Objective: To explore the influencing factors for the primary and secondary school students with abnormal angle of trunk rotation for the prevention. Methods: The students of Grade Four to Nine in Jiashan County of Zhejiang Province were selected by cluster sampling method. A self-designed questionnaire was used to collect social demographic data, diet behaviors, physical activities, reading and writing habits. The angle of trunk rotation was measured by scoliometer. Logistic regression model was used to investigate the influencing factors for abnormal angle of trunk rotation. Results: This study included 2 942 schoolchildren, with 1 582 ( 53.78% ) boys and 1 360 ( 46.23% ) girls. The incidence rate of abnormal angle of trunk rotation was 7.82%. The incidence rate of abnormal angle of trunk rotation in girls was 10.74%, which was higher than 5.31% in boys ( P<0.05 ). Grade ( OR=1.485, 95%CI: 1.058-2.085 ), gender ( OR=2.084, 95%CI: 1.536-2.828 ), frequency of eating fresh vegetables in the past week ( OR=0.749, 95%CI: 0.633-0.887 ) and watching electronic screen in the dark ( OR=1.188, 95%CI: 1.002-1.408 ) were the influencing factors for abnormal angle of trunk rotation in primary and secondary school students. Grade ( OR=2.664, 95%CI: 1.481-4.791 ) and watching electronic screen in the dark ( OR=1.325, 95%CI: 1.030-1.704 ) were influencing factors for abnormal angle of trunk rotation in boys. Frequency of eating fresh vegetables in the past week ( OR=0.714, 95%CI: 0.574-0.887 ) and uncorrected eyesight less than 5.0 ( OR=1.795, 95%CI: 1.164-2.767 ) were influencing factors for abnormal angle of trunk rotation in girls. Conclusion The abnormal angle of trunk rotation in primary and secondary school students is related to gender, grade, reading and writing behaviors as well as diets; and the influencing factors are different in male and female students.

Diabetes Mellitus, Type 2 ; drug therapy ; Humans ; Hypoglycemic Agents ; adverse effects ; Neoplasms ; chemically induced ; Risk Factors

OBJECTIVETo develop an HPLC method for determining flavoniod-glycosides in Yixintong pills.

METHODAgilent ZORBAX Extend-C18 (4.6 mm x 250 mm, 5 microm) was used with tetrahydrofuran-methanol-acetonitrile-water-acetic acid (30:3:4:120:3) as mobile phase. Detection wavelength was 330 nm.

RESULTGood linearities of rhamnosylvitexin and vetexin-glucoside were obtained within the range of 0.044-1.78 microg (r = 0.9999) and 0.042-0.85 microg (r = 0.9999); the average recoveries were 100.0% and 99.2%; RSD were 0.92% and 1.8%, respectively.

CONCLUSIONThis method can be used for quality control of Yixintong pills.

Apigenin ; analysis ; Chromatography, High Pressure Liquid ; Crataegus ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control

OBJECTIVETo evaluate the influence of Tinglizi on collagen volume fraction (CVF) and perivascular collagen volume area (PVCA ) in left ventricle tissue of cardiac hypertrophy induced by abdominal aortic banding in rats.

METHODVentricular remodeling was induced by abdominal aortic banding (AAB) in rats. After 30 day treatment, the systolic blood pressure (SBP), diastolic blood pressure (DBP); heart rate (HR) were measured. The histological assay consisted of the HE stain for determining the myo-cardium cell cross section and collagen stain (Van Gieson' method) for determining collagen content, including collagen volume fracton (CVF) and perivascular collagen volume area (PVCA).

RESULTThe experimental data demonstrated that Tinglizi decreased SBP, DBP, HR and could significantly reduce the total collagen content (CVF, PVCA) and lessen the myocardium cell cross section (P < 0.05).

CONCLUSIONTinglizi may decrease the total collagen content of ventricle and attenuate the ventricular remodeling induced by abdominal aortic banding.

Animals ; Blood Pressure ; drug effects ; Cardiomegaly ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Heart Rate ; drug effects ; Heart Ventricles ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Ventricular Remodeling ; drug effects

OBJECTIVETo construct a eukaryotic expression vector encoding the gene of extracellular region of mouse B7-H4, to express it in yeast cell line and to determine its biological activity.

METHODSThe extracellular region of the mouse B7-H4 gene was amplified with Xho I and EcoR I by PCR from a mouse B7-H4 chimeric plasmid. Digested with Xho I and EcoR I, the mB7-H4 gene was inserted into the yeast expression plasmid Ppic9. The DNA sequence was confirmed by double digestion and the Ppic9-mB7-H4 plasmid was transfected into the yeast cells. The expression of mB7-H4 was confirmed by PCR, Western Blot and ELISA analysis, and its biological function was determined.

RESULTPpic9-mB7-H4 transfectants expressed mB7-H4 in yeast cells, and mB7-H4 effectively inhibited the proliferation of T lymphocytes with a fractional inhibition rate of 28.3 % and inhibited IL-2, IL-4, IL-10 and IFN-gamma production with fractional inhibition rates of 68.8%, 78.8%, 67.6% and 77.7%, respectively.

CONCLUSIONThe eukaryotic expression plasmid mouse B7-H4 has been successfully constructed and the expressed products of B7-H4 possess biological activity.

Animals ; B7-1 Antigen ; biosynthesis ; genetics ; Mice ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; T-Lymphocytes ; immunology ; V-Set Domain-Containing T-Cell Activation Inhibitor 1

OBJECTIVETo construct the recombinant adenovirus containing B7-H4 gene with AdEasy XL system and to identify its biological activities.

METHODSThe full-length mouse B7-H4 gene was amplified by RT-PCR from C57 mouse lung and put into T vector, then verified by sequencing. Digested with Xhol I and EcoR V the B7-H4 gene was inserted into pshuttle-CMW(PSC). Pme I linearized shuttle plasmid was transformed into E.coli BJ5183-AD-1 to obtain the recombinant adenoviral plasmid pAd-mB7-H4 by efficient homologous recombination. Then the recombinant adenovirus-mB7-H4/Ad was obtained by packaging Pac I linearized in D-293 cells. The mRNA and protein expression of B7-H4 in mB7-H4/Ad infected AD-293 cells were detected by RT-PCR and Western blot, respectively. mB7-H4/Ad was used to infect L929 cells, the bioactivity of expressed B7-H4 in stimulation of T lymphocytes proliferation and cytokine production were tested.

RESULTSThe full-length of mB7-H4 was cloned from mouse lung tissue cDNA and verified by sequencing. The recombinant plasmid pAd-m B7-H4 was successfully generated by homologous recombination, and the primary mB7-H4/Ad was obtained by packaging pAd-B7-H4 in AD-293 cells. Compared with the negative control, L929 cells infected with mB7-H4/Ad effectively inhibited the proliferation of T lymphocytes and cytokines production.

CONCLUSIONThe bioactive recombinant adenovirus mB7-H4/Ad has been successfully constructed.

Adenoviridae ; genetics ; Animals ; B7-1 Antigen ; biosynthesis ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Mice ; Mice, Inbred C57BL ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; T-Lymphocytes ; immunology ; Transfection ; V-Set Domain-Containing T-Cell Activation Inhibitor 1

BACKGROUNDGrowing evidence from population and clinic based studies showed that obstructive sleep apnea (OSA) and its characterizing chronic intermittent hypoxia (IH) were independently associated with the development of type 2 diabetes mellitus. However, the pathogenesis by which OSA induces glucose metabolic disorders is not clear. We determined changes in pancreatic β cell mass and the mammalian target of rapamycin (mTOR)/hypoxia inducible factor 1 (HIF-1)/vascular endothelial growth factor A (VEGF-A) pathway following IH exposure.

METHODSA controlled gas delivery system regulated the flow of nitrogen and oxygen into a customized cage housing mice during the experiment. Twenty-four male wild C57BL/6J mice were either exposed to IH (n = 12) or intermittent air as a control (n = 12) for 56 days. Mice were anaesthetized and sacrificed after exposure, pancreas samples were dissected for immunofluorescent staining. Insulin and DAPI staining labelled islet β cells. Insulin positive area and β cell number per islet were measured. P-S6, HIF-1α and VEGF-A staining were performed to detect the activation of mTOR/HIF-1/VEGF-A pathway.

RESULTSAfter eight weeks of IH exposure, insulin positive area increased by an average of 18.5% (P < 0.05). The β cell number per islet increased (92 vs. 55, respectively for IH and the control groups, P < 0.05) with no change in the size of individual β cells. Islet expression of HIF-1α and VEGF-A were higher in IH group than control group, and percentage of p-S6 positive β cell also increased after IH exposure (16.8% vs. 4.6% respectively for IH and the control groups, P < 0.05).

CONCLUSIONThe number of pancreatic β cells increased as did the activity of the mTOR/HIF-1/VEGF-A pathway after exposure to IH.

Animals ; Hypoxia ; pathology ; Hypoxia-Inducible Factor 1 ; physiology ; Insulin-Secreting Cells ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Signal Transduction ; TOR Serine-Threonine Kinases ; physiology ; Vascular Endothelial Growth Factor A ; physiology

OBJECTIVETo study the methylation status of p73 gene promoter in patients with myelodysplastic syndrome (MDS) and explore its significance with clinical prognosis.

METHODSMethylation of p73 promoter was detected in bone marrow cells from 135 MDS patients and 13 healthy controls by methylation-specific PCR (MSP). The results of MSP were confirmed by bisulfite sequencing. The expression of p73 mRNA was detected by real-time quantitative PCR. Primary bone marrow cells from MDS patients were treated with decitabine, the changes of p73 methylation status and p73 mRNA expression were measured. The role of p73 methylation in the prognosis of MDS and the correlated clinical data were explored.

RESULTSp73 hypermethylation was present in 37.04% of MDS cases and patients with high risk MDS (RAEB-1 and RAEB-2) exhibited a significantly higher frequency of p73 methylation than that of low risk MDS (58.8% vs 29.7%, P = 0.002). The expression of p73 mRNA in the methylated group was decreased compared to that of the unmethylated group (P = 0.032). Decitabine treatment decreased the level of p73 methylation and increased the level of p73 transcripts. Patients with p73 methylation progressed rapidly to AML (P < 0.001) and had shorter survival (P = 0.002) than those who did not have p73 methylation. In the multivariate Cox regression model, BM blast and p73 methylation status emerged as independent prognostic factor for overall survival and leukemia free survival.

CONCLUSIONp73 gene methylation is common in patients with MDS and may indicate poor prognosis. p73 may be a therapeutic target in MDS.

Aged ; Case-Control Studies ; DNA Methylation ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Prognosis ; Promoter Regions, Genetic ; Tumor Protein p73 ; Tumor Suppressor Proteins ; genetics

Objective To screen the critical genes related to the development of esophageal squamous cell carcinoma ( ESCC ) by weighted gene co-expression network analysis ( WGCNA ) and to verify by experiments.Methods Gene expression data of ESCC were downloaded from gene expression omnibus (GEO) database based on gene chip platform ( GPL) 570, GPL571, GPL96/97 or GPL14613 platform, respectively. Meanwhile, the obtained differentially expressed genes together with gene expression data of 81 ESCC patients from the cancer genome atlas ( TCGA ) and clinical data were analyzed by WGCNA to set up co-expression networks including mRNA and microRNA ( miRNA ) . The expression of miRNA in ESCC tissues and paracancerous tissues was examined by quantitative real-time polymerase chain reaction ( RT-PCR ) .And the expression of target protein Kruppel like factor 4 ( KLF4 ) and desmocollin 2 ( DSC2 ) were detected by immunohistochemistry .After ESCC cell line ECA-109 cells were transfected with miRNA-92b-3p mimic, cell cycle was tested by flow cytometry ,the cell invasion and migration ability was measured by Transwell chamber assay and scratch-wound assay.The expression of KLF4 and DSC2 was observed by confocal laser scanning microscopy and Western blotting .The target genes were verified by luciferase assay .T-test, rank sum test, chi-square test and Pearson correlation analysis were performed for statistical analysis .Results A total of 4023 differential expression gene ( DEG) and 328 differential expression miRNA ( DEM) were screened and 11 gene modules were set up by WGCNA .Among them, the brown modules were negatively associated with tumor grade and T stage (r=-0.340 and -0.268, P=0.002 and 0.016).Meanwhile, has-miR-92b and the potential target genes KLF4 and DSC2 were all in the brown module .Furthermore, the results of RT-PCR showed the expression of miRNA-92b-3p in ESCC tissues was higher than that in paracancerous tissues (3.052(1.652, 5.371) vs.0.985(0.558, 2.032)), and the difference was statistically significant (Z=-4.021,P<0.01). The results of immunohistochemistry demonstrated that the positive rates of KLF 4 and DSC2 in ESCC tissues were 43.3％(13/30) and 20.0％(6/30), respectively, which were lower than those of paracancerous tissues (70.0％(21/30) and 63.3％(19/30)), and the differences were statistically significant (χ2 =4.344 and 1.589, both P<0.05).After ECA-109 cells were transfected with miRNA-92b-3p mimics, the percentage of cells at G0/G1 phase decreased ((63.71 ±2.83)％vs.(54.62 ±4.00)％) and the percentage of cells at the S phase and G2/M phase increased ((31.81 ±2.88)％vs.(41.20％±2.87)％, and (3.87 ±1.75)％vs. (8.10 ±1.71)％, respectively), and the differences were statistically significant (t =3.215, 4.000 and 2.998;P=0.032, 0.016 and 0.040).The invasion and migration ability of the cells were significantly improved (79.67 ±27.54 vs.280.33 ±46.18, (69.72 ±3.91)％ vs.(84.90 ±5.25)％), and the differences were statistically significant (t=6.465 and 4.019, P=0.003 and 0.016).The results of Western blotting indicated that, compared with control mimic group , the expression of KLF4 and DSC2 was both dramatically downregulated after transfected with miRNA-92b-3p mimics transfected (1.00 ±0.23 vs.0.42 ±0.03, 1.00 ±0.20 vs.0.55 ± 0.21), and differences were statistically significant (t=4.470 and 5.493, P=0.042 and 0.032).The results of luciferase assay demonstrated that miRNA-92b-3p could directly bind KLF4 and DSC2. Conclusion WGCNA is an efficient systemic biological approach by which miRNA-92b-3p is identified as a new cancer-promoting gene .

OBJECTIVETo evaluate the roles of enteric nervous system neurotransmitters, nitric oxide (NO), substance P (SP) and vasoactive intestinal polypeptide (VIP), and interstitial cells of Cajal (ICC) in the colon in slow transit constipation in rats.

METHODSThirty-two healthy Wistar rats were randomly assigned to control and constipated groups. In the constipated group, the rats were daily administered with diphenoxylate (8 mg/kg) to develop slow transit constipation, while the control rats were fed with water. The number and the weight of fecal granule and the body weight of rats were recorded every 5 days for 90 days. Transit functions of intestinal movement were examined by an activated charcoal suspension pushing test one week after stopping the administration of diphenoxylate. The levels of NO and SP in the colonic mucosa were measured by nitrate reductase methods and ELISA respectively. The distribution of VIP and ICC positive cells confirmed with symbolic c-kit+ cells in the colonic wall were observed by immunohistochemical methods.

RESULTSThe daily number of fecal granule in the constipated group was significantly less than that in the control group (P<0.01). The mean weight of each fecal granule in the constipated group was significantly higher than that in the control group (P<0.01). The discharge time of the first granule of black faeces in the constipated group (430.2+/- 132.1 min) was significantly longer than that in the control group (337.2+/- 74.7 min; P<0.05). There were no significant differences in NO and SP levels and the density of VIP positive cells in the distal colonic segment between the two groups. The number of c-kit+ cells in the distal colonic wall in the constipated group was significantly reduced compared with that in the control group (P<0.05).

CONCLUSIONSThe reduction of ICC number in the distal colon may be contributed to the pathogenesis of slow transit constipation in rats.

Animals ; Body Weight ; Coiled Bodies ; Colon ; cytology ; innervation ; Constipation ; etiology ; Male ; Neurotransmitter Agents ; physiology ; Nitric Oxide ; analysis ; physiology ; Proto-Oncogene Proteins c-kit ; analysis ; Rats ; Rats, Wistar ; Substance P ; analysis ; physiology ; Vasoactive Intestinal Peptide ; analysis ; physiology