AIM: To analyze the relationship between genetic polymorphisms of metabolizing enzymes CYP2E1, GSTM1 andKazakh's esophageal squamous cell cancer in China.METHODS: The genotypes of cytochromes P450 (CYP) 2E1 and gl...AIM: To analyze the relationship between genetic polymorphisms of metabolizing enzymes CYP2E1, GSTM1 andKazakh's esophageal squamous cell cancer in China.METHODS: The genotypes of cytochromes P450 (CYP) 2E1 and glutathione S-transferase (GST) M1 were investigated by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) following PCR in 104 Kazakh's patients with esophageal cancer (EC) and 104 non-cancer controls.RESULTS: The frequency of CYP2E1 c1/c1 genotype was significantly higher in patients with cancer (77.9%) thanin control subjects (24.0%) (P＜0.05; OR, 11.13; 95%CI,5.84-21.22). The difference of GSTM1 null was significantly more frequent in the cancer (34.6%) vsthe control group (3.8%) (P＜0.05; OR, 13.24; 95%CI, 4.50-38.89). On the other hand, the combination of GSTM1 presence and CYP2E1 c1/c1 genotypes increased the risk for cancer (P＜0.05;OR, 13.42; 95%CI, 6.29-28.3).CONCLUSION: The CYP2E1 c1/c1, GSTM1 deletion genotypes are genetically susceptible biomarkers for ESCC in Kazakh population. Individuals with allele c1 of RsaI polymorphic locus for CYP2E1 may increase the risk of ESCC. Moreover, CYP2E1 wild type (c1/c1) increased thesusceptibility to ESCC risk in Kazakh individuals with GSTM1 presence genotype.展开更多
AIM: To investigate the effects and molecular mechanismsof recombinant human growth hormone (rhGH) onprotecting liver function and alleviating portal hypertensionof liver cirrhotic rats.METHODS: Liver cirrhosis of mal...AIM: To investigate the effects and molecular mechanismsof recombinant human growth hormone (rhGH) onprotecting liver function and alleviating portal hypertensionof liver cirrhotic rats.METHODS: Liver cirrhosis of male Sprague-Dawley ratswas induced by administration of thioacetamide. The ratswith or without liver cirrhosis were randomly divided intofour groups. Group A consisted of the normal rats wastreated with normal saline (NS), group B consisted of thenormal rats was treated with rhGH, group C consisted ofcirrhotic rats was treated with NS, and group D consistedof cirrhotic rats was treated with rhGH. The rats of differentgroups were subcutaneously injected with 0.5 mL of NS or333 ng/kg of rhGH daily for 7 d. After treatments, thefollowing parameters were examined, including GH-bindingcapacity (RO by 12SI-hGH binding, growth hormone receptormRNA(GHR mRNA) expression by RT-PCR, relative contentof collagen (RCC) by histomorphomertry, and level ofmalon-dialdehyde (MDA) and superoxide dismutase (SOD)in liver tissue by thiobarbituric acid reaction and pyrogallicacid self-oxidation, respectively. Serum albumin (ALB),alanine transaminase (ALT) and portal vein pressure (PVP)were also examined.RESULTS: rhGH up-regulated both the GH-binding capacity(R0 and the expression of GHR mRNA in vivo. RT in groupA (72+12 fmol/mg protein) was significantly higher thanthat in group C (31+4 fmol/mg protein) (P<0.05). RT ingroup B (80+9 fmol/mg protein) increased markedlycompared to group A (P<0.05). RT in group D (40+7 fmol/mgprotein) raised remarkably compared with group C (P<0.05),but less than that in group A, and there was no significantGH binding affinity contrast (Kd) change. The GHR mRNAlevel (iOD, pixel) in group A (29+3) was significantly higherthan that in group C (23+3) (P<0.05). GHR mRNA levelswere significantly raised in group B (56+4) and group D(42+8) compared with groups A and C (29+3 and 23+3,respectively) (P<0.05). Compared with the normal liver,MDA level was higher and SOD level was lower in cirrhoticlivers. After rhGH treatment, MDA level was significantlydeclined to 12.0+2.2 nmol/mg protein and SOD was raisedto 1029+76 U/rag protein in group D (P<0.05). ALB levelsin groups B and D (42+7 g/L and 37+7 g/L, respectively)were significantly raised compared with those in groups Aand C (35+5 g/L and 29+4 g/L, respectively) (P<0.05).ALT level was markedly lower in group D (69+7 U/L)compared to group C (89+15 U/L) (P<0.05), and close togroup A (61+10 U/L). RCC in group C (22.30+3.86%) wassignificantly higher than that in group A (1.14+0.21%) andgroup D (14.70+2.07%) (P<0.05). In addition, rhGHmarkedly alleviated portal hypertension in liver cirrhoticrats (group D vsC, 9.3+1.5 cmH20 vs 14.4+2.0 cmH20)(P<0.05).CONCLUSION: Pharmacological doses of rhGH canincrease RT and GHR mRNA expression, ameliorate liverfunctions, repress fibrosis and decline portal hypertension,suggesting it has potentially clinical usage as a hepatotropicfactor.展开更多
AIM: To investigate the pathological effect of Helicobacter pylori (H pylori) on human hepatic cells, proteomic methods were used to find and to identify proteins that were overexpressed in HepG2 cells treated by H py...AIM: To investigate the pathological effect of Helicobacter pylori (H pylori) on human hepatic cells, proteomic methods were used to find and to identify proteins that were overexpressed in HepG2 cells treated by H pylori. METHODS: H pylori was co-cultured with HepG2 for 6 h. Two-dimensional gel electrophoresis was used to gain the protein expression pattern of untreated and H pylori-treated HepG2. After staining and image analysis, spots of interest were isolated and subjected to mass spectrometry. RESULTS: Seven proteins, which were up-regulated in H pylori -treated HepG2 cells, were identified. These proteins included integrin beta-1, protein kinase C alpha, LIM/ homeobox protein Lhx1, eIF-2-beta, MAP kinase kinase 3, PINCH protein and Ras-related protein Rab-37, which involved in transcription regulation, signal transduction, metabolism and so on, CONCLUSION: H pylori may exert the pathological effect on HepG2 cells by up-regulating the expression of some proteins.展开更多
Objective: To explore the correlation between structure domains and functions of chemokine receptor CXCR4. Methods: After the establishment of wild type chemokine.receptor CXCR4 and CXCR2 expressing cell lines, 5 CXCR...Objective: To explore the correlation between structure domains and functions of chemokine receptor CXCR4. Methods: After the establishment of wild type chemokine.receptor CXCR4 and CXCR2 expressing cell lines, 5 CXCR4/CXCR2 chimeras, 2 CXCR4 mutants were stably expressed on CHO cell line. Binding activities of all variants with the ligand, recombinant human SDF-1β, signal transduction ability after stimulation and their function as coreceptor for HIV-1 were studied with ligand-binding assay, Cytosensor/ microphysiometry and cell-cell reporter gene fusion assay. Results: Among all 7 changed CXCR4 receptors, 3 chimeras (2444a, 4442, 4122), and 1 mutant (CXCR4-Tr) bond with SDF-1β in varying degrees, of which only 2444a totally and CXCR4-Tr partially maintain signaling. All changed receptors except for 4222 could act as coreceptors for HIV-1 (LAI) in varying degrees. Conclusion: Several structure domains of CXCR4 are involved in the binding with SDF-1β, among which, N-terminal extracellular domain has high affinity of binding with SDF-1β, and the 3rd extracellular loop contributes to the binding, too. Although the C-terminal in-tracellular domain has no association with the maintenance of the overall structure of the receptor and ligand binding capability, the signaling is decreased when this domain is truncated. For CXCR4 signaling, not only is the conserved motif DRY box needed, but also the characterized conformation of the whole molecule must be formed when activation is required. There are some overlaps between SDF-1β binding domains and coreceptor function domains in molecular structure of CXCR4.展开更多
Differential proteome profiles of human lung squamous carcinoma tissue compared to paired tumor-adjacent normal bronchial epithelial tissue were established and analyzed by means of immobilized pH gradient-based two-d...Differential proteome profiles of human lung squamous carcinoma tissue compared to paired tumor-adjacent normal bronchial epithelial tissue were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and matrix-assisted laser desorp-tion/ionization time of flight mass spectrometry (MALDI-TOF-MS). The results showed that well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were obtained under the condition of 0.75-mg protein-load. The average deviation of spot position was 0.733±0.101 mm in IEF direction, and 0.925±0.207 mm in SDS-PAGE direction. For tumor tissue, a total of 1241±88 spots were detected, 987±65 spots were matched with an average matching rate of 79.5%. For control, a total of 1190±72 spots were detected, and 875±48 spots were matched with an average matching rate of 73.5%. A total of 864±34 spots were matched between tumors and controls. Forty-three differential proteins were characterized: some proteins were related to oncogenes, and others involved in the regulation of cell cycle and signal transduc-tion. It is suggested that the differential proteomic approach is valuable for mass identification of differentially expressed proteins involved in lung carcinogenesis. These data will be used to establish human lung cancer proteome database to further study human lung squamous carcinoma.展开更多
Lipopolysaccharide(LPS),the principal component of the outer membrane of Gram-negative bacteria,stimulates various cell types to release numerous proinflammatory mediators such as TNF-α,IL-6 and IL-12,which may damag...Lipopolysaccharide(LPS),the principal component of the outer membrane of Gram-negative bacteria,stimulates various cell types to release numerous proinflammatory mediators such as TNF-α,IL-6 and IL-12,which may damage cells and lead to organ injury,even sepsis and septic shock.Toll-like receptor 4(TLR4) has been identified as the receptor involved in the recognition of LPS,but the role of LPS uptake in activating signal transduction remains controversial.In the present study,TNF-α was used as a marker of macrophages/ monocytes activated by LPS,and CQ was used as an inhibitor of endosome mature in order to definitude what stage of the signal transduction elicited by LPS was interrupted.We found that there indeed existed internalization of LPS and internalization partially participated in LPS signaling since CQ inhibited cytokine release,and decreased accumulation of FITC-LPS in hPBMCs.In contrast,anti-hTLR4 antibody could decrease cytokine release,but had no inhibition on accumulation of FITC-LPS.This result revealed that inhibition of cytokine release was related to reduction of FITC-LPS accumulation in the cells.But TLR4 on the cell surface couldn't participate in internalization of LPS.Thus,LPS signaling and internalization couldn't be viewed as mutually independent processes.Cellular & Molecular Immunology.2004;1(5):373-377.展开更多
基金Supported by the Xinjiang Key Lab Fund, XJDX0202-2003-05
文摘AIM: To analyze the relationship between genetic polymorphisms of metabolizing enzymes CYP2E1, GSTM1 andKazakh's esophageal squamous cell cancer in China.METHODS: The genotypes of cytochromes P450 (CYP) 2E1 and glutathione S-transferase (GST) M1 were investigated by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) following PCR in 104 Kazakh's patients with esophageal cancer (EC) and 104 non-cancer controls.RESULTS: The frequency of CYP2E1 c1/c1 genotype was significantly higher in patients with cancer (77.9%) thanin control subjects (24.0%) (P＜0.05; OR, 11.13; 95%CI,5.84-21.22). The difference of GSTM1 null was significantly more frequent in the cancer (34.6%) vsthe control group (3.8%) (P＜0.05; OR, 13.24; 95%CI, 4.50-38.89). On the other hand, the combination of GSTM1 presence and CYP2E1 c1/c1 genotypes increased the risk for cancer (P＜0.05;OR, 13.42; 95%CI, 6.29-28.3).CONCLUSION: The CYP2E1 c1/c1, GSTM1 deletion genotypes are genetically susceptible biomarkers for ESCC in Kazakh population. Individuals with allele c1 of RsaI polymorphic locus for CYP2E1 may increase the risk of ESCC. Moreover, CYP2E1 wild type (c1/c1) increased thesusceptibility to ESCC risk in Kazakh individuals with GSTM1 presence genotype.
基金Supported by the Natural Science Foundation of Guangdong Province,No.984213 and Academic Foundation of Sun Yat -Sen University and Ministry of Public Health for Project 211,No.F000099075
文摘AIM: To investigate the effects and molecular mechanismsof recombinant human growth hormone (rhGH) onprotecting liver function and alleviating portal hypertensionof liver cirrhotic rats.METHODS: Liver cirrhosis of male Sprague-Dawley ratswas induced by administration of thioacetamide. The ratswith or without liver cirrhosis were randomly divided intofour groups. Group A consisted of the normal rats wastreated with normal saline (NS), group B consisted of thenormal rats was treated with rhGH, group C consisted ofcirrhotic rats was treated with NS, and group D consistedof cirrhotic rats was treated with rhGH. The rats of differentgroups were subcutaneously injected with 0.5 mL of NS or333 ng/kg of rhGH daily for 7 d. After treatments, thefollowing parameters were examined, including GH-bindingcapacity (RO by 12SI-hGH binding, growth hormone receptormRNA(GHR mRNA) expression by RT-PCR, relative contentof collagen (RCC) by histomorphomertry, and level ofmalon-dialdehyde (MDA) and superoxide dismutase (SOD)in liver tissue by thiobarbituric acid reaction and pyrogallicacid self-oxidation, respectively. Serum albumin (ALB),alanine transaminase (ALT) and portal vein pressure (PVP)were also examined.RESULTS: rhGH up-regulated both the GH-binding capacity(R0 and the expression of GHR mRNA in vivo. RT in groupA (72+12 fmol/mg protein) was significantly higher thanthat in group C (31+4 fmol/mg protein) (P<0.05). RT ingroup B (80+9 fmol/mg protein) increased markedlycompared to group A (P<0.05). RT in group D (40+7 fmol/mgprotein) raised remarkably compared with group C (P<0.05),but less than that in group A, and there was no significantGH binding affinity contrast (Kd) change. The GHR mRNAlevel (iOD, pixel) in group A (29+3) was significantly higherthan that in group C (23+3) (P<0.05). GHR mRNA levelswere significantly raised in group B (56+4) and group D(42+8) compared with groups A and C (29+3 and 23+3,respectively) (P<0.05). Compared with the normal liver,MDA level was higher and SOD level was lower in cirrhoticlivers. After rhGH treatment, MDA level was significantlydeclined to 12.0+2.2 nmol/mg protein and SOD was raisedto 1029+76 U/rag protein in group D (P<0.05). ALB levelsin groups B and D (42+7 g/L and 37+7 g/L, respectively)were significantly raised compared with those in groups Aand C (35+5 g/L and 29+4 g/L, respectively) (P<0.05).ALT level was markedly lower in group D (69+7 U/L)compared to group C (89+15 U/L) (P<0.05), and close togroup A (61+10 U/L). RCC in group C (22.30+3.86%) wassignificantly higher than that in group A (1.14+0.21%) andgroup D (14.70+2.07%) (P<0.05). In addition, rhGHmarkedly alleviated portal hypertension in liver cirrhoticrats (group D vsC, 9.3+1.5 cmH20 vs 14.4+2.0 cmH20)(P<0.05).CONCLUSION: Pharmacological doses of rhGH canincrease RT and GHR mRNA expression, ameliorate liverfunctions, repress fibrosis and decline portal hypertension,suggesting it has potentially clinical usage as a hepatotropicfactor.
文摘AIM: To investigate the pathological effect of Helicobacter pylori (H pylori) on human hepatic cells, proteomic methods were used to find and to identify proteins that were overexpressed in HepG2 cells treated by H pylori. METHODS: H pylori was co-cultured with HepG2 for 6 h. Two-dimensional gel electrophoresis was used to gain the protein expression pattern of untreated and H pylori-treated HepG2. After staining and image analysis, spots of interest were isolated and subjected to mass spectrometry. RESULTS: Seven proteins, which were up-regulated in H pylori -treated HepG2 cells, were identified. These proteins included integrin beta-1, protein kinase C alpha, LIM/ homeobox protein Lhx1, eIF-2-beta, MAP kinase kinase 3, PINCH protein and Ras-related protein Rab-37, which involved in transcription regulation, signal transduction, metabolism and so on, CONCLUSION: H pylori may exert the pathological effect on HepG2 cells by up-regulating the expression of some proteins.
文摘Objective: To explore the correlation between structure domains and functions of chemokine receptor CXCR4. Methods: After the establishment of wild type chemokine.receptor CXCR4 and CXCR2 expressing cell lines, 5 CXCR4/CXCR2 chimeras, 2 CXCR4 mutants were stably expressed on CHO cell line. Binding activities of all variants with the ligand, recombinant human SDF-1β, signal transduction ability after stimulation and their function as coreceptor for HIV-1 were studied with ligand-binding assay, Cytosensor/ microphysiometry and cell-cell reporter gene fusion assay. Results: Among all 7 changed CXCR4 receptors, 3 chimeras (2444a, 4442, 4122), and 1 mutant (CXCR4-Tr) bond with SDF-1β in varying degrees, of which only 2444a totally and CXCR4-Tr partially maintain signaling. All changed receptors except for 4222 could act as coreceptors for HIV-1 (LAI) in varying degrees. Conclusion: Several structure domains of CXCR4 are involved in the binding with SDF-1β, among which, N-terminal extracellular domain has high affinity of binding with SDF-1β, and the 3rd extracellular loop contributes to the binding, too. Although the C-terminal in-tracellular domain has no association with the maintenance of the overall structure of the receptor and ligand binding capability, the signaling is decreased when this domain is truncated. For CXCR4 signaling, not only is the conserved motif DRY box needed, but also the characterized conformation of the whole molecule must be formed when activation is required. There are some overlaps between SDF-1β binding domains and coreceptor function domains in molecular structure of CXCR4.
基金This work was supported by a grant from National 973 Project (2001CB5102) for Outstanding Scholars of New Era from the Ministry of Education of China (2002-48)+1 种基金 National Natural Science Foundation of China (30000028) the key research program from Sc
文摘Differential proteome profiles of human lung squamous carcinoma tissue compared to paired tumor-adjacent normal bronchial epithelial tissue were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and matrix-assisted laser desorp-tion/ionization time of flight mass spectrometry (MALDI-TOF-MS). The results showed that well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were obtained under the condition of 0.75-mg protein-load. The average deviation of spot position was 0.733±0.101 mm in IEF direction, and 0.925±0.207 mm in SDS-PAGE direction. For tumor tissue, a total of 1241±88 spots were detected, 987±65 spots were matched with an average matching rate of 79.5%. For control, a total of 1190±72 spots were detected, and 875±48 spots were matched with an average matching rate of 73.5%. A total of 864±34 spots were matched between tumors and controls. Forty-three differential proteins were characterized: some proteins were related to oncogenes, and others involved in the regulation of cell cycle and signal transduc-tion. It is suggested that the differential proteomic approach is valuable for mass identification of differentially expressed proteins involved in lung carcinogenesis. These data will be used to establish human lung cancer proteome database to further study human lung squamous carcinoma.
基金This work was supported by a grant from the National Natural Science Foundation of China 30271512(to Zhou Hong)by a grant from National Kcy Technologies R&D Program G1999054203(to Zheng Jiang and Zhou Hong).
文摘Lipopolysaccharide(LPS),the principal component of the outer membrane of Gram-negative bacteria,stimulates various cell types to release numerous proinflammatory mediators such as TNF-α,IL-6 and IL-12,which may damage cells and lead to organ injury,even sepsis and septic shock.Toll-like receptor 4(TLR4) has been identified as the receptor involved in the recognition of LPS,but the role of LPS uptake in activating signal transduction remains controversial.In the present study,TNF-α was used as a marker of macrophages/ monocytes activated by LPS,and CQ was used as an inhibitor of endosome mature in order to definitude what stage of the signal transduction elicited by LPS was interrupted.We found that there indeed existed internalization of LPS and internalization partially participated in LPS signaling since CQ inhibited cytokine release,and decreased accumulation of FITC-LPS in hPBMCs.In contrast,anti-hTLR4 antibody could decrease cytokine release,but had no inhibition on accumulation of FITC-LPS.This result revealed that inhibition of cytokine release was related to reduction of FITC-LPS accumulation in the cells.But TLR4 on the cell surface couldn't participate in internalization of LPS.Thus,LPS signaling and internalization couldn't be viewed as mutually independent processes.Cellular & Molecular Immunology.2004;1(5):373-377.
