AIM: To study the effect of human endostatin mediated by retroviral gene transfer on the growth of human hepatocarcinoma cell line SMMC7721 in nude mice. METHODS: Human endostatin gene together with rat serum albumin ...AIM: To study the effect of human endostatin mediated by retroviral gene transfer on the growth of human hepatocarcinoma cell line SMMC7721 in nude mice. METHODS: Human endostatin gene together with rat serum albumin signal peptide was transferred into human liver carcinoma SMMC7721 cells by retroviral vector pLncx to build a stable transfectant (SMMC-endo). PCR and Western blot analysis were used to verify the transfection and secretion of human endostatin gene in SMMC7721 cells. The endothelial cell proliferation assay in vitro was conducted to test the biological activity of the expressed human endostatin.The inhibitory effect of endostatin expressed by transfected 5MMC7721 on the growth rates of tumor cells in vivo was observed. The mean microvessel density in the specimen was also counted.RESULTS: PCR amplification proved that the genome of SMMC-endo cells contained a 550bp specific fragment of endostatin gene. Western blot analysis confirmed the secretion of human endostatin gene in the conditioned medium of transfected SMMC-endo cells. The endothelial proliferation assay showed that the conditioned medium of SMMC-endo cells significantly inhibited the proliferation of human umbilical vein endothelial cells by 48 %, significantly higher than that of SMMC-pLncx (10.2 %, P＜0.01). In vivo experiments revealed that only in 3 out of 5 mice tumors were formed and the mean size of flank tumors from SMMC-endo cells was 94.5 % smaller than that from the control SMMC-pLncx cells 22 days after tumor inoculation (P＜0.001).The mean microvessel density in tumor samples from SMMC-endo cells was only 8.6±1.1, much fewer than that of 22.6±4.5 from SMMC-pLncx cells (P＜0.01).CONCLUSION: Human endostatin mediated by retroviral gene transfer can inhibit human liver carcinoma cell SMMC7721 growth in nude mice.展开更多
BACKGROUND： Some reports indicate that electric and/or chemical stimulation at various brain sites of experimental animals can raise regional cerebral blood flow and improve cerebral circulation; however, its mechani...BACKGROUND： Some reports indicate that electric and/or chemical stimulation at various brain sites of experimental animals can raise regional cerebral blood flow and improve cerebral circulation; however, its mechanism is still unclear.OBJECTIVE： To observe the effects of electric stimulation at cerebellar fastigial nucleus on serum C-reactive protein of patients with acute cerebral infarction.DESIGN： Non-randomized synchronized contrast study.SETTING： The Second People＇s Hospital of Xinxiang City.PARTICIPANTS： A total of 54 patients with acute cerebral infarction were selected from the Department of Neurology, the Second People＇s Hospital of Xinxiang from December 2005 to December 2006. There were 31 males and 23 females, and their ages ranged from 56 to 80 years. All patients met the diagnostic criteria of the Fourth National Cerebrovascular Academic Meeting, were finally diagnosed by using CT examination,and provided the confirmed consent. Based on therapeutic demands, patients were divided into electric stimulation group and routine treatment group with 27 cases in each group. In addition, 21 healthy subjects,including 11 males and 10 females and aging 53 - 78 years, were selected as the control group. All the subjects in the control group did not have any histories of cerebrovascular diseases and severe body diseases.METHODS： Based on routine drug therapy, patients in the electric stimulation group were also treated by using CVFT-010M cerebral circulation function therapeutic device （made in Shanghai）. Electrode was fixed at bilateral mastoid in the first group and at extensible sides of upper limbs in the second group. Electric stimulation was given twice a day and lasted for 30 minutes each time. Ten days were regarded as a course.Parameters of device： mode Ⅲ, frequency 198%, and intensity 90% - 110% （bionic current）. Patients in the routine treatment group received the routine drug treatment. Content of serum C-reactive protein was measured in both electric stimulation group and routine treatment group before treatment and at 20 days after treatment, while in the control group on the exact day of health examination by using immunization.MAIN OUTCOME MEASURES： Level of serum C-reactive protein in the three groups.RESULTS： All 54 patients with acute cerebral infarction and 21 healthy subjects were involved in the final analysis. Level of serum C-reactive protein was higher in both electric stimulation group and routine treatment group than that in the control group before treatment （P 〈 0.01）. While, level of serum C-reactive protein was lower in the electric stimulation group than that in the routine treatment group after electric stimulation at cerebellar fastigial nucleus （P 〈 0.01）.CONCLUSION： Electric stimulation at cerebellar fastigial nucleus can decrease level of serum C-reactive protein in patients with acute cerebral infarction, and this may be one of the therapeutic mechanisms for curing acute cerebral infarction.展开更多
文摘AIM: To study the effect of human endostatin mediated by retroviral gene transfer on the growth of human hepatocarcinoma cell line SMMC7721 in nude mice. METHODS: Human endostatin gene together with rat serum albumin signal peptide was transferred into human liver carcinoma SMMC7721 cells by retroviral vector pLncx to build a stable transfectant (SMMC-endo). PCR and Western blot analysis were used to verify the transfection and secretion of human endostatin gene in SMMC7721 cells. The endothelial cell proliferation assay in vitro was conducted to test the biological activity of the expressed human endostatin.The inhibitory effect of endostatin expressed by transfected 5MMC7721 on the growth rates of tumor cells in vivo was observed. The mean microvessel density in the specimen was also counted.RESULTS: PCR amplification proved that the genome of SMMC-endo cells contained a 550bp specific fragment of endostatin gene. Western blot analysis confirmed the secretion of human endostatin gene in the conditioned medium of transfected SMMC-endo cells. The endothelial proliferation assay showed that the conditioned medium of SMMC-endo cells significantly inhibited the proliferation of human umbilical vein endothelial cells by 48 %, significantly higher than that of SMMC-pLncx (10.2 %, P＜0.01). In vivo experiments revealed that only in 3 out of 5 mice tumors were formed and the mean size of flank tumors from SMMC-endo cells was 94.5 % smaller than that from the control SMMC-pLncx cells 22 days after tumor inoculation (P＜0.001).The mean microvessel density in tumor samples from SMMC-endo cells was only 8.6±1.1, much fewer than that of 22.6±4.5 from SMMC-pLncx cells (P＜0.01).CONCLUSION: Human endostatin mediated by retroviral gene transfer can inhibit human liver carcinoma cell SMMC7721 growth in nude mice.
文摘BACKGROUND： Some reports indicate that electric and/or chemical stimulation at various brain sites of experimental animals can raise regional cerebral blood flow and improve cerebral circulation; however, its mechanism is still unclear.OBJECTIVE： To observe the effects of electric stimulation at cerebellar fastigial nucleus on serum C-reactive protein of patients with acute cerebral infarction.DESIGN： Non-randomized synchronized contrast study.SETTING： The Second People＇s Hospital of Xinxiang City.PARTICIPANTS： A total of 54 patients with acute cerebral infarction were selected from the Department of Neurology, the Second People＇s Hospital of Xinxiang from December 2005 to December 2006. There were 31 males and 23 females, and their ages ranged from 56 to 80 years. All patients met the diagnostic criteria of the Fourth National Cerebrovascular Academic Meeting, were finally diagnosed by using CT examination,and provided the confirmed consent. Based on therapeutic demands, patients were divided into electric stimulation group and routine treatment group with 27 cases in each group. In addition, 21 healthy subjects,including 11 males and 10 females and aging 53 - 78 years, were selected as the control group. All the subjects in the control group did not have any histories of cerebrovascular diseases and severe body diseases.METHODS： Based on routine drug therapy, patients in the electric stimulation group were also treated by using CVFT-010M cerebral circulation function therapeutic device （made in Shanghai）. Electrode was fixed at bilateral mastoid in the first group and at extensible sides of upper limbs in the second group. Electric stimulation was given twice a day and lasted for 30 minutes each time. Ten days were regarded as a course.Parameters of device： mode Ⅲ, frequency 198%, and intensity 90% - 110% （bionic current）. Patients in the routine treatment group received the routine drug treatment. Content of serum C-reactive protein was measured in both electric stimulation group and routine treatment group before treatment and at 20 days after treatment, while in the control group on the exact day of health examination by using immunization.MAIN OUTCOME MEASURES： Level of serum C-reactive protein in the three groups.RESULTS： All 54 patients with acute cerebral infarction and 21 healthy subjects were involved in the final analysis. Level of serum C-reactive protein was higher in both electric stimulation group and routine treatment group than that in the control group before treatment （P 〈 0.01）. While, level of serum C-reactive protein was lower in the electric stimulation group than that in the routine treatment group after electric stimulation at cerebellar fastigial nucleus （P 〈 0.01）.CONCLUSION： Electric stimulation at cerebellar fastigial nucleus can decrease level of serum C-reactive protein in patients with acute cerebral infarction, and this may be one of the therapeutic mechanisms for curing acute cerebral infarction.
