AIM: To investigate the inhibitory effect of in vivoexpression of expressing plasmid pCH510 of recombinant fibronectin polypeptide (CH50) on hepatocellular carcinoma and the improved therapeutic effect of pCH510 in co...AIM: To investigate the inhibitory effect of in vivoexpression of expressing plasmid pCH510 of recombinant fibronectin polypeptide (CH50) on hepatocellular carcinoma and the improved therapeutic effect of pCH510 in combination with chemotherapeutic agents and Hsp70-H22 hepatocarcinoma antigen peptide on tumor.METHODS: Mice were inoculated with H22 hepatccarcinoma cells. The chemotactic effect of the expression of plasmid pCH510 on immunocytes was observed after in vivo transfection, tissue slicing and HE staining. Inhibitory effect of transfection with pCH510 on routine tumor originatedfrom different inoculative doses was observed. The inhibitory effect of immediate transfection with pCH510 after chemotherapy on tumor was compared with that of transfection 5 days after chemotherapy. The change of function and amount of mouse peritoneal macrophages and the peripheral blood immunocytes resulted from administration of chemotherapeutic agents were detected. The peptides mixture was prepared from H22 hepatocarcinoma cells, pCH510 + Hsp70-H22 antigen peptides were injected into tumor-bearing mice with or without chemotherapy, to observe the inhibitory effects on tumor.RESULTS: At the tumor tissue site injected with pCH510,there were a great number of immunocytes which mainly were macrophages, lymphocytes and neutrophils.Transfection of plasmid pCH510 inhibited significantly the murine tumor induced by different inoculative doses. The inhibitory effect was negatively correlated with the inoculative dose. The therapeutic effect was not improved by immediate transfection with pCH510 after chemotherapy, but was significantly improved by transfection with pCH510 5 days after chemotherapy. Chemotherapeutic agent decreased the number of immunocytes and suppressed their activation in vivo. After injection of drug, the amount of immunocytes was the lowest from d 1 to d 3 and returned to normal level on the 10th day. Transfection with plasmid pCH510 alone could inhibit tumor induced by the inoculation with 10^4 H22 cells. The tu展开更多
The expression of CH50 polypoptide in bladder cancer cell line BIU-87 and the effects on the invasion ability of BIU-87 were investigated. The eukaryotic expressing vector pCH510 of polypeptide CH50 was introduced int...The expression of CH50 polypoptide in bladder cancer cell line BIU-87 and the effects on the invasion ability of BIU-87 were investigated. The eukaryotic expressing vector pCH510 of polypeptide CH50 was introduced into BIU-87 cells by gene transfection in vitro. The expression of CH50 polypeptide was detected by using immunohistochemical S-P method. The expression of the transfected gene was identified by RT-PCR. Cell invasion assay kit was applied to detect the effect of CH50 polypeptide on the invasion ability of BIU-87. The results showed that the BIU-87 cells transfected with pCH510 could express the CH50 polypeptide, while in the control group, no CH50 polypeptide was detectable. In the transfection group, the invasion ability of BIU-87 in vitro was lower than in control group (P<0.05). It was concluded that CH50 polypeptide was successfully expressed in BIU-87 cells by gene transfection, by which the in vitro invasion ability of BIU-87 was inhibited.展开更多
Background:The aim of this study was to analyze the state of activity and levels of complement in the cerebrospinal fluid(CSF)of patients with various prion diseases(PrDs).Findings:The proteomic data emphasized the le...Background:The aim of this study was to analyze the state of activity and levels of complement in the cerebrospinal fluid(CSF)of patients with various prion diseases(PrDs).Findings:The proteomic data emphasized the levels of 20 known complement components found in the CSF of the sCJD panel that were lower than those found in the non-PrD panel.50%of the complement hemolytic activity(CH50)assays revealed significantly lower activity of complement in the CSF of the sCJD panel.The decreased levels of three key complement subunits,C3a/α,C4β,and C9 in the CSF of the sCJD panel were verified by Western blots.Furthermore,the CH50 values in the CSF of 136 sCJD,39 gCJD,22 FFI and 145 non-CJD patients were individually tested.Compared with the control of non-PrD,the CH50 value in the CSF specimens of various PrDs,especially in three subtypes of inherited PrDs,were significantly lower.Relationship analysis identified that the CH50 activity in the CSF was negatively associated with the protein 14–3–3 positive in the CSF.Conclusion:These results indicate a silent complement system in the CSF of PrD patients.展开更多
基金National Development Program(973)for Key Basic Research(No.2002CB513100)the National Natural Science Foundation of China(No.39870763)
文摘AIM: To investigate the inhibitory effect of in vivoexpression of expressing plasmid pCH510 of recombinant fibronectin polypeptide (CH50) on hepatocellular carcinoma and the improved therapeutic effect of pCH510 in combination with chemotherapeutic agents and Hsp70-H22 hepatocarcinoma antigen peptide on tumor.METHODS: Mice were inoculated with H22 hepatccarcinoma cells. The chemotactic effect of the expression of plasmid pCH510 on immunocytes was observed after in vivo transfection, tissue slicing and HE staining. Inhibitory effect of transfection with pCH510 on routine tumor originatedfrom different inoculative doses was observed. The inhibitory effect of immediate transfection with pCH510 after chemotherapy on tumor was compared with that of transfection 5 days after chemotherapy. The change of function and amount of mouse peritoneal macrophages and the peripheral blood immunocytes resulted from administration of chemotherapeutic agents were detected. The peptides mixture was prepared from H22 hepatocarcinoma cells, pCH510 + Hsp70-H22 antigen peptides were injected into tumor-bearing mice with or without chemotherapy, to observe the inhibitory effects on tumor.RESULTS: At the tumor tissue site injected with pCH510,there were a great number of immunocytes which mainly were macrophages, lymphocytes and neutrophils.Transfection of plasmid pCH510 inhibited significantly the murine tumor induced by different inoculative doses. The inhibitory effect was negatively correlated with the inoculative dose. The therapeutic effect was not improved by immediate transfection with pCH510 after chemotherapy, but was significantly improved by transfection with pCH510 5 days after chemotherapy. Chemotherapeutic agent decreased the number of immunocytes and suppressed their activation in vivo. After injection of drug, the amount of immunocytes was the lowest from d 1 to d 3 and returned to normal level on the 10th day. Transfection with plasmid pCH510 alone could inhibit tumor induced by the inoculation with 10^4 H22 cells. The tu
文摘The expression of CH50 polypoptide in bladder cancer cell line BIU-87 and the effects on the invasion ability of BIU-87 were investigated. The eukaryotic expressing vector pCH510 of polypeptide CH50 was introduced into BIU-87 cells by gene transfection in vitro. The expression of CH50 polypeptide was detected by using immunohistochemical S-P method. The expression of the transfected gene was identified by RT-PCR. Cell invasion assay kit was applied to detect the effect of CH50 polypeptide on the invasion ability of BIU-87. The results showed that the BIU-87 cells transfected with pCH510 could express the CH50 polypeptide, while in the control group, no CH50 polypeptide was detectable. In the transfection group, the invasion ability of BIU-87 in vitro was lower than in control group (P<0.05). It was concluded that CH50 polypeptide was successfully expressed in BIU-87 cells by gene transfection, by which the in vitro invasion ability of BIU-87 was inhibited.
基金supported by Chinese National Natural Science Foundation Grants(81401670)China Mega-Project for Infectious Disease(2011ZX10004-101,2012ZX10004215)the SKLID Development Grant(2012SKLID102).
文摘Background:The aim of this study was to analyze the state of activity and levels of complement in the cerebrospinal fluid(CSF)of patients with various prion diseases(PrDs).Findings:The proteomic data emphasized the levels of 20 known complement components found in the CSF of the sCJD panel that were lower than those found in the non-PrD panel.50%of the complement hemolytic activity(CH50)assays revealed significantly lower activity of complement in the CSF of the sCJD panel.The decreased levels of three key complement subunits,C3a/α,C4β,and C9 in the CSF of the sCJD panel were verified by Western blots.Furthermore,the CH50 values in the CSF of 136 sCJD,39 gCJD,22 FFI and 145 non-CJD patients were individually tested.Compared with the control of non-PrD,the CH50 value in the CSF specimens of various PrDs,especially in three subtypes of inherited PrDs,were significantly lower.Relationship analysis identified that the CH50 activity in the CSF was negatively associated with the protein 14–3–3 positive in the CSF.Conclusion:These results indicate a silent complement system in the CSF of PrD patients.
