Objective To investigate the influences of 4 kinds of collagenase with different degrees of activity on the survival rate,contraction and relaxation functions of isolated rats' myocardiocytes.Methods Rats' myocardiocytes were isolated by enzymolysis with 4 kinds of collagenase with different degrees of activity.The survival rate of myocardiocytes was observed immediately after isolation(D),one hour after loading with calcium(E time point) and 10 minutes after electric stimulation(F time point).The contraction and relaxation functions of myocardiocytes,including contraction amplitude(ph),the proportionality of ph/single cardiocyte length(bl),maximal velocity of contraction(+dL/dt) and maximal velocity of relaxation(-dL/dt),were measured with IonOptix video edge tracker.Results From 203U/mg to 299U/mg,with lowering of the activity of collagenase,the isolation time became longer,the survival rate of myocardiocytes declined 1 hour after loading calcium and 10 minutes after electric stimulation(P

Objective To investigate the inhibitory effect on enhancement of contraction and relaxation functions of cardiomyocytes induced by of MN9202 TEA, Forskolin, and MB. Methods The twitch amplitude was measured with a video edge tracker method. The following indexes, including ph (peak height), peak height/baseline percent (ph/bl, %), maximal velocity of contraction (+dL/dt) and maximal velocity of diastolization (-dL/dt), were recorded by a computer. Results TEA, Forskolin and MB increased electrically-induced contraction as shown by the following indexes: ph, ph/bl%, +dL/dt and -dL/dt. Forskolin and MB markedly augmented function of cardiac myocyte contraction and diastolization. Comparing with the control group, Forskolin (10-8mol/L) increased ph from 0.12?0.03(um)to 0.24?0.06 (um), ph/bl % from 12?3% to 27?6%, +dL/dt from 1.8?0.5 (um/s) to 3.8?0.9 (um/s), -dL/dt from 1.8?0.22 (um/s) to 3.5?0.7 (um/s). These indexes increased by 91%, 123%, 110% and 89%, respectively. MB also increased significantly; ph, ph/bl %, +dL/dt and -dL/dt were 0.14?0.04um, 11?4%, 2.2?0.3um/s and 2.2?0.6um/s, respectively. MN9202 (3?10~ -6mol/L) decreased the indexes (+dL/dt and -dL/dt ) which were increased by Forskolin significantly. Conclusion TEA, Forskolin and MB increased function of cardiac myocyte contraction and diastolization. These drugs might elevate intracellular calcium content indirectly and directly. The effects of TEA, Forskolin and MB were attenuated by MN9202. MN9202 might not only block Dihydropyridine Receptor, but might also inhibit calcium influx. Further study is needed to elucidate exact mechanism of MN9202.

BACKGROUND: How to choose traditional Chinese remedy to treat cerebrovascular disease, not only improving the brain blood supply but also not affecting the blood pressure and heart rate, has been a promising research.OBJECTIVE: To study the role of yangyintongnao granule on average blood pressure and heart rate in anaesthetized dog.DESIGN: Complete grouping design and randomized controlled study based on hybrid dog.SETTING: Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University of Chinese PLA and Department of Pharmacology, Faculty of Preclinical Medicine, Fourth Military Medical University of Chinese PLA MATERIALS: The experiment had finished by Cardiovascular Laboratory of Physiology Department in the Fourth Military Medical University of Chinese PLA from March to June 2003. Totally 23 healthy hybrid dogs in either sex were selected. These dogs were divided randomly into 4 groups: high dose group of yangyintongnao( n = 8), moderate dose group of yangyintongnao ( n = 6), low dose group of yangyintongnao( n = 5) and saline group( n = 4).METHODS: Anesthetic dogs in high, moderate and low dose group of yangyintongnao granule were given different doses of yangyintongnao granule: 2 g/kg, 1 g/kg and 0. 5 g/kg respectively. All doses of drug were calculated based on the body mass of dogs, and the drugs were dissolved in 100 mL saline and given through the gastric canal. Dogs in saline group were perfused with equal saline. Aortic average blood pressure was measured by femoral artery intubation via a piezometric transducer. Heart rate was obtained from R-R intervals of a standard Ⅱ lead of electrocardiograph(ECG). The mean blood pressure and heart rate were recorded 0. 5, 1, 1.5, 2.0,3.0, 4. 0, 5.0, 6. 0 hours after treatment.MEAN OUTCOME MEASURES: The changes of blood pressure and heart rate at different time before and after medication.RESULTS: Totally 29 dogs were brought into the final analysis. Blood pressure: The mean blood pressure was reduced -5.4% to -6. 2% respectively in high dose group and moderate dose group after treatment. But in low dose group the average blood pressure sometimes increased sometimes decreased, mainly decreased. It increased by 6.6% ( P ＞ 0. 05) and decreased by -4. 1% ( P ＞ 0.05) at maximum. The average blood pressure in saline group changed by - 9.6% ( P ＞ 0.05). Heart rate: The heart rate in high and medium dose group gradually reduced as time went on. It reduced by - 4. 4%, - 12.2% and - 9.5% respectively in high, moderate and low doses group. The change in each group was not statistically significant as the same in saline group( P ＞ 0.05).CONCLUSION: Yangyintongnao granule has no significant influence on average blood pressure and heart rate.

Objective To compare the efficacy and survival of patients with malignant obstructive jaundice using either endoscopic self-expandable metallic stents or surgery,and to evaluate the compounding factors influencing prognosis.Methods 56 patients with malignant obstructive jaundice treated with endoscopic self-expandable metallic stents (the endoscopic group) were compared with 90 patients who received surgery (the surgery group) during the same study period.Clinical data and survival of the 2 groups of patients were retrospectively analyzed.Results The success rate was 100％ in the endoscopic group.The serum bilirubin,alkaline phosphatase (ALP) and γ-glutamyl transferase (γ-GT) decreased significantly by using either therapeutic endoscopy or surgery (P＜0.01).There was no significant difference between the two groups in the reduction of serum total bilirubin.The mean survival of the endoscopic and surgery groups were 340 d and 795 d respectively.The accumulative survivals of the endoscopic group at 3,6 and 12 months as evaluated by the Kaplan-Meier method were 82.6 ％,61.1％ and 46.6 ％,respectively,and for the surgery group were 97.0％,90.9 ％ and 65.4％ respectively. There was a significant difference in survival between the two groups (P＜0.01).Survival after therapeutic endoscopy was similar to surgery for patients with metastasis and hilar biliary obstruction.Conclusions Self-expandable metallic stents gave similar palliation in the relief of jaundice in patients with malignant biliary obstruction.The stents had no effect on the primary tumor.Therapeutic endoscopy with self-expandable metallic stents is a safe and effective method for the relief of jaundice in patients with obstructive jaundice caused by non-resectable malignant tumors.

Objective To compare the operation modes of AP-960 full automated ELISA system.Methods Two models were tested: beginning with adding specimens(model 1) and beginning with adding enzymes(model 2).The following factors were compared: the consumption of suckers,usage of time,result of test and the usual alarm during the test.Results The consumptions of suckers were 564 and 84 respectively.The needing time of the former mode was 125 minutes and that of the latter mode was 90 minutes.Conclusion The mode beginning with adding enzymes(model 2) is better for the current status of our clinical laboratory.

Objective To screen the optimal dose of Tangtong Formula in vitro. Methods RSC96 Schwann Cells were cultivate by DMEM mediums which contains different concentrations of glucose (5–125 mmol/L). The prevention effects of Tangtong Formula at different concentrations (0.25, 0.5, 0.75, 1.0, 1.5, 2.0, 3.0, 5.0 mg/mL) on the proliferation of RSC96 Schwann Cells induced by high glucose were detected. After the RSC96 Schwann Cells were cultivated in 100 mmol/L and 125 mmol/L high glucose mediums for 72 h, the apoptosis of RSC96 Schwann Cells was detected by flow cytometry Annexin V/PI, and the apoptosis rate was calculate; the proliferation situation of RSC96 Schwann cells in different times was detected by CCK-8 method. Results RSC96 Schwann cells were in apoptosis after being intervened by 100 mmol/L and 125 mmol/L high glucose mediums. The apoptosis rates were respectively(7.46±0.96)％ and(16.53±1.01)％, with statistical significance compared with control group (P<0.01). Different concentrations of Tangtong Formula could alleviate the inhibitory effect of high glucose on the proliferation of RSC96 Schwann cells, and the threshold concentration of Tangtong Formula in 24 h was 0.25 mg/mL. The concentrations of Tangtong Formula in 0.5 mg/mL, 1.0 mg/mL, and 1.5 mg/mL could inhibit the apoptosis of RSC96 Schwann cells induced by high glucos, compared with 125 mmol/L high glucose group, the apoptosis rate of RSC96 Schwann cells decreased significantly (P<0.05, P<0.01). Conclusion Among three different doses, when the dose of Tangtong Formula is in 1.5 mg/mL, the effects on inhibiting apoptosis are the best.

Purpose:To Explore the requirements and accuracy for applying Microwave-ELISA Two-Steps-Test to the factory physical examination.Methods:Do the Microwave-ELISA Two-Steps-Test for 1,128 factory physical examination specimens that need the HBsAg and HBeAg inspection;comparing the result to the conventional ELISA test;and retest the specimens with the HBsAg-positive results.Results:There are 119 HBsAg-positive and 32 HBeAg-positive results for the ELISA test;there are 122 HBsAg-positive and 34 HBeAg-positive results for the microwave test;all the HBsAg-positive results are proved to be positive by using the colloid gold HbsAg rapid testing paper.Conclusion:Microwave-ELISA Two-Steps-Test is timesaving,simple and high sensitive for testing Hepatitis-B signs,which can reduce the hook phenomenon and can be used to do the large number of HBsAg and HBeAg checks in factory physical examinations.It can hasten the result report and greatly improve the efficiency.

Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of heme oxygenase-1 ( HO-1 ) in THP-1 cells and its possible mechanism .Methods Human monocyte cells THP-1 were cultured in vitro and then were incubated with various concentrations (0, 0.01, 0.1, 1.0 or 5.0 ng/ml) of MALP-2 for 16 h, or were stimulated by 5.0 ng/ml MALP-2 for different length of time (0 h, 4 h, 8 h, 12 h, 16 h or 24 h).The expression of HO-1 at mRNA and protein levels were detected by real-time PCR analysis and Western blot assay .The enzyme activity of HO-1 was detected by colorimetric analysis.THP-1 cells were pre-incubated with 30 μmol/L of SB203580, PD98059 and SP600125 for 30 min and then were cultured with 5.0 ng/ml MALP-2 for 16 h to investigate the role of mito-gen-activated protein kinases (MAPKs) signaling pathway in HO-1 production.After incubating THP-1 cells with 5.0 ng/ml MALP-2 for different periods of time, NF-E2-related factor 2 (Nrf2) protein was detected by Western blot assay to study the effects of Nrf2 pathway on MALP-2-induced HO-1 expression.Nrf2 and HO-1 proteins were measured by Western blot assay after transfecting THP-1 cells (1×106/well) with Nrf2 siRNA at a final concentration of 100 nmol/L.Results MALP-2 enhanced the expression of HO-1 at mRNA and protein levels as well as the enzyme activity of HO-1 in THP-1 cells in a concentration-dependent manner.The expression of HO-1 protein induced by MALP-2 was significantly inhibited by 30 μmol/L MAPKs specific inhibitors ( SB203580 , PD98059 and SP600125 ) .MALP-2 induced Nrf2 translocation at a concentration of 5.0 ng/ml.The expression of Nrf2 and HO-1 proteins were significantly decreased in Nrf 2 siRNA-transfected THP-1 cells.Conclusion MAPKs and Nrf2 signaling pathways were involved in the MALP-2 induced HO-1 expression .

Objective:To observe the molecular mechanism involved in expression of hemeoxygenase -1 (HO-1) induced by a macrophage-activating lipopeptide-2 (MALP-2).Methods:THP-1 cells were cultured in vitro and stimulated by MALP-2 for 12 h, expression of HO-1 was detected by Western blot .TLR2 and TLR6 neutralizing antibodies incubation , dominant negative plasmids transfection were used to assess the functional of TLR 2,6 in mediating HO-1 expression.Phosphorylation of c-Src and Akt were detec-ted by Western blot, and c-Src siRNA and PI3K inhibitor LY294002 were used to investigate the role of c-Src and PI3K in HO-1 ex-pression.Results:MALP-2 induced c-Src phosphorylation , and TLR2 and TLR6 neutralizing antibodies , or their dominant negatively plasmids could abrogate this effect .In addition, siRNA of c-Src could decrease the phosphorylation level of Akt , and the PI3K inhibi-tor could inhibit HO-1 expression.Conclusion: MALP-2 can induce THP-1 cells expression of HO-1 through TLR2,6/c-Src/PI3K pathways .