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OBJECTIVE： To investigate the effects of chelidonine on proliferation， collagen synthesis and TGF-β1 receptor of activated hepatic stellate cells CFSC-8B. METHODS： CFSC-8B cells in logarithmic phase were collected and then divided into normal control group， model group， solvent group （ethanol）， positive control group （1 μg/mL colchicine ethanol solution）， chelidonine low， medium and high concentration groups （2.1， 4.2， 8.4 μg/mL chelidonine ethanol solution）. Except for normal control group， other groups were activated with 20 μg/L TGF-β1 for 24 h； the latter 5 groups were intervened with relevant medicine for 24 h. Cell proliferation of activated cells was assayed by CCK-8 assay. Hydroxyprolin （Hyp） content was assayed by enzyme digestion； the levels of typeⅠ collagen （Col-Ⅰ） and type Ⅲ collagen （Col-Ⅲ） were assayed by ELISA； the expressions of TβR-Ⅰ and TβR-Ⅱ protein were assessed by Western blot； mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ in hepatic stellate cells were assessed by RT-PCR. RESULTS： Compared with normal control group， cell proliferation rate， Hyp content， the levels of Col-Ⅰ and Col-Ⅲ， the protein expressions of TβR-Ⅰ and TβR-Ⅱ as well as mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ were increased significantly （P＜0.05）. Compared with model group， there were no significant difference in above indexes of hepatic stellate cells in solvent group （P＞0.05）； there were no significant difference in the proliferation rate of hepatic stellate cells in chelidonine low concentration group （P＞0.05）， above indexes of hepatic stellate cells were decreased significantly in positive control group and chelidonine high concentration group （P＜0.05）. The decrease of Hyp and Col-Ⅲ levels were not significant in chelidonine medium concentration， but other above indexes were decreased significantly （P＜0.05）. Compared with chelidonine medium concentration group， the rate of cell proliferation， Col-Ⅰ level， protein and mRNA expressions of TβR-Ⅰ and TβR-Ⅱ were decreased significantly in chelidonine high concentration group （P＜0.05）. CONCLUSIONS： Chelido- nine can inhibit the proliferation， collagen synthesis as well as the protein and mRNA expressions of TβR-Ⅰand TβR-Ⅱ in activated CFSC-8B cells.
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Objective:To study effects of neuroligin (NLG) on the proliferation and apoptosis in human neuroblastoma SH-SY5Y cells.Methods:The SH-SY5Y cells were cultured in vitro for 24 hours,and then transfected with NLG siRNA at dose of 50,100,200 μmol/L,respectively.MTT procedure was used to detect the cell proliferation,and expression levels of apoptosis gene including Bax or Bcl-2 and Bcl-xl were measured by RT-PCR.Results:Compared to control groups proliferation of SH-SY5Y cells were distinctly inhibited after NLG siRNA transfection accompany with a dose-dependent,which was caused by activation of apoptosis.Conclusions:NLG protect neuron by inhibiting apoptosis.
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Objective To investigate the expressions of Stat3, CyclinD1 and proliferating cell nuclear antigen (PCNA) in non-small cell lung cancer (NSCLC), and analyze their relationships with carcinogenesis and progression of NSCLC. Methods SP immunohistochemistry was used to detect the expressions of Stat3, CyclinD1 and PCNA in 45 cases of NSCLC. Results The positive rate of Stat3 was 71. 1 % in NSCLC. The expression intensities of Stat3 was not associated with different tumor sizes and histological type, but it was associated with the expression intensities of CyclinDi and PCNA (r= 0.531, P
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Purpose The aim is to study the transduction of m urine stem cell factor(SCF) into umbilical blood cells by LipofectinRMmediated and expression in them.Methods Murine SCF cDNA encoding extra cellu lar domain was isolated using PCR from plasmid pRC/CMV(containing SCF gene), and then recombined into the expression vector pcDNA3,and transferred into enrichme nt cultural umbilical blood cells. Semi-quantitative RT-PCR was used to examin e the expression on mRNA level.And the effects of supernatant of transfected umb ilical blood cells were investigated alone or in coordinate with GM-CSF by colo ny formation test of human bone marrow cells in semisolid culture.Results SCF mRNA was expressed in transfected umbilical blood cells.The su pernat ant of transfected umbilical blood cells could increase the number of CFU-GM, s ynergizing with GM-CSF.Conclusion The supernatant of umbil ical b lood cells transfected with vector containing mSCF gene can stimulate the colony formation of human bone marrow cells in combination with GM-CSF.
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Objective:To study the relationship betwen the mechanism of autoimmune disease and CD4+CD25+ T cell population in NIK mutated mice-aly mice.Methods:NIK mutated mice-aly/aly mice were used as model,aly/+mice as NIK normal control;cell populations were determined by FACS and the thymus structure were analyzed by immunohistochemistry.Results:The CD4+CD25+CD8- population were remarkably decreased in aly mice;and the UEA-1 positive cells were absent in aly mice.Conclusion:The autoimmune disease in aly mice might be the result of deceased the CD4+CD25+ population;the UEA-1 positive cells might play an important role in the development of CD4+CD25+ population. [
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Objective:Identification of dominant epitope of P2 in EAN and studying of similarity between the dominant epitope and LPS of Campylobacter jejuni.Methods:Screening peptide library with EAN serum.Identified positive clone by ELISA and compared similarity between positive clone and LPS of Campylobacter jejuni.Results:Affinity of all indentified positive clone increased remarkbly compared to negative control(P