AIM: To gain cdcSTBX25A-fas chimeric gene bearing the regulative fragment cdcSTBX25A and opening read frame of fas in order to construct and identify eukaryotic expression vectors, pAdTrack-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas, which have the potential to transfer the tumor proliferative signal to promoting-apoptosis signal through up-regulate the fas expression by c-myc. METHODS: A pair of primers were designed according to the known sequences of cdcSTBX25A and fas. The cdcSTBX25A-fas chimeric gene was obtained by PCR reaction and inserted into the two plasmids pAdTrack-CMV and pAdTrack, respectively. The two recombinant plasmids were transferred into E. coli DH5?. The positive clones were screened in LB media with 50 mg/L kanamycin and identified by agarose gel electrophoresis, endonuclease digestion and PCR. The nucleotide sequence of inserted cdcSTBX25A-fas was determined by dideoxy chain termination method. Using software, BLAST was conducted to analyze the structure and sequence of the target fragments and compared with GenBank. RESULTS: The chimeric target gene, cdcSTBX25A-fas, of 1 603 bp was obtained. The positive host bacteria E. coli DH5? of recombinant plasmids were screened and amplified. The double-enzyme digestion showed the pAdTrack-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas presenting 9.2 kb, 1.6 kb bands, and 8.3 kb, 1.6 kb bands respectively. The sequence analysis confirmed that the two shuttle plasmids containing 1 597 bp cdcSTBX25A-fas with the ORF of fas. CONCLUSION: The eukaryotic expression plasmids pAd-Track-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas were successfully constructed.

AIM: To detect the mRNA and protein level of HIF-1 alpha in the tissues of rat's radiated mucosa. METHODS: The left buccal mucosa was irradiated and excised. The right buccal mucosa was excised to serve as own control tissue. The mRNA of HIF-1 alpha was determined by using the semi quantitative RT-PCR. SABC method was employed to immunostain and to elucidate the localization, intensity and distribution of HIF-1 alpha protein.RESULTS: A Sprague-Dawley rat's model of radiation-induced oral mucositis (ROM) was successfully established. The results of RT-PCR indicated that the left buccal mucosa expressed HIF-1 alpha mRNA while the right buccal mucosa did not or seldom expressed it. Immunohistochemical analysis of HIF-1 alpha demonstrated that the left side mucosa expressed HIF-1 alpha protein.CONCLUSION: The mucosa of ROM expresses the mRNA and protein of HIF-1 alpha. The expressions of HIF-1 alpha are correlated with the severity of ROM.

Objective:To evaluate the residual risk (RR) of transfusion transmitted HIV (TT-HIV) after the implementation of nucleic acid amplification test (NAT) in blood screening test among blood centers in China.Methods:The data of blood donors and HIV infection markers from 2017 to 2020 were collected from 28 blood centers via the Platform of Comparison of blood establishments Practice in Chinese Mainland. The new infection rate/window period mathematical model was used for two types of blood screening strategies, namely, two rounds ELISA plus individual NAT take turn with pooling NAT (2ELISA+ ID-NAT/MP-NAT) and two ELISA plus one round pooling NAT (2ELISA+ MP-NAT), and the RR of HIV infection was estimated also based on first donors (FDs) and repeated donors (RDs) in different blood donation years. T-test analyses were conducted for comparing TT HIV RR among FDs and RDs in different blood donation years with two blood screening strategies, and the variation trend of RR in HIV test was observed.Results:From 2017 to 2020, the RR of FDs in 2ELISA+ ID-NAT/MP-NAT blood screening strategy was 2.869/10 6 person-year, 3.795/10 6 persons-year, 3.879/10 6 person-year, and 2.890/10 6 person-year respectively. The RR of RDs was 1.797/10 6 person-year, 1.502/10 6 person-year, 1.857/10 6 person-year, and 1.483/10 6 person-year respectively. Significant difference exists between RR of FDs and RDs, with F=9.898 and p<0.05. In 2ELISA+ MP-NAT strategy, the RR of FDs was 3.508/10 6 person-year, 1.868/10 6 person-year, 2.204/10 6 person-year, and 1.765/10 6 person-year respectively. The RR of RDs was 0.948/10 6 person-year, 0.926/10 6 person-year, 0.748/10 6 person-year, and 0.682/10 6 person-year respectively. Statistical difference existed between RR of FDs and RDs, with F=17.126 and P<0.05. There was no significant difference between the RR of FDs in these two strategies with F=3.493 and P>0.05, while there was a difference between the RR of RDs in these two strategies with F=24.516 and P<0.05, and a difference between the RR of total donors (TDs) in these two strategies F=20.216 and P<0.05. Conclusions:The RR of TT HIV significantly decreased after the introduction of NAT into blood test among blood centers in China. There were some differences in the RR of HIV testing among different blood screening strategies. There could be significant differences in the RR of HIV testing among different groups of blood donors. Compared with FDs, RDs is the low risk group for HIV.