Circ_0067934 correlates with poor prognosis and promotes laryngeal squamous cell cancer progression by sponging miR-1324.

OBJECTIVE: As a kind of non-coding RNA, circular RNA (circRNA) plays a regulatory role in many tumors. Our intention was to investigate the clinical significance, biological function, and molecular regulation mechanisms of circ_0067934 in laryngeal squamous cell cancer (LSCC) progression. PATIENTS AND METHODS: The expression levels of circ_0067934 in LSCC tumor samples and cells were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The associations of circ_0067934 expression with clinicopathological features and overall survival in LSCC patients were statistically analyzed. The biological function of circ_0067934 in LSCC cells was analyzed by CCK-8, colony formation, EdU, and transwell assays. Dual-Luciferase reporter assay was conducted to verify that circ_0067934 may sponge miR-1324 to regulate proliferation and migration of LSCC. RESULTS: Circ_0067934 expression was remarkably upregulated in LSCC tissues and cells. High level of circ_0067934 was significantly related to tumor size, lymph node status, and distant metastasis of LSCC. Furthermore, highly expressed circ_0067934 could result in notably worse survival. In vitro experiments showed that down-regulation of circ_0067934 inhibited proliferation and migration of LSCC cells. Mechanism analysis revealed that circ_0067934 regulated LSCC development by sponging miR-1324. CONCLUSIONS: Circ_0067934 may function as an oncogene in LSCC, which provided a feasible prognostic biomarker and therapeutic target for LSCC.

MiR-16 inhibits hepatocellular carcinoma progression by targeting FEAT through NF-κB signaling pathway.

OBJECTIVE: MicroRNA-16 (miR-16) expression has been proved to take part in the initiation and development of several cancers, including hepatocellular carcinoma (HCC). However, its role and its molecular mechanism in HCC cells remain unclear. Our study aimed to elucidate miR-16 probable role and potential mechanism in HCC cells. PATIENTS AND METHODS: MiR-16 expression in HCC was measured by Real Time-Polymerase Chain Reaction (RT-PCR). MiR-16 mimic or inhibitor was applied to increase or decrease miR-16 expression in Huh7 cells separately. The cell viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). The invaded cells and migrated cells were detected by the transwell assay. The epithelial-mesenchymal transition (EMT) and the nuclear factor-κB (NF-κB) were performed using Western blot. The tumor growth was measured via xenograft tumor formation assay. Moreover, bioinformatical methods and luciferase reporter assay were carried out to confirm the miR-16 target gene. RESULTS: MiR-16 expression was downregulated in HCC tissues and cells. Furthermore, the increasing miR-16 expression was suppressed, whereas the decreasing miR-16 expression promoted cell proliferation, invasion, and migration in Huh7 cells. Moreover, miR-16 targeted FEAT in regulating HCC progression. FEAT was associated with a poor prognosis of HCC patients. Especially, miR-16 suppressed EMT and NF-κB pathway in HCC and inhibited the tumor growth in vivo. CONCLUSIONS: We stated that miR-16 suppressed HCC cell progression by targeting FEAT and inhibiting EMT and NF-κB pathway. MiR-16 may be clinically utilized as a factor for the clinical diagnosis and prognosis of HCC.

[Prevalence and distribution of hypertension in population aged 15 years and over in Guizhou province].

Objective: To understand the prevalence and distribution of hypertension in population aged 15 years and over in Guizhou province and provide evidence for the prevention and management of hypertension. Methods: Face to face interviews using national standard questionnaire were conducted among the study subjects selected in Guizhou through multi-stage random sampling. Blood pressure measurement for them was done with Omron HEM-1300 professional portable blood pressure monitor. SPSS 19.0 software was used for statistical analysis. The ratio was compared by the χ(2) test. The influencing factors of hypertension was analyzed by multivariate logistic regression analysis. Results: A total of 13 480 participants were investigated, including 5 509 (40.8%) men and 7 971 (59.2%) women; 6 558 (48.6%) urban residents and 6 922 (51.4%) rural residents. Among the subjects surveyed, 3 232 (23.9%) were smokers, 2 412 (17.9%) were alcoholic and 4 859 (36.0%) were obese or overweight. A total of 3 937 (29.2%) hypertension patients were found. The prevalence of hypertension was 29.2%. The standardized prevalence of hypertension were 18.97% (compared with national population composition) and 21.16% (compared with Guizhou province population composition), respectively. The hypertension prevalence in men and women were 29.8% and 28.8%, respectively. The hypertension prevalence in rural population (35.8%) was higher than that in urban population (22.2%). The difference was statistically significant (P<0.001). The hypertension prevalence in people aged 65 years and over was 56.2%. The prevalence of hypertension were 34.3% and 27.6% in smokers and non-smokers, 39.2% and 27.0% in alcoholic and non-alcoholic and 40.7% and 22.7% in obese or overweight group and normal or less weight group, respectively. There were significant statistical differences in prevalence of hypertension among the population in urban area and rural area, with different age, education levels, smoking status, drinking status and BMI (P<0.001). Conclusions: The prevalence of hypertension in Guizhou was at a high level. The hypertension prevalence in rural area was higher than that in urban area. Hypertension prevalence increased significantly with age. The prevalence of hypertension was negatively associated with the education level of the people. Older age, living in rural area, smoking, drinking, obesity were the risk factors for hypertension.

[Multiple-factor analysis of serum allergen distribution of patients with allergic rhinitis and level of main allergen IgE in Shenyang area].

Objective:To discuss multiple-factor analysis of serum allergen distribution of patients with allergic rhinitis and level of main allergen IgE in Shenyang area, and to provide a scientific basis for the prevention of allergic rhinitis in this area.Method:Serum IgE was detected in 749 cases［501 cases of male (66.8%), and 248 cases of female (33.2%)］, with allergic rhinitis.The age range was from 3 to 65 years old, and they were divided into 5 groups based on age. A questionnaire survey was conducted to analyze the distribution of serum allergens and to carry out a multiple-factor analysis of level of the main allergen IgE in patients.Result:The primary allergen was house dust mite/dust mite in each age group, and the differences in the positive rate of elm, mold, cat/dog fur scurf, cockroach and ragweed among different groups were statistically significant（P<0.01 or P<0.05）.The ingested allergen sequencing for all age groups: the distribution of 7-14 years old group and 15-35 years old group were consistent with that of the whole, among the under 6 years old group, mango and pineapple was ranked 1st,beef and mutton was ranked 2nd,the occurrence rate of shrimp and crab rose to the 3rd place, among the 36-60 years old group, mango, pineapple and milk was ranked 1st, egg was ranked 2nd, and beef and mutton ranked 3rd, the differences in positive rate of mango and pineapple, beef and mutton, crab and nut among all groups were statistically significant(P<0.01 or P<0.05).The IgE level of cockroach was impacted by the allergic history, home cultivation of flowers and plants and animal domestication. The IgE level of mold was impacted by sex, allergy history, home rearing of pet and furniture updates. The IgE level of wormwood was impacted by allergy history and asthma history. The IgE level of peanut was impacted by age and allergy history. The IgE level of egg was impacted by history of food and drink allergy, home cultivation of flowers and plants and home rearing of pet. The IgE level of crab was impacted by the allergy history and home rearing of pet.Conclusion: The primary inhaled allergen in all groups is house dust mite/dust mite, and the ingested allergen varies in each group, which has provided a reference basis for prevention of ingested allergy.The varying influence factors for IgE level of primary allergens could be used to prevent the patients from contacting the allergen,and each risk factor has become a focus of prevention and control for patients, offering a major help to the prevention and treatment of allergic rhinitis.

Effect of FGF2 on the activity of SPRYs/DUSP6/ERK signaling pathway in endometrial glandular epithelial cells of endometriosis.

OBJECTIVE: We aimed at exploring the positive feedback loop in eutopic and ectopic endometrial glandular epithelial cells (EuECs and EECs) in endometriosis. MATERIALS AND METHODS: Normal epithelial cells (NECs), EuECs and EECs were treated with fibroblast growth factor (FGF)2, FGF2 neutralizing antibody, mitogen-activated protein kinases (MAPKs) inhibitors U0126 and PD98059. FGF2 protein level was detected by enzyme-linked immunosorbent assay (ELISA). The expressions of FGF2, FGF receptor 1 (FGFR1), extracellular signal-regulated kinase (ERK)1/2/pERK1/2 and Sproutys (SPRYs) (Sprouty1, Sprouty2, Sprouty4) and dual specificity phosphatase 6 (DUSP6) were detected by Western blot. The mRNA levels of FGF2, FGFR1 (FGF receptor 1), SPRYs (Sprouty1, Sprouty2, Sprouty4) and DUSP6 mRNA were detected by RT-PCR. RESULTS: Among treatment groups, the content of FGF2 in EuECs and EECs was significantly higher than that in NECs (p < 0.05). The mRNA and protein levels of FGF2, FGFR1, SPRYs (Sprouty1, Sprouty2, Sprouty4) and DUSP6 in EuECs and EECs were increased after adding FGF2 (p < 0.05), but decreased after adding FGF2 neutralizing antibody, no significant change was found in NECs (p > 0.05). The inhibitory effect of PD9805 on NECs was not significantly different from that of U0126 (p > 0.05); however, the inhibitory effects of PD9805 on EuECs and EECs were significantly lower than those of U0126 (p< 0.05). CONCLUSIONS: The positive feedback loop existed in EuECs and EECs, but maybe not in NECs. The results may provide the guideline to treat endometriosis patients.

Electricity production from municipal solid waste using microbial fuel cells.

The organic content of municipal solid waste has long been an attractive source of renewable energy, mainly as a solid fuel in waste-to-energy plants. This study focuses on the potential to use microbial fuel cells to convert municipal solid waste organics into energy using various operational conditions. The results showed that two-chamber microbial fuel cells with carbon felt and carbon felt allocation had a higher maximal power density (20.12 and 30.47 mW m(-2) for 1.5 and 4 L, respectively) than those of other electrode plate allocations. Most two-chamber microbial fuel cells (1.5 and 4 L) had a higher maximal power density than single-chamber ones with corresponding electrode plate allocations. Municipal solid waste with alkali hydrolysis pre-treatment and K3Fe(CN)6 as an electron acceptor improved the maximal power density to 1817.88 mW m(-2) (~0.49% coulomb efficiency, from 0.05-0.49%). The maximal power density from experiments using individual 1.5 and 4 L two-chamber microbial fuel cells, and serial and parallel connections of 1.5 and 4 L two-chamber microbial fuel cells, was found to be in the order of individual 4 L (30.47 mW m(-2)) > serial connection of 1.5 and 4 L (27.75) > individual 1.5 L (20.12) > parallel connection of 1.5 and 4 L (17.04) two-chamber microbial fuel cells . The power density using municipal solid waste microbial fuel cells was compared with information in the literature and discussed.

Glycolysis recycling of rigid waste polyurethane foam from refrigerators.

Rapid growth of rigid waste polyurethane (WPUR) foam from refrigerators attracts the attention all over the world. In this study, glycolysis was chosen to treat WPUR from scrapped refrigerators collected in Shanghai, China. Glycolysis reagents and catalysts were selected. The results indicated that the glycolysis efficiency of ethylene glycol (EG) was higher than that of diethylene glycol, and the catalytic efficiency of alkali metal salts (NaOH) was more excellent than that of triethanolamine and organic salts of alkali metal (NaAc). When EG was 100%WPUR as a glycolysis reagent and NaOH was 1%WPUR as a catalyst at a constant temperature of 197.85°C for 2 h, the glycolysis product had the highest glycolysis conversion rate. In order to maximize the recycling of WPUR, regenerative Polyurethane was performed by adding 10% distilled mixed polyol, which conformed to the QB/T 26689-2011 requirements.

Transient expressions of doppel and its structural analog prionDelta32-121 in SH-SY5Y cells caused cytotoxicity possibly by triggering similar apoptosis pathway.

Doppel (Dpl) is a recently identified prion (PrP)-like protein due to the structural and biochemical similarities, however, its natural function and pathogenic role in neurodegenerative diseases remains unclear. To investigate the possible pathogenic pathway of Dpl and its structural analog for cell apoptosis, mammalian expressing recombinant plasmids containing human PRND gene encoding the full-length Dpl and a truncated human PRNP gene deleting the sequences encoding the peptide from aa 32 to 121 (PrPDelta32-121) were generated. MTT assays showed the cell viabilities of the human neuroblastoma cell line SH-SY5Y receiving Dpl and PrPDelta32-121 expressing plasmids were remarkably lower. Obvious apoptosis phenomena were observed to be associated with the cells transient expressing Dpl and PrPDelta32-121, including reduced mitochondrial transmembrane potential (psim), decreased pro-caspase-3 quantity, more numbers of annexin V- and annexin V/PI-double-stained cells and depressed Bcl-2 level. Moreover, we also found that the Dpl- and PrPDelta32-121-induced cytotoxicities and relevant apoptotic events in SH-SY5Y cells could be fully antagonized by co-expression of the human full-length PrP. These data highly indicate that cytotoxicity induced by the expression of Dpl and truncated PrP in neural derived cells are closely related with the apoptosis process, probably triggering the mitochondrial pathway. It also implies that the cell-benefit activity of the full-length PrP may result from its anti-apoptosis capacity.

Treatment with peginterferon plus ribavirin vs. interferon plus ribavirin for 48 weeks in Chinese patients with chronic hepatitis C.

INTRODUCTION: Virological responses to interferon (IFN) treatment for chronic hepatitis C (CHC) have been found to be correlated with racial differences. Furthermore, higher sustained virological response (SVR) rates were obtained in many randomised clinical trials (RCT) in Caucasian patients with CHC after treating with peg-IFN than those with IFN. However, it is not clear whether this conclusion can be adapted to patients of Chinese descent. METHODS: A search of MEDLINE and China National Knowledge Infrastructure between 1966 and 2008 was performed. Trials comparing the use of peg-IFN plus ribavirin vs. IFN plus ribavirin in treating Chinese patients with CHC were assessed. RESULTS: Of the 94 studies screened, seven RCTs including 398 patients (peg-IFN therapy 232, IFN therapy 166) were analysed. The SVRs obtained in patients treated with peg-IFN plus ribavirin were significantly higher than in patients treated with IFN plus ribavirin [70% vs. 35%, relative risk, 1.76; 95% confidence interval: 1.21-2.56; p (0.01)]. Withdrawal rates were similar between patients treated with peg-IFN plus ribavirin and IFN plus ribavirin. CONCLUSION: Chinese patients with CHC also have a greater likelihood of achieving an SVR with peg-IFN plus ribavirin.

Immunization of mice with plasmids coexpressing HPV16 E5 and the novel oncogene hWAPL.

The novel human oncogene hWAPL is associated with uterine cervical cancer. The HPV16 E5 oncoprotein could induce genomic instability in normal human cells. However, the mechanism of E5 interaction with hWAPL still awaits definition. In our present studies, the eukaryotic expression plasmids, pcDNA3-hWAPL and pcDNA3-hWAPL-E5 were constructed and carried out to vaccinate mice directly. The result that indicated the polyclonal antibody titer in immunized mice sera was increased by enzyme-linked immunosorbent assay. In addition, the proliferative responses of immunized mice spleen cells showed the optical densities values in vaccinated group remarkably higher than that in the control group. In conclusion, the recombinant plasmids could induce strong humoral and cellular immune response and exhibited great potential as therapeutic targets in the treatment of cervical cancer. However, the result didn't show significant difference in group with coexpression of HPV16 E5-hWAPL and group with only hWAPL expression. Consistent with these observations, we demonstrated that HPV16 E5 was not the optimal factor to cooperate with hWAPL in gene therapy.

[Codon usage of Chinese hamster ovary cells.].

BACKGROUND: To investigate the optimal codons of dihydrofolate reductase (dhfr) minus Chinese hamster ovary cells (CHO dhfr-). METHODS: A cDNA library of CHO dhfr- containing high abundence mRNA was constructed and protein-coding sequences were obtained after identification and analysis. Codon frequence of CHO dhf- was compared with that of Chinese hamster in CUTG database. Then codon usage variation among cDNA was investigated using correspondence analysis (COA). RESULTS: Fifty qualified cDNAs from CHO dhfr- were selected, which encodes proteins of high abundence. Comparing with the codon frequence of Chinese hamster, the highest frequence of synonymous codons for amino acids in CHO dhfr- cells were the same as Chinese hamster except that of Arg and Pro. This method of COA identifies the first main factor which can account for the largest fractions (14.7%) of variation among cDNAs. Twenty-two synonymous codons were identified as the optimal codons of CHO cell. CONCLUSION: CHO dhfr- cell has its own optimal codons, it is suggested that codon bias is one of reasons for functional diversity of different mammal cells and it is an effective stratagy to modification of the codon usage of the foreign gene according to the optimal codons of CHO dhfr- to increase the production of foreign gene.

Identification of differentially expressed genes induced by angiotensin II in rat cardiac fibroblasts.

1. Cardiac fibroblasts play an important regulatory role in cardiac remodelling by undergoing proliferation, differentiation and upregulating various gene products, including some cytokines and extracellular matrix (ECM) proteins. A highly potent mediator of cardiac remodelling is angiotensin (Ang) II. 2. In the present study, the suppression subtractive hybridization method was used to identify differentially expressed cDNAs in adult rat cardiac fibroblasts induced by AngII. 3. Following mRNA isolation of non-stimulated and AngII-stimulated cells, cDNAs of both populations were prepared and subtracted by suppression polymerase chain reaction. Sequencing of the partially enriched cDNAs identified 36 genes differentially expressed, including ECM proteins (pro-alpha(1) collagen type III, fibronectin), structural protein (spectrin), enzyme (GTP-specific succinyl-CoA synthetase), transcriptional regulators (glucocorticoid-induced leucine zipper, inhibitor of DNA binding 3) and proteins involved in cell division control (cdc2) or cell signalling (insulin-like growth factor binding protein-3, mutant p53-binding protein, grp75, CGI-121, protein phosphatase type 2A, tspan-2 and Sam68). 4. The diversity of genes identified in the present study further emphasises the central role of AngII in the regulation of cardiac remodelling.

Effect of Chinese Herb Extracts on Spore Germination of Oidium murrayae and Nature of Inhibitory Substance from Chinese Rhubarb.

More than 21% of the Chinese herbs tested contained substances in their aqueous extracts inhibitory to conidial germination of the powdery mildew fungus Oidium murrayae. Extracts from Chinese rhubarb and Japanese knotweed were very effective in controlling powdery mildew on cucumber, pumpkin, and eggplant. The inhibitory substance in Chinese rhubarb was soluble in polar solvents and less soluble in nonpolar solvents. The inhibitor in the aqueous extract was not dialyzable in the membrane tubing with molecular weight cut-off of 14,000, but was exchangeable by anion but not cation exchange resins, indicating that the inhibitor has a molecular weight larger than 14,000 and negative charge on its molecule.

The relationship between HPV16 and expression of CD44v6, nm23H1 in esophageal squamous cell carcinoma.

The esophageal squamous cell carcinoma (ESCC) has high incidence in Shaanxi Province of China. More and more researches indicated that human papillomavirus type 16 (HPV16) might play an important role in carcinogenesis of ESCC but the relationship between HPV16 and CD44v6, nm23H1 has not been elucidated. HPV16 was detected by amplifying HPV16 E6 gene through polymerase chain reaction (PCR) method and the expression of CD44v6, nm23H1 in 40 ESCCs and fifteen normal esophageal mucosa (NEM) from Shaanxi Province was examined by Streptavidin-Peroxidase (SP) method using monoclonal antibody specific to CD44v6 and nm23H1. The positive rates of HPV16 E6 gene, CD44v6 and nm23H1 were 60% (24/40), 65% (26/40) and 45% (18/40) respectively in ESCCs and 26.67% (4/15), 33.33% (5/15) and 86.67% (13/15) respectively in NEMs. There exited statistical difference for HPV16, CD44v6 and nm23H1 between NEMs and ESCCs respectively (p < 0.05). The relationship between HPV16 and the expression of CD44v6 in ESCCs was statistical significance (P = 0.021), but no significant correlation was found between HPV16 and the expression of nm23H1 (P = 0.436) in ESCCs. The infection rate of HPV16 had no statistical difference in all pathological features we observed, but the expression rates of CD44v6 and nm23H1 had statistical correlation with invasion (p = 0.001, 0.013) and lymph nodes metastasis (p = 0.014, 0.002) respectively. In different histology grade of ESSCs, the relationship between HPV16 and CD44v6 was statistical significance in grade I (p = 0.044) but was not in grade II (p = 0.165) and grade III (p = 0.658), however as to the expression of nm23H1 there exited no statistical significance in all histology grades of ESCC (p > 0.05). The expression rates of CD44v6 and nm23H1 were statistically different between grade I and II (p = 0.004, 0.016) respectively and between grade I and grade III (p = 0.014, 0.020), but not statistically different between grade II and III (p = 0.792, 0.943) respectively. Our data firstly suggested that there existed the statistical relationship between the infection of HPV16 and the expression of CD44v6 in ESCCs and that HPV16 may be involved in invasion and metastasis of ESCC.

[Dynamic changes of angiotensin II type-1 receptor mRNA expression in volume-overloaded ventricular hypertrophy].

Using reverse transcriptase-polymerase chain reaction (RT-PCR) the changes of the angiotensin II receptor subtype 1-a (AT1a) and 1-b (AT1b) mRNA levels were examined in hypertrophied ventricles caused by ventral aorta-caval fistula of SD rats. The results show as follows. (1) Three days after operation, the ventricular weight increased significantly, whereas the level of AT1a and AT1b mRNA in both ventricles remained unchanged. (2) Twelve days after operation, the weight increase of both ventricles was more evident, the levels of AT1a and AT1b mRNA in the right ventricle increased (by 62.6% and 72.0%), as compared with control. In the left ventricle, on the other hand, the level of AT1a mRNA increased by 79.0%, while the level of AT1b mRNA showed no obvious increase. (3) Thirty-five days after operation, the levels of AT1a and AT1b mRNA in both ventricles increased still more significant, (i.e., 70.0% and 83.9% in the right and 96.5% and 86.9% in the left). Moreover, 12 d and 35 d after operation, the level of AT1a mRNA was higher than that of AT1b mRNA in both ventricles. (4) There was a positive correlation between the degree of ventricular hypertrophy with the level of AT1 mRNA in ventricular myocardium (r = 0.6168-0.8223). The results suggest that the increased mRNA expression of AT1 in myocardial hypertrophy caused by volume overload may be a mechanism underlying the increased responsiveness of hypertrophic myocardial cells to Ang II, and a difference in the expression between AT1a and AT1b mRNAs in myocardial hypertrophy may be related to the different physiological or pathophysiological roles of Ang II, mediated by the two subtypes of AT1.

The role of labetalol and propofol in the management of HELLP syndrome--a case report.

The acronym HELLP describes a syndrome of (H) hemolysis, (EL) elevated liver enzymes, and (LP) low platelets, which symptomatically defines a unique group of preeclamptic/eclamptic women. Pregnancies complicated by this syndrome are associated with poor maternal and fetal outcome. This poses a considerable challenge to the anesthesiologists as many of these patients require emergent Cesarean section. We reported a 30-year-old woman with gestation of 32 wk who sustained Cesarean section under the impression of pregnancy with HELLP syndrome. General anesthesia was performed for this patient. The preoperative evaluation, general management, and intraoperative monitoring system were described. The individual role and use of labetalol and propofol were also discussed in this report.

Long-term propofol infusion and airway management in a patient with Goldenhar's syndrome.

A 2-year-old patient of Goldenhar's syndrome received an operation for corneal transplantation. Difficult endotracheal intubation from the congenital anomaly was treated with laryngeal mask airway and pediatric fiberoptic laryngoscope. Long-term propofol infusion (> 10 h) for anaesthetic maintenance in this small child was used with rapid and smooth recovery.