Objective To establish a method for culturing neonatal mice cardiomyocytes, and construct an in vitro model of cardiomyocytes for assessing the cardiac toxicity of chemicals. Methods Hearts of neonatal mice of 1 day old were digested with enzyme mixture of trypsin/collagenase type Ⅱ/dispase and the cell suspensions were pre-plated to flask for a short time and then seeded on coated dishes. The cultured cells were treated with 5-fiuorine-deoxy-uridine(15 μg/ml)and uridine(35 μg/ml)to enrich cardiomyocytes that were identified according to the morphology and immunocytochemistry of α-actin antigen. Cardiac myocytes were incubated with 0-64 μg/ml of a fusarium mycotoxin butenolide(BUT) for 12 h. Cell viability was then evaluated by MTT assay. Microscopic observation showed that BUT induced significant morphological changes including cellular swelling, vacuolation and breakage of muscle fibers. Results It was found that 96% of the cultured cells were cardiomyocytes and the myocytes kept beating after 90 days of culture. Concentration-dependent decreases in cell viability following exposure of cardiac myocytes to BUT were observed. Conclusion The results indicated that relatively pure primary culture of neonatal mice cardiomyocytes is successfully established. BUT possesses the potential to induce myocardial toxicity.

Objective To investigate the effect of midazolam combined with fentanyl as adjuvant therapy on inflammatory factors and biochemical indexes in children with serious hand-foot-mouth disease with mechanical ventilation .Methods One hundred and thirty children with serious hand-foot-mouth disease treated with mechanical ventilation were selected at Maternal and Child Health Care Hospital from January 2010 to January 2014 . The patients were divided into two groups according to the treatment regimen :58 cases treated with midazolam for sedation and analgesia as control group and 72 cases treated with midazolam combined with fentanyl as observation group . Inflammatory factors (interleukin-6 , interferon-γ ,high-sensitivity C-reactive protein and tumor necrosis factor-α) and biochemical indices (albumin ,alanine transaminase [ALT] ,aspartate transaminase [AST] ,alkaline phosphatase ,glutamate transpeptidase [γ-GT] and fasting blood glucose) before drug exposure and on withdrawal were compared between two groups .Adverse reactions were analyzed in the two groups .Continuous variables were compared using two-sample t-test , while categorical variables were compared using chi-square test . Results Interleukin-6 ,interferon-γ, high sensitive C reactive protein and tumor necrosis factor-α on withdrawal decreased significantly in both groups than those before drug exposure (all P< 0 .05) .All observation indices on drug withdrawal in observation group were significantly lower than control group (all P< 0 .05) .Levels of albumin ,ALT ,AST ,alkaline phosphatase and γ-GT were not significantly different between two time points in both groups (all P>0 .05) .However ,in observation group ,fasting blood glucose level decreased significantly on drug withdrawal compared with that before drug exposure ([5 .17 ± 0 .28] vs [10 .31 ± 1 .39] mmol/L ,t=46 .237 ,P=0 .000) ,and that was also lower than control group ([5 .17 ± 0 .28] vs [5 .85 ± 0 .34] mmol/L ,t=4 .372 ,P=0 .000) .Incidence of adverse reactions in observation group was significantly lower than control group (15 .3% vs 32 .8% ,χ2=4 .707 ,P=0 .030) . Conclusions Midazolam combined with fentanyl as adjuvant therapy is helpful to improve blood glucose , stabilize biochemical indices and reduce inflammation factor secretions in children with serious hand-foot-mouth disease with mechanical ventilation .This therapy is safe and worthy of clinical use .

OBJECTIVETo study the effect of deoxynivalenol (DON), a mycotoxin produced by Fusarium graminearum, on action potentials of cultured cardiomyocytes and the possible protective effects of sodium selenite.

METHODSVentricular myocytes from neonatal Wistar rats were cultured, and the transmembrane action potentials were recorded with glass microelectrodes before and after addition of DON at different concentrations. The cultured cardiomyocytes were pretreated with 0.5 mg/L selenium (as sodium selenite) to observe the protective effects of selenium against the effects of DON.

RESULTSDON at concentrations of 50, 100 and 200 mg/L decreased the action potential parameters including action potential amplitude (APA), overshoot (OS), threshold potential (TP), maximum rate of depolarization (Vmax) and action potential discharging frequency (APF), and prolonged the action potential duration of 10%, 50% and 90% repolarization (APD(10), APD(50) and APD(90)). Some of the parameters, such as APA, Vmax, APD and APF, changed in a concentration-dependent manner. The cultured cardiomyocytes pretreated with 0.5 mg/L of selenium for about 16 h presented only slight changes in action potential parameters induced by 200 mg/L DON.

CONCLUSIONSDON inhibit the membrane action potentials of cardiomyocytes, suggesting DON may interfere with the transmembrane movement of Ca(2+) and K(+), and sodium selenite may decrease the toxic effect of DON on cultured cardiomyocytes.

Action Potentials ; drug effects ; Animals ; Animals, Newborn ; Antioxidants ; pharmacology ; Cells, Cultured ; Female ; Male ; Myocytes, Cardiac ; drug effects ; physiology ; Rats ; Rats, Wistar ; Selenium ; pharmacology ; Trichothecenes ; toxicity

OBJECTIVETo investigate the changes of gene expression profile of rat liver tissue by cDNA microarrays.

METHODSTwenty Wistar rats in control group (n = 10) and isoniazid (INH) group (n = 10) were orally administrated with normal saline and 400 mg/kg INH for 14 days, respectively. The differentially expressed genes significantly correlated with liver injury were screened and analyzed. The mechanisms of liver injury caused by INH were specifically analyzed at level of gene expression based on the biological functions of those differentially expressed genes.

RESULTSThirty-seven differentially expressed genes were found in the rats administrated with INH. Among them, 25 genes were up-regulated, while the other 12 genes were down-regulated. These differentially expressed genes were functionally related to the changes of CYP450-related genes, fatty acid metabolism, and protein metabolism.

CONCLUSIONINH can cause the remarkable changes of the gene expression profiles in rat liver cells, which is important for further elucidating the mechanisms of liver injury caused by INH.

Animals ; Antitubercular Agents ; toxicity ; Chemical and Drug Induced Liver Injury ; etiology ; metabolism ; Disease Models, Animal ; Gene Expression Profiling ; Isoniazid ; toxicity ; Liver ; drug effects ; metabolism ; pathology ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Wistar ; Transcriptome

This study is to evaluate the therapeutic effect of fibroblast growth factor 21 (FGF21) on type 2 diabetic mice model and to provide mechanistic insights into its therapeutic effect. Type 2 diabetic animal model was established with high calorie fat diet and low dose streptozotocin (STZ) injection. Mice were then randomized into 5 groups: model control, FGF21 0.25 and 0.05 μmol x kg(-1) x d(-1) groups, insulin treatment group. Ten age-matched normal KM mouse administered with saline were used as normal controls. Serum glucose, insulin, lipid products and the change of serum and liver tissue inflammation factor levels between five groups of mouse were determined. The results showed that blood glucose, insulin, free fatty acids (FFAs), triglycerides, and inflammatory factor average FGF-21 of type 2 diabetes model group and normal control group were significantly higher (P < 0.01), while compared with insulin group, no difference was significant. Average blood glucose, insulin, blood lipid and inflammatory factor of FGF-21 treatment group compared with type 2 diabetes group was significantly lower (P < 0.01) and insulin group has no difference with the model control group. The results of OGTT and HOMA-IR showed that insulin resistance state was significantly relieved in a dose-dependent manner. Thus, this study demonstrates that FGF-21 significantly remits type 2 diabetic mice model's insulin resistance state and participates in the regulation of inflammatory factor levels and type 2 diabetes metabolic disorders.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetes Mellitus, Type 2 ; drug therapy ; Diet, High-Fat ; Fatty Acids, Nonesterified ; blood ; Fibroblast Growth Factors ; pharmacology ; Insulin ; blood ; Insulin Resistance ; Mice ; Streptozocin ; Triglycerides ; blood

OBJECTIVESTo investigate the influencing factors of nonalcoholic fatty liver disease (NAFLD).

METHODSA hospital-based case-control study was conducted in patients with NAFLD and controls without NAFLD in a hospital from January to August in 2007. All data were analyzed by SPSS 13.0 software.

RESULTSOne-way analysis of variance found that the two groups were significantly different in cigarette smoking, alcohol and tea comsumption, movement index, speed of food intake, frequency of social engagement, kinds of edible oil, marine products, family history of NAFLD, hypertension, higher blood sugar, abnormality of blood fat, higher level of ALT, higher level of AST, hyperuricemia, obesity, decrease of high density lipoprotein (HDL), and increase of low density lipoprotein. By non-conditional logistic stepwise regression analysis, 12 of 18 factors were used to construct a model, ten of which were the risk factors and two were protective factors of NAFLD. Risk factors included obesity (OR=6.35), hypertension(OR=3.82), dyslipidemia (OR=2.95), decrease of HDL (OR=2.85), hyperglycemia (OR=2.82), increase of ALT (OR=2.80), hyperuricemia (OR=2.35), HBsAg positive (OR=1.99), family history of fatty liver (OR=1.79) and frequently intake of marine products (OR=1.58), and protective factors included tea drinking (OR=0.72) and exercise (OR=0.90).

CONCLUSIONSThere are many influencing factors of NAFLD, and life styles are the key factors. Genetic background may also play some roles in NAFLD.

Adult ; Aged ; Alcohol Drinking ; adverse effects ; Case-Control Studies ; Cholesterol ; blood ; Fatty Liver ; blood ; epidemiology ; etiology ; prevention & control ; Feeding Behavior ; Female ; Hepatitis B ; complications ; Humans ; Hypertension ; complications ; Life Style ; Male ; Middle Aged ; Obesity ; complications ; Odds Ratio ; Regression Analysis ; Risk Factors ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo study the effect of butenolide (BUT) on cultured chondrocytes differentiation and the possible protective effects of selenium (Se).

METHODSEx-vivo cultured chondrocytes were divided into six groups: (1) Control group (without BUT and Se); (2) Se 0.1 microg/ml control group; (3) BUT 0.1 microg/ml group; (4) BUT 1.0 microg/ml group; (5) BUT 5.0 microg/ml group; and (6) BUT 1.0 microg/ml + Se 0.1 microg/ml group. The expression of collagen II (Col II), collagen X (ColX), basic fibroblast growth factor (bFGF), and parathyroid hormone-related peptide (PTHrP) in (or around) chondrocytes in all groups were analyzed by immunohistochemistry.

RESULTSThe expressions of Col II in 1.0 microg/ml BUT group and 5.0 microg/ml BUT group were significantly lower than those in the control group (P < 0.05). The expression of Col II in 1.0 microg/ml BUT + Se group were significantly higher than those in the 1.0 microg/ml BUT group and 5.0 microg/ml BUT group (P < 0.05). The expressions of bFGF and PTHrP of BUT groups were significantly higher than those in the Se and control groups (P < 0.05). No expression of ColX was observed in all groups.

CONCLUSIONBUT can affect the collagen II synthesis of the chondrocytes. Selenium supplementation may play a protective role.

4-Butyrolactone ; analogs & derivatives ; pharmacology ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Humans ; Protective Agents ; pharmacology ; Selenium ; pharmacology ; T-2 Toxin ; toxicity

OBJECTIVE: To construct the recombinant adeno-associated virus(rAAV) vector plasmid pSNAV2.0-TK containing HSV1-TK gene, to produce recombinant adeno-associated virus rAAV2/HSV1-TK, and to detect the integration and expression of HSV1-TK gene in lens epithelial cells transfected by rAAV2/HSV1-TK, and to provide foundation for gene therapy of posterior capsular opacification. METHODS: The recombinant vector plasmid constructed by gene recombinant technology was analyzed by PCR and restriction enzyme digestion. The cell strain BHK-21/TK was screened by G418 after the plasmid was transfected into BHK-21 cells,with the helper virus HSV1-rc/UL2 to produce the recombinant virus rAAV2/HSV1-TK. The purity of rAAV2/HSV1-TK was detected by SDS-PAGE and HPLC, and the titre of rAAV2/HSV1-TK was observed by dot blot hybridization. The HSV1-TK gene in lens epithelial cells transfected by rAAV2/HSV-TK was investigated by PCR and RT-PCR. RESULTS: The recombinant plasmid proved successful by PCR and restriction enzyme digestion. The recombinant virus rAAV2/HSV1-TK was produced successfully and its titre was 1 x 10(12) v.g./mL by dot blot hybridization. The HSV1-TK gene was integrated and expressed in lens epithelial cells. CONCLUSION The recombinant adeno-associated virus vector plasmid containing HSV1-TK gene is successfully constructed, and high titre recombinant adeno-associated virus (rAAV2/HSV1-TK) is obtained. The HSV1-TK gene in lens epithelial cells is expressed after being transfected by rAAV2/HSV1-TK.
Animals ; Cloning, Molecular ; Cricetinae ; Dependovirus ; genetics ; metabolism ; Epithelium, Corneal ; cytology ; metabolism ; Genetic Vectors ; Herpesvirus 1, Human ; enzymology ; genetics ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Thymidine Kinase ; biosynthesis ; genetics ; Transfection

OBJECTIVETo study the effect of different treatment period, of isoniazid (INH) on the metabonomic profile of rat urine and its relationship with traditional toxicity evaluation of blood biochemical indicators and histopathology and to explore the feasibility of metabonomics in the application of drug toxicity.

METHODSSixty male Wistar rats were orally administrated with 0, 50, 100, 200, and 400 mg x kg(-1) INH for 3, 7, and 14 days, respectively. Rat urine was then collected and its 1H nuclear magnetic resonance (NMR) spectra were acquired. All animals underwent traditional toxicity evaluation.

RESULTSHepatotoxicity was revealed by traditional toxicity evaluation in rats treated with higher dosage and longer treatment of INH. Time-response relationship existed during the treatment. Time-dependent metabonomics changes conformed with the results of traditional toxicity evaluation. The urine metabonomics showed a trajectory bias from those of the controls or pre-administration, and such bias exaggerated along with the prolongation of treatment, indicating a severer toxic injury. Along with the increase of the concentrations of urinary taurine and glucose and the decrease of the concentrations of urinary citrate and 2-oxoglutarate, the 1H NMR spectra of urine in rats treated with INH also changed.

CONCLUSIONSThe metabonomics technique can distinguish the onset and development of toxicity, which helps track and identify biomarkers. The hepatic toxicity induced by INH is related to the injury of mitochondrial function, reduction of energy metabolism in tricarboxylic acid cycle, and perturbations in the metabolism of glucose and lipid. The effect of INH on the rat urine metabonomic profile is related with INH toxicology. Therefore, metabonomics can be recognized as an ideal technique to explore and evaluate the drug toxicities.

Animals ; Antitubercular Agents ; toxicity ; Biomarkers ; chemistry ; urine ; Chemical and Drug Induced Liver Injury ; metabolism ; urine ; Citric Acid ; chemistry ; urine ; Citric Acid Cycle ; drug effects ; Glucose ; chemistry ; metabolism ; Isoniazid ; toxicity ; Ketoglutaric Acids ; chemistry ; urine ; Lipids ; Magnetic Resonance Spectroscopy ; Male ; Metabolome ; drug effects ; Mitochondria ; drug effects ; Rats ; Rats, Wistar ; Taurine ; chemistry ; urine ; Toxicity Tests ; methods

A novel rapid method for detection of the illicit beta2-agonist additives in health foods and traditional Chinese patent medicines was developed with the desorption corona beam ionization mass spectrometry (DCBI-MS) technique. The DCBI conditions including temperature and sample volume were optimized according to the resulting mass spectra intensity. Matrix effect on 9 beta2-agonists additives was not significant in the proposed rapid determination procedure. All of the 9 target molecules were detected within 1 min. Quantification was achieved based on the typical fragment ion in MS2 spectra of each analyte. The method showed good linear coefficients in the range of 1-100 mg x L(-1) for all analytes. The relative deviation values were between 14.29% and 25.13%. Ten claimed antitussive and antiasthmatic health foods and traditional Chinese patent medicines from local pharmacies were analyzed. All of them were negative with the proposed DCBI-MS method. Without tedious sample pretreatments, the developed DCBI-MS is simple, rapid and sensitive for rapid qualification and semi-quantification of the illicit beta2-agonist additives in health foods and traditional Chinese patent medicines.
Adrenergic beta-2 Receptor Agonists ; analysis ; Drugs, Chinese Herbal ; chemistry ; Food, Organic ; analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Substance Abuse Detection ; methods ; Tandem Mass Spectrometry