AIM: To study the effect of TNP-470 on cell growth,proliferation and apoptosis in human colon cancer xenograftsin nude mice.METHODS: Human colon cancer xenografts weretransplanted into 20 nude mice. Mice were randomly...AIM: To study the effect of TNP-470 on cell growth,proliferation and apoptosis in human colon cancer xenograftsin nude mice.METHODS: Human colon cancer xenografts weretransplanted into 20 nude mice. Mice were randomly dividedinto two groups. TNP-470 treated group received TNP-470(30 mg/kg, s.c) every other day and the control groupreceived a sham injection of same volume saline solution.They were sacrificed after 4 weeks and their tumors wereprocessed for histological examination. The expression ofproliferating cell nuclear antigen (PCNA) in tumors wasdetected using immunohistochemical method with imageanalysis, and apoptosis in tumor cells was measured by TdT-mediated biotinyated-dUTP nick end labeling (TUNEL)staining.RESULTS: Comparing with controls, tumor growth wassignificantly inhibited in TNP-470 treated group, the inhibitoryrate being 54.4 %. Expression of PCNA in tumors of TNP-470 treated group (PI 54.32±11.47) was significantly lowerthan that of control group (PI 88.54+12.36), P＜0.01.Apoptosis index (AI) of TNP-470 treated group (18.95±1.71)was significantly higher than that of control group(7.26+1.44),P＜0.001, typical morphological change of apoptosis in tumorcells was observed in TNP-470 treated group.CONCLUSION: Besides the anti-angiogenic effects, TNP-470 can inhibit tumor growth by inhibiting the proliferationand inducing apoptosis of tumor cells.展开更多
AIM: Heme oxygenase (HO)-I catalyzes the conversion of heme to biliverdin, iron and carbon monoxide. HO-1 is induced by many stimuli including heme, Hb, heat stress,lipopolysaccharide (LP5) and cytokines. Previous stu...AIM: Heme oxygenase (HO)-I catalyzes the conversion of heme to biliverdin, iron and carbon monoxide. HO-1 is induced by many stimuli including heme, Hb, heat stress,lipopolysaccharide (LP5) and cytokines. Previous studies demonstrated that LP5 induced HO-1 gene activation and HO-1 expression in liver. However, the mechanisms of LPS-induced HO-1 expression in liver remain unknown. The effect of toll-like receptor-4 (TLR4) on LPS-induced liver HO-1 expression and the role of TNF-α and IL-1β in this condition were determined.METHODS: HO-1 expression was determined by immunofluorescent staining and immunoblotting. Double immunofluorescent staining was performed to determine the cell type of HO-1 expression in liver.RESULTS: A low dose of LPS significantly increased HO-1 expression in the liver which was localized in Kupffer cells only. Furthermore, HO-1 expression was enhanced by three doses of LPS. HO-1 expression was significantly inhibited in the liver of TLR4 mutant mice. While the liver HO-1 expression in TNF KO mice was much lower than that in C57 mice following the same LPS treatment, IL-1β KO had a slight influence on liver HO-1 expression following LPS treatment.CONCLUSION: The preserfl: results confirm that macrophages are the major source of HO-1 in the liver induced by LPS.This study demonstrates that TLR4 plays a dominant role in mediating HO-1 expression following LPS. LPS-induced HO-1 expression is mainly mediated by endogenous TNF-α, but only partially by endogenous IL-1β.展开更多
AIM: To study the distribution and expression of non-muscle myosin light chain kinase (nmMLCK) in rabbit livers.METHODS: Human nmMLCK N-terminal cDNA was amplified by polymerase chain reaction (PCR) and was inserted i...AIM: To study the distribution and expression of non-muscle myosin light chain kinase (nmMLCK) in rabbit livers.METHODS: Human nmMLCK N-terminal cDNA was amplified by polymerase chain reaction (PCR) and was inserted into pBKcmv to construct expression vectors. The recombinant plasmid was transformed into XL1-blue. Expression protein was induced by IPTG and then purified by SDS-PAGE and electroelution, which was used to prepare the polycolonal antibody to detect the distribution and expression of nmMLCK in rabbit livers with immunofluorescene techniques.RESULTS: The polyclonal antibody was prepared, by which nmMLCK expression was detected and distributed mainly in peripheral hepatocytes.CONCLUSION: nmMLCK can express in hepatocytes peripherally, and may play certain roles in the regulation of hepatic functions.展开更多
To investigate whether the polymorphisms of β2-adrenergic receptor (β2-AR) at position 16, 27, 164 are asso-ciated with asthma in Northern Chinese subjects. Methods Genomic DNA was collected from unrelated Northern ...To investigate whether the polymorphisms of β2-adrenergic receptor (β2-AR) at position 16, 27, 164 are asso-ciated with asthma in Northern Chinese subjects. Methods Genomic DNA was collected from unrelated Northern Chinese population of Han ethnicity, including 125 un-related asthmatic individuals and 96 healthy controls. β2-AR genes at position 16, 27, 164 were amplified by using restriction fragment length polymorphism (RFLP) and allelic specific polymerase chain reaction methods. All asthmatics had their serum IgE (total and specific) antibody or skin-prick test measured, bronchial reactivity to methacholine (Mch) and bronchial rever-sibility by β2-agonist evaluated. Results (1) The frequency of Gly 16 homozygous was significantly higher in the asthmatic group than that in healthy controls (22.4% vs. 8.3%, P < 0.05), OR was 2.9 with 95% CI 1.26-6.78. The proportion of Gly 16 allele was also higher in asthmatics than that in control (0.46 vs. 0.36, P < 0.05); Gly16 homozygous was not independently associated with asthma pathogenesis (P = 0.21, OR 0.42 with 95% CI 0.11-1.61). (2) Of 51 night attack patients, 18 carrying Gly16 homozygosity, if compared with 10 of 74 nonnocturnal asthmatics carrying this genotype, there was significant difference between these two groups (35.3% vs. 13.5%, P < 0.01). (3) The average dose of PD20-Mch was significantly lower in patients carrying Gln 27 homozygous than those carrying homozygous Glu 27 and Gln/Glu 27 heterozygous (0.2 ± 0.3, 1.6 ± 0.8, and 2.1 ± 3.0 μmol/L,P < 0.05). Conclusion β2-AR gene polymorphisms might confer susceptibility to asthma in Chinese Northern patients. β2-AR gene, coordinated with other candidate loci, plays a role in the development of asthma.展开更多
AIM: To determine the expressions of cysteinyl leukotriene receptors, CysLT, and CysLT2 , in airway eosinophilic inflammation of OVA-induced asthmatic mice and the modulation by montelukast, a CysLT1 receptor antagoni...AIM: To determine the expressions of cysteinyl leukotriene receptors, CysLT, and CysLT2 , in airway eosinophilic inflammation of OVA-induced asthmatic mice and the modulation by montelukast, a CysLT1 receptor antagonist. METHODS: Asthma model was induced by chronic exposure to ovalbumin (OVA) in C57BL/6 mice. The eosinophils展开更多
基金the Natural Science Foundation of Guangdong Province,No.013072
文摘AIM: To study the effect of TNP-470 on cell growth,proliferation and apoptosis in human colon cancer xenograftsin nude mice.METHODS: Human colon cancer xenografts weretransplanted into 20 nude mice. Mice were randomly dividedinto two groups. TNP-470 treated group received TNP-470(30 mg/kg, s.c) every other day and the control groupreceived a sham injection of same volume saline solution.They were sacrificed after 4 weeks and their tumors wereprocessed for histological examination. The expression ofproliferating cell nuclear antigen (PCNA) in tumors wasdetected using immunohistochemical method with imageanalysis, and apoptosis in tumor cells was measured by TdT-mediated biotinyated-dUTP nick end labeling (TUNEL)staining.RESULTS: Comparing with controls, tumor growth wassignificantly inhibited in TNP-470 treated group, the inhibitoryrate being 54.4 %. Expression of PCNA in tumors of TNP-470 treated group (PI 54.32±11.47) was significantly lowerthan that of control group (PI 88.54+12.36), P＜0.01.Apoptosis index (AI) of TNP-470 treated group (18.95±1.71)was significantly higher than that of control group(7.26+1.44),P＜0.001, typical morphological change of apoptosis in tumorcells was observed in TNP-470 treated group.CONCLUSION: Besides the anti-angiogenic effects, TNP-470 can inhibit tumor growth by inhibiting the proliferationand inducing apoptosis of tumor cells.
文摘AIM: Heme oxygenase (HO)-I catalyzes the conversion of heme to biliverdin, iron and carbon monoxide. HO-1 is induced by many stimuli including heme, Hb, heat stress,lipopolysaccharide (LP5) and cytokines. Previous studies demonstrated that LP5 induced HO-1 gene activation and HO-1 expression in liver. However, the mechanisms of LPS-induced HO-1 expression in liver remain unknown. The effect of toll-like receptor-4 (TLR4) on LPS-induced liver HO-1 expression and the role of TNF-α and IL-1β in this condition were determined.METHODS: HO-1 expression was determined by immunofluorescent staining and immunoblotting. Double immunofluorescent staining was performed to determine the cell type of HO-1 expression in liver.RESULTS: A low dose of LPS significantly increased HO-1 expression in the liver which was localized in Kupffer cells only. Furthermore, HO-1 expression was enhanced by three doses of LPS. HO-1 expression was significantly inhibited in the liver of TLR4 mutant mice. While the liver HO-1 expression in TNF KO mice was much lower than that in C57 mice following the same LPS treatment, IL-1β KO had a slight influence on liver HO-1 expression following LPS treatment.CONCLUSION: The preserfl: results confirm that macrophages are the major source of HO-1 in the liver induced by LPS.This study demonstrates that TLR4 plays a dominant role in mediating HO-1 expression following LPS. LPS-induced HO-1 expression is mainly mediated by endogenous TNF-α, but only partially by endogenous IL-1β.
基金National Natural Science Foundation of China,No.39870324Grant for Excellent Young Teachers of Ministry of education of China,No.39870324National Science Foundation of AnHui Province,No.9904312
文摘AIM: To study the distribution and expression of non-muscle myosin light chain kinase (nmMLCK) in rabbit livers.METHODS: Human nmMLCK N-terminal cDNA was amplified by polymerase chain reaction (PCR) and was inserted into pBKcmv to construct expression vectors. The recombinant plasmid was transformed into XL1-blue. Expression protein was induced by IPTG and then purified by SDS-PAGE and electroelution, which was used to prepare the polycolonal antibody to detect the distribution and expression of nmMLCK in rabbit livers with immunofluorescene techniques.RESULTS: The polyclonal antibody was prepared, by which nmMLCK expression was detected and distributed mainly in peripheral hepatocytes.CONCLUSION: nmMLCK can express in hepatocytes peripherally, and may play certain roles in the regulation of hepatic functions.
文摘To investigate whether the polymorphisms of β2-adrenergic receptor (β2-AR) at position 16, 27, 164 are asso-ciated with asthma in Northern Chinese subjects. Methods Genomic DNA was collected from unrelated Northern Chinese population of Han ethnicity, including 125 un-related asthmatic individuals and 96 healthy controls. β2-AR genes at position 16, 27, 164 were amplified by using restriction fragment length polymorphism (RFLP) and allelic specific polymerase chain reaction methods. All asthmatics had their serum IgE (total and specific) antibody or skin-prick test measured, bronchial reactivity to methacholine (Mch) and bronchial rever-sibility by β2-agonist evaluated. Results (1) The frequency of Gly 16 homozygous was significantly higher in the asthmatic group than that in healthy controls (22.4% vs. 8.3%, P < 0.05), OR was 2.9 with 95% CI 1.26-6.78. The proportion of Gly 16 allele was also higher in asthmatics than that in control (0.46 vs. 0.36, P < 0.05); Gly16 homozygous was not independently associated with asthma pathogenesis (P = 0.21, OR 0.42 with 95% CI 0.11-1.61). (2) Of 51 night attack patients, 18 carrying Gly16 homozygosity, if compared with 10 of 74 nonnocturnal asthmatics carrying this genotype, there was significant difference between these two groups (35.3% vs. 13.5%, P < 0.01). (3) The average dose of PD20-Mch was significantly lower in patients carrying Gln 27 homozygous than those carrying homozygous Glu 27 and Gln/Glu 27 heterozygous (0.2 ± 0.3, 1.6 ± 0.8, and 2.1 ± 3.0 μmol/L,P < 0.05). Conclusion β2-AR gene polymorphisms might confer susceptibility to asthma in Chinese Northern patients. β2-AR gene, coordinated with other candidate loci, plays a role in the development of asthma.
文摘AIM: To determine the expressions of cysteinyl leukotriene receptors, CysLT, and CysLT2 , in airway eosinophilic inflammation of OVA-induced asthmatic mice and the modulation by montelukast, a CysLT1 receptor antagonist. METHODS: Asthma model was induced by chronic exposure to ovalbumin (OVA) in C57BL/6 mice. The eosinophils
