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Objective@# To investigate the AIDS-related knowledge, attitudes and practice among university students in a pilot site of AIDS health education in Zhoushan City, so as to provide the reference for AIDS health education in universities.@*Methods@#University students were recruited from Zhejiang Ocean University using a stratified cluster sampling method in 2018 and 2020, and the AIDS-related knowledge, attitudes and practice were collected using a questionnaire survey. @*Results@#A total of 2 862 and 2 850 students were surveyed in 2018 and 2020, including 1 429 ( 49.93% ) and 1 414 ( 49.61% ) male students, respectively. The overall awareness of AIDS-related knowledge was 86.62% and 94.11% in 2018 and 2020, respectively. There were 544 ( 19.01% ) and 394 ( 13.82% ) students that were in favor of one-night stand, 308 ( 10.76% ) and 198 ( 6.95% ) students that were in favor of commercial sexual behaviors, and there were 59 ( 2.06% ) and 34 ( 1.19% ) students that had casual sexual behaviors within one year, and 20 ( 0.70% ) and 8 students ( 0.28% ) with commercial sexual behaviors within one year. There was a significant difference in the awareness of AIDS-related knowledge among students with different years at the university ( P<0.05 ). The lowest awareness of AIDS-related knowledge was seen in freshmen in 2018 ( 81.74% ), and the highest awareness was found in freshmen in 2020 (97.17% ). The proportions of being in favor of one-night stand ( 2018: 31.35% vs. 6.70%; 2020: 22.07% vs. 5.71%; P<0.05 ), being in favor of commercial sexual behaviors ( 2018: 19.91% vs. 2.23%; 2020: 12.09% vs. 1.88%; P<0.05), having casual sexual behaviors within one year (2018: 3.71% vs. 0.42%; 2020: 2.19% vs. 0.21%; P<0.05), and having commercial sexual behaviors within one year ( 2018: 1.33% vs. 0.07%; 2020: 0.50% vs. 0.07%; P<0.05 ) were significantly greater in male students than in female students.@*Conclusions@#The pilot AIDS health education is effective to increase the awareness of AIDS-related knowledge, possess the correct attitudes towards sexual behaviors and reduce high-risk sex behaviors among university students in Zhoushan City. Intensified AIDS health education is recommended among senior and male university students.
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Objective To analyze the effect of GC-rich DNA fragments on the level of transgenic expression in Chinese hamster ovary (CHO) celts and its position effect.Methods The synthetic DNA fragment with GC-rich was cloned into the 5'or 3'or both 5'and 3'ends of expression cassette of expression vector.Three new expression vectors (pIRES-G1,pIRES-G2 and pIRES-G3) which was inserted with the GC-rich DNA fragments in different position were transfected CHO ceils,respectively,and then was observed under fluorescence microscope;the control vector was pIRES-EGFP.Stable transfected cell lines were screened under G418,and enhanced green fluorescent protein(EGFP) expression was analyzed by flow cytometry and the transgenic copy number was detected by quantitative real-time quantitative polymerase chain reaction (qRT-PCR).Results Three expression vectors with a GC-rich DNA fragments in different position were constructed successfully.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector could obviously improve the expression level of vector in CHO cells;and the expression level of the stably transfected CHO cells increased 1.39 fold and 1.32 fold compared to the control vector,respectively;the transgene copy number increased 1.32 fold and 1.24 fold compared with the control vector.While the insertion of GC-rich DNA fragments at 5'end of expression cassette had no obvious effect on the level of gene expression.Conclusion The role of DNA fragment with GC-rich in improving the transgenic expression of CHO cells is related to its position in the vector.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector can improve transgenic expression.
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BACKGROUND:Matrix attachment region(MAR)are DNA elements that bound to the nuclear matrices after chromatin digested with restriction endonuclease.Plenty of studies have shown that MAR considered as initiaI point of DNA replication or transcription of regulatory gene.Thereby,construction of MAR expression vector can elevate the overall level of transgene expression,enhance stability of exogenous gene.as welI as increase frequency of stable transfectant cells.OBJECTIVE:To construction pLXSN-CAT recombinant retrovirus vector that containing chloramphenicol acetyltransferase(CAT)via cloning MAR sequence of human.and to explore the influence of MAR on the gene expression.METHODS:An open experiment was performed at the Department of Biochemistry and Molecular Biology.Xinxiang Medical College from September 2007 to December 2007 The PLXSN-CAT vector of CAT was constructed by the laboratory.TaqDNA polymerase,T_4 DNA ligase,DNA Marker,restriction enzyme BamH I,agarose gel DNA purification kit,as well as plasmid purification kit were purchased fromTakara Biotechn0Iogy(Dalian)Co.,Ltd.The sequence of csp-B MAR was amplified by polymerase chain reaction(PCR)method applied to human DNA.The fragment was inserted into retrovirus vector PLXSN-CAT plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.RESULTS AND CONCLUSION:The length of specific fragment applied by PCR was 931 bp,and the recombinant plasmid PLXSN-CAT-MAR presented two bands:5.9 kb and 931 bp using respective restriction enzymes BamH I The sequence of MAR was confirmed by blasting to Genbank(serial numobr:M6271 6).It suggested that MAR had been cloned into PLXSN-CATR vector correctly.The recombinant retrovirus vector PLXSN-MAR was successfully constructed.
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BACKGROUND: Matrix attachment region (MAR), a DNA sequence, is still bound to the nuclear matrices after chromatindigested with restriction endonuclease, not only affects expression of endogenous gene, but also overcames transgenic silence andimproves transcription and expression of exogenous gene. OBJECTIVE: To investigate the influence of ?-interferon MAR of CHO cells on the transgenic expression of chloramphenicolacetyltransferase (CAT). DESIGN, TIME AND SETTING: The opening experiment was performed at the Department of Biochemistry and MolecularBiology, Molecular Institute, Xinxiang Medical College from October 2006 to April 2007. MATERIALS: CHO cell lines were obtained from China Center for Type Culture Collection. The pCATG vector of CAT and G418screening markers were constructed by this laboratory. METHODS: Human ?-interferon MAR by PCR was digested with SacI/KpnI and BamHI/SalI, and was inserted into pCATGvector, which was propagated in Escherichia coli JM109, then extracted and purified followed by enzyme digestion analysis. Vectorof CAT expression cassette and human ?-interferon MAR by the two sides was successfully constructed, and christened aspCAT-MAR. Two methods were compared between CHO cells of pCATG transformation and CHO cells of pCATG-MARtransformation. After G418 selecting, genome DNA of cell lines of G418 was extracted, then primers for PCR to amplify the CATtarget gene fragment was designed. MAIN OUTCOME MEASURES: The activity of CAT was analyzed by ELISA method. It was also tested to see if thepCATG-MAR was stably integrated into genomic DNA in the transfected cells. RESULTS: CHO cells of pCATG transformation was screened to have 16 strains of positive cell, and CHO cells of pCATG-MARtransformation was screened to have 17 strains of positive cell. Human ?-interferon MAR could increase the CAT gene expressionby 2.8 fold. The coefficient of variation of CHO cells of pCATG transformation was 2.065 0, and coefficient of variation of CHOcells of pCATG-MAR transformation was 0.813 1. Genome DNA of stable transformation cell lines was amplified by a fragment of437 bp. The results confirmed the pCAT-MAR vector was stably integrated into genomic DNA. CONCLUSION: Human ?-interferon MAR can increase transgenic expression in CHO cells and decrease the transgenicexpression variation in different transfected cells.
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A method of observation, examination and record was introduced in this paper, which was applicable for microorganism materials (example : bacteria, yeast moulds), animal and plant materials (example: animal tissue, flowers), bio-masromolecule (example: starch), industrial crystals, metallograph tests and general physics tests, through the hyphae, spores, spore sacs, expanding hyphae in fermentation and riboflavin crystals of E. ashbyii T30 (a riboflavin high-yield strain) had been observed.