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.

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 investigate the prognosis and influencing factors of secondary cytoreductive surgery (SCS) combined with chemotherapy in the treatment of recurrent ovarian cancer.Methods:A total of 102 patients with recurrent ovarian cancer admitted to our hospital from Jun. 2012 to Jun. 2015 were selected and grouped according to treatment methods. 31 patients who received paclitaxel/carboplatin (TC) chemotherapy were included in the control group, and 71 patients who received SCS combined with TC chemotherapy were included in the observation group. Clinical efficacy and 5-year survival outcome of the two groups after treatment, were compared and factors affecting the prognosis of the observation group were analyzed.Results:The total effective rate, 1-year survival rate, 3-year survival rate, and 5-year survival rate of the observation group were significantly higher than those of the control group. The median survival time of the observation group was 52 months and was significantly longer than that of the control group by 17 months ( P<0.05) ; There was no statistical difference between the death group and the survival group in terms of age, pathological type, tissue differentiation, recurrence tumor size, or location of recurrence tumors. The number of patients with FIGO stage IV, more than 3 recurrent tumors, ascites and residual lesion size >1 cm in the death group were significantly larger than those in the survival group. The serum CA125 level of patients in the death group was significantly higher than that in the survival group. Logistic regression analysis showed that the number of recurring tumors>3, with ascites, and residual lesions>1 cm, and high level of CA125 were independent risk factors for death after SCS combined with TC chemotherapy ( P<0.05) . Conclusions:SCS combined with chemotherapy can effectively improve the therapeutic effect, relieve the clinical symptoms, improve the survival rate of patients, and prolong the survival time of patients. The prognosis of SCS combined with chemotherapy is affected by the number of recurrent tumors, the presence or absence of ascites, the size of residual lesions, and CA125 level. The prognosis and survival of patients can be improved by adopting appropriate treatment.

Objective:To analyze the early identification, clinical characteristics, diagnosis and treatment of gestational hypertension crisis combined with adrenal disease.Methods:The clinical data of 23 patients of HCP complicated with adrenal disease admitted from Jul. 2009 to Jul. 2019 were retrospectively studied. The clinical characteristics, imaging characteristics, treatment and clinical transfer were studied.Results:The occurrence of all the 23 cases were acute. Among them, 16 cases had eclampsia combined with Cushing’s syndrome, 4 cases were pregnancy combined with primary aldosteronism (PA) and extreme hypokalemia, and 3 cases had eclampsia combined with pheochromocytoma (PHEO) . After admission, the patients were given symptomatic support treatment for sedation, analgesia, blood pressure control, dehydration, cranial pressure reduction, electrolyte balance and spasmolysis, and patients with severe preeclampsia and preeclampsia terminated their pregnancy in time. After treatment, 3 patients gave live birth, 12 received postpartum surgical treatment and 10 received drug treatment. The clinical symptoms improved and imaging examination suggested the lesions in the brain narrowed and disappeared, except one patient had major cerebral hemorrhage and died of multiple organ failure.Conclusions:In case of HCP and severe hypokalemia, relevant examinations should be improved in combination with symptoms to comprehensively diagnose whether it is complicated with adrenal diseases. The treatment methods and process of HCP with adrenal diseases need to be optimized, so as to judge the timing of termination of pregnancy, and minimize the impact on mother and fetus.

Background: Interferon lambda receptor 1 (IFNLR1) is a type II cytokine receptor that clings to interleukins IL-28A, IL29B, and IL-29 referred to as type III IFNs (IFN-λs). IFN-λs act through the JAK-STAT signaling pathway to exert antiviral effects related to preventing and curing an infection. Although the immune function of IFN-λs in virus invasion has been described, the molecular mechanism of IFNLR1 in that process is unclear. Objectives: The purpose of this study was to elucidate the role of IFNLR1 in the pathogenesis and treatment of porcine reproductive and respiratory syndrome virus (PRRSV). Methods: The effects of IFNLR1 on the proliferation of porcine alveolar macrophages (PAMs) during PRRSV infection were investigated using interference and overexpression methods. Results: In this study, the expressions of the IFNLR1 gene in the liver, large intestine, small intestine, kidney, and lung tissues of Dapulian pigs were significantly higher than those in Landrace pigs. It was determined that porcine IFNLR1 overexpression suppresses PRRSV replication. The qRT-PCR results revealed that overexpression of IFNLR1 upregulated antiviral and IFN-stimulated genes. IFNLR1 overexpression inhibits the proliferation of PAMs and upregulation of p-STAT1. By contrast, knockdown of IFNLR1 expression promotes PAMs proliferation. The G0/G1 phase proportion in IFNLR1-overexpressing cells increased, and the opposite change was observed in IFNLR1-underexpressing cells. After inhibition of the JAK/STAT signaling pathway, the G2/M phase proportion in the IFNLR1-overexpressing cells showed a significant increasing trend. In conclusion, overexpression of IFNLR1 induces activation of the JAK/STAT pathway, thereby inhibiting the proliferation of PAMs infected with PRRSV. Conclusion Expression of the IFNLR1 gene has an important regulatory role in PRRSVinfected PAMs, indicating it has potential as a molecular target in developing a new strategy for the treatment of PRRSV.

Background: Interferon lambda receptor 1 (IFNLR1) is a type II cytokine receptor that clings to interleukins IL-28A, IL29B, and IL-29 referred to as type III IFNs (IFN-λs). IFN-λs act through the JAK-STAT signaling pathway to exert antiviral effects related to preventing and curing an infection. Although the immune function of IFN-λs in virus invasion has been described, the molecular mechanism of IFNLR1 in that process is unclear. Objectives: The purpose of this study was to elucidate the role of IFNLR1 in the pathogenesis and treatment of porcine reproductive and respiratory syndrome virus (PRRSV). Methods: The effects of IFNLR1 on the proliferation of porcine alveolar macrophages (PAMs) during PRRSV infection were investigated using interference and overexpression methods. Results: In this study, the expressions of the IFNLR1 gene in the liver, large intestine, small intestine, kidney, and lung tissues of Dapulian pigs were significantly higher than those in Landrace pigs. It was determined that porcine IFNLR1 overexpression suppresses PRRSV replication. The qRT-PCR results revealed that overexpression of IFNLR1 upregulated antiviral and IFN-stimulated genes. IFNLR1 overexpression inhibits the proliferation of PAMs and upregulation of p-STAT1. By contrast, knockdown of IFNLR1 expression promotes PAMs proliferation. The G0/G1 phase proportion in IFNLR1-overexpressing cells increased, and the opposite change was observed in IFNLR1-underexpressing cells. After inhibition of the JAK/STAT signaling pathway, the G2/M phase proportion in the IFNLR1-overexpressing cells showed a significant increasing trend. In conclusion, overexpression of IFNLR1 induces activation of the JAK/STAT pathway, thereby inhibiting the proliferation of PAMs infected with PRRSV. Conclusion Expression of the IFNLR1 gene has an important regulatory role in PRRSVinfected PAMs, indicating it has potential as a molecular target in developing a new strategy for the treatment of PRRSV.

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 .

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