Objective To construct short-hairpin RNA (shRNA) eukaryotic express vectors targeting of insulin like growth factor-1 receptor (IGF1R) gene, and to explore the changes of adhesion, invasion and FAK protein expression of MHCC-97 H hepatocellular carcinoma cells with RNA interference. Methods The shRNA oligonucleotide fragments were designed and synthesized based on the sequence of IGF1R mRNA. Double strands were formed after annealing and inserted into pGCsi-U6-Neo-GFP vector. The recombinant was transformed into Stable 3, then plasmids were extracted and identified by restriction enzyme and sequencing analysis. The most effective vectors were selected by RT-PCR and Western blotting after transfecting 293T cells. The best one was used to transform MHCC-97H cells which were selected with G418 to obtain positive colons. The changes of adhesion, invasion and FAK protein expression in MHCC-97H cells were studied. Results The restriction enzyme analysis and sequencing analysis demonstrated that shRNA had been inserted into vectors, and their sequences were the same as the design. It was indicated by RT-PCR and Western blotting that the silencing efficacy of IGF1R was 88%. The ability of adhesion and invasion significantly decreased after IGF1R silencing in MHCC-97H cells, and so was the expression of FAK protein. Conclusion IGF1R pGCsi-U6-Neo-GFP shRNA can significantly decrease the abilities of adhesion and invasion in MHCC-97H cells, and inhibit the expression of FAK protein.

Objective:To understand the prevention knowledge of brucellosis among permanent residents in the agricultural area of Hulunbuir City, Inner Mongolia Autonomous Region, and to provide a scientific basis for formulating brucellosis prevention strategies and measures.Methods:From May to December 2018, Daur Autonomous Banner of Morin Dawa and Arun Banner in the agricultural area of Hulunbuir City were selected as the survey sites. Three townships were selected from each banner, and one administrative village from each township was selected as the survey village. The permanent residents aged ≥14 who had lived in the survey village for at least one year were selected as the survey subjects. Serum was separated by the examiner and the rose-bengal plate agglutination test (RBPT) was conducted. The positive sera were further tested by tube agglutination test (SAT).Questionnaires were conducted by investigators in the form of face-to-face case interview, including general demographic information, family information, brucellosis prevention knowledge awareness status, epidemiological contact history, prevention status, general behavior, etc., to calculate the awareness rate of brucellosis prevention knowledge.Results:A total of 880 sera were isolated, of which 185 were positive for RBPT. Further examination of SAT showed that the positive rate of SAT was 17.27% (152/880). The overall awareness rate of brucellosis prevention knowledge was 46.73% (16 450/35 200). The awareness rates of males and females were 47.82% (8 473/17 720) and 45.64% (7 977/17 480), respectively. The 21 - 40 years old group had the highest awareness rate [60.94% (3 705/6 080)]. The awareness rates of primary school and below, junior high school, senior high school and above were 45.96% (11 969/26 040), 48.79% (3 962/8 120) and 49.90% (519/1 040), respectively. The awareness rate of people with livestock raising behavior in the year before the survey was 47.35% (8 011/16 920), while the awareness rate of people without such behavior was 46.17% (8 439/18 280).Conclusions:The overall awareness rate of brucellosis prevention knowledge among permanent residents in the agricultural area of Hulunbuir City is low. It is necessary to strengthen the training of relevant knowledge and strengthen the publicity and education of people with low awareness rate.

OBJECTIVE: To explore molecular etiology and clinical characteristics of two pedigrees affected with hereditary factor VII(FVII) deficiency. METHODS: The nine exons and flanking sequences of the F7 gene of the probands were amplified by PCR. The amplicons were analyzed by direct sequencing. Suspected mutations were subjected to SWISS-MODEL modeling and analysis of protein structure change by Pymol software and conservation of amino acids across various species. RESULTS: For proband of pedigree 1, the prothrombin time (PT), FVII activity (FVII:C) and FVII antigen (FVII:Ag) were 36.3 s, 3%, 53.56%, respectively. Sequencing revealed a compound heterozygous variants of c.80_81delCT and c.1371G>T(p.Arg439Ser). His son carried a heterozygous c.1371G>T (p.Arg439Ser) variant. For proband of pedigree 2, the PT, FVII:C and FVII:Ag were 22.3 s, 4%, 1.58%, respectively. Sequencing has revealed a compound heterozygous c.278G>T(p.Arg75Met) missense variant in exon 3 and c.1278T>G (p.His408Gln) in exon 9 of the F7 gene. His mother and son both carried a heterozygous c.278G>T(p.Arg75Met) variant. Three-dimensional simulation and homology analysis revealed that the p.Arg439Ser and p.Arg75Met can respectively alter part of hydrogen bonds and two highly conserved amino acids. CONCLUSION Two novel heterozygous missense variants of the F7 gene [c.1371G>T(p.Arg439Ser) and c.278G>T(p.Arg75Met)] probably account for the decrease of factor VII in the two pedigrees.
Asian Continental Ancestry Group ; Factor VII ; Factor VII Deficiency ; Genotype ; Heterozygote ; Humans ; Mutation ; Pedigree