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 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.

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.

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 the genotypes of β-lactamases produced by Klebsiella pneumoniae.Methods Plasmid conjugation,PCR amplification,gene cloning and DNA sequencing,isoelectric focusing electrophoresis and extended-spectrum β-lactamase(ESBLs)confirmatory test were carried out for analyzing the encoding gene of β-lactamases in clinical strains of Klebsiella pneumoniae collected from hospital wards.Results Totally 75 clinical strains of Klebsiella pneumoniae were collected,in which 48 strains were confirmed to produce genotype of β-laetamases(64.0%),including 39 ESBLs-producing Btraim(52.0%).Among 48 strains,17 isolates(35.4%)carried 2 types of ESBLs genes,7(14.6%)carried 3 types of ESBL8 genes,and 5(10.4%)carried 4 types of ESBLs genes.CTX-M was the most comon type(30/48,62.5%),followed by TEM(26/48,54.2%)and SHV(25/48,52.1%).Among 9 isolates with DHA-1 AmpC β-laetamase,8 produced AmpC β-lactamases and ESBLs.Class A carbapenemase KPC-2 was produced in 3 isolates.False negative rate of ESBLs confirmatory test was 23.1%(9/39).Condusion Genotypes of β-lactamases produced by Klebsiella pneumoniae are complicated,which results in multi-drug resistance in clinic.

Objective To provide experimental evidence for the development of multi-epitope-baseded marker vaccines through investigating the humoral and cellular immune responses in BALB/c mice induced by the multiple antigen peptides (MAPs) with the mimic epitope.Methods Three types of MAPs in eight branched forms containing the mimic epitope of Mycoplasma genitalium adhesion protein (MgPa) were prepared using poly-lysine as the core matrix.The purity of MAPs was analyzed by reverse phase high performance liquid chromatography (RP-HPLC).The molecular weights of MAPs were characterized by Mass Spectrometry.The BALB/c mice were immunized intramuscularly for four times with single or mixed MAPs.The specific IgG antibody and the subtype of IgG antibody in serum of the immunized mice were detected by indirect ELISA.The proliferative responses of the spleen lymphocytes were detected using MTT assay.The ELISA were used to detect IFN-γ and IL-4 levels in the cultured supematant of spleen lymphocytes.Results The three types of MAPs containing the mimic epitopes were successfully prepared with high purity.They,could stimulate mice to produce specific IgG antibodies,of which,the major antibody isotype was Th1 immune response-associated IgG2a.Compared with the single MAP immunization group,the mixed-MAPs immunized mice produced more IgG,IgG1 and IgG2a antibody (P＜0.05).Furthermore,these MAPs could enhance the specific proliferation of spleen lymphocytes in immunized mice and induce the production of IFN-γ and IL-4.The levels of IFN-γand IL-4 in mixed-MAPs group were significantly higher than those of the single MAPs group (P＜0.01).Conclusion The three types of MAPs could induce strong specific cellular and humoral immune responses.The immunological competence of the mixed-MAPs was stronger than those of the single MAP.