Background and purpose:Arsenic trioxide,verified as a breakthrough in the management of acute promyelocytic leukemia,has been applied to a variety of solid tumors.Gall bladder carcinoma,lacking specific clinical manifestations,is usually diagnosed at advanced stages of the diseases and few cases can be resected by operation.Chemotherapy has not shown significant activity in gall bladder carcinoma.This study was to investigate the biological effect of As2O3 on the growth of human gall bladder carcinoma cell and its mechanism.Methods:GBC cells were cultured with different concentrations of As2O3,the proliferative activity of the cells was detected by MTT methods,and the cell cycle status was carried out by flow cytometry(FCM).Western blot and RT-PCR were performed to analyse the expression of cyclin D1,D2,D3,CDK4 and CDK6.GBC cells were transient transfected with cyclin D1 promoter construct pGL3 and then treated by different doses of As2O3.The luciferase activity was measured.Results:The treatment of As2O3 in gall bladder carcinoma cells could inhibit the growth of cells in a time and dose dependent manner,make cells arrest in G1 phase and down regulate the expression of cyclin D1.In addition,the activity of cyclin D1 promoter was down-regulated by As2O3 in a dose-dependent manner and decreased about 70 percent when treated with 4 ?mol/L As2O3.Conclusions:As2O3 can significantly inhibit the growth of human gall bladder carcinoma cells as well as down-regulate the expression of cyclin D1 in vitro.

Objective: To observe the effects of acupoint thread-embedding therapy and low-carbohydrate diet therapy on obese patients with food addiction. Methods: Sixty-five eligible patients were randomized into a thread-embedding group of 33 cases and a diet group of 32 cases to respectively receive 12-week treatment. Before treatment, after treatment and at 6-month follow-up, the two groups were observed and compared in terms of body mass (BM), waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), body mass index (BMI), body fat rate (BFR), basal metabolic rate (BMR) and Yale food addiction scale version 2.0 (YFAS 2.0). Results: At the end of treatment, there were no significant differences in the general efficacy, and the improvements in BM, BMI, WC, HC, WHR and BFR between the thread-embedding group and diet group (all P>0.05). At follow-up, the thread-embedding group showed more significant improvements in all the aforementioned indicators compared with the diet group except HC (all P<0.05). At the end of treatment and follow-up, BMR and YFSA 2.0 had more significant improvements in the thread-embedding group than in the diet group (all P<0.05). Conclusion: Acupoint thread-embedding therapy can produce significant efficacy in treating obese patients with food addiction; it can improve the food addiction state and work better in maintaining the efficacy compared with low-carbohydrate diet therapy.

Recent data have revealed that inhibiting autophagy exacerbates lipid accumulation in hepatocytes and nitrite treatment reduces total triglyceride levels in the high-fat diet mice. Therefore, the present study aimed to determine the effects of nitrite on simple hepatic steatosis and the possible role of autophagy. Firstly, steatotic L-02 cells were induced by incubating L-02 cells with 1.2 mmol · L(-1) oleic acid (OA) for 24 h. Secondly, steatotic L-02 cells were treated with 0.2 mmol · L(-1) sodium nitrite (SN) plus 3-methyladenine (3-MA), or chloroquine (CQ) for 24 h, and then lipid accumulation was measured with oil red O staining and triglyceride quantification. The notable steatosis could be observed in L-02 cells following exposure to 1.2 mmol · L(-1) OA for 24 h. Treatment with 0.2 mmol · L(-1) sodium nitrite reduced lipid accumulation in steatotic L-02 cells. 3-MA weakened the ability of sodium nitrite to ameliorate hepatic steatosis. Additionally, the sodium nitrite increased number of LC3-II immunostaining puncta and LC3-II protein expression was confirmed by immunofluorescence or Western blot analysis, and the effects were enhanced by CQ treatment. The number of increased cytoplasm vacuoles and lysosomes increased was confirmed by phase contrast and fluorescence microscope respectively. The increased autolysosome was detected by electron microscopy, this phenomenon could be reversed by CQ treatment. These data demonstrated that sodium nitrite enhanced the autophagic flux and decomposition of triglycerides in steatotic L-02 cells.
Adenine ; analogs & derivatives ; Autophagy ; Blotting, Western ; Cells, Cultured ; Chloroquine ; Cytoplasm ; Fatty Liver ; Hepatocytes ; drug effects ; Humans ; Lipid Metabolism ; drug effects ; Microscopy, Fluorescence ; Microtubule-Associated Proteins ; metabolism ; Oleic Acid ; Sodium Nitrite ; pharmacology ; Triglycerides

AIM: To establish an animal model of experimental autoimmune myocarditis (EAM) in BALB/c mice and to investigate the expression and significance of Toll-like receptor 3 in mouse EAM. METHODS: BALB/c mice were immunized with cardiac myosin extracted from porcine ventricular myocardium covered by complete freund's adjuvant (CFA) on 0 d and 7 d, then divided into immunized with CFA only. Serum and myocardium samples were collected at 14 d and 21 d after the first immunization. HE staining was used to identify the areas of inflammation. The myosin IgG antibody was examined by indirect ELISA assay. The changes of TLR3 protein and mRNA expression in myocardial tissue were measured by immunohistochemistry and real time-PCR. RESULTS: Compared to control group, immunohistochemistry results showed that there was positive expression of TLR3 in the myocardium of mice with EAM and the mRNA of TLR3 were more than 20 times (P<0.05). The expression of interferon beta mRNA in EAM group was more than 14 times as many as basal expression, that of tumor necrosis factor alpha was more than 18 times (P<0.05). CONCLUSION: The expression of Toll-like receptor 3 in myocardium is up-regulated in experimental autoimmune myocarditis. The inflammatory response to cardiac myosin may associate with the TLR3 signal transduction pathway.

OBJECTIVETo investigate the effects of sodium metavanadate (SMV) on blood sugar and glucose phosphorylation in mice, and to discuss the possible mechanism of its hypoglycemic effects.

METHODSDiabetic mice (D) and control mice (V) were randomly allocated to drink SMV (0.2 mg/ml) (CV and DV groups) or NaCl (80 mmol/L) (C and V groups) respectively. The study lasted for 5 weeks. Liver glucokinase, muscle hexokinase, blood glucose and insulin were assayed at the end of each week.

RESULTSBlood glucose was higher in the diabetic groups before the administration of SMV, and the blood glucose level of group DV decreased from (18.77 +/- 1.28) to (8.94 +/- 0.94) mmol/L (P < 0.01) after oral administration of SMV for one week. While liver glucokinase increased from (1.29 +/- 0.64) to (15.36 +/- 1.57) mIU/min/mg protein and muscle hexokinase increased from (1.93 +/- 0.50) to (18.62 +/- 1.71) mIU/min/mg protein (P < 0.01) respectively. There was no continuous change of these parameters during the later weeks. No significant change of serum insulin was observed in the diabetic mice. There was a remarkable negative correlation of blood glucose level with liver glucokinase and muscle hexokinase levels.

CONCLUSIONThe hypoglycemic effects of SMV was independent of insulin level. In consideration of the close relations of the activities of liver glucokinase and muscle hexokinase with diabetes, and the improving of impaired glucose phosphorylation in diabetic mice by oral sodium metavanadate, which might be the mechanism of hypoglycemic effects of SMV.

Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; blood ; Female ; Glucokinase ; metabolism ; Hexokinase ; metabolism ; Hypoglycemic Agents ; pharmacology ; Insulin ; blood ; Liver ; metabolism ; Mice ; Mice, Inbred ICR ; Muscle, Skeletal ; metabolism ; Phosphorylation ; Random Allocation ; Vanadates ; pharmacology

OBJECTIVETo analyze the prevalence of asthma and asthma related symptoms among children aged 0-14 years in three Chinese cities and to obtain a crude estimation of the trend of childhood asthma prevalence in China.

METHODSA cross-sectional, population-based survey of prevalence of asthma was conducted in children aged from 0 to 14 years in 3 major cities of China (Beijing, Chongqing, and Guangzhou) with different geographic locations. All the subjects were randomly selected by a multi-stage sampling method. Three to five schools and kindergartens in 2 urban districts in each city were randomly selected for the survey, and a validated questionnaire that included the core questions of the International Study of Asthma and Allergies in Childhood, Phase III questionnaire and several additional questions were used. All questionnaires were completed by parents or guardians of the selected children. Children whose parents responded affirmatively to the question "Has your child ever been diagnosed as asthma by a doctor" were recognized as victims of asthma.

RESULTSThe prevalence of asthma in Beijing, Chongqing, and Guangzhou was 3.15%, 7.45%, and 2.09%, respectively. These values were significantly higher than those obtained 10 years ago in the national epidemiological survey in 2000 which used the same method of investigation and the same diagnotic criteria (χ²=3.938, P=0.047; χ²=73.506, P≤0.001; χ²=11.956, P=0.001, in each city). Of the asthmatic children 57.21%, 69.91%, and 60.00% had their first attack before the age of 3 in Beijing, Chongqing, and Guangzhou, respectively. Wheezing was the primary clinical manifestation for all asthmatic children, followed by persistent cough and repeated respiratory infections. Both the prevalence of asthma and asthma-related symptoms were statistically higher in males than in females.

CONCLUSIONThe prevalence of childhood asthma is statistically higher than that 10 years ago in the three Chinese cities.

Asthma ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Prevalence

OBJECTIVETo examine the expression of nestin and neurogenin 3 (Ngn3), the markers of pancreatic stem cells, in the human fetal pancreas.

METHODSThe human fetal pancreas tissue of 12 and 14 weeks were examined for the expression of nestin and Ngn3 using the techniques of immunofluorescence dye and RT-PCR.

RESULTSBoth nestin and Ngn3 expressed widely in 12 and 14 weeks before in human fetal pancreatic tissue. In these positive cells there was no co-expressing insulin or glucagon. There were nestin and Ngn3 co-expressing cells in ducts but not in the islets. The results of RT-PCR also indicated the expression of nestin and Ngn3.

CONCLUSIONSThere was no expression of the markers of mature endocrine cells in the nestin and Ngn3 positive cells, and they were the marks of no-differentiation cells in the human fetal pancreatic tissue.

Basic Helix-Loop-Helix Transcription Factors ; biosynthesis ; genetics ; Fluoroimmunoassay ; Humans ; In Vitro Techniques ; Intermediate Filament Proteins ; biosynthesis ; genetics ; Microscopy, Fluorescence ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Nestin ; Pancreas ; cytology ; embryology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

The effects of the Smad3- knockout on the hematopoiesis of mouse were investigated in this work. Five pairs of wild type and Smad3- null mice were studied. White blood cell(WBC), red blood cell(RBC) and platelet (PLT) counting of peripheral blood cells were performed with blood obtained from tails. And white blood cells were classified by their morphology. Bone marrow nucleated cells (BMNCs) were counted and classified. The CFU-GM, BFU-E, CFU-GEMM yields were measured in each pair of mice. CFU-S yield of each mouse was measured by injecting bone marrow cells into lethally irradiated 8-10 weeks old wild type female mice. And the pathomorphism of their bone marrows, spleens and livers were observed. As a result, WBC and PLT of Smad3- null mice were significantly higher than those in wild type mice. Smad3- null mice had much more proportion of granulocytes in classification. There wasn't any difference in RBC counting and BFU-E measurement. The yield of CFU-GM increased, while the yields of CFU-GEMM and CFU-S markedly reduced. Bone marrows are actively proliferative, with granulocytosis. The granulocyte/erythrocyte ratio increased. There were no obviously alterative in spleen and liver. Thus Smad3- knockout results in a decreased number of stem and progenitor cells. Moreover hematopoietic differentiation is abnormal with a tendency to forming more granulocytes and platelets. The effect of Smad3 on hematopoiesis is correlative to that of TGF-beta.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Erythrocytes ; cytology ; metabolism ; Erythroid Precursor Cells ; cytology ; metabolism ; Female ; Granulocyte-Macrophage Progenitor Cells ; cytology ; metabolism ; Granulocytes ; cytology ; metabolism ; Hematopoiesis ; genetics ; Mice ; Mice, Knockout ; Myeloid Progenitor Cells ; cytology ; metabolism ; Smad3 Protein ; genetics

OBJECTIVEIn recent years, the incidence of allergic diseases in China is increasing. With serious influence on the patients' quality of life and even life-threatening, now allergic diseases have become an important public health problem in our country. This survey aimed to obtain a general epidemic outline of children's major allergic diseases, i.e., childhood asthma, allergic rhinitis and eczema in the downtown of Beijing, Chongqing and Guangzhou.

METHODChildhood asthma questionnaire was established by Delphi method based on guidelines for childhood asthma diagnosis and prevention made by Chinese medical association in 2008 with reference to the International Study of Asthma and Allergies in Childhood questionnaire (ISAAC). This study was carried out from Oct. 2008-Mar. 2009, in Beijing, Chongqing and Guangzhou by a multi-stage sampling method, covering a sample of 24 290 children aged 0 - 14 years by a multi-stage sampling method, the population of children surveyed in the downtown of Beijing Chongqing and Guangzhou was 10 372, 9874 and 4072 respectively; of whom males were 5545, 5258, 2159, and females were 4917, 4588, 1877; the ratio of males to females in the three cities were 1.11:1, 1.15:1, 1.15:1; the average age of surveyed children were 7.63, 8.74 and 8.30 years old. All the questionnaire was filled in by the parents of children or other guardian of children.

RESULTThe self-reported prevalence of asthma in Beijing, Chongqing and Guangzhou was 3.15%, 7.45%, 2.09%, the self-reported prevalence of allergic rhinitis was 14.46%, 20.42%, 7.83%, and the prevalence of eczema was 20.64%, 10.02%, 7.22%. In the survey, asthma combined with allergic rhinitis had the highest rate, 49.54%, 50.14%, 34.83%, respectively in the 3 cities. The self-reported prevalence of asthma and allergic rhinitis was significantly higher in male than female. Besides, the self-reported prevalences of asthma and allergic rhinitis were relatively lower in children under 1 year and over 10 years of age.

CONCLUSIONThe prevalence of asthma, allergic rhinitis and eczema and other allergic diseases in children 0 - 14 years of age in the three cities gradually increased, and the prevalence of allergic diseases in Chongqing was relatively higher.

Adolescent ; Asthma ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Cities ; Eczema ; epidemiology ; Female ; Humans ; Hypersensitivity ; epidemiology ; Incidence ; Infant ; Male ; Prevalence ; Surveys and Questionnaires

BACKGROUNDValproic acid (VPA) improves early survival and organ function in a highly lethal poly-trauma and hemorrhagic shock model or other severe insults. We assessed whether VPA could improve organ function in a rat model of septic shock and illustrated the possible mechanisms.

METHODSForty Sprague-Dawley rats were randomly assigned to four groups (n = 10): control group, VPA group, LPS group, and LPS + VPA group. Lipopolysaccharide (LPS) (10 mg/kg) was injected intravenously to replicate the experimental model of septic shock. Rats were treated with VPA (300 mg/kg, i.v.) or saline. Six hours after LPS injection, blood was sampled for gas analysis, measurement of serum alanine aminotransferase, aspartate aminotransferase, urine nitrogen, creatinine and tumor necrosis factor-alpha. Lung, liver and kidney were collected for histopathological assessment. In addition, myeloperoxidase activity and tumor necrosis factor-a in pulmonary tissue were measured. Acetylation of histone H3 in lung was also evaluated by Western blotting.

RESULTSLPS resulted in a significant decrease in PaO2, which was increased by VPA administration followed LPS injection. In addition, LPS also induced an increase in the serum levels of alanine aminotransferase, aspartate aminotransferase, urine nitrogen, creatinine, and tumor necrosis factor-alpha. However, these increases were attenuated in the LPS + VPA group. The lungs, liver and kidneys from the LPS group were significantly damaged compared with the control group. However, the damage was attenuated in the LPS + VPA group. Myeloperoxidase activity and tumor necrosis factor-alpha levels in pulmonary tissue increased significantly in the LPS group compared with the control group. These increases were significantly inhibited in the LPS + VPA group. Acetylation of histone H3 in lung tissue in the LPS group was inhibited compared with the control. However, the level of acetylation of histone H3 in the LPS + VPA group was markedly elevated in contrast to the LPS group.

CONCLUSIONSTreatment with VPA can attenuate multiple organ damage caused by LPS induced septic shock. Our data also suggest that the beneficial effects are in part due to the decrease in inflammatory cytokines and restoration of normal acetylation homeostasis.

Acute Kidney Injury ; drug therapy ; metabolism ; Animals ; Blotting, Western ; Chemical and Drug Induced Liver Injury ; drug therapy ; metabolism ; Lung Injury ; drug therapy ; metabolism ; Male ; Multiple Organ Failure ; drug therapy ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Septic ; drug therapy ; metabolism ; Valproic Acid ; therapeutic use