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 investigate the efficiency of double moxibustion combined with intermittent catheterization on urination diary in patients with neurogenic bladder (NB) after spinal cord injury (SCI).MethodsFrom August 2014 to August 2016, hospitalized patients with NB after SCI were reviewed retrospectively and classified into the moxibustion group and the routine group. The urination diary related indicators, International Urinary Incontinence Advisory Committee Urinary Incontinence Questionnaire Short Form (ICI-SQ-SF) score, and routine urine leucocyte count were collected.ResultsA total of 278 patients were enrolled in the study, in whom, the data of 96 were incomplete, and 26 got lost to follow-up. The routine group and the moxibustion group finally contained 70 and 86 respectively. There was no significant difference between the two groups in sex, age, average course of the disease, SCI segment or grading of injury (P>0.05). After 2 months treatment, the urine routine leucocyte count decreased to (5.72± 0.36)/μL in the routine group and (3.22±0.27)/μL in the moxibustion group, and the ICI-Q-SF score decreased to 8.61±0.45 in the routine group and 5.07±0.38 in the moxibustion group. The urine routine leucocyte count and ICI-Q-SF score were lower than those before treatment, and were lower in the moxibustion group than those in the routine group (P<0.05). In the 7th to 8th week, in the routine group, the average single urine volume was (300±70) mL, the interval between two micturition was (3.5±0.6) hours, the frequency of urinary incontinence was (3.3±0.4) times per week, and the average residual urine volume was (125±42) mL; in the moxibustion group, they were (326±78) mL, (3.8±1.1) hours, (2.3±0.3) times per week, and (103±37) mL, respectively. The indexes were not statistically significant different from those in the 1st to 2nd week (P>0.05), except the average single urine volume in the routine group and all the indexes in the moxibustion group (P<0.05). They were all better in the moxibustion group than those in the routine group (P<0.05).ConclusionThe treatment of moxibustion and intermittent catheterization may improve bladder function, reduce residual urine, reduce urinary incontinence and improve the quality of life for patients with NB after SCI.