10.3969/j.issn.1007-3969.2013.04.002

A pair of specific primers was designed based on the reported Bm86 gene of Boophilus microplus,the Bm86 gene was cloned by PCR using the plasmid pMD18-T-Bm86 as templates,and subcloned into the prokaryotic plasmid pGEX-4T-1.The recombined plasmid was transformed into E.coli BL21(DE3) and followed by expression of the protein induced by different concentration of IPTG for different time.SDS-PAGE showed that the recombinant plasmid pGEX-4T-1/Bm86 expressed a fusion protein Bm86-GST(Mr 94 000) after being induced with IPTG.High level expre-ssion of Bm86-GST was found at 1 mmol/L IPTG condition after incubation for 8 h at 37 ℃,and the expression level of the recom-binant Bm86-GST reached up to 29% of total E.coli proteins.Western-blotting analysis showed that the recombinant Bm86-GST was recognized by the rabbit anti-B.microplus positive serum.

Through this research a lentiviral vector expressing the gene of folate-binding protein-1 (FOLR1) was constructed and the corrsponding expression products were identified. Firstly, full-length of the FORL1 gene was amplified by PCR and cloned into the plasmid pWPI. Then it was further confirmed by PCR and sequencing. Secondly, after the recombinant pWPI and its helper plasmid co-transfected the virus packaging 293T cells, SKOV3 cells were infected with the virus particles and sorted by flow cytometry. Thirdly, the FOLR1 gene was detected by RT-PCR and its protein expression was detected by Western blot. Finally, the recombinant expression vector was successfully constructed, and lentiviruses were successfully packaged by the 293T cells. A great quantity of green fluorescent cells could be seen after the SKOV3 cells were effectively infected with the lentiviruses carrying the FOLR1 gene. The sorting could be done and detected by cytometrying the FORL1 gene and its stable expression by the two methods above, which laid experimental foundation for exploring its biological function in ovarian cancers.
Cell Line ; Cell Line, Tumor ; Cloning, Molecular ; Female ; Folate Receptor 1 ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Kidney ; cytology ; Lentivirus ; genetics ; metabolism ; Ovarian Neoplasms ; pathology ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

Species identification using DNA sequences is the basis for DNA taxonomy. In this study, we sequenced the ribosomal large-subunit RNA gene sequences (3,037-3,061 bp) in length of 13 Chinese Theileria stocks that were infective to cattle and sheep. The complete 28S rRNA gene is relatively difficult to amplify and its conserved region is not important for phylogenetic study. Therefore, we selected the D2-D3 region from the complete 28S rRNA sequences for phylogenetic analysis. Our analyses of 28S rRNA gene sequences showed that the 28S rRNA was useful as a phylogenetic marker for analyzing the relationships among Theileria spp. in ruminants. In addition, the D2-D3 region was a short segment that could be used instead of the whole 28S rRNA sequence during the phylogenetic analysis of Theileria, and it may be an ideal DNA barcode.
Animals ; Base Sequence ; China ; DNA, Ribosomal/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 28S/genetics ; Ruminants ; Sequence Alignment ; Sequence Analysis, DNA/veterinary ; Theileria/*classification/genetics/isolation & purification ; Theileriasis/*parasitology

In order to determine the effect of various hosts on feeding performance of Rhipicephalus (Boophilus) microplus, we used 3 mammalian species as hosts, cattle (Qinchuan), sheep (T an), and rabbits (Japanese white rabbit) for infest-ing ticks. Five hundreds of R. microplus larvae were exposed to each animal (3 animals/host species). Tick recoveries were 11.0%, 0.47%, and 5.5% from cattle, sheep, and rabbits, respectively. The averages of tick feeding periods were not significantly different on cattle, sheep, and rabbits, 28.8, 25.3, and 26.7 days, respectively. The average weights of individual engorged female from cattle, sheep, and rabbits were 312.5, 219.1, and 130.2 mg, respectively and those of egg mass weights each to 85.0, 96.6, and 17.8 mg. The highest egg hatching rate was in the ticks from cattle (96.0%), fol-lowed by those from rabbits (83.0%) and sheep (19.2%). These data suggest that rabbits could be as an alternative host to cultivate R. microplus for evaluating vaccines and chemical and biological medicines against the tick in the laboratory, although the biological parameters of ticks were less than those from cattle.
Animals ; Cattle* ; Female ; Humans ; Larva ; Ovum ; Rabbits* ; Rhipicephalus* ; Sheep* ; Ticks ; Vaccines ; Weights and Measures

Background Occupational stress and depressive symptoms of disease prevention and control personnel are serious. Objective To investigate the relationship between occupational stress, psychological capital, and depressive symptoms of disease prevention and control personnel, and analyze the potential mediating effect of psychological capital on the relationship between occupational stress and depressive symptoms. Methods From July to September 2020, a cluster random sampling method was used to select 2201 employees from 21 centers for disease control and prevention as study subjects covering all levels of administrative divisions in Jiangsu Province. A total of 2036 valid questionnaires were collected with a recovery rate of 92.5%. The Core Occupational Stress Scale, Patient Health Questionnaire, and Psychological Capital Questionnaire were used to investigate their occupational stress, depressive symptoms, and psychological capital. Stratified regression analysis was used to explore the effects of occupational stress and psychological capital on depressive symptoms. A mediating effect model was used to analyze and verify the potential mediating effect of psychological capital on the relationship between occupational stress and depressive symptoms. Results The total scores in M (P₂₅, P₇₅) of occupational stress, depressive symptoms, and psychological capital in the target population were 42.0 (37.0, 48.0), 8.0 (4.0, 9.0), and 4.6 (4.0, 5.0) respectively. The positive rate of occupational stress was 31.0% (631/2036), and the positive rate of depressive symptoms was 22.0% (448/2036). The dimensional scores of organization and reward, and demand and effort of occupational stress were positively correlated with the total score of depressive symptoms [Spearman correlation coefficients (r_s) were 0.371 and 0.269, P<0.05]. The dimensional scores of social support and autonomy of occupational stress and the score of psychological capital were negatively correlated with the total score of depressive symptoms (r_s=−0.373, −0.112, −0.494, P<0.05). The organization and reward, and demand and effort had positive effects on depressive symptoms (b=0.188, 0.177, P<0.05), while social support and autonomy had negative effects on depressive symptoms (b=−0.290, −0.078, P<0.05), and associated with a 22.5% increase of explanatory variance. Psychological capital had a negative effect on depressive symptoms (b=−0.368, P<0.05), and associated with an 11.0% increase of explanatory variance. Psychological capital had mediating effects on the associations of social support, organization and reward, and autonomy with depressive symptoms, and the mediating effect values were −0.210 (95%CI: −0.253-−0.171), 0.096 (95%CI: 0.071-0.122), and −0.164 (95%CI: −0.229-−0.103), respectively. The corresponding mediating effect percentages were 40.23%, 26.97%, and 45.56%, respectively. Conclusion Occupational stress of disease prevention and control personnel can directly affect depressive symptoms, but also indirectly through psychological capital. Psychological capital plays a partial mediating role in the associations of social support, organization and reward, and autonomy of occupational stress with depressive symptoms. The occurrence of depressive symptoms can be reduced by decreasing occupational stress and increasing psychological capital.