To investigate the effect of miR-331-3p on the proliferation of porcine renal epithelial cells (PK15) and its mechanism, the pcDNA 3.1(+) overexpression vector of miRNA-331-3p (pcDNA 3.1(+)-miR-331-3p) was constructed. PK15 cells were divided into four groups, including experimental group, experimental control group, inhibitor group and inhibitor control group. Experimental group and experimental control group were transfected with pcDNA 3.1(+)-miR-331-3p and pcDNA 3.1(+), respectively. Inhibitor group and inhibitor control group were transfected with miR-331-3p inhibitor and miR-331-3p negative control (miR-331-3p NC), respectively. Above all, CCK-8 reagent was used to plot the cell proliferation curve and Propidium (PI) staining was used to detect the proportion of cell stages. Secondly, its expression change were detected by quantitative real-time PCR that included the growth inhibitory protein family member 5 (ING5), cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 3 (CDK3), cyclin-dependent kinase 4 (CDK4), Cyclin B and cyclin-dependent kinase inhibitor 1A (CDKN1A). The results showed that the expression of miRNA-331-3p was significantly increased in the experimental group. The cell proliferation curve showed that the number of cells in experimental group was significantly higher than that in experimental control group or inhibitor control group at 48 h and 72 h (P<0.05). Simultaneously, Inhibitor group was significantly lower than experimental control group or inhibitor control group in the number of cells at 48 h and 72 h (P<0.05), but there was no significant difference between the experimental group and the control group. Compared with the experimental control group, the proportion of cells of experimental group in G0/G1 phase decreased, the proportion of S phase and G2/M phase increased, and the inhibitor control group showed the opposite trend. Simultaneously, the expression levels of CDK2, CDK3, CDK4 and Cyclin B genes in the experimental group were significantly increased, while ING5 and CDKN1A genes inhibiting proliferation showed a significant downward trend. These results demonstrate that the miR-331-3p overexpression vector was successfully constructed, and miR-331-3p has the ability to promote the proliferation of PK15 cells. The study lays a solid foundation for further research for its role in pig growth and development.
Animals ; Cell Line ; Cell Proliferation ; genetics ; Epithelial Cells ; cytology ; Genetic Vectors ; MicroRNAs ; genetics ; Swine

Objective structured clinical examination (OSCE) is an important assessment method for examining clinical thinking and clinical skills of medical students. OSCE has been applied in the quarterly phase examination in our department. According to the characteristics of otolaryngology, three stations of common items and five stations for personalized tests were set in the assessment. According to the different training that residents received during their stay in the previous department, different contents were assessed in the station for personalized examination. During the OSCE, residents took turns entering into each examine station and teachers gave scores according to the marking criteria of different station. After the examinations, two-way assessment and feedback were conducted between students and teachers. Through the practice in standardized training of residents in recent years, we found that OSCE was more comprehensive and flexible than others, which gained good application effects.

Objective To investigate the prevalence of Mycoplasma pneumoniae ( Mp) infection in children in Hengyang from 2013 to 2016 and to analyze the p1 genotypes of the isolated Mp strains by using polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP) , nested polymerase chain reaction (nPCR) and rapid-cycle polymerase chain reaction (Rapid-Cycle PCR).Methods Throat swab samples of children with acute respiratory tract infection were collected from four hospitals in Hengyang , Hu-nan Province from 2013 to 2016.Mp strains in these samples were identified by PCR amplification of the 16S rRNA gene.PCR-RFLP, nPCR and Rapid-Cycle PCR were performed for Mp p1 genotyping in order to fur-ther analyze the genotypes of Mp strains circulating in Hengyang .Results A total of 109 clinical strains of Mp were identified from the 984 throat swab samples .The sensitivities of PCR-RFLP and nPCR for genoty-ping MP strains were both 100％, while that of rapid-Cycle PCR was 98 .17％.All of the three methods showed 100％specificity for genotyping.Of all isolated Mp strains, 78.90％ were p1 gene type Ⅰ and 21.10％were p1 gene typeⅡ(t=93.239, P=0.01).From 2013 to 2016, the annual isolation rates of p1 gene type Ⅰ and type Ⅱ strains were 93.10％, 87.5％, 76.92％, 65.79％ and 6.90％, 12.5％, 23.08％, 34.21％, respectively.The rate of Mp p1 gene type Ⅰinfection decreased over year , while that of p1 gene type Ⅱinfection increased gradually .Conclusion PCR-RFLP, nPCR and rapid-Cycle PCR are reliable for genotyping of Mp p1 gene.The predominant genotype of Mp strains circulating in Hengyang is p 1 gene type Ⅰ, but the incidence of p 1 gene type Ⅱinfection gradually increases from 2013 to 2016 .

OBJECTIVE: To evaluate the diagnostic value of modified Berlin questionnaire on predicting obstructive sleep apnea-hypopnea syndrome (OSAHS)in Chinese adults. METHOD: Differ from the original version, BMI cut-off point was adjusted to 25.0 in modified Berlin questionnaire according to the Asia -Pacific obesity definition. A total number of 244 samples who experienced polysomnography (PSG) were included. After well informed, each patient finished questionnaire by an interview. The results of the original and modified questionnaires and polysomnography reports were compared with polysomnography reports to evaluate the diagnostic accuracy of the modified Berlin questionnaire. RESULT: In male population, the sensitivity and specificity of original Berlin questionnaire were 74.03% and 65.71%, respectively. The percentage of diagnostic consistency was 72.49%, and Kappa coefficient was 0.304 (P < 0.01). In aspect of modified version, the sensitivity and specificity were 92.21% and 48.57%. The percentage of diagnostic consistency was 84.13%, and Kappa coefficient was 0.437 (P < 0.01). In female population, the sensitivity and specificity of original version were 50.00% and 61.90%, respectively. The percentage of diagnostic consistency was 54.55%, and Kappa coefficient was 0.110 (P > 0.01). When comes to the modified Berlin questionnaire, the sensitivity and specificity were 76.47% and 47.62% in female subjects, and the percentage of consistency agreement was 65.45%, with a Kappa coefficient 0.248 (P > 0.01). CONCLUSION Compared to the original version, the modified Berlin questionnaire has a better diagnostic consistency with a considerable sensitivity and specificity in male population. However, in female subjects group, the consistency of the modified Berlin questionnaire was still not significant remains unsatisfactory, although P value has been improved. Modified Berlin questionnaire could be. used as a primary screening tool for male OSAHS patients. However, but the screening questionnaire for female should still be need to be further explored.
Adult ; Female ; Humans ; Male ; Middle Aged ; Obesity ; Polysomnography ; Sensitivity and Specificity ; Sleep Apnea Syndromes ; diagnosis ; Sleep Apnea, Obstructive ; diagnosis ; Surveys and Questionnaires

Objective To construct a fusion protein that used for treatment of resistance and palindromia in leukemia and studied its biological activity. Methods IL-3 and LP gene fragments were amplified by PCR. After enzymatic digestion and T4 ligation, the fusion gene was cloned into expression vector pAYZ. The product was purified by exchange chromatography and anti-Etag affinity chromatography. IL3-G4SLP fusion protein was analyzed by SDS-PAGE and Western blot. Protein biological activity was detected by FACS. Results The fusion protein was expressed as soluble protein by E.Coli 16C9. The protein expression level was about 1 mg/L, its purity was over 95 %, and the expression level was about 1 mg/L. The fusion protein can combined specificely with CD123 on leukemia stem cells. Conclusion Fusion protein IL-3-G4S-LP can target on leukemia stem cells and maybe as a potential drug used for treatment of resistance and palindromia in leukemia.