Objective To construct an expression vector containing esp1 in order to investigate the effects on the chromatid separation. Methods A conditional plasmid vector containing esp1 was constructed, and was transfected into spc cells. The expression level of esp1 was detected by fluorescence microscopy, and the chromosomes were observed. Results EGFP-N1-esp1 was expressed in spc cells, and improper chromosome segregation was reduced. The plasmid of pTRE-d2EGFP-esp1 was constructed successfully. Conclusion Up-regulation of esp1 has inhibitory effects on the abnormal chromatid separation of tumors and may be potential in the treatment of cancer.

Adult ; Aged ; Antihypertensive Agents ; therapeutic use ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypertension ; drug therapy ; Male ; Middle Aged ; Phytotherapy

Objective to develop a rotation monitoring device to monitor animal rotation during experiments.Methods Use serial port controls,USB and rotary encoder et al.modern computer cience,with the combination of traditionally experimental methods and the software designed in VC language.Results A mini animal rotation monitor was developed,which could continuously monitor animal rotation and intelligently display the rotaion.conclusion Statistic analysis of experimental data proofs the rationality and effectiveness of the apparatus.The rotational behavior monitor can play an importance role in research into the pathogenesis of Parkinson's Disease(PD)and screening the anti-PD drugs.

Objective To explore the function of the baseline model for end-stage liver disease (MELD) scores,MELD-Na scores and iMELD scores in short-term prognosis in the initial treatment of hepatitis B virus (HBV) related acute-on-chronic liver failure (ACLF) patients.Methods 232 HBV-related ACLF patients who received initial treatment in 302 Military Hospital of China from January 2011 to January 2013 were enrolled in this prospective clinical follow-up.The relationship between the baseline MELD scores,MELD-Na scores,iMELD scores and clinical outcomes were analyzed,and the value of these three models for short term prognosis was assessed.Results Finally the 12-week clinical follow-up was completed in 191 patients,with the completion rate of 82.33％.Eighty-five patients died,with the fatality rate of 44.50％.Compared with the survival group,in non-survival group,the baseline of MELD scores (26.65 ± 7.75 vs.21.19 ± 5.42,t=-5.720,P=0.000),MELD-Na scores (29.16 ± 11.35 vs.21.72 ± 6.33,t=-5.729,P=0.000),iMELD scores (47.19 ± 10.96 vs.38.02 ±7.01,t=-7.011,P=0.000),total bilirubin [TBil (μmol/L):374.3 ± 150.1 vs.305.5 ± 147.1,t=-3.182,P=0.002],creatinine [Cr (μmol/L):110.7 ±90.1 vs.71.1 ± 35.1,t=-4.157,P=0.000] and international normalized ratio (INR:2.3 ± 0.9 vs.2.0 ± 0.6,t=-2.754,P=0.006) were significantly increased,but the baseline of serum Na+ (mmol/L:132.8 ± 6.1 vs.136.7 ± 5.1,t=4.861,P=0.000) was significantly lowered.It was shown by Spearman correlation analysis thai the baseline MELD scores,MELD-Na scores and iMELD scores all had positive correlation with the short-term prognosis of patients (r value was 0.398,0.404,and 0.470,respectively,all P=0.000),the baseline of serum Na+ had a negative correlation with the short-term prognosis of patients (r=-0.365,P=0.000).It was shown by receiver operating characteristic curve (ROC curve) that the cut-off scores of the baseline of MELD scores,MELD-Na scores and iMELD scores were 25.07,25.43 and 43.11 respectively,and the area under ROC curve (AUC) of the baseline of MELD scores,MELD-Na scores and iMELD scores were 0.731,0.735 and 0.773,respectively.The sensitivity of the three models was 55.3％,57.7％,63.5％,and the specificity was 84.9％,84.0％,84.9％ respectively.The value of the three models had no difference in short-term prognostic prediction.According to the respective cut-off score,the three prediction models were divided into four groups,and all of them had differences in fatality rate on the whole (x2 for MELD scores was 34.740,P=0.000; x2 for MELD-Na scores was 36.861,P=0.000; x2 for iMELD scores was 50.127,P=0.000).The mortality was elevated gradually as the equation scores increased.Conclusion The baseline of MELD scores,MELD-Na scores and iMELD scores can predict well the short-term prognosis of the initial treatment in HBV-related ACLF patients,and have relatively good clinical value for guiding therapy.

To study the effect of Gegen Qinlian decoction and its major effective components on five hepatic microsomal CYP450 isozymes in rats. The in vitro hepatic microsomal incubation technique was used to co-culture Gegen Qinlian decoction and its major effective components together with each probe substrate. HPLC-MS/MS was used to establish the analytical method for metabolites of the five isoform probe substrates of CYP450 isozymes, detect the linearity among micoromal protein concentration, incubation time and metabolite formation amount. And HPLC-MS/MS was applied to determine the formation rate (V) of corresponding metabolites (acetaminophen, 4-OH-chlorzoxazone, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone) specific probe substrates of the five isoform probe substrates of CYP450 isozymes (phenacetin, polbutamide, dextromethorphan, chlorzoxazone, testosterone), in order to determine the activity of each isozyme. The result showed good linearity among acetaminophen, 4-OH-tolbutamide, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone, satisfactory precision, stability and average recovery, suggesting the method was feasible. The optimized in vitro microsomal incubation conditions conformed to the requirements in the guideline of drug-drug interaction. Gegen Qinlian decoction showed different degrees of inhibitor effect on 5 CYP450 isoforms (CYP1A2, CYP2C11, CYP2D2, CYP2E1, CYP3A1/2). Its major effective component berberine could inhibit each CYP450 isoform at high concentrations (except for CYP1A2, CYP3A1/2).
Animals ; Chromatography, High Pressure Liquid ; methods ; Cytochrome P-450 Enzyme Inhibitors ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Isoenzymes ; antagonists & inhibitors ; Liver ; enzymology ; Rats ; Tandem Mass Spectrometry ; methods

OBJECTIVETo observe the clinical effect of Huikangling Tablet (HT, extracted from Scabrous Patrinia root) on peripheral blood micrometastasis of differentiated thyroid carcinoma (DTC) patients.

METHODSTotally 87 DTC patients with positive micrometastasis were randomly assigned to the treatment group (45 cases) and the control group (42 cases). DTC endocrine inhibition treatment standards were executed in all patients. They all took levothyroxine sodium (50 microg/tablet, from low dose, 25 microg each time, once per day, 0.5 h before breakfast), and its dosage was gradually added one week later. The dosage was adjusted according to tested results of TSH combined recurrence risk stratification and endocrine suppression induced adverse reactions risk stratification. Patients in the treatment group took HT (0.4 g per tablet, 3 tablets each time, three times per day for a total of 12 weeks) combined TSH suppression therapy, while those in the control group only received TSH suppression therapy. Peripheral micrometastatic cytokeratin 19 (CK19) and polymorphic epithelial mucin1 (MUC1) were detected by FCM at week 4 and 12. Meanwhile, distant metastasis and adverse reactions were observed.

RESULTSAfter 4-week treatment positive micrometastasis was shown in 18 cases (40%) of the treatment group and 29 cases (69%) in the control group with statistical difference (chi2 = 5.68, P < 0.05). After 12-week treatment positive micrometastasis was shown in 7 cases (15.6%) of the treatment group and 17 cases (44.7%) in the control group with statistical difference (chi2 = 8.49, P < 0.01). Pulmonary metastasis occurred in 2 cases and bone metastasis in 1 case of the control group at follow-ups. Cervical lymph node metastasis without accompanied recurrence of thyroid cancer occurred in one case of the treatment group. No obvious liver or renal abnormalities occurred.

CONCLUSIONHT inhibited peripheral blood micrometastasis of DTC patients and its mechanism needed to be further studied.

Antineoplastic Agents ; pharmacology ; therapeutic use ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Neoplasm Micrometastasis ; drug therapy ; Neoplasm Recurrence, Local ; Tablets ; Thyroid Neoplasms ; drug therapy

The clinical values of acetoacetate ( AcAA ) and β hydroxybutyrate ( βHBA ) determination in classification of type 1 and2 diabetes were explored. 102 normal control subjects,33 cases of type 1 diabetes, and 104cases of type 2 diabetes were enrolled. Serum AcAA, βHBA, fasting plasma glucose ( FPG), C-peptide, and insulin levels were measured. The results showed that serum AcAA, βHBA, total ketone tody (TKB) levels in the diabetic groups were significantly higher than those of the normal group( P＜0. 01 ). AcAA, βHBA, TKB levels in type 1diabetes were higher as compared with those of type 2 diabetes( P＜0.01 ). The AcAA, βHBA, and TKB levels were negatively related with C-peptide and insulin in diabetic patients( P＜0. 01 ). All the type 1 diabetic patient were found to have TKB and lower C-peptide levels. TKB positive and lower C-peptide in type 2 diabetes were found in 47% and 26% respectively. Receiver operating characteristic (ROC) curve suggested that the area under the ROC curve of type 1 and type 2 diabetes was 0.926. The optimal operating point of the total ketone body was 0. 532 mmol/L with higher sensitivity and specificity. Enzymatic determination of acetoacetate and β hydroxybutyrate seems to have important clinical values for classification of type 1 and 2 diabetes.

OBJECTIVETo study the flavone constituents in Chrozophora sabulosa (Xinjiang origin).

METHODThe compounds were extracted with 95% ethyl alcohol, isolated by various column chromatography and identified by spectroscopic methods.

RESULTSeven flavanoids were isolated and identified as quercetin (I), kaempferol (II), apigenin (III), chrysoerid (IV), isoquercitrin (V), chrysoerin-7-O-beta-D-glucoside (VI) and quercetin-3-O-alpha-D-arabinfuranoside (VII).

CONCLUSIONAll of these seven compounds were obtained from this genus for the first time.

Apigenin ; chemistry ; isolation & purification ; Euphorbiaceae ; chemistry ; Flavones ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Quercetin ; chemistry ; isolation & purification

Administration, Inhalation ; Animals ; Female ; Male ; Pesticides ; analysis ; toxicity ; Rats ; Rats, Wistar

Objective Intercellular adhesion molecule-1 (ICAM-1) plays an important role in mediating pulmonary infiltration of neutrophils .The aim of the study was to observe the expression of ICAM-1 and its potential regulators MK 2/HuR in pulmonary micro-vascular endothelial cells ( PMVEC ) in mice with acute respiratory distress syndrome ( ARDS) induced by lipopolysaccharide ( LPS) . Methods Ten 6-8 weeks old healthy C57BL/6 mice were randomly divided into an LPS and a control group of equal number , the former injected intraperitoneally with LPS diluted in 100 μL PBS while the latter with PBS only , both at 5 mg per kg of the body weight .At 24 hours after injection , all the mice were sacrificed .Real-time PCR was used to determine the mRNA expressions of HuR and ICAM-1 in the PMVECs, Western bolt employed to detect the protein expressions of MK2, HuR and ICAM-1, and flow cytometry adopted to measure the ICAM-1 expression on the surface of the PMVECs and pulmonary infiltration of neutrophils . Results The W/D ratio in the lung tissue of the mice was significantly lower in the LPS than in the control group (3.61 ±0.28 vs 6.16 ±0.40, P<0.05), while the rate of neutrophil infiltration markedly higher in the former than in the latter ([13.92 ±3.23]%vs [3.24 ±1.24]%, P<0.05).The mRNA and protein expressions of ICAM-1 in the PMVECs were significantly elevated in the LPS group as compared with that in the control (P<0.05), and so was the mRNA expression of HuR (P<0.05).No remarkable changes were observed in the expressions of total MK 2 and HuR proteins, but phosphorylated MK2 (p-MK2) and cytoplasmic HuR were increased in the LPS-stimulated mice. Conclusion Specific blockage or reduction of the HuR expression in PMVECs may lower the expression of ICAM-1, reduce neutrophil infiltration , and lessen pathophysiological changes in mice with ARDS .