For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

To evaluate the regulating effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression in hippocampus tissues of rats with cerebral ischemia, SD rats were divided into the sham-operation group, the model group, the Buyang Huanwu decoction group (ig, 3.15 g · kg⁻¹) and the simple prescription group (ig, 2.41 g · kg⁻¹). Each group was further divided into five subgroups based on time points after the administration, i. e. 1 d, 3 d, 7 d, 14 d and 28 d, respectively. CDK4/Cyclin D1 expressions of the group at different time points were examined by using immunohistochemistry and real-time qPCR. According to the results, the cerebral ischemia model group showed higher CDK4/Cyclin D1 expression than the sham-operation groups (P < 0.05), suggesting that the cell cycle signal pathway would be activated by the cerebral ischemic injury. Both Buyang Huanwu decoction and simple prescription groups showed significantly lower cyclin expression than the model group at 3 d, 7 d, 14 d, 28 d (P < 0.05), indicating both Buyang Huanwu decoction and its simple prescription could play the neuroprotective effect through the cell cycle signal pathway.
Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Cycle ; drug effects ; Cyclin D1 ; genetics ; metabolism ; Cyclin-Dependent Kinase 4 ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley

Objective To determine if Shenfu injectio(SFI)has any analeptic action in patients emerging from general anesthesia.Methods Eighty-six ASAⅠorⅡadult patients undergoing elective abdominal surgery under general anesthesia were randomly divided into 2 groups(n=43 each):SFI group and control group.The patients were premedicated with intramuscular phenobarbital 0.1g and atropine 0.5mg.Anesthesia was induced with propofol 2 mg?kg~(-1),fentanyl 4-5?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with propofol infusion(2-4 mg?kg~(-1)?h~(-1)),0.5%-1.0% isoflurane inhalation and intermittent i.v.boluses of fentanyl and vecuronium.The patients were intubated and mechanically ventilated.The propofol infusion and isoflurane inhalation were stopped during skin closure.The patients were still unconscious and on mechanical ventilation at the end of surgery and transferred to PACU with a tube in trachea.As soon as the patients reached the PACU,SFI 1 ml?kg~(-1) in Ringer's solution 100 ml was infused over 10 min.In control group the patients received Ringer's solution 100 ml without SFI.The following times were recorded:(1)the time when the patients opened their eyes on command;(2)the time when mechanical ventilation was stopped;(3)the time when oxygen inhalation was stopped;(4)the extubation time;(5)the time of staying in PACU.Venous blood samples were taken before(T_0) and 5,15 and 45 min(T_(1,2,3))after SFI infusion for determination of plasma?-endorphin concentration.Results The awakening time,the mechanical ventilation time,oxygen inhalation time,extubation time and duration of PACU stay were significantly shorter in SFI group than in control group.There were no significant differences in MAP and HR after SFI between the two groups.The plasma?-endorphin concentration was significantly higher in group SFI than in control group.Conclusion Shenfu injectio can make patients emerging from general anesthesia faster.

Objective To explore the relationship of vascular endothelial growth factor(VEGF),IFN-? and IL-4 and effect of Bude-sonide on their in asthmatic rats.Methods Thirty-six male SD rats were randomly divided into 3 groups:asthma group,Budesonide-treatment group and control group.On the first day of the experiment and the 8th day,the rat models of the asthma group and Budesonid treatment group were allergized by the OVA/Al(OH)3 through intraperitoneal injection,respectively.And starting from the 15th day,they were challenged by the OVA through atomization for 2 weeks.Control group was allergized and challenged by NS atomization.Budesonid treatment group was interfered in Budesonide inhalation before suscitation in 0.5 h.After 12 h the same inhal done was again in Budesonide group.Twenty-four hours after the last challenge,the rats in 3 groups were sacrificed,and blood and bronchoalveolar lavage fluid(BALF) were collected.The concentrations of IL-4,IFN-? and VEGF in serum and BALF were measured by enzyme-linked immunosorbent assay.Results The concentrations of IL-4 in serum and BALF in asthma group and Budesonide treatment group were significantly increased than those in control group(P

Objective To investigate whether the left ventricular delayed contraction site determined by tissue Doppler imaging might be an optimal left ventricular lead position for improved outcomes of cardiac resynchronization therapy (CRT) in patients with non-ischemic cardiomyopathy. Methods Thirty-three patients subjected to CRT were selected, and all were performed conventional ultrasound cardiography and tissue Doppler examinations before operation. The left ventricular delayed contraction site was determined according to the interval between the onset of QRS and the peak systolic velocity. Retrograde coronary venography was performed during operation, and the left ventricular lead site was selected according to the left ventricular delayed contraction site determined by tissue Doppler examination before operation. The coronary sinus lead site was determined under the guidance of X ray of dorsaventral, lateral, right anterior oblique and left anterior oblique positions. Patients were divided into group A(n=20, the left ventricular lead site was in line with the delayed contraction site) and group B (n=13, the left ventricular lead site was not in line with the delayed contraction site). Results There was no significant difference in age, NYHA grading, left ventricular end-systolic volume(LVESV), left ventricular ejection fraction(LVEF), pulmonary arterial systolic pressure, QRS width and Ts-SD between the two groups before operation(P> 0.05). Six months after CRT, there was no significant difference in NYHA grading, LVESV and mitral regurgitation(MR) grading between the two groups(P>0.05), while the increase in LVEF and decrease in LVESV of group A were more significant than those of group B (P<0.01). Conclusion In patients with non-ischemic cardiomyopathy, CRT significantly improves left ventricular performance, and the more favourable outcomes are achieved in those pace at the delayed contraction site. Tissue Doppler imaging may help to guide the implant of left ventricular lead.

Flow cytometry was previously applied for analysis of Rh(D) antigen density, therefore it was suggested that the flow cytometry may be used for routine detection of weak D positive phenotypes. This study was purposed to evaluate its practicability. Six weak D positive and 7 DEL individuals were detected by using saline, IAT and absorption/elution test from 2010 to 2011 years. By RHD genotyping, zygosity analysis and sequencing, 3 cases of weak D type 15, 3 cases of partial D type DVI-III and 7 cases of DEL carrying RHD1227A alleles were identified. Taking 2 normal Rh(D)-positive and 2 D-negative samples as controls, all the samples were tested by using flow cytometry, and the median fluorescence intensities were observed as well. The results indicated that all weak D type 15 and partial D type DVI samples were detected to be positive by flow cytometry, as compared with 2 Rh(D)-negative samples (P < 0.05). Seven 7 DEL samples were tested to be negative (P > 0.05), although one of 7 DEL was tested as "±" in IAT and strong positive in absorption/elution. The RHD zygosity analysis showed this DEL individual as RHD(+)/RHD(+) homozygote. It is concluded that the sensitivity of detecting D antigen by flow cytometry is similar to that of IAT, but lower than absorption/elution test. As for detecting weak D or partial D antigens, IAT is easier than flow cytometry; as for identifying DEL, the flow cytometry is not sensitive enough.
Adult ; Alleles ; Blood Donors ; Blood Grouping and Crossmatching ; Flow Cytometry ; methods ; Genotype ; Humans ; Phenotype ; Rh-Hr Blood-Group System ; blood ; genetics ; immunology

The purpose of the present study was to investigate the effect of 17beta-estradiol (17beta-E(2)) on the structure and relaxation and contraction activity of thoracic aortas in ovariectomized rats with insulin resistance induced by fructose. Ovariectomized mature female Sprague-Dawley rats were fed with high fructose diet for 8 weeks to induce insulin resistance. Physiological dose of 17beta-E(2) (30 mug/kg) was injected subcutaneously every day for 8 weeks. Systolic blood pressure (SBP) was measured by use of tail-cuff. Serum nitric oxide (NO), estradiol (E(2)), fasting blood sugar (FBS) and fasting serum insulin (FSI) were measured respectively in each group. The insulin sensitive index (ISI) was calculated. The thoracic aortas were fixed in formalin, sliced and HE dyed. The structure of thoracic aortas, lumen breadth, media thickness, media thickness/lumen breadth ratio and media cross-section area were measured. The contraction response of thoracic aorta rings induced by L-phenylephrine (PE) and the relaxation response of thoracic aorta rings induced by ACh and sodium nitroprusside (SNP) were measured. To explore the mechanism, nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methyl ester (L-NAME) was used. The results obtained are as follows: (1) 17beta-E(2) protected against the effect of high fructose diet, which caused an increase in SBP, hyperinsulinemia and a decrease in ISI in ovariectomized rats. (2) The structure of thoracic aortas had no significant difference among the groups. (3) Compared with the ovariectomized group (OVX) or fructose fed group (F), serum nitric oxide was significantly reduced, the contraction response of thoracic aorta rings to PE was enhanced and the relaxation response to ACh was depressed significantly in ovariectomized+fructose fed group (OVX+F). The effect of high fructose was reversed by 17beta-E(2). After pretreatment with L-NAME, the effect of 17beta-E(2), which enhanced the relaxation response of thoracic aorta rings to ACh in ovariectomized+fructose+17beta-E(2) group (OVX+F+E(2)), was partly blocked. (4) The relaxation response of thoracic aorta rings to SNP had no significant difference among the groups. (5) The contraction response of thoracic aorta rings without endothelium to PE had no significant difference among the groups. These findings suggest that 17beta-E(2) may provide protection against the effect of high fructose diet, which causes hypertension, dysfunction of endothelial cells and insulin resistance. The mechanism of this effect of 17beta-E(2) could be partly associated with the increase of NO by NOS pathway, or associated with the decrease in the level of systolic blood pressure and serum insulin, and the improvement of insulin resistance.
Animals ; Aorta ; physiology ; Estradiol ; pharmacology ; Female ; Fructose ; Insulin Resistance ; physiology ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Vasoconstriction ; drug effects ; Vasodilation ; drug effects ; Vasomotor System ; drug effects

OBJECTIVETo investigate the mechanism of norcantharidin (NCTD)-induced SMMC-7721 hepatoma cell apoptosis.

METHODSSMMC-7721 cell growth inhibition was measured by the MTT method. Apoptosis was detected by Annexin V/propidium iodide staining. The mitochondrial membrane potential was measured by flow cytometry. Western blot analysis was used to evaluate the level of cytochrome c, caspase-3, AIF, Bcl-2 and Bax expression.

RESULTSNCTD inhibited SMMC-7721 cell growth in a time- and dose-dependent manner. The cells treated with NCTD showed the loss of mitochondrial membrane potential. The activities of caspase-3, cytochrome c, AIF, and Bax were up-regulated after NCTD treatment at different doses. The expression of Bcl-2 was decreased after treatment with NCTD.

CONCLUSIONSNCTD could induce SMMC-7721 cell apoptosis. The activation of the mitochondrial pathway was involved in the process of NCTD-induced SMMC-7721 cell apoptosis.

Apoptosis ; drug effects ; Apoptosis Inducing Factor ; metabolism ; Blotting, Western ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cytochromes c ; metabolism ; Flow Cytometry ; Humans ; Liver Neoplasms ; enzymology ; metabolism ; pathology ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the cognition and health education needs on child neglect among parents/carers to the preschool children in Guangdong Province.

METHODSA total of 621 parents/carers of preschool children aged 3-6 years from four cities (Dongguan, Huizhou, Zhongshan, and Foshan) in Guangdong Province completed self-designed questionnaires concerning the cognition and health education needs on child neglects.

RESULTSOnly 5.4% of the parents/carers had a good knowledge of child neglect, and 55.9% had never heard about this term. About 90.5%-90.7%of the surveyed parents/carers were willing to learn more about child neglect and 94.6%-97.4% of them agreed that education on child neglect was necessary. It was found that normal brochures, books, newspapers, and magazines were the preferred way of education for parents/carers (77.0%); also nearly half of the parents/carers welcomed courses (55.6%) or lectures (49.1%). Most of them believed that awareness raising activities should be carried out in kindergarten (70.5%), public places (61.4%) or by providing collective training for teachers in kindergarten (59.6%).

CONCLUSIONSMost parents/carers of preschool children in Guangdong Province have a poor knowledge of child neglect. Efforts should be made to strengthen awareness raising activities on this topic via appropriate pathways.

Caregivers ; Child Abuse ; Child, Preschool ; Cognition ; Cross-Sectional Studies ; Health Education ; Humans ; Parents

OBJECTIVETo formulate the standard measuring tool for the evaluations on fire-heat syndrome in oral cavity by means of Chinese medicine (CM).

METHODSThe measuring scale for fire-heat syndrome in the oral cavity by means of CM was investigated by symptom collection, item pool formulation, item selection, pre-investigation, evaluations on the reliability, validity and reactivity of the measuring scale, according to the principles for measuring scale design and under the guidance of CM theories.

RESULTSThe measuring scale was composed of two integrative parts: the self-filling section and the interview section. As far as the reliability was concerned, the total Cronbach α coefficient of the measuring scale was 0.866, the total test-retest reliability coefficient was 0.726 and the split-half reliability coefficient was 0.851. As far as the validity was concerned, the scores for the subjects of fire-heat syndrome in oral cavity and healthy people in their oral cavity in the items of symptoms were statistically different (P<0.01); three common divisors were extracted according to the theoretical dimensions, the accumulated contribution rate was 63.468%. As far as the reactivity was concerned, the difference between the symptom scores before and after the test in which 31 subjects used the Chinese herb toothpaste was statistically significant (P<0.01).

CONCLUSIONSThis measuring scale has relatively good reliability, validity and reactivity, and it can be used in an objective quantitative evaluation on patients suffering from fire-heat syndrome in oral cavity, and thus lay the foundations for the evaluations on the therapeutic effects of Chinese herb toothpaste on fire-heat syndrome in oral cavity.

Drugs, Chinese Herbal ; pharmacology ; Evaluation Studies as Topic ; Humans ; Mouth ; drug effects ; pathology ; Statistics as Topic ; Surveys and Questionnaires ; Syndrome ; Tooth ; drug effects ; Toothpastes ; pharmacology