ObjectiveTo explore the mechanism by which the tumor suppressor gene Testin affects the proliferation, migration, and invasive biological activity of lung adenocarcinoma cell lines by regulating the RhoA pathway. MethodThe cbioportal tumor gene expression was used to screen for genes with high correlation with TES gene expression in lung adenocarcinoma, and the 200 genes with the highest correlation were selected for pathway enrichment analysis. Upload these 200 genes to the David gene annotation tool for GO_Biological Process pathway analysis, GO Molecular Function pathway analysis, KEGG pathway analysis, and Reactome pathway analysis. The lung adenocarcinoma cell line H1299 was cultured, and an overexpression Testin plasmid was constructed and transfected into H1299 cells. The mRNA and protein expression of RhoA, Rac1, and Cdc42 were detected using qRT PCR and western blot. On the basis of downregulating RhoA expression through overexpression of Testin, the overexpression plasmid of RhoA (TES+RhoA) was transfected simultaneously to induce a downregulation of RhoA expression, and the changes in malignant phenotype of lung adenocarcinoma cells were detected. The biological activity changes of adenocarcinoma cell lines after the above intervention were verified through CCK-8 experiment, Transwell experiment, and Matrigel experiment. Results The results of pathway analysis prediction showed that Testin may be involved in regulating the Rho GTPase signaling pathway. Overexpression of Testin did not affect the mRNA levels of RhoA, Rac1, and Cdc42 (all P>0.05), nor did it affect the protein expression levels of Rac1 and Cdc42 (all P>0.05), but it significantly reduced the protein level of RhoA (P<0.05). Knocking down RhoA in lung adenocarcinoma cell H1299 can significantly inhibit cell proliferation, migration, and invasion ability (all P<0.05). Simultaneously transfecting RhoA overexpression plasmid on the basis of overexpression of Testin can downregulate RhoA expression, but does not affect Testin expression. ConclusionsRhoA plays a pro-cancer role in lung adenocarcinoma, and Testin can inhibit RhoA expression. Overexpression of RhoA can rescue Testin's effect on lung adenocarcinoma cell proliferation, migration, and invasion. Testin exerts its anti-cancer biological activity by regulating RhoA.
ObjectiveTo summarize the results of testing and analysis of antigen and antibody for diseases under the frame of children's immunization program, in order to know the effects of prevention and control of such diseases in this area.
MethodsA total of 150 children from each of the 5 communities or administrative villages in Yongning District of Nanning City were selected for our survey between January and December 2012. The 150 children were composed of 30 children (residents, 1-6 years old, 5 children from each different age group) randomly selected from each of the four directions (east, south, west and north) and the mid-area of each community or village. The serum samples were collected to analyze the existence of poliomyelitis antibody, measles antibody, hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), and diphtheria antibody.
ResultsAmong the 150 school-age children, antibody immune qualified rate was high for hepatitis B (HBV) antigen, in which the HBsAg immune qualified rate was 99.3%, and HBsAb immune qualified rate was 88.0%, showing no significant difference between boys and girls (P>0.05). All poliomyelitis Ⅰ, Ⅱ, and Ⅲ antibody positive rates reached 100.0%. Measles antibody test results were also satisfying for each age group, among whom the 2 and 3 year-olds reached a positive rate of the highest, 100%, and the 1, 4, 5, and 6 year-old children had a measles antibody positive rate of 96.0%, 84.0%, 88.0%, and 96.0%, respectively. The positive rate for diphtheria antibody was 100%.
ConclusionThe antibody and antigen detection and analysis results for the children's immune program targeted diseases are generally satisfying in this area. Especially, the prevention of poliomyelitis and diphtheria is the best. However, prevention of HBV and measles is not as good. Therefore, tracking immunization coverage, promoting public awareness on immune planning, actively participating in the vaccination of children should be enhanced for further disease prevention.
Objective To perform rapid antimicrobial susceptibility testing (RAST) on positive blood cultures of Enterobacterales using a total laboratory automation (TLA) system following both Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) standards, and to evaluate the two RAST methods. Methods Positive blood culture bottles growing Enterobacterales [54 Escherichia coli (E. coli) and 60 Klebsiella pneumoniae (K. pneumoniae) isolates] were collected at West China Hospital of Sichuan University between April and August 2022. CLSI RAST (8 and 16 h) and EUCAST RAST (4, 6, and 8 h) were performed using the TLA system, and results were compared with Vitek 2 antimicrobial susceptibility testing. Results CLSI RAST demonstrated lower categorical agreement with Vitek 2 (E. coli: 66.7% at 8 h, 81.9% at 16 h; K. pneumoniae: 72.8% at 8 h, 84.0% at 16 h) and tended to overcall resistance. EUCAST RAST showed increasing zone readability over time and high categorical agreement with Vitek 2 (E. coli: 97.1%, 96.2%, and 96.1% at 4, 6, and 8 h, respectively; K. pneumoniae: 96.1%, 97.1%, and 97.9% at 4, 6, and 8 h, respectively), as well as low error rates (major errors <3%). With the exception of ciprofloxacin, both the readability and categorical agreement for each antimicrobial agent against E. coli at 8 h were favorable (>90%). Similarly, for K. pneumoniae, the readability and categorical agreement for each antimicrobial agent at 8 h also demonstrated good performance (>90%). Conclusions According to CLSI-M52 criteria, EUCAST RAST at 4, 6, and 8 h shows equivalent performance to Vitek 2. Except for ciprofloxacin against E. coli, all antimicrobials tested at 8 h exhibit good readability and categorical agreement (>90%). CLSI RAST has lower categorical agreement and a higher tendency to interpret isolates as resistant.
