Objective To study the effect of microRNA-204 (miR-204) on the biological characteristics of breast cancer cells.Methods Real-time PCR was used to detect the expression of miR-204 in human breast cancer cell MDA-MB-231 after transfection of miR-204 mimics and inhibitor for 48 h.Flow cytometry was used to analyse the effect of miR-204 on the proliferation and apoptosis of MDA-MB-231 cells.The effect of miR-204 on the migration of MDA-MB-231 cells was detected by Transwell migration assay.Results Real-time PCR analysis showed that miR-204 mimics and inhibitors had significant effect compared with normal control group(P<0.01).Flow cytometry analysis showed that compared with normal control group,the number of G1 phase cells of miR-204 mimics group was significantly decreased(P<0.01),while the number of G2/M cells of miR-204 mimics group was significantly increased(P<0.01).In contrast,the number of G1 phase cells of miR-204 inhibitor group was significantly increased(P<0.01),while the number of G2/M cells of miR-204 inhibitor group was significantly decreased(P<0.01).miR-204 mimics group significantly promoted apoptosis,while the inhibitor group significantly inhibited apoptosis(P<0.01).Transwell migration analysis showed that the number of cells of miR-204 mimics group were significantly reduced,while the number of cells was significantly increased in the inhibitor group(P<0.01).Conclusion We find miR-204,which can promote cell apoptosis and inhibit cell proliferation and migration,is a negative factor in the breast cancer cell line MDA-MB-231.

In this study, we successfully expanded a full length gene encoding the monooxygenase eytochrome P450 2C9 gene from human liver of Chinese Han by RT-PCR. Our findings indicated that except G-->T mutation at the 190th nucleotide site, the other nucleotide sequences are completely consistent with CYP2C9 (NM017460) in GenBank. The SDS-PAGE and Western-Blot analysis showed that the CYP 2C9 gene was successfully expressed in the host cell E. coli BL21 (DE3). Our current study lays the foundation for the evaluation of pre-clinical drug metabolism and safety in the future.
Aryl Hydrocarbon Hydroxylases ; biosynthesis ; genetics ; China ; ethnology ; Cloning, Molecular ; Cytochrome P-450 CYP2C9 ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Point Mutation ; Polymorphism, Genetic ; Recombinant Proteins ; biosynthesis ; genetics

The aim of this study was to identify antigens for a vaccine or drug target to control rabbit coccidiosis. A combination of 2-dimensional electrophoresis, immunoblotting, and mass spectrometric analysis were used to identify novel antigens from the sporozoites of Eimeria stiedae. Protein spots were recognized by the sera of New Zealand rabbits infected artificially with E. stiedae. The proteins were characterized by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) analysis in combination with bioinformatics. Approximately 868 protein spots were detected by silver-staining, and a total of 41 immunoreactive protein spots were recognized by anti-E. stiedae sera. Finally, 23 protein spots were successfully identified. The proteins such as heat shock protein 70 and aspartyl protease may have potential as immunodiagnostic or vaccine antigens. The immunoreactive proteins were found to possess a wide range of biological functions. This study is the first to report the proteins recognized by sera of infected rabbits with E. stiedae, which might be helpful in identifying potential targets for vaccine development to control rabbit coccidiosis.
Coccidiosis* ; Computational Biology ; Eimeria ; Electrophoresis ; HSP70 Heat-Shock Proteins ; Immunoblotting ; Mass Spectrometry ; Rabbits ; Sporozoites

Objective To analyze the risk factors that affect the early recurrence (recurrence occurring within 3 months after surgical resection) of hepatocellular carcinoma (HCC), and to discuss the risk factors influencing the survival after hepatectomy. Methods The clinical data of 257 HCC patients, who were admitted to authors' hospital during the period from January 1, 2007 to March 31, 2014 to receive cTACE within 3 months after surgical resection of hepatocellular carcinoma, were retrospectively analyzed. According to DSA findings (lipiodol CT scan was performed in part of patients with undetermined diagnosis), the patients were divided into recurrence group and non-recurrence group. By using univariate analysis and multiple logistic regression analysis, the correlation of the clinical and pathological data with the early recurrence was analyzed. The patients were followed up, the survival time was recorded. The relationship between patient's clinical data and postoperative survival was evaluated. Results ① Of the 257 patients, early recurrence was detected in 150 patients (58. 4％, recurrence group) and no recurrence was observed in 107 patients (41. 6％, non-recurrence group). ②The presence of satellite nodules and the integrity of tumor encapsulation were two independent factors associated with the postoperative residual lesions. ③The maximum diameter of the tumor, Edmondson grade and the vascular cancer thrombus were the independent risk factors affecting survival. ④The median survival time of patients in recurrence group was markedly shortened than that of patients in non-recurrence group (39 months vs. 93 months). Conclusion The early recurrence (within 3 months after resection) of hepatocellular carcinoma is associated with the presence of satellite nodules and the integrity of tumor encapsulation. The survival of patients after hepatectomy is related to the maximum diameter of the tumor, Edmondson grade and the vascular cancer thrombus. The median survival time in patients having early recurrence is significantly shortened than that in patients having no early recurrence. (J Intervent Radiol, 2018, 27: 215-222)

Objective: To analyze the relationship between daily behavior and self-perceived academic performance among students in Jiangxi, and to provide a scientific basis for developing health behavior among adolescents and children. Methods: By using stratified cluster sampling, 12 primary and secondary schools from six counties of Jiangxi Province were selected, and a total of 2 284 students were investigated with questions including breakfast consumption, number of days participating in sports activities (≥45 min) and the average screen time during the past 7 days. Regression analysis was carried out by using ordered multi-classification logistic. Results: Of the 2 284 students surveyed, 1 374(60.16%) perceived their academic performance as very good or good, 613 (26.84%) and 297(13.00%) perceived as moderate or poor. Primary school, having breakfast every day, not playing on mobile or computer during weekday was positively associated with higher self-perceived academic performance (OR=0.50, 0.57 and 0.64, respectively, P<0.05), no sports activities ≥45 min during the past 7 d was associated with poor(OR=1.57, P<0.01). Among the 2 284 students, 67.43% students ate breakfast every day in the past 7 d, 75.39% students did not play mobile phones or computers at school, 40.89% students did not participate in sports activities (≥45 min). Conclusion Breakfast skipping and insufficient physical exercise shows adverse effects, and no/limited screen time during weekdays shows positive effects on academic performance. Intervention of visky behaviors should to carried out from different perspectives to promote the develpment of the health habits among students.

OBJECTIVETo understand the epidemiologic and etiologic characteristics of diarrheagenic Escherichia (E.) coli infections in Shenzhen.

METHODSStool samples were collected from acute diarrheal patients in four sentinel hospitals in Shenzhen and diarrheagenic E. coli strains were isolated and identified with multiplex real-time PCR. Serotyping and pulsed-field gel electrophoresis (PFGE) typing were conducted for the diarrheagenic E. coli isolates.

RESULTSA total of 74 diarrheagenic E. coli strains were isolated from 1 823 stool samples (4.06%). The patients were mainly young children aged <3 years and adults aged 20-39 years, and the infections mainly occurred during May-September of a year. Enterotoxigenic E. coli (ETEC) and enteropathognic E. coli (EPEC) were predominant (45.9% and 31.1%). Serogroups and PFGE patterns varied among the diarrheagenic E. coli isolates. However, serogroup O159 were predominant in ETEC and there were 5 clusters with ≥2 strains sharing same PFGE patterns.

CONCLUSIONSETEC and EPEC were predominant in diarrheagenic E. coli strains isolated from diarrheal patients in Shenzhen. Age and season specific characteristics of diarrheagenic E. coli infections were observed. The serotypes and PFGE patterns of diarrheagenic E. coli strains varied. Close attention should be paid to the possible ETEC outbreak.

Adult ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; microbiology ; Disease Outbreaks ; Electrophoresis, Gel, Pulsed-Field ; Enterotoxigenic Escherichia coli ; classification ; isolation & purification ; Escherichia coli Infections ; epidemiology ; Humans ; Real-Time Polymerase Chain Reaction ; Serotyping ; Young Adult

Sodium-glucose cotransporter 2 (SGLT2) inhibitors reduce cardiovascular mortality in patients with diabetes mellitus but the protective mechanism remains elusive. Here we demonstrated that the SGLT2 inhibitor, Empagliflozin (EMPA), suppresses cardiomyocytes autosis (autophagic cell death) to confer cardioprotective effects. Using myocardial infarction (MI) mouse models with and without diabetes mellitus, EMPA treatment significantly reduced infarct size, and myocardial fibrosis, thereby leading to improved cardiac function and survival. In the context of ischemia and nutritional glucose deprivation where autosis is already highly stimulated, EMPA directly inhibits the activity of the Na⁺/H⁺ exchanger 1 (NHE1) in the cardiomyocytes to regulate excessive autophagy. Knockdown of NHE1 significantly rescued glucose deprivation-induced autosis. In contrast, overexpression of NHE1 aggravated the cardiomyocytes death in response to starvation, which was effectively rescued by EMPA treatment. Furthermore, in vitro and in vivo analysis of NHE1 and Beclin 1 knockout mice validated that EMPA's cardioprotective effects are at least in part through downregulation of autophagic flux. These findings provide new insights for drug development, specifically targeting NHE1 and autosis for ventricular remodeling and heart failure after MI in both diabetic and non-diabetic patients.
Animals ; Diabetes Mellitus ; Diabetes Mellitus, Type 2/drug therapy* ; Glucose ; Humans ; Mice ; Myocardial Infarction/metabolism* ; Sodium-Glucose Transporter 2 Inhibitors/therapeutic use* ; Ventricular Remodeling

Sodium-glucose cotransporter 2 (SGLT2) inhibitors have been reapproved for heart failure (HF) therapy in patients with and without diabetes. However, the initial glucose-lowering indication of SGLT2i has impeded their uses in cardiovascular clinical practice. A challenge of SGLT2i then becomes how to separate their anti-HF activity from glucose-lowering side-effect. To address this issue, we conducted structural repurposing of EMPA, a representative SGLT2 inhibitor, to strengthen anti-HF activity and reduce the SGLT2-inhibitory activity according to structural basis of inhibition of SGLT2. Compared to EMPA, the optimal derivative JX01, which was produced by methylation of C2-OH of the glucose ring, exhibited weaker SGLT2-inhibitory activity (IC₅₀ > 100 nmol/L), and lower glycosuria and glucose-lowering side-effect, better NHE1-inhibitory activity and cardioprotective effect in HF mice. Furthermore, JX01 showed good safety profiles in respect of single-dose/repeat-dose toxicity and hERG activity, and good pharmacokinetic properties in both mouse and rat species. Collectively, the present study provided a paradigm of drug repurposing to discover novel anti-HF drugs, and indirectly demonstrated that SGLT2-independent molecular mechanisms play an important role in cardioprotective effects of SGLT2 inhibitors.