This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Base Sequence ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, KIR ; biosynthesis ; genetics

Objective To analyze the factors influencing the occurrence of macrosomia so as to provide a theoretical basis for the prevention of macrosomia.Methods The data were selected from the findings of Survey on Prevalence and Risk Factors for Birth Defects in Shaanxi Province.This study intended to select data of macrosomia in Xianyang City and its influencing factors for statistical analysis.The rates were compared by χ2 ;the influencing factors were analyzed by univariate and multivariate Logistic regression.Results The incidence of fetal macrosomia among single live birth neonates in Xianyang was 5.2%.The incidence of macrosomia was 6.5%in males and 4.2% in females,with a significant difference (P <0.001).From 2010 to 2013 the annual incidence rate of macrosomia was 5.4%,6.2%,5.9% and 4.4%,with no significant difference (P =0.151).The incidence rate of macrosomia in urban and rural areas was 6.3% and 5.5%,respectively,without significant difference (P =0.556).The incidence of macrosomia was 3.2% in Binxian County,5.6% in Changwu County,6.7% in Liquan County,5.4% in Qianxian County,8.0% in Wugong County,and 4.2% in Xunyi County,with significant differences (P <0.001).Logistic regression analysis showed that the occurrence of macrosomia was related to the baby's sex,mother's age,the times of pregancy,the time of antenatal care,alcohol intake during pregnancy and father's occupation.Conclusion The incidence of macrosomia of Xianyang from 2010 to 2013 was close to the national average.The sex of baby,mother's age,the times of pregnancy,the times of antenatal care,drinking history during pregnancy and father's occupation may affect the occurrence of macrosomia.

Objective The aim of this study is to investigate the macrolide resistance rate and molecular type withmultiple-locus variable-number tandem-repeat analysis (MLVA) of Mycoplasma pneumoniae of Beijing in 2016 in pediatric patients. Methods Real-time quantitative polymerase chain reaction (PCR) was used to identify M. pneumoniae, and MLVA was performed. The domain V of the 23S rRNA was sequenced to detect macrolide-resistant point mutations. We also investigated the activities of antibiotics against M. pneumoniae isolates in vitro. Results The PCR detection rate of M. pneumoniae in children in Beijing was 40％, and the macrolide resistance rate was 66％. The A2063G mutation in the 23S rRNA V region is the dominant mutation (137/146, 93.84％), whereas the A2064G mutation is rare (9/146, 6.16％). Seventy-three samples were typed successfully by MLVA typing, including 86.3％ (63/73) were MLVA type 4-5-7-2, and 13.7％ (10/73) were MLVA type 3-5-6-2. No other types were found. No strains were resistant to levofloxacin or tetracycline. Conclusion In 2016, a specific decrease in the macrolide resistance rate occurred in Beijing. The detection rate and macrolide resistance rate of outpatients are lower than those of inpatients. The A2063G mutants M. pneumoniae have high levels of resistance to erythromycin and azithromycin. The primary MLVA type is 4-5-7-2, followed by 3-5-6-2. No other MLVA types were detected. No strains resistant to tetracycline or levofloxacin were found in vitro.

Objective: To evaluate the predictive value of PARIS bleeding score on in-hospital bleeding of acute myocardial infarction (AMI) patients after drug-eluting stent (DES) implantation with dual-antiplatelet therapy (DAPT). Methods: There were 27 594 AMI patients enrolled in China acute myocardial infarction (CAMI) registry between 2013-01-01 to 2014-09-30 from 107 hospitals, and 14 625 of them had successful in-hospital DES implantation with DAPT were studied. Based on BARC (bleeding academic research consortium definition) criteria, the end point major bleeding (MB) events were defined by both BARC type 3, 5 and BARC type 2, 3, 5; the incidence of in-hospital bleeding, clinical features and predictive value of PARIS bleeding score according to different BARC type were evaluated. Results: Compared with non-MB patients, MB patients had the higher PARIS bleeding score, P<0.001. Based on PARIS score risk stratification, taking BARC type 3, 5 as endpoint, 77/14 625 (0.53%) patients had bleeding events, PARIS scores were different among high risk, mid risk and low risk patients, P<0.001; bleeding risk in mid risk patients was 2.38 times higher than low risk patients, P=0.006 and bleeding risk in high risk patients was 4.78 times higher than low risk patients, P<0.001.Taking BARC type 2,3,5 as endpoint,223(1.52%)patients had bleeding events,bleeding risk in mid risk patients was 1.64 times higher than low risk patients, P=0.002 and bleeding risk in high risk patients was 2.23 times higher than low risk patients, P=0.001. ROC analysis showed that PARIS score had predictive value on both BARC type 3, 5 and BARC type 2, 3, 5 bleeding, area under curve (AUC) of BARC type 3, 5 (AUC: 0.672) was higher than AUC of BARC type 2, 3, 5 (AUC:0.596) (z=2.079, P=0.038), which implied that PARIS score had better predictive value in severe bleeding events. Conclusion: PARIS bleeding score had predictive value on in-hospital bleeding in AMI patients after DES implantation with DAPT, it can also be used in bleeding risk stratification. PARIS bleeding score had better predictive value on severe bleeding.

Objective: We investigated changes in the intestinal flora of children with Mycoplasma pneumoniae pneumonia (MPP). Methods: Between September 2019 and November 2019, stool samples from 14 children with MPP from The Fourth Hospital of Baotou city, Inner Mongolia Autonomous Region, were collected and divided into general treatment (AF) and probiotic (AFY) groups, according to the treatment of "combined Bifidobacterium, Lactobacillus, Enterococcus, and Bacillus cereus tablets live". High-throughput 16S rDNA sequencing was used to identify intestinal flora. Results: Intestinal flora abundance and diversity in children with MPP were decreased. Both Shannon and Simpson indices were lower in the AF group when compared with healthy controls ( P < 0.05). When compared with healthy controls, the proportion of Enterorhabdus was lower in the AF group, while the proportion of Lachnoclostridium was higher ( P < 0.05). The proportion of Bifidobacteria and Akkermansia was lower in the AFY group but Enterococcus, Lachnoclostridium, Roseburia, and Erysipelatoclostridium proportions were higher. The proportion of Escherichia coli- Shigella in the AFY group after treatment was decreased ( P < 0.05). Conclusions The intestinal flora of children with MPP is disturbed, manifested as decreased abundance and diversity, and decreased Bifidobacteria. Our probiotic mixture partly improved intestinal flora disorders.
Child ; DNA, Ribosomal ; Escherichia coli ; Gastrointestinal Microbiome ; Humans ; Mycoplasma pneumoniae ; Pneumonia, Mycoplasma ; Technology