significantly correlated with tumor tissue MVD (/=0.508 and /=0.423, P<0.01 and P<0.05 respectively). Both the pattern of time-density curve and the gross enhancement morphology of HCC lesions were also correlated with tumor MVD, and reflected the distributive features of tumor microvessels within HCC lesions. Correlation between the likelihood of intrahepatic metastasis of HCC lesions with densely enhanced pseudocapsules and rich pseudocapsular tumor MVD was found.CONCLUSION: Enhancement imaging features of HCC lesions on DSCT scanning are correlated with tumor MVD,and reflect the intra-tumor distribution characteristics of tumor microvessels. DSCT is valuable in assessing the angiogenic activity and tumor neovascularity of HCC patients in vivo.展开更多
AIM: To explore the new target genes transactivated by hepatitis C virus (HCV) core protein and to elucidate the pathogenesis of HCV infection.METHODS: Reverse transcribed cDNA was subjected tomicroarray assay. The co...AIM: To explore the new target genes transactivated by hepatitis C virus (HCV) core protein and to elucidate the pathogenesis of HCV infection.METHODS: Reverse transcribed cDNA was subjected tomicroarray assay. The coding gene transactivated by HCV core protein was cloned and analyzed with bioinformatics methods.RESULTS: The expressive vector of pcDNA3.1(-)-core was constructed and confirmed by restriction enzyme digestion and DNA sequencing and approved correct. mRNA was purified from HepG2 and HepG2 cells transfected with pcDNA3.1(-)-core, respectively. The cDNA derived was subjected to microarray assay. A new gene namedHCTP4 was cloned with molecular biological method in combination with bioinformatics method.CONCLUSION: HCV core is a potential transactivator.Microarray is an efficient and convenient method for analysis of differentially expressed genes.展开更多
AIM: To investigate the transactivating effect of hepatitis C virus (HCV) core protein and to screen genes transactivated by HCV core protein. METHODS: pcDNA3.1(-)-core containing full-length HCV core gene was constru...AIM: To investigate the transactivating effect of hepatitis C virus (HCV) core protein and to screen genes transactivated by HCV core protein. METHODS: pcDNA3.1(-)-core containing full-length HCV core gene was constructed by insertion of HCV core gene into EcoRI/BarnHI site. HepG2 cells were cotransfected with pcDNA3.1(-)-core and pSV-lacZ. After 48 h, cells were collected and detected for the expression of β-gal by an enzyme-linked immunosorbent assay (ELISA) kit. HepG2 cell swere transiently transfected with pcDNA3.1(-)-core using Upofectamine reagent. Cells were collected and total mRNA was isolated. A subtracted cDNA library was generated and constructed into a pGEM-Teasy vector. The library was amplified with E. coil strain JM109. The cDNAs were sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR). RESULTS: The core mRNA and protein could be detected in HepG2 cell lysate which was transfected by the pcDNA3.1(-)-core. The activity of β-galactosidase in HepG2 cells transfected by the pcDNA3.1(-)-core was 5.4 times higher than that of HepG2 cells transfected by control plasmid. The subtractive library of genes transactivated by HCV core protein was constructed successfully. The amplified library contained 233 positive clones. Colony PCR showed that 213 clones contained 100-1 000 bp inserts. Sequence analysis was performed in 63 clones. Six of the sequences were unknown genes. The full length sequences were obtained with bioinformatics method, accepted by Genl3ank. It was suggested that six novel cDNA sequences might be target genes transactivated by HCV core protein. CONCLUSION: The core protein of HCV has transactivating effects on SV40 early promoter/enhancer. A total of 63 clones from cDNA library were randomly chosen and sequenced. Using the BLAST program at the National Center for Biotechnology Information, six of the sequences were unknown genes. The other 57 sequences were highly similar to known genes.展开更多
A modified chitosan adsorbent was synthesized through a simple preparation procedure, and it demonstrated good adsorption performance for selective removal of low density lipoprotein in human plasma. Phase inversion ...A modified chitosan adsorbent was synthesized through a simple preparation procedure, and it demonstrated good adsorption performance for selective removal of low density lipoprotein in human plasma. Phase inversion technique was employed to form chitosan beads, to which epoxy groups were then introduced by reacting with ethyleneglycol diglycidylether, and tryptophan was subsequently coupled to the epoxy-activated beads.展开更多
AIM: To clone, identify and study new NSSATP2 gene and its spliced variant transactivated by hepatitis C virus nonstructural protein 5A. METHODS: On the basis of subtractive cDNA library of genes transactivated by NS5...AIM: To clone, identify and study new NSSATP2 gene and its spliced variant transactivated by hepatitis C virus nonstructural protein 5A. METHODS: On the basis of subtractive cDNA library of genes transactivated by NS5A protein of hepatitis C virus, the coding sequence of new gene and its spliced variant were obtained by bioinformatics method. Polymerase chain reaction (PCR) was conducted to amplify/VSSATP2 gene. RESULTS: The coding sequence of a new gene and its spliced variant were cloned and identified successfully. CONCLUSION: A new gene has been recognized as the new target transactivated by HCV NS5A protein. These results brought some new clues for studying the biological functions of new genes and pathogenesis of the viral proteins.展开更多
AIM: To investigate the biological function of complete S protein and to look for proteins interacting with complete S protein in hepatocytes.METHODS: We constructed bait plasmid expressing complete S protein of HBV b...AIM: To investigate the biological function of complete S protein and to look for proteins interacting with complete S protein in hepatocytes.METHODS: We constructed bait plasmid expressing complete S protein of HBV by cloning the gene of complete S protein into pGBKT7, then the recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After extracting and sequencing of plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics.RESULTS: Nineteen colonies were selected and sequenced.Among them, five colonies were Homo sapiens solute carrier family 25, member 23 (SLC25A23), one was Homo sapiens calreticulin, one was human serum albumin (ALB)gene, one was Homo sapiens metallothionein 2A, two were Homo sapiens betaine-homocysteine methyltransferase,three were Homo sapiensNa+ and H+ coupled amino acid transport system N, one was Homo sapiens CD81 antigen (target of anti-proliferative antibody 1) (CD81), three were Homo sapiens diazepam binding inhibitor, two colonies were new genes with unknown function.CONCLUSION: The yeast-two hybrid system is an effective method for identifying hepatocyte proteins interacting with complete S protein of HBV. The complete S protein may bind to different proteins i.e., its multiple functions in vivo.展开更多
A silica-supported zirconium based solid acid (ZS) has been used as catalyst for the Fries rearrangement of phenyl acetate (PA). The catalyst showed a higher PA conversion activity and a much higher selectivity for o...A silica-supported zirconium based solid acid (ZS) has been used as catalyst for the Fries rearrangement of phenyl acetate (PA). The catalyst showed a higher PA conversion activity and a much higher selectivity for o-hydroxyacetophenone (o-HAP) than for strongly acidic zeolite catalysts. The supported catalyst was characterized by XRD, IR, XPS, pyridine-TPD and the surface area measurements. The catalytic properties were influenced significantly by pretreatment temperature.展开更多
In this paper, the properties of carbon deposited on hexaaluminateLaNiAl_(11)O_(19) catalyst were characterized by X-ray photoelectron spectroscopy (XPS), and in themeantime, the amount of carbon deposited on the cata...In this paper, the properties of carbon deposited on hexaaluminateLaNiAl_(11)O_(19) catalyst were characterized by X-ray photoelectron spectroscopy (XPS), and in themeantime, the amount of carbon deposited on the catalyst, after both CH_4 decomposition and CO_2reforming of CH_4, was determined by means of thermogravimetric analysis (TGA), respectively. Therates of carbon deposited on the catalyst were also investigated and the apparent kinetic equationof CO_2 reforming of CH_4: ν_c = kp^(0.72)(CH_4)·p^(-0.55)(CO_2), was established by analyzing therelation between the rates of deposited carbon and the pressure ratio of CH_4 and CO_2.展开更多
A graph G is called a (g, f)-uniform graph if for each edge of G, there is a(g, f)-factor containing it and another (g, f)-factor excluding it. In this paper a necessary andsufficient condition for a graph to be a (g,...A graph G is called a (g, f)-uniform graph if for each edge of G, there is a(g, f)-factor containing it and another (g, f)-factor excluding it. In this paper a necessary andsufficient condition for a graph to be a (g, f)-uniform graph is given and some applications of thiscondition are discussed. In particular, some simple sufficient conditions for a graph to be an [a,b]-uniform graph are obtained for a b.展开更多
基金Supported by the Grants From Sasakawa Medical Foundation of International Cooperation Department of Ministry of Public Health of China(054)and Science Foundation of West China University of Medical Sciences(L99016)
文摘AIM: To investigate the potential involvement of leptin in carcinogenesis of hepatocellular carcinoma (HCC) and to elucidate the etiology, carcinogenesis and progress of HCC.METHODS: Expressions of Ob gene product, leptin and its receptor, Ob-R were investigated in 36 cases of HCC spedmens and corresponding adjacent non-tumorous liver tissues with immunohistochemical staining. The effect of leptin on proliferation of Chang liver cell line and liver cancer cell line SMMC-7721 was studied with cell proliferation assay (MTT).RESULTS: Leptin expression was detected in 36 cases of adjacent non-tumorous liver tissues (36/36,100%) with moderate (++) to strong (+++) intensity; and in 72.22%(26/36) of HCC with weaker (+) intensity (P<0.05). Thirty of 36 (83.33%) cases of adjacent non-tumorous liver tissues were positive for Ob-R, with moderate (++) to strong (+++) intensity. In HCC, 11/36 (30.56%) cases were positive, with weak (+) intensity (P<0.05). In cell proliferation assay, leptin inhibited the proliferation of Chang liver cells. The cell survival rate was 10-13% lower than that of the untreated cells (P>0.05). Leptin had little effect on the proliferation of liver cancer cells (/)>0.05).CONCLUSION: High level expression and decreased or absent expression of leptin and its receptor in adjacent non-tumorous liver cells and HCC cells, inhibitory effect of leptin on the proliferation of normal Chang liver cells and no effect of leptin on proliferation of liver cancer cells,may provide new insights into the carcinogenesis and progression of human HCC. It could be assumed that leptin acting as an inhibitor and/or promoter, is involved in the process of carcinogenesis and progress of human HCC.
文摘AIM: To study the effect of cholecystokinin-octapeptide (CCK-8) on lipopolysaccharide (LPS) -induced pulmonary artery smooth muscle cell (PASMCs) injury and the role of heine oxygenase-1 (HO-1), and to explore the regulation mechanism of c-Jun N-terminal kinase (JNK) and activator protein-1 (AP-1) signal transduction pathway in inducing HO-1 expression further. METHODS: Cultured PASMCs were randomly divided into 4 or 6 groups: normal culture group, LPS (10 mg/L), CCK-8 (10^-6 mol/L) plus LPS (10 mg/L) group, CCK-8 (10^-6 mol/L) group, zinc protoporphyrin 9 (ZnPPIX) (10^-6 mol/L) plus LPS (10 mg/L) group, CCK-8 (10^-6 mol/L) plus ZnPPIX and LPS (10 mg/L) group. Seven hours after LPS administration, ulterstructrual changes and content of malondialdehyde (MDA) of PASMCs in each group were investigated by electron microscopy and biochemical assay respectively. HO-1 mRNA and protein of PASMCs in the former4 groups were examined by reverse transcriptase polymerase chain reaction (RT-PCR) and immunocytochemistry staining. Changes of c-fos expression and activation of JNK of PASMCs in the former 4 groups were detected with immunocytochemistry staining and Western blot 30 min after LPS administration. RESULTS: The injuries of PASMCs and the increases of MDA content induced by LPS were alleviated and significantly reduced by CCK-8 (P<0.05). The specific HO-1 inhibitor-ZnPPIX could worsen LPS-induced injuries and weaken the protective effect of CCK-8. The expressions of c-fos, p-JNK protein and HO-1 mRNA and protein were all slightly increased in LPS group, and significantly enhanced by CCK-8 further (P<0.05). CONCLUSION: HO-1 may be a key factor in CCK-8 attenuated injuries of PASMCs induced by LPS, and HO-1 expression may be related to the activation of JNK and activator protein (AP-1).
基金Supported by the National Natural Science Foundation of China, No. C03011402, No. C30070690 the Science and Technique Foundation of PLA during the 9th Five-year Plan period, No. 98D063the Launching Foundation for Students Studying Abroad of PLA, No. 98H038the Youth Science and Technique Foundation of PLA during the 10th Five-year plan period, No. 01Q138the Science and Technique Foundation of PLA during the 10th Five-year Plan period, No. 01MB135
文摘AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtractive hybridization (SSH) technique and to clone genes associated with its transactivation activity, and to pave the way for elucidating the pathogenesis of hepatitis B virus infection.METHODS: pcDNA3.1(-)-complete S containing full-length HBV S gene was constructed by insertion of HBV complete S gene into BarmH-I/Kpn I sites. HepG2 cells were cotransfected with pcDNA3.1(-)-complete S and pSV-lacZ.After 48 h, cells were collected and detected for the expression of β-galactosidase (β-gal). Suppression subtractive hybridization and bioinformatics techniques were used.The mRNA of HepG2 cells transfected with pcDNA3.1(-)-complete S and pcDNA3.1(-) empty vector was isolated,and detected for the expression of complete S protein by reverse transcription polymerase chain reaction (RT-PCR)method, and cDNA was synthesized. After digestion with restriction enzyme RcaI, cDNA fragments were obtained.Tester cDNA was then divided into two groups and ligated to the specific adaptors 1 and 2, respectively. After tester cDNA had been hybridized with driver cDNA twice and underwent nested PCR twice, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out within E. coli strain DH5α. The cDNA was sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR) amplification.RESULTS: The complete S mRNA could be detected by RT-PCR in HepG2 cells transfected with the pcDNA3.1(-)-complete S. The activity of β-gal in HepG2 cells transfected with the pcDNA3.1(-)-complete S was 6.9 times higher than that of control plasmid. The subtractive library of genes transactivated by HBV complete S protein was constructed successfully. The amplified library contains 86 positive clones. Colony PCR showed that 86 clones contained DNA inserts of 200-1 000 bp, respectively.Sequence analysis was performed in 35 clones randomly,and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 33 coding sequences were obtained. These cDNA sequences might be target genes transactivated by complete S protein of HBV. Moreover, two unknown genes were discovered, full length coding sequences were obtained by bioinformatics techniques,one of them was named complete S transactivated protein 1 (CSTP1) and registered in GenBank (AY553877).CONCLUSION: The complete S gene of HBV has a transactivating effect on SV40 early promoter. A subtractive cDNA library of genes transactivated by HBV complete S protein using SSH technique has been constructed successfully. The obtained sequences may be target genes transactivated by HBV complete S protein among which some genes coding proteins are involved in cell cycle regulation, metabolism, immunity, signal transduction, cell apoptosis and formation mechanism of hepatic carcinoma.
文摘AIM: To investigate the correlation of enhancement features of hepatocellular carcinoma (HCC) revealed by single-level dynamic spiral CT scanning (DSCT) with tumor microvessel density (NVD), and to determine the validity of DSCT in assessing in vivo tumor angiogenic activity of HCC. METHODS: Twenty six HCC patients were diagnosed histopathologically. DSCT was performed for all patients according to standard scanning protocol. Time-density curves were generated, relevant curve parameters were measured,and gross enhancement morphology was analyzed. Operation was performed to remove HCC lesions i to 2 weeks following CT scan. Histopathological slides were carefully prepared for the standard F8RA immunohistochemical staining and tumor microvessel counting. Enhancement imaging features of HCC lesions were correlatively studied with tumor MVD and its intra-tumor distribution characteristics. RESULTS: On DSCT images of HCC lesions, three patterns of time-density curve and three types of gross enhancement morphology were recognized. Histomorphologically, the distribution of positively stained tumor endothelial cells within tumor was categorized into 3 types. Curve parameters such as peak enhancement value and contrast enhancement ratio were significantly correlated with tumor tissue MVD (/=0.508 and /=0.423, P<0.01 and P<0.05 respectively). Both the pattern of time-density curve and the gross enhancement morphology of HCC lesions were also correlated with tumor MVD, and reflected the distributive features of tumor microvessels within HCC lesions. Correlation between the likelihood of intrahepatic metastasis of HCC lesions with densely enhanced pseudocapsules and rich pseudocapsular tumor MVD was found.CONCLUSION: Enhancement imaging features of HCC lesions on DSCT scanning are correlated with tumor MVD,and reflect the intra-tumor distribution characteristics of tumor microvessels. DSCT is valuable in assessing the angiogenic activity and tumor neovascularity of HCC patients in vivo.
基金Supported by the National Natural Science Foundation of China, No. 39970674
文摘AIM: To explore the new target genes transactivated by hepatitis C virus (HCV) core protein and to elucidate the pathogenesis of HCV infection.METHODS: Reverse transcribed cDNA was subjected tomicroarray assay. The coding gene transactivated by HCV core protein was cloned and analyzed with bioinformatics methods.RESULTS: The expressive vector of pcDNA3.1(-)-core was constructed and confirmed by restriction enzyme digestion and DNA sequencing and approved correct. mRNA was purified from HepG2 and HepG2 cells transfected with pcDNA3.1(-)-core, respectively. The cDNA derived was subjected to microarray assay. A new gene namedHCTP4 was cloned with molecular biological method in combination with bioinformatics method.CONCLUSION: HCV core is a potential transactivator.Microarray is an efficient and convenient method for analysis of differentially expressed genes.
基金Supported by the National Natural Science Foundation of China,No.39970674
文摘AIM: To investigate the transactivating effect of hepatitis C virus (HCV) core protein and to screen genes transactivated by HCV core protein. METHODS: pcDNA3.1(-)-core containing full-length HCV core gene was constructed by insertion of HCV core gene into EcoRI/BarnHI site. HepG2 cells were cotransfected with pcDNA3.1(-)-core and pSV-lacZ. After 48 h, cells were collected and detected for the expression of β-gal by an enzyme-linked immunosorbent assay (ELISA) kit. HepG2 cell swere transiently transfected with pcDNA3.1(-)-core using Upofectamine reagent. Cells were collected and total mRNA was isolated. A subtracted cDNA library was generated and constructed into a pGEM-Teasy vector. The library was amplified with E. coil strain JM109. The cDNAs were sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR). RESULTS: The core mRNA and protein could be detected in HepG2 cell lysate which was transfected by the pcDNA3.1(-)-core. The activity of β-galactosidase in HepG2 cells transfected by the pcDNA3.1(-)-core was 5.4 times higher than that of HepG2 cells transfected by control plasmid. The subtractive library of genes transactivated by HCV core protein was constructed successfully. The amplified library contained 233 positive clones. Colony PCR showed that 213 clones contained 100-1 000 bp inserts. Sequence analysis was performed in 63 clones. Six of the sequences were unknown genes. The full length sequences were obtained with bioinformatics method, accepted by Genl3ank. It was suggested that six novel cDNA sequences might be target genes transactivated by HCV core protein. CONCLUSION: The core protein of HCV has transactivating effects on SV40 early promoter/enhancer. A total of 63 clones from cDNA library were randomly chosen and sequenced. Using the BLAST program at the National Center for Biotechnology Information, six of the sequences were unknown genes. The other 57 sequences were highly similar to known genes.
文摘A modified chitosan adsorbent was synthesized through a simple preparation procedure, and it demonstrated good adsorption performance for selective removal of low density lipoprotein in human plasma. Phase inversion technique was employed to form chitosan beads, to which epoxy groups were then introduced by reacting with ethyleneglycol diglycidylether, and tryptophan was subsequently coupled to the epoxy-activated beads.
基金Supported by the National Natural Science Foundation of China,No.C03011402,No.C30070690 and the 9th Five-year Plan Period Research and Development Foundation of PLA,No.98D063 and the Start-up for Students Studying Overseas of PLA,No.98H038 and the 10th F
文摘AIM: To clone, identify and study new NSSATP2 gene and its spliced variant transactivated by hepatitis C virus nonstructural protein 5A. METHODS: On the basis of subtractive cDNA library of genes transactivated by NS5A protein of hepatitis C virus, the coding sequence of new gene and its spliced variant were obtained by bioinformatics method. Polymerase chain reaction (PCR) was conducted to amplify/VSSATP2 gene. RESULTS: The coding sequence of a new gene and its spliced variant were cloned and identified successfully. CONCLUSION: A new gene has been recognized as the new target transactivated by HCV NS5A protein. These results brought some new clues for studying the biological functions of new genes and pathogenesis of the viral proteins.
基金Supported by the National Natural Science Foundation of China, No. C03011402, No. C30070690the Science and Technique Foundation of PLA during the 9th Five-year plan period, No. 98D063 the Launching Foundation for Students Studying Abroad of PLA, No. 98H038 the Youth Science and Technique Foundation of PLA during the 10lh Five-year plan period, No. 01Q138and the Science & Technique Foundation of PLA during the 10th Five-year plan period, No. 01MB135
文摘AIM: To investigate the biological function of complete S protein and to look for proteins interacting with complete S protein in hepatocytes.METHODS: We constructed bait plasmid expressing complete S protein of HBV by cloning the gene of complete S protein into pGBKT7, then the recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After extracting and sequencing of plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics.RESULTS: Nineteen colonies were selected and sequenced.Among them, five colonies were Homo sapiens solute carrier family 25, member 23 (SLC25A23), one was Homo sapiens calreticulin, one was human serum albumin (ALB)gene, one was Homo sapiens metallothionein 2A, two were Homo sapiens betaine-homocysteine methyltransferase,three were Homo sapiensNa+ and H+ coupled amino acid transport system N, one was Homo sapiens CD81 antigen (target of anti-proliferative antibody 1) (CD81), three were Homo sapiens diazepam binding inhibitor, two colonies were new genes with unknown function.CONCLUSION: The yeast-two hybrid system is an effective method for identifying hepatocyte proteins interacting with complete S protein of HBV. The complete S protein may bind to different proteins i.e., its multiple functions in vivo.
文摘A silica-supported zirconium based solid acid (ZS) has been used as catalyst for the Fries rearrangement of phenyl acetate (PA). The catalyst showed a higher PA conversion activity and a much higher selectivity for o-hydroxyacetophenone (o-HAP) than for strongly acidic zeolite catalysts. The supported catalyst was characterized by XRD, IR, XPS, pyridine-TPD and the surface area measurements. The catalytic properties were influenced significantly by pretreatment temperature.
文摘In this paper, the properties of carbon deposited on hexaaluminateLaNiAl_(11)O_(19) catalyst were characterized by X-ray photoelectron spectroscopy (XPS), and in themeantime, the amount of carbon deposited on the catalyst, after both CH_4 decomposition and CO_2reforming of CH_4, was determined by means of thermogravimetric analysis (TGA), respectively. Therates of carbon deposited on the catalyst were also investigated and the apparent kinetic equationof CO_2 reforming of CH_4: ν_c = kp^(0.72)(CH_4)·p^(-0.55)(CO_2), was established by analyzing therelation between the rates of deposited carbon and the pressure ratio of CH_4 and CO_2.
基金Financial supports from the Natural Science Foundation of Shandong Province(Y2000B09)Shandong Provincial Science and Technology Program(03C05)are gratefully acknowledged
基金Supported by National Natural Science Foundation (10471078, 10201019) and RSDP (20040422004) of China
文摘A graph G is called a (g, f)-uniform graph if for each edge of G, there is a(g, f)-factor containing it and another (g, f)-factor excluding it. In this paper a necessary andsufficient condition for a graph to be a (g, f)-uniform graph is given and some applications of thiscondition are discussed. In particular, some simple sufficient conditions for a graph to be an [a,b]-uniform graph are obtained for a b.