1.Effect of PP60c-Src on Ang Ⅱ- induced signal transduction in rat vascular smooth muscle cells
Chinese Journal of Pathophysiology 2005;21(4):685-689
AIM: The aim of the present study was to clarify the mechanism of intracellular signal transduction in Ang Ⅱ- induced proliferation of vascular smooth muscular cells (VSMC) by observing the effect of c- Src on Ang Ⅱ- mediated mitogen- activated protein kinase (MAPK) activation and c- Fos protein expression in cultured VSMC of rats. METHODS: Cultured aortic VSMCs from SD rats were transfected with anti-sense c-Src oligodeoxynucleotides (ODNs) wrapt with lipofectin to inhibit c- Src activity and protein production. Untransfected VSMCs were used as control. We observed the role of Ang Ⅱ stimulation in MAPK activation and c- Fos protein expression. c- Src kinase activity was measured by protein immunoprecipitation and kinase autophosphorylation. The phosphorylation rate of the substrate myelin basic protein (MBP) was employed to assess MAPK activity.Western immunoblot was used to detect protein expression of c- Src and c-Fos. RESULTS: c-Src protein expression in VSMC transfected with different concentrations of anti- sense ODNs significantly decreased in a negative dose- effect manner. c- Src kinase activity was also markedly inhibited. Following the stimulation of Ang Ⅱ on transfected VSMCs with anti-sense ODNs, the increase rate of c- Src activity was 8.7% of that in control, the activity of MAPK was 1.6% compared with control and c- Fos protein expression was as 30.0% as that of control. CONCLUSION: Ang Ⅱ induces c- Src activation. MAPK activation and c - Fos protein expression by Ang Ⅱ is dependent on c- Src activation. These findings indicate that c- Src is an important signal factor in Ang Ⅱ-induced VSMC proliferation.
2.Effect of PP~(60c-Src) on Ang II- induced signal transduction in rat vascular smooth muscle cells*
Chinese Journal of Pathophysiology 1986;0(04):-
AIM: The aim of the present study was to clar ify the mechanism of intracellular signal transduction in Ang II-induced prolife ration of vascular smooth muscular cells (VSMC) by observing the effect of c-Src on Ang II-mediated mitogen-activated protein kinase (MAPK) activation and c-Fos protein expression in cultured VSMC of rats. METHODS: Cultured aortic VSMCs from SD rats were transfected wit h anti-sense c-Src oligodeoxynucleotides (ODNs) wrapt with lipofectin to inhibit c-Src activity and protein production. Untransfected VSMCs were used as control . We observed the role of Ang II stimulation in MAPK activation and c-Fos protei n expression. c-Src kinase activity was measured by protein immunoprecipitation and kinase autophosphorylation. The phosphorylation rate of the substrate myeli n basic protein (MBP) was employed to assess MAPK activity. Western immunoblot w as used to detect protein expression of c-Src and c-Fos. RESULTS: c-Src protein expression in VSMC transfected with diffe rent concentrations of anti-sense ODNs significantly decreased in a negative dos e-effect manner. c-Src kinase activity was also markedly inhibited . Following t he stimulation of Ang II on transfected VSMCs with anti-sense ODNs, the increase rate of c-Src activity was 8.7% of that in control, the activity of MAPK was 1 .6% compared with control and c-Fos protein expression was as 30.0% as that of control. CONCLUSION: Ang II induces c-Src activation. MAPK activation and c-Fos protein expression by Ang II is dependent on c-Src activation. These find ings indicate that c-Src is an important signal factor in Ang II -induced VSMC proliferati on.
3.Survey on Pervasive Developmental Disorder in 2-6 Year-old Children in Beijing
Jing LIU ; Xiaoling YANG ; Meixiang JIA
Chinese Mental Health Journal 1991;0(05):-
Objective: Exploring the epidemiological state of pervasive developmental disorder in 2-6 year-old children in Beijing. Methods: A total of 21866 children aged 2-6 year-old were recruited from permanent residents of Beijing by 2-phase cluster sampling.The Clancy Autism Behavior Scale was used for screening.The Childhood Autism Rating Scale and DSM-Ⅳ were used for diagnosis.Results: 16 children were diagnosed as pervasive developmental disorder (14 with autistic disorder, 1 with atypical autistic disorder, 1 with Rett syndrome).The false negative rate was 0.80 ‰.The prevalence of pervasive developmental disorder was 0.73 ‰, the adjusted prevalence was 1.53 ‰,the average annual detective rate was 0.11‰.The prevalence rate had no difference in distribution between urban-rural, among different age groups, sex and nations(?2=0.11~1.85,P=0.739-0.173).The low family income was related to high prevalence significantly ( tendency ?2=4.70,P=0.030 ).25% children with pervasive developmental disorder had not been identified by parents or had not been to clinics.80% children with pervasive developmental disorder had not get intervention.All children had demand to get intervention. Conclusions: The prevalence of pervasive developmental disorder in 2-6 year-old children in Beijing was not low.Low family income is the risk factor.The rate of intervention was low.The government should pay close attention to the children with pervasive developmental disorder and their needs.
4.Risk factor for children with autism during perinatal period
Changmin ZHAO ; Jiancong LIU ; Jing LIU ; Meixiang JIA ; Wenhua SANG
Clinical Medicine of China 2011;27(7):774-776
Objective To compare the risk factor of chilldren with autism and ordinary children during perinatal period. Methods One hundred and fourty children with autism and 82 ordinary children were reviewed by self-written general circumstance questionnaire and risk factor questionnaire. Results Viral influenza during pregnancy (x2 =15.29) ,bom suffocate( x2 =6. 04) , premature delivery (x2 =6. 48) , dystocia (x2 =2. 83) and artificial feeding ( x2 = 6. 02 ) were risk factors for children autism (P < 0. 05 ). Conclusion Childeren autism is associated with risk factors in rinatal period. Early dectecion and early prevention and treatment may improve the outcome.
5.The anxiety evaluation of parents of children with autism
Changmin ZHAO ; Jiancong LIU ; Jing LIU ; Meixiang JIA ; Wenhua SANG
Clinical Medicine of China 2011;27(8):878-879
Objective To evaluate the relationship between the anxiety condition and educational background of the autistic children's parents. Methods Questionnaires for the self-evaluation of anxiety were collected from the parents of 140 children with autism. Results The autistic children's mothers had significantly higher score of anxiety than the fathers (42. 73 ±8. 25) (t =6. 783,P <0. 05). The autistic boy's parents had significantly higher anxiety pressure than autistic girl's parents ( 51.38 ± 11.24 vs. 43.23 ± 6. 12) ( t = 4. 894,P <0. 05). The anxiety intensity of the autistic children's parents was negatively correlated with the parents'educational background ( F = 10. 788, P < 0. 05 ). Conclusion The autistic children' s parents had certain anxiety,which is correlated with the their educational background and genders of the autistic children. It is necessary to interfere the negative mood to facilitate the treatment of the autistic children.
6.Experimental Study on Liver Regeneration Following Portal Branch Ligation in Rats
Meixiang GUO ; Lihua GAO ; Libo LIU ; Zhaohua MENG ; Xiumei GONG
Chinese Journal of Bases and Clinics in General Surgery 2003;0(05):-
Objective To study liver regeneration of the non-ligated liver lobes following portal branch ligation (PBL). Methods Sixty male Wistar rats were randomly divided into PBL group and sham operation (SO) group. Under ether anesthesia, the rats were subjected to PBL and sham operation, respectively. The animals were sacrificed on the 1st, 2nd, 3rd, 7th and 14th day respectively. The blood sample was collected from heart and the livers were harvested to determine serum alanine aminotransferase (ALT) levels and total liver weight, respectively. The hepatic histopathology was studied through light microscopy. The number of liver cell nuclear mitosis index was counted. The number of proliferative cell nuclear antigen (PCNA) index was counted by immunohistochemistry. The hepatic ultrastructural changes were studied under electron microscope. Results Elevated serum ALT level was observed in the first postoperative day in PBL group compared with SO group (P
7.Effect of P1k3 on the transcriptional activity of p73 in H1299 cells
Meixiang SANG ; Lihua LIU ; Chunyan DING ; Jun MENG ; Baoen SHAN
China Oncology 2010;20(1):6-11
Background and purpose: Protein kinase P1k3 could increase the transcriptional activity of p53. However, the effect of P1k3 on the transcriptional activity of p73 is still unknown. Our study was to investigate the effect of P1k3 on the transcriptional activity of p73. Methods: Luciferase reporter assay, RT-PCR and colony formation assay were adopted to study the effect of P1k3 and kinase-deficient P1k3 (K52R) on the transcriptional activity of p73 in p53-deficient human lung carcinoma H1299 cells. Results: Luciferase reporter assay showed that p73 increased the luciferase activities induced by p21~(WAF1) and Bax promoters (P<0.05). After co-transfection with p73 and P1k3, the luciferase activities induced by p21~(WAF1) and Bax promotors were significantly decreased in a dose-dependent manner as compared with the group that transfected p73 only (P<0.05). However, kinase-deficient Plk3 (K52R) had no significant effect on the luciferase activities induced by p21~(WAF1) and Bax promoters (P>0.05). RT-PCR showed that p73 increased the mRNA expressions of p21~(WAF1) and BAX (P<0.05). P1k3 decreased the expressions of p21~(WAF1) and Bax induced by p73 (P<0.05). Kinase-deficient P1k3 (K52R) had no significant effect on the expressions of p21~(WAF1) and Bax induced by p73 (P>0.05). Colony formation assay revealed that p73 decreased the colony formation of H1299 cells (P<0.05). P1k3 decreased the inhibitory effect of p73 on the colony formation of H1299 cells (P<0.05). Kinase-deficient P1k3 (K52R) had no significant effect on the inhibitory effect of p73 on the colony formation of H1299 cells (P>0.05).Conclusion: P1k3 can inhibit the transcriptional activity of p73, where as kinase-deficient P1k3 has no effect on the transcriptional activity of p73.
8.Relationship between HBsAg level and HBV DNA level as well as illness severity in patients with HBV-related acute-on-chronk liver failure
Meixiang LI ; Dongying XIE ; Hong DENG ; Qiong LIU ; Zhiliang GAO
Chinese Journal of Postgraduates of Medicine 2011;34(28):20-23
Objective To analyze the relationship between HBsAg level and HBV DNA level as well as illness severity in patients with HBV-related acute-on-chronic liver failure (ACLF).Methods One hundred and nineteen patients with chronic hepatitis B (CHB group) and 98 patients with ACLF(ACLF group) were enrolled.HBsAg and HBeAg were assayed with Roche electrochemical luminescence method.HBV DNA was quantified using a real-time polymerase chain reaction (PCR) assay.HBsAg and HBV DNA levels were compared between two groups and between HBeAg-positive and HBeAg-negative patients in ACLF group respectively,also the correlationbetween HBsAg and HBV DNA was studied.Results The proportions of HBeAg-negative patients were 68.4%(67/98) and 42.9%(51/119) in ACLF group and CHB group respectively,and there was significant difference between two groups (P <0.01).There was no significant difference in HBV DNA between two groups (P > 0.05).HBV DNA in HBeAg-positive patients was higher than that in HBeAg-negative patients in two groups(P < 0.05).There was no significant difference in HBsAg between HBeAg-positive patients and HBeAg-negative patients in ACLF group (P > 0.05 ),but they were higher than that in HBeAg-positive patients in CHB group (P< 0.05).HBsAg was correlated to ALT,AST in HBeAg-positive patients (P < 0.05).No significant correlation was found among HBsAg and HBV DNA as well as biochemical changes (P> 0.05).There was no significant difference in the ratio of different HBsAg levels among the patients of different HBV DNA in ACLF group (P> 0.05).Conclusion The level of HBsAg does not directly correlate with serum HBV DNA level,and has no directly correlation with the severity of the disease in patients with HBV-related ACLF.
9.Construction of Recombinant Adenoviral Vector Carrying the Human BMP-2 Gene with the Technology of in vitro Ligation
Lei WANG ; Xun QU ; Fengcai WEI ; Jinbo FENG ; Shanzhen SUN ; Shaohua LIU ; Meixiang YANG
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Objective: To provide an efficeut protocol for constructing recombinant adenovirus, an in vitro ligation was used instead of homologous recombination. Methods: Gene BMP-2 was ligated into pShuttle 2 vector ( pShuttle 2-BMP-2 ) and then fragment containing BMP-2 gene and promoter pcmvie excised by PI-SCe Ⅰ and Ⅰ-Ceu Ⅰ endonuclease. The fragment was further combined with adenovirus vector (Adeno-X-BMP-2) , which was finally linearized with Pac Ⅰ and trans-fered to HEK293 to package adenovirus particles. Results: Both PCR assay and restiction analysis showed that the recombined rectors pShuttle2-BMP-2 and Adeno-X-BMP-2 contains the target BMP-2 gene. THe packaged adenovirus was also i-dentified by PCR assay with specific primers for BMP-2. Conclusions: The BMP-2 incorporated recombinant adenovirus was obtained and this laid a foundation for further study on BMP-2 mediated gene therapy. The in vitro ligation method de-scinbed here for constructing recombined adenovirus was more efficient than traditional homologous recombination.
10.EFFECT OF SODIUM NITROPRUSSIDE AND N-NITRO-L-ARGININE-MYTHEL-ESTER ON APOPTOSIS OF SPERMATOGENIC CELLS IN RAT TESTIS
Meixiang LI ; Liping HE ; Yuanjie XIE ; Xiaohong ZHANG ; Nan WEN ; Zhifeng LONG ; Yueshun LIU
Acta Anatomica Sinica 1953;0(01):-
Objective To evaluate the effect of sodium nitroprusside(SNP)and N-nitro-l-arginine-mythel-ester (L-NAME) on apoptosis of spermatogenic cells in rats. Methods Fourty adult male Sprague-Dawley rats (60-70days) were divided into four groups.Each group was injected intraperitoneally with one of the following agents, once a day, for 12 days: 1. SNP; 2.L-NAME;3.SNL+L-NAME;4.Normal saline NS group.Two hours after the last time injection the rats were sacrificed.TUNEL staining and flow cytometry analysis were used to detect the apoptosis of spermatogenic cells. Results Sub-monoploid and apoptosis index (AI) in SNP group was significantly higher than that of NS group and sub-monoploid and apoptosis index (AI) in L-NAME group were significantly lower than that of NS group by FCM and TUNEL (P0.5) was found.Conclusion SNP can accelerate the apoptosis of spermatogenic cells and L-NAME can inhibite the apoptosis of spermatogenic cells,The effect of SNP and L-NAME on apoptosis of spermatogenic cells probably occurs through the action of nitric oxide.