Objective: To establish a method for the determination of oleanolic acid(OA)in Pfaffia glomerata to compare the contents of OA in different parts of P. glomerata from Guangxi, China, with different growth years and different habitats. Methods: HPLC method was adopted. The determination was performed on Phenomenex Luna C18(250 mm×4.6 mm, 5 μm)column with mobile phase consisting of methanol-water-glacial acetic acid-triethylamine(285:15: 0.2:0.1, V/V) at the flow rate of 1.0 ml/min. The column temperature was set at 30℃. The detection wavelength was set at 210 nm, and the injection volume was 20 μl. The content of OA in P. glomerata was determined by the established method, and the extraction process of OA was optimized by the orthogonal test. Results: The linear range of OA was 0.0206-2.060 mg/ml(r2=1.0000). RSDs for the precision, repeatability and stability tests were all lower than 2%(n=6 or n=7). The average recovery of OA was 100.89%(RSD=1.69%, n=9). The optimum extraction conditions of OA were as follows: 20-fold ethanol(80%, V/V), extracting for three times, refluxing 1.5 hour each time, and processing acid hydrolysis with 3.0% sulfuric acid(g/g)for 1 hour. Under these conditions, the OA had the highest extraction efficiency. The contents of OA in reed head, root, old stem, tender stem, leaf and flower of P. glomerata were 4.87, 4.61, 2.67, 0.99, 0.24 and 1.13 mg/g, respectively. The average contents of OA in the roots of P. glomerata aged 1, 2, 3, 4 and 5 years in Guangxi were 3.08, 4.07, 4.71, 4.62 and 4.46 mg/g, respectively. Conclusion: The established extraction process and detection method is suitable for the extraction and content determination of OA in P. glomerata. Although OA is distributed in all parts of P. glomerata, the contents significantly vary in different parts. The content of OA is highest in reed head and lowest in leaves. The OA content in P. glomerata becomes stable after 3 years of growth in Guangxi.

Objective To observe the effects of epidermal growth factor (EGF) on cell proliferation and cell cycle of cultured bovine corneal endothelial cells. Methods Bovine corneal endothelial cells were cultured with different concentrations of hEGF (1, 10 and 100 ng/mL). MTT test was performed to evaluate the cell proliferation. The bovine corneal endothelial cells were divided into two groups:control group (cultured in DMEM) and EGF-stimulated group (cultured in DMEM with EGF). Flow cytometry was performed to determine the cell cycle phases on the third and seventh day. Results Compared with the control, EGF enhanced the cell proliferation in a dose-related response. 10 ng/mL and 100 ng/mL EGF were much more effective than 1 ng/mL.On the third day, S phase cells accounted for 24.5% and G_2-M phase cells 0.08% in the control group,while 24.6% and 0.06%, respectively in the EGF-stimulated group. However, on the seventh day, those came to 20.8% and 0.41% in the control group, and 18.2% and 1.55% in the EGF-stimulated group,indicating a significant change in the cell cycle (P

This article describes the progress of developing the training base and training methods for bronchoscopists at Changhai hospital in recent years,and then discusses the potential issues and solutions that might occure in the course of training,and finally explores the model and methodology to optimize the training program for Chinese bronchosocpists.

OBJECTIVETo investigate the protective effect of hydrogen against hyperoxia-induced oxidative stress injury in premature rat type II alveolar epithelial cells (AECs).

METHODSThe type II AECs isolated from premature rats were randomly divided into air (21% oxygen) control group, hyperoxia (95% oxygen) control group, air + hydrogen group, and hyperoxia+ hydrogen group. The cells with hydrogen treatment were cultured in the presence of rich hydrogen. After the corresponding exposure for 24 h, the cell morphology was observed microscopically. MTT assay was used to evaluated the cell proliferation ability, and JC-1 fluorescence probe was used to detect the mitochondrial membrane potential (δφ) changes of the type II AECs. The concentration of maleic dialdehyde (MDA) and superoxide dismutase (SOD) activity in the cell supernatant were detected using colorimetric method.

RESULTSNo significant differences were found in cell growth or measurements between air control and air + hydrogen groups. Compared with air control group, the cells exposed to hyperoxia showed significantly suppressed proliferation, reduced mitochondrial membrane potential, increased MDA content, and decreased SOD activity. Intervention with hydrogen resulted in significantly increased cell proliferation and SOD activity and lowered MDA content, and restored the mitochondrial membrane potential in the cells with hyperoxia exposure (P<0.05).

CONCLUSIONHydrogen can significantly reduce hyperoxia-induced oxidative stress injury in premature rat type II AECs, improve the cellular antioxidant capacity, stabilize the mitochondrial membrane potential, and reduce the inhibitory effect of hyperoxia on cell proliferation.

Animals ; Animals, Newborn ; Antioxidants ; metabolism ; Cell Proliferation ; Cells, Cultured ; Epithelial Cells ; drug effects ; Female ; Hydrogen ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Membrane Potential, Mitochondrial ; Oxidative Stress ; drug effects ; Oxygen ; adverse effects ; Pulmonary Alveoli ; cytology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

Objective To investigate the supply of iodized salt in non-excessive iodine counties and iodine-free salt in excessive iodine counties at household level in Hebei province so as to provide a basis for the prevention and control of iodine deficiency disorders(IDD). Methods According to the national project of IDD surveillance,the county was taken as the elementary sampling unit. The towns and villages were selected by systematic and random sampling in every county and households were chosen by random sampling to collect their edible salt in Hebei province in 2008. The salt iodine content in non-and excessive iodine regions was detected by direct titrition method and semiquantitative method respectively. Results all 48 448 salt samples were collected from 167 non-excessive iodine counties. Weighed by the population of counties,the rate of non-iodized salt was 4.73%. Iodized salt accounted for 95.27%,out of which,96.13% were qualified and the consuming rate of qualified iodized salt was 91.96%. Eighty point eighty three percent(135/167) of the counties covered by iodized salt above 95%,92.81% (155/167) passing rate of iodized salt above 90% and 82.04 (137/167) consuming rate of qualified iodized salt. All 1466 salt samples were collected in 5 counties with excessive water iodine content and the coverage rate of iodine-free salt was 93.25%(1367/1466). Conclusions In a nutshell,the national targets for preliminary elimination of IDD have been achieved in regions of non-excessive iodine of Hebei province. Nevertheless,the coverage rate of iodized salt and qualified iodize salt rate in some counties are still below the national standard. Therefore the prevention and control of IDD need to be strengthened. The supply of iodized salt in excessive iodine regions should be timely stopped.

Objective To understand the status of drinking-water-borne endemic fluorosis and the effect of preventive measure in Hebei province, so as to provide a basis to prevent and cure the disease. Methods Thirtyeight affected counties(cities, districts) with drinking-water-borne endemic fluorosis were sampled by random sampling in Hebei in 2009. All affected villages in every county were divided into mild, moderate and severe endemic fluorosis areas and a village was randomly selected from each category of the area to carry out the monitoring of endemic fluorosis. Dental fluorosis of children aged 8 - 12 were examined and 6 copies of urine samples were randomly collected in each age group in the above-mentioned villages. Clinical skeletal fluorosis was diagnosed among adults aged 16 and over and 20 copies of urine samples were tested for fluorosis in every village.Results A total of 112 affected villages were investigated, among which the drinking water quality of 66 villages were improved and 46 villages were not improved. A total of 236 copies of water samples from the 66 villages were measured and the fluoride content ranged from 0.1 to 4.3 mg/L, among which 20 copies of water samples exceeded the fluorine standard of 1.2 mg/L, accounting for 33.3%. A total of 230 copies of water samples were collected in the 46 villages and the fluoride content ranged from 0.2 to 4.6 mg/L, among which 76.1% (35/46) of the water samples exceeded the fluorine standard of 1.2 mg/L. A total of 5169 children aged 8 - 12 were examined of dental fluorosis, the dental fluorosis rate was 36.43%(1883/5169) and the dental fluorosis index was 0.81. A sum of 71 497 adults aged over 16 years were examined, and the rate of skeletal fluorosis was 4.81%(3438/71 497), moderate or severe clinical detection rate of skeletal fluorosis was 1.56%( 1114/71 497). A total of 2876 copies of children urine samples and 2021 copies of adult urine samples were tested and the geometric mean of fluoride content was 2.30,3.32 mg/L, respectively. Conclusions The prevalence of dental fluorosis of children in the areas with improved water is less than 30% and the rate of dental fluorosis and skeletal fluorosis decline gradually with time.The rate of dental fluorosis and skeletal fluorosis increases with the increase of water fluoride in the water quality not improved areas. The endemic fluorosis is still comparatively serious in Hebei. The progress of improving water quality in the areas with endemic fluorosis should be accelerated and the acceptability of improved water should be enhanced.

Objective To explore the effect of long non-coding RNA(lncRNA)SOCS2-AS1 on the proliferation,invasion and migration of gastric cancer cells and its potential mechanism.Methods 40 cases of gastric cancer tissues and 40 cases of adjacent tissues were selected as experimental specimens.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of lncRNA SOCS2-AS1 in gastric cancer tissues and adjacent tissues.Human gastric cancer SGC-7901 cells were divided into vector group,pcDNA-SOCS2-AS1 group,NC-siRNA group,SOCS2-AS1-siRNA group,pcDNA-SOCS2-AS1+vector group and pcDNA-SOCS2-AS1+pcDNA-YAP1 group.Vector group,pcDNA-SOCS2-AS1 group,NC-siRNA group and SOCS2-AS1 siRNA group were transfected with vector,pcDNA-SOCS2-AS1,NC-siRNA and SOCS2-AS 1 siRNA plasmids,respectively;the pcDNA-SOCS2-AS1+vector group were transfected with the pcDNA-SOCS2-AS1+vector plasmids;the pcDNA-SOCS2-AS1+pcDNA-YAP1 group were transfected with both pcDNA-SOCS2-AS1+pcDNA-YAP1 plasmids simultaneously.RT-qPCR was used to detect the expression level of lncRNA SOCS2-AS1 in cells;Western blotting was used to detect the expression of Hippo-YAP pathway-related proteins LATS1 and YAP1 in gastric cancer cells;EDU staining was used to detect the proliferation of gastric cancer cells;Transwell assay was used to detect cell migration and invasion.Results The relative expression levels of SOCS2-AS1 in gastric cancer tissues,adjacent tissues,normal gastric epithelial cells GES-1 and gastric cancer cells SGC-7901 were 1.00±0.18,0.38±0.09,1.03±0.21 and 0.42±0.11.Compared with the adjacent tissues and normal gastric epithelial cell line GES-1,SOCS2-AS1 was significantly down-regulated in gastric cancer tissues and gastric cancer cell line SGC-7901,and the differences were statistically significant(P＜0.05).The cell proliferation rates in the vector group,pcDNA-SOCS2-AS1 group,NC-siRNA group and SOCS2-AS1-siRNA group were(36.23±2.53)％,(18.15±1.04)％,(35.83±2.15)％and(56.11±6.57)％;the invasive cell numbers were 168.15±19.11,54.36±6.27,124.47±14.53 and 238.62±21.34;the numbers of migrating cells were 194.22±16.33,91.61±8.47,148.14±16.25 and 279.31±24.68;the relative expression levels of LATS1 protein were 0.93±0.08,2.86±0.21,0.96±0.09 and 0.31±0.03;the relative expression levels of YAP1 protein were 0.95±0.11,0.28±0.04,0.92±0.12 and 3.16±0.28;the pcDNA-SOCS2-AS1 group was significantly lower than the vector group,and the SOCS2-AS1-siRNA group was significantly higher than the NC-siRNA group,and the differences were statistically significant(all P＜0.05).The cell proliferation rates of pcDNA-SOCS2-AS1+vector group and pcDNA-SOCS2-AS1+pcDNA-YAP1 group were(33.62±5.73)％and(88.46±9.17)％;the number of invasive cells were 53.61±6.44 and 131.73±15.29;the number of migrating cells were 68.35±7.63 and 159.81±16.47,the differences were statistically significant(all P＜0.05).Conclusion LncRNA SOCS2-AS1 inhibits the proliferation,invasion and migration of gastric cancer cells by activating the Hippo-YAP signaling pathway.

Objective:To investigate the effects of Clostridium butyricum on colitis and intestinal microbiota in mice with or without antibiotic pretreatment. Methods:Thirty specific pathogen free BALB/c mice were randomly divided into the blank control group, dextran sulfate sodium (DSS) group, antibiotic + DSS group, Clostridium butyricum + DSS group and antibiotic+ Clostridium butyricum + DSS group, with 6 mice in each group. After the mice were pretreated with quadruple antibiotics (ampicillin 1 g/L, neomycin 1 g/L, metronidazole 1 g/L, and vancomycin 0.5 g/L) in normal drinking water for 30 d, the mice colitis model was induced with DSS. At the same time, the mice in Clostridium butyricum + DSS group and antibiotics+ Clostridium butyricum + DSS group were given 1×10 6colony-forming unit (CFU) Clostridium butyricum by gavage. The effect of Clostridium butyricum on mice with colitis was evaluated by disease activity index (DAI), colon length and histopathological score. The level of serum inflammatory factors was detected by enxyme linked immunosorbent assay, and the effect of Clostridium butyricum on gut microbita in mice was determined by fecal 16S rRNA sequencing. Results:The general condition of mice of the blank control group were good, and their DAI scores fluctuated around 0. Since the fourth day after DSS drinking water was given, the mice of the DSS group showed signs of colitis such as weight loss, unformed stools and bloody stools. On the fourth day after intervention, the DAI score of Clostridium butyricum + DSS group was lower than that of DSS group (0.000±0.000 vs. 0.444±0.111), and the difference was statistically significant ( t=4.000, P=0.016 1). On the tenth and twelfth day after the intervention, the DAI scores of antibiotic+ Clostridium butyricum + DSS group were both lower than those of antibiotic+ DSS group (0.000±0.000 vs. 1.111±0.222, 0.667±0.000 vs. 1.889±0.222), and the differences were statistically significant ( t=5.000 and 5.500, both P<0.05). The histopathological score of mice colon tissue of Clostridium butyricum + DSS group was lower than that of DSS group (2.50±1.73 vs. 5.50±1.00), and the histopathological score of mice colon tissue of antibiotic+ Clostridium butyricum+ DSS group was lower than that of antibiotic+ DSS group (1.25±0.96 vs. 5.00±0.82), and the differences were statistically significant ( t=3.000 and 5.960, both P<0.05). The serum level of interleukin (IL)-1β Clostridium butyricum+ DSS group was higher than that of blank control group ((4.464±0.075) ng/L vs. (3.907±0.080) ng/L), the serum levels of tumor necrosis factor-α, IL-6 and IL-1β of Clostridium butyricum+ DSS group and antibiotic+ Clostridium butyricum + DSS group were all lower than those of DSS group ((2.402±0.383) ng/L , (1.845±0.345) ng/L vs. (6.958±1.084) ng/L, (1.752±0.146) ng/L, (1.307±0.048) ng/L vs. (3.537±0.608) ng/L, (4.464±0.075) ng/L, (4.066±0.190) ng/L vs. (7.477±0.339) ng/L), and the differences were statistically significant ( t=5.005, 3.964, 4.495, 4.693, 6.294, 8.674 and 8.774 , all P<0.05). The results of 16S rRNA sequencing showed that there were a significantly large number of anti-inflammatory or short-chain fatty acid producing bacteria in the gut microbiota of mice intervened by Clostridium butyricum, among which the dominant bacteria genus in Clostridium butyricum + DSS group and antibiotic+ Colstridium butyicum+ DSS group were Mucispirillum (linear discriminant analysis (LDA)=3.667 log10, P=0.004) and Stenotrophomonas (LDA=2.778 log10, P=0.044). In the antibiotic+ Clostridium butyricum+ DSS group, the dominant bacteria genus were Peptococcus (LDA=2.685 log10, P=0.018), Butyricimonas (LDA=2.712 log10, P=0.011), Bilophila (LDA=3.204 log10, P=0.014), Intestinimonas (LDA=3.346 log10, P=0.010), Candidatus- Saccharimonas (LDA=3.363 log10, P=0.029), Desulfovibrio (LDA=3.402 log10, P=0.025), Oscillibacter (LDA=2.870 log10, P=0.019) and Akkermansia (LDA=4.031 log10, P=0.005). Conclusions:Clostridium butyricum can effectively improve colitis in mice and regulate the intestinal microbial structure of mice, whlie antibiotic pretreatment can strengthen its regulation of intestinal microbiota to and enhance the efficacy of Clostridium butyricum.

OBJECTIVETo evaluate the association between two single nucleotide polymorphisms located in the promoter of transforming growth factor-β1 receptor 2 (TGFBR2) gene and hypertension in Han Chinese population.

METHODSThe subjects were recruited from the population of cluster sampling survey for essential hypertension (EH) in two townships of Yixing city, Jiangsu province in 2009. Overall, 2012 patients with hypertension and 2116 age (± 2 years) and sex-matched unrelated controls were selected. Epidemiological data, physical measurements results and serum glucose and lipid biomarker were collected and detected. Linkage disequilibrium (LD) analysis were applied and two tagging single nucleotide polymorphisms (tagSNP) in 5' upstream of TGFBR2 gene (rs6785358, -3779A/G; rs764522, -1444C/G) were selected for genotyping and analyzing for the association with hypertension.

RESULTSThe frequencies of AA, AG, GG in case and control of rs6785358 were 1455 (72.3%), 517 (25.7%), 40 (2.0%) and 1582 (74.8%), 490 (23.2%), 43 (2.0%) respectively, and CC, CG, GG of rs764522 were 1524 (75.7%), 464 (23.1%), 24 (1.2%) and 1654 (78.2%), 436 (20.6%), 26 (1.2%) respectively. SNP rs764522 was significantly associated with EH and OR (95%CI) were 1.17 (1.01 - 1.36) (P < 0.05) in dominant model after adjustment for confounding factors such as age, sex, glucose, lipids, smoking and alcohol drinking. Further stratification analysis by age, sex, smoking and alcohol drinking indicated that individuals carrying G allele (CG/GG genotype) of SNP rs764522 had higher susceptibility to EH than CC genotype (OR = 1.21, 95%CI: 1.01 - 1.45) (P < 0.05) in ≥ 55 years group. No statistical significance was detected in the distribution of genotypes and allele frequencies for SNP rs6785358 between cases and controls (P > 0.05). Haplotype analysis showed that no significant frequency difference of haplotype structured by rs6785358 and rs764522 was found between cases and controls (P > 0.05), and no significant blood pressure change was found between genotype variations of rs6785358 and rs764522 (P > 0.05).

CONCLUSIONSNP rs764522 of TGFBR2 gene is associated with increased risk of EH in elderly Han Chinese population.

Aged ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Hypertension ; epidemiology ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Protein-Serine-Threonine Kinases ; genetics ; Receptors, Transforming Growth Factor beta ; genetics