1.Effective measures to strengthen the honesty and credit education for postgraduates in scientif-ic research platform
Cong LUO ; Faping YI ; Fangzhou SONG
Chinese Journal of Medical Education Research 2013;(10):996-998
Molecular medicine and cancer research center in Chongqing Medical University adhered to academic honesty and credit education ,established original laboratory records system , regularly carried out seminar and improved paper submission program thus to reject academic miscon-ducts from the source,guarantee the authenticity of the data and improve the academic moral level of postgraduates.
2.Construction of biochemistry and molecular biology teaching system
Youquan BU ; Faping YI ; Xianjun LIU ; Ying ZHANG ; Fangzhou SONG
Chinese Journal of Medical Education Research 2013;(5):470-474
In this paper,the recent advances in both biomedical sciences and higher medical education reform were reviewed and analyzed.Furthermore,we proposed and reconstructed the teaching system of biochemistry and molecular biology course in our university,including its teaching content,teaching methods,teacher team,teaching management,etc.The preliminary practice of this system has obtained significant positive effects on teaching quality and student performance.
3.Screening and Identification of The Proteins Interacting with The Transcription Factor X-box Binding Protein 1
Fengjin GUO ; Fangzhou SONG ; Faping YI ; Haien CHENG
Progress in Biochemistry and Biophysics 2006;33(8):745-753
X-box binding protein1 (XBP1) is an important transcription factor, which participates in many signal transduction procession.To investigate the biological function of XBP1, yeast two-hybrid system to screen proteins interacting with XBP1 in hepatocytes was performed. The XBP1 coding sequence was amplified by polymerase chain reaction (PCR) method, and was cloned in pGEM-T vector. After the target region was sequenced, it was subcloned into the bait plasmid pGBKT7, then was transformed into yeast AH109(a type). After the expression of bait plasmid pGBKT7-XBP1 in AH109 yeast strains were proved by Western blot. The transformed yeast AH109 was mated with yeast Y187(α type) containing hepatocyte cDNA library plasmids pACT2 in 2 ×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medilium (SD/-Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After sequencing and verifying ORF of positive colonies,7 different kinds of proteins were obtained. In order to further testify the interaction between the screened proteins and XBP1, one of positive colonies MT1E was cloned. The interaction between MT1E and XBP1 in vitro/in vivo were examined successfully with GST pulldown and coimmunoprecipitations. It was shown that MT1E would be a new regulatory protein of XBP1. These screened proteins by yeast two-hybird system were closely correlated with liver fundamental metabolism,protein synthesis and transport,cell proliferation and apoptosis. The results mentioned above contributed to reveal the XBP 1 biological function, and brought some new clues for further exploration of the expressing and regulating mechanism of XBP1.
4.Exploration on graduate management mode based on scientific research innovation platform
Faping YI ; Youquan BU ; Geli LIU ; Luyu ZHANG ; Yingxiong WANG ; Fangzhou SONG
Chinese Journal of Medical Education Research 2013;(2):127-130
In order to strengthen the graduate management in scientific research platform and to ensure the quality of graduate training,the ideological and moral education was invigorated through establishing virtual party branch,the behavior was regulated through establishing and amplifying the daily management system,the student interests were protected through establishing financial management system and the cultivation quality was guaranteed through perfecting the academic management system.Satisfactory results were achieved in molecular medicine and cancer research center in Chongqing Medical University.
5.Stable expression of MCHR2 in CHO cell line and analysis of its signal transduction pathways
Junxia YANG ; Chengfu YUAN ; Hua SHI ; Lili WEI ; Ji CHEN ; Faping YI ; Yongping MA ; Fangzhou SONG
Chinese Pharmacological Bulletin 1987;0(03):-
Aim To construct a eukaryotic expression vector of human MCHR2 and to analyze the stable expression and signal transduction pathways of MCHR2 in CHO cells. Methods The full-length MCHR2 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and inserted into eukaryotic expres-sion vector pcDNA3.1(+), resulted in the recombinant expression vector pcDNA3.1-MCHR2. The recombinant plasmid was confirmed by the restriction enzymatic digestion and DNA sequencing analysis, and then transfected into CHO cells by lipofectamine. A stably-transfected cell line was obtained by the dominant G418 selection, and the expression of the MCHR2 gene on transcription and translation levels were identified by RT-PCR and Western blot. Signal transduction pathways mediated by MCHR2 were analyzed by measurement of intracellular cAMP and calcium. Results The eukaryotic expression vector pcDNA3.1-MCHR2 was successfully constructed and the MCHR2 gene was stably transfected into CHO cells. A stably-transfected cell line was established and the MCHR2 gene was efficiently transcribed and translated. MCH stimulation had no effect on the production of cAMP, however, it could induce a clear and transient increase of intracellular calcium, suggesting that MCHR2 was only coupled to Gq protein. Conclusion The stable expression of MCHR2 and analysis of its signal transduction pathways in CHO cell line provided a solid experimental foundation for further studies on the function of the MCHR2 gene in vitro.
6.Inhibition effect of silencing HPV16 E6 gene on human cervical cancer CaSki cell carcinoma xenograft in nude mice
Haien CHENG ; Yi ZHANG ; Cuili ZHANG ; Weiwei PAN ; Hua SHI ; Faping YI ; Fengjin GUO ; Fangzhou SONG
Journal of Third Military Medical University 1988;0(06):-
Objective To learn the effect on carcinoma xenograft in nude mice by inhibiting human papillomavirus 16(HPV16) E6 gene expression in CaSki cell. Methods The recombinant plasmids expressing HPV16 E6 small interference RNA (siRNA) were transfected into CaSki cell. The cells expressing recombinant plasmid was screened out with G418. The expression of E6 mRNA was determined by RT-PCR. The cells were inoculated into BALB/c nude mice subcutaneouly and the growth of the xenograft carcinoma was observed. After the pGensil-CH2 recombinant was injected into the carcinoma, the growth of carcinoma and pathological changes of carcinoma were observed. Results The CaSki cell expressing E6 siRNA was obtained, and HPV16 E6 mRNA expression in CaSki cell was down-regulated. The oncogenicity of the CaSki cell expressing E6 siRNA was degraded, the inhibition rate was up to 71.4% as compared with that of control group. The growth of tumor in nude mice was inhibited after the E6 siRNA plasmids were injected into the nude mice. The volume and weight of the tumor treated by siRNA were smaller than that of control group significantly. More necrotic area and less cell division phase were observed under light microscope in the E6 siRNA treated tumor. Conclusion The oncogenicity of the CaSki cell was degraded after silencing HPV16 E6 gene in CaSki cell by E6 siRNA.
7.Preliminary study on the effect of Pinggan huoxue powder on cerebral ischemia-reperfusion injury in rats and its mechanism
Faping SONG ; Jie ZHANG ; Yong TANG ; Zongmei LIU ; Xiaoling WANG
China Pharmacy 2022;33(24):2999-3004
OBJECTIVE To investigate the effect and mechanism of Pinggan huoxue powder on cerebral ischemia-reperfusion injury (CIRI) in rats. METHODS CIRI model was induced by suture-occluded method. SD rats were divided into sham operation group, model group, positive control group (Tongxinluo capsule 0.325 g/kg), Pinggan huoxue powder low-dose, medium-dose and high-dose groups (3.02, 6.04, 12.08 g/kg), with 12 rats in each group. Each group was given constant volume of water or drug intragastrically, once a day, for consecutive 14 d. Longa score was used to evaluate neurological function of rats on the 1st, 7th and 14th day of intervention. After the last administration, the percentage of cerebral infarction volume in rats was measured, and neuronal injury (neuronal density) and neuronal apoptosis were observed; the protein expressions of p53, B cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), Caspase-3 and activated Caspase-3 (Cleaved caspase-3) in brain tissue were detected. RESULTS Compared with sham operation group, Longa score at the 1st, 7th and 14th day of intervention and the percentage of cerebral infarction volume in model group significantly increased (P<0.05), and the arrangement of cortical cells in ischemic side was disordered, Nissl bodies were absent and vacuolated, and a large number of cell apoptosis occurred. The neuronal density and protein expression of Bcl-2 at the lesion side were significantly decreased (P<0.05), while the protein expressions of p53, Bax, Caspase-3 and Cleaved Caspase-3 were significantly increased (P<0.05 or P<0.01). Compared with model group, the expressions of Bcl-2 in cerebral ischemia cortex of rats in Pinggan huoxue powder high-dose group were significantly increased (P<0.05). Longa scores of rats in Pinggan huoxue powder medium-dose and high-dose groups on the 7th and 14th day of intervention were significantly decreased (P<0.05), and other indexes were also significantly reversed (P<0.05 or P<0.01). CONCLUSIONS Pinggan huoxue powder can protect cerebral nerves and inhibit apoptosis of CIRI model rats, the mechanism of which may be related to down-regulating the protein expressions of p53, Bax and Caspase-3 and up-regulating the protein expression of Bcl-2.