circRNA CRIM1 regulates the migration and invasion of bladder cancer by targeting miR182/Foxo3a axis

PurposeTo explore the molecular mechanism of circRNA CRIM1 in the regulation of bladder cancer by targeting the miR182/Foxo3a axis.Methods50 pairs of cancer tissues and para-cancerous tissues of patients with bladder cancer were collected. RT-PCR method was used to detect the expression of CRIM1 and miR-182. The association between circRNA CRIM1 and clinical data was analyzed. qPCR was used to measure the expression of circRNA CRIM1 and miR-182 in bladder cancer cell UMUC3 and endothelial cell line HUVEC. CRIM1 genes and miR-182 in UMUC3 cell lines were overexpressed and silenced, respectively, to investigate their effects on invasion and migration of bladder cancer, and to detect the changes of miR182/Foxo3a expression. The association between circRNA CRIM1 and miR182/Foxo3a was determined by bioinformatics analysis.ResultsThe results showed that there was a significant association between the expression of circRNA CRIM1 and distal migration. The expression of CRIM1 in adjacent tissues was significantly down-regulated and negatively correlated with distal migration. The overexpression of circRNA CRIM1 reduced migration and invasion processes in bladder cancer cells. After circRNA CRIM1 was overexpressed, the miR-182 was significantly down-regulated. The expression levels of Foxo3a mRNA and proteins were up-regulated after miR-182 silencing of bladder cancer cell line UMUC3. miR-182 silencing inhibited invasion and migration of cancer cells to some extent. In bladder cancer cells and tissues, CRIM1 and Foxo3a were significantly down-regulated, miR-182 was significantly up-regulated.ConclusioncircRNA CRIM1 regulated the migration and invasion of bladder cancer by targeting the miR182/Foxo3a axis.

circRNA CRIM1 regulates the migration and invasion of bladder cancer by targeting miR182/Foxo3a axis.

PURPOSE: To explore the molecular mechanism of circRNA CRIM1 in the regulation of bladder cancer by targeting the miR182/Foxo3a axis. METHODS: 50 pairs of cancer tissues and para-cancerous tissues of patients with bladder cancer were collected. RT-PCR method was used to detect the expression of CRIM1 and miR-182. The association between circRNA CRIM1 and clinical data was analyzed. qPCR was used to measure the expression of circRNA CRIM1 and miR-182 in bladder cancer cell UMUC3 and endothelial cell line HUVEC. CRIM1 genes and miR-182 in UMUC3 cell lines were overexpressed and silenced, respectively, to investigate their effects on invasion and migration of bladder cancer, and to detect the changes of miR182/Foxo3a expression. The association between circRNA CRIM1 and miR182/Foxo3a was determined by bioinformatics analysis. RESULTS: The results showed that there was a significant association between the expression of circRNA CRIM1 and distal migration. The expression of CRIM1 in adjacent tissues was significantly down-regulated and negatively correlated with distal migration. The overexpression of circRNA CRIM1 reduced migration and invasion processes in bladder cancer cells. After circRNA CRIM1 was overexpressed, the miR-182 was significantly down-regulated. The expression levels of Foxo3a mRNA and proteins were up-regulated after miR-182 silencing of bladder cancer cell line UMUC3. miR-182 silencing inhibited invasion and migration of cancer cells to some extent. In bladder cancer cells and tissues, CRIM1 and Foxo3a were significantly down-regulated, miR-182 was significantly up-regulated. CONCLUSION: circRNA CRIM1 regulated the migration and invasion of bladder cancer by targeting the miR182/Foxo3a axis.

[Differentiation between hyperintense hepatocellular carcinoma and focal nodular hyperplasia in hepatobiliary phase with multimodal parameters of magnetic resonance imaging].

Objective: To differentiate hyperintense hepatocellular carcinoma (HCC) with focal nodular hyperplasia (FNH) in the hepatobiliary phase by MRI multimodal parameters. Methods: A retrospective cross-sectional study method was adopted. Clinical data on 15 cases with hyperintense HCC and 15 cases with FNH in the hepatobiliary phase admitted to the First Affiliated Hospital of the Army Medical University from January 2012 to December 2019 were collected. All patients with solitary lesions who underwent Gd-EOB-DTPA-enhanced MRI examinations were included. Surgically resected specimens were verified by pathological and immunohistochemical examination. HCC and FNH imaging features were analyzed by two radiologists. Results: (1) HCC and FNH apparent diffusion coefficient (ADC) values were 1 205.07±239.65×10-3 mm2/s and 1 434.73±217.6×10-3 mm2/s, respectively, and the SIADC difference was statistically significant (P<0.05) between the two groups. (2) In the dynamic contrast-enhanced MRI sequence, 15 cases of HCC were significantly enhanced in the arterial phase, of which 13 cases were characterized by continuous enhancement, and 2 cases were characterized by wash-in and wash-out enhancement. There was no statistically significant difference (P>0.05) between the two groups. SIenhancement rate between HCC and FNH (1.39±0.60 vs. 1.33±0.50, P>0.05) had no significant difference. (3) HCC and FNH morphological features in the hepatobiliary phase included: annular hypointensity: HCC (8 cases) vs. FNH (0 cases); contrast filling defects: HCC (8 cases) vs. FNH (0 cases); linear hyposignal separation: HCC (10 cases) vs. FNH (0 cases); and stellate scars: HCC (0) vs. FNH (5 cases), and there were statistically significant differences (P<0.05) between the two groups . Conclusion: Multimodal MRI have significant value for differentiating hyperintense HCC and FNH in the hepatobiliary phase.

[Effect of occupational factors on pre-diabetes mellitus among iron and steel workers].

Objective: To investigate the prevalence of pre-diabetes mellitus (PDM) and the impact of occupation-related factors on PDM, among workers from a steel company in Tangshan city, Hebei province. Methods: Clustering sampling method was used to select a steel company and to carry out occupational health-related physical checkup programs for eligible workers who had working in this company for longer than one year. The study began in February and ended up in June, 2017. Workers who were with FPG level as ≤6.9 mmol/L, and free from diabetes, were selected as the subjects for this study. Questionnaires were used and physical examinations and FPG testing conducted. Results: The total number of subjects in this study was 4 173, of which 2 648 appeared as pre-diabetic, with the prevalence rate as 63.4%. Increase of the PDM prevalence was in parallel with the length of service, among the workers. The risk for the pre-diabetes in those who worked more than 8 hours per day was 1.696 times higher than those who worked less than or equal to 8 h/d (95%CI:1.517-1.937). Compared with those workers without exposures to heat, noise or carbon monoxides, the proportion of pre-diabetes appeared higher in workers exposed to heat, noise or CO with OR=1.782 (95%CI: 1.205-2.636), 1.815 (95%CI: 1.209-2.794) and 1.653 (95%CI: 1.158-2.361), respectively. Risks for those who were exposed to heat or noise were higher than those who were free from exposure to any occupational hazards (OR=2.098, 95%CI: 1.296-3.397). Prevalence rates of pre-diabetes in those who were exposed to heat, noise or CO, were higher than that those who were not. Conclusion: Working hours and exposures to heat, noise or CO appeared as influential factors on PDM.

The aPKCι blocking agent ATM negatively regulates EMT and invasion of hepatocellular carcinoma.

Epithelial-to-mesenchymal transition (EMT) has an important role in invasion and metastasis of hepatocellular carcinoma (HCC). To explore the regulatory mechanism of atypical protein kinase C ι (aPKCι) signaling pathways to HCC development, and find an agent for targeted therapy for HCC, immortalized murine hepatocytes were employed to establish an EMT cell model of HCC, MMH-RT cells. Our study showed that EMT took place in MMH-R cells under the effect of transforming growth factor-ß1 (TGF-ß1) overexpressing aPKCι. Furthermore, we showed that the aPKCι blocking agent aurothiomalate (ATM) inhibited EMT and decreased invasion of hepatocytes. Moreover, ATM selectively inhibited proliferation of mesenchymal cells and HepG2 cells and induced apoptosis. However, ATM increased proliferation of epithelial cells and had little effect on apoptosis and invasion of epithelial cells. In conclusion, our result suggested that aPKCι could be an important bio-marker of tumor EMT, and used as an indicator of invasion and malignancy. ATM might be a promising agent for targeted treatment of HCC.

Preparation and characterization of an antiserum against truncated UL54 protein of pseudorabies virus.

Pseudorabies virus (PRV) early protein UL54 is a homolog of herpes simplex virus 1 immediate-early protein ICP27, which is a multifunctional protein essential for the virus replication. However, the precise role of the PRV UL54 protein in the virus life cycle is still poorly understood. To shed more light on this problem, we considered it essential to have available an antiserum specifically detecting this protein. Since it was known that a full-length UL54 protein is a too big molecule for efficient expression in prokaryotic systems, it was truncated from 1 to 66 N-terminal amino acids, fused to EYFP-His tag and expressed in Escherichia coli through an appropriate expression vector. The truncated protein was purified by Ni-NTA affinity chromatography and used for raising an antiserum in rabbits. Western blot analysis showed that this antiserum specifically recognized the purified truncated as well as full-length UL54 protein in PRV-infected cells. Immunofluorescence assay confirmed the latter finding and also demonstrated localization of this protein first in nucleoli and later in whole nuclei of PRV-infected cells. These results indicate that the prepared antiserum could serve as a valuable tool in further studies of PRV UL54 protein function.

Recovery of pyruvic acid from biotransformation solutions.

The aim of this investigation was to separate pyruvic acid of biotransformation solutions from lactic acid through complex extraction. For this purpose, complex extraction was investigated from model solutions. Tri-n-octanylamine (TOA) was used as the extractant. The effects of various diluents, the stoichiometry of pyruvic acid to TOA, and the initial pH of the aqueous phase on the extraction process were investigated in this study. The effects of sodium hydroxide (NaOH) and trimethylamine (TMA) on the back extraction process were also studied, respectively. The optimal conditions attained from the model solutions proved efficient on the biotransformation solutions of different concentrations. A total recovery of 71-82% of pyruvic acid was obtained, whereas 89-92% of lactic acid was removed. The purity of pyruvic acid reached 97% after the removal of TMA by a simple distillation.

An enzymatic route to produce pyruvate from lactate.

A bacterial strain of Acinetobacter sp., which was capable of enzymatic production of pyruvate from lactate, was cultured in a 5-l reactor with a basal salt medium. After 14 h of fed-batch fermentation, 9.56 g l(-1) cell concentration in the broth was obtained with 20 g l(-1) (178 mM) sodium lactate and 4 g l(-1) NH(4)Cl in the medium; and the biotransformation ability was 2.51 units ml(-1). The cells were harvested from one reactor and then used for pyruvate production from lactate in the same reactor. l-lactate at a concentration about 527 mM was almost stoichiometrically converted to pyruvate in 28 h. After a total 42 h of cell culture and biotransformation, the transformative yield was about 0.72 g g(-1) pyruvate from lactate and the rate of pyruvate production was calculated as 1.33 g l(-1) h(-1) during the process. The results suggested this simple enzymatic production of pyruvate from lactate should be a promising process and may bring a yield higher than that by microbial fermentation. By this process, the recovery of pyruvate from such a simple reaction liquid is relatively easy and inexpensive to perform.

Rayleigh light scattering study on the reaction of nucleic acids and methyl violet.

A new quantitative determination method for nucleic acids in aqueous solutions, based on the enhancement of Rayleigh light scattering of methyl violet by nucleic acids, has been developed. The sensitivity of the assay allows amounts of nucleic acids as little as 100 ng/ml to be quantitated reliably. In addition to its high sensitivity, this method has other advantages: rapidity of reaction (<5 min), simplicity of operation (one-step assay), commonality of spectrofluorimeter and reagents, stability of mixtures formed, and reproducibility. Under the experimental conditions, there is little or no interference from proteins, nucleosides, and most metal ions. Interference by a few metal ions, detergents, and some salts can be minimized by dilution. The method can also be used to determine the total amount of nucleic acids without the arduous choice of standard and difficult separation of DNA and RNA.

Enhancement of Rayleigh light scattering of acid chrome blue K by proteins and protein assay by the scattering technique.

Rayleigh light scattering of Acid Chrome Blue K (ACBK) is enhanced greatly by proteins. Based on this, a method for protein assay in aqueous solution was developed. This assay matches the sensitivity of the colorimetric dye-binding method with a linear range of 0.136-10.88 micrograms ml-1. The measurements can be made easily on a common fluorimeter. The reaction between ACBK and proteins is completed in 2 min and the scattered light signal is stable for at lest 3 h. Protein-to-protein variability is encountered in this method as in many other protein assays. There is little or no interference from amino acids, most metal ions and complexing agents (e.g., EDTA). Interferences from salts, urea and detergents can be minimized by dilution.

Microdetermination of proteins by resonance light scattering spectroscopy with bromophenol blue.

Reaction of bromophenol blue with proteins results in an enhanced resonance light scattering at 334 nm. Based on this, a new quantitative determination method for proteins in the aqueous solution is established. This assay is characterized by high sensitivity (0.34-18.7 microg/ml), short reaction time (>2 min), and simplicity (a one-step assay). Due to protein-to-protein variability, this method gave results higher than that of the bromocresol green assay in detection of human serum albumin.