With the rapid development of life science and technology and advent of knowledge explosion age,changes have taken place in modern educational ideas and goals.These changes require reform in the contents and methods of anatomy teaching to meet the needs of the new era.Philosophy provides a scientific world outlook and methodology for anatomy teaching reform.Teaching reform guided by philosophy is helpful in improving teaching efficiency and lays a solid foundation for the growth of medical students.

It was construct EGFP and HSV1TK co-expression vector and detect its expression and function in eukaryocyte ovarian cancer cells SKOV3.The HSV1TK gene were inserted into cloning vector pMD18-T to construct the plasmid of pMD18/ HSV1TK.The HSV1TK gene fragment obtained from pMD18T/HSV1TK that was digested,and then inserted into pcDNA3.1-EGFP.The recombinant pcDNA3.1-EGFP/ HSV1TK was identified with restriction analysis and DNA sequence.The expression plasmid pcDNA3.1-EGFP/ HSV1TK was transfected into ovarian cancer cells SKOV3 mediated by liposome reagent,then the expressions of EGFP in cells were observed by fluorescence microscopy and HSV1TK was detected with RT-PCR.The killing SKOV3 effect and bystander effect of HSV1TK/GCV was observed by MTT and light microscope.Results showed that the sequence of the cloned DNA fragment was identical to HSV1TK that was reported on GenBank,and the expression of vector pcDNA3.1-EGFP was correct.The recombinant expression plasmid was successfully transferred into ovarian cancer cells SKOV3,and effective expression of HSV1TK was also testified by RT-PCR.Thus,the recombinant eukaryotic co-expression vector of EGFP and HSV1TK was successfully constructed and effectively expressed in ovarian cancer cells SKOV3.HSV1TK/GCV has better killing effects and bystander effect in SKOV3 cells.

AIM:To investigate the ultrasound parameters and transfection efficiency of microbubble carrying herpes simplex virus thymidine kinase suicide gene(hsv1tk)for transfecting SKOV3 cells mediated by the ultrasound.METHODS:Under the combination conditions of different ultrasound exposure times(8,15,30,and 60 s with the interval as 1 s)and microbubbles at different concentrations,SKOV3 cells were exposed to the ultrasound so as to screen out the optimal ultrasound exposure time and microbubble concentration by 3-(4,5-dimethyhhizol-2-y1)2,5-diphenyl tetrazolium bromide(MTT)assays.The SKOV3 cells were divided into 6 groups and treated by ultrasound,ultrasound combined with liposome,ultrasound combined with microbubbles and liposome,and ultrasound combined with microbubbles respectively.The groups of negative control and positive contro1 were treated by naked plasmid and liposome respectively,and then they were used to transfect plasmids pcDNA3.1-EGFP/hsv1tk.The transfection efficiencies were observed qualitatively by fluorescence microscope and quantitatively by flow cytometry(FCM).The expression of hsv1tk was detected by reverse transcription polymerase chain reaction(RT-PCR).RESULTS:MTT assays showed that the transfection had no significant inhibition to cell viability under the conditions of the microbubble concentration(0.5 W/cm2),frequency(1 MHz),microbubble concentration(0.56×1011/L),and the interval of every 8 s ultrasound exposure time(1s).Under the fluorescence microscope,the green fluorescence intensity for the group of ultrasound combined with microbubbles and liposome was the greatest.The analysis of FCM showed that the transfection efficiency in group of ultrasound and microbubbles was higher than that in group of ultrasound[(11.74±0.19)% vs(2.19±0.22)%].Furthermore,the transfection efficiency in group of ultrasound combined with microbubbles and liposome was(25.62±0.08)%,which was the highest among all groups(P<0.05).The detection of RT-PCR showed the expression of hsv1tk in group of ultrasound combined with microbubbles and liposome was the highest among the groups(P<0.05).CONCLUSION:Under the conditions of theoptimal ultrasound parameters,ultrasound-mediated microbubbles can not only independently promote the hsv1tk gene transfection,but also strengthen the transfection efficiency of liposome.Meanwhile it can also be effectively expressed in SKOV3.

It was to assess the role of ErbB-4 in proliferation and invasion of human ovary cancer (OC) cells. Erb-4 receptor expression in human OC cells ovcar-3 was tested by immunohistochemisty (IHC). The effect of monoclonal antibody to ErbB-4(Ab-3) on the growth of ovcar-3 cells was evaluated by MTT assay. Following Ab-3(2.0 μg/ml) treatment, cell cycle and apoptosis were measured by flow cytometry (FCM). Results showed that OVCAR-3 cells expressed ErbB-4 level obviously. Ab-3(0.625~10 μg/ml)could inhibit the proliferation of OVCAR-3 cells in dosage and time dependent manner. The inhibition concentration (IC50) was 4.4836 μg/ml after 72 hour' treatment. Treated by Ab-3(2.0 μg/ml), cell cycle was arrested in S phase in OVCAR-3 cells, and apoptosis of OVCAR-3 cells was induced. In the membrane invasion culture system, the invasion number of Ab-3 treatment (78.0±6.1) was lower than the number of control groups (132.0±17.2) (P＜0.01). Therefore, Ab-3 can inhibit proliferation, invasion and induce apoptosis of OVCAR-3 expressing exogenous ErbB4.