A high performance liquid chromatographic method with hypersil C_(18) column, 0.020 mol/L potassium dihydrogen phosphate solution(which contains 0.25% triethylamine, pH 5.1)-methanol(20∶ 80, V/V) as mobile phase and fluorescence detection with 280 nm as excitation wavelength and 320 nm as emission wavelength has been developed for the determination of terbutaline sulfate and propranolol hydrochloride in Erythrocyte suspensions. Erythrocyte surface receptor oscillating reacted 1 h with terbutaline in 37 ℃ water bath. Free terbutaline and propranolol were separated from the membrane receptor-binding complex through 1500 r/min centrifugal in 5 min. The linear range was 0.010-3.000 mg/L in Alsever's solutions and the correlation coefficients were 0.9999 and 0.9998. The relative standard deviations(RSDs) of terbutaline and propranolol were 0.8% and 1.2%, respectively. The limits of detection(LODs) of both were 1.00 and 3.00 mg/L. This simple, safety and sensitive method was suitable for the determination of terbutaline and propranolol in erythrocyte suspensions and for the study of receptor-ligand binding.

In order to study the expression of melatonin receptors Mel1a and RORβ in the human brain,and to furthe investigate the melatonin's function in Alzheimer's disease,two melatonin receptor cDNAs were cloned and introduced into pBluescript Ⅱ (KS+)vector.The recombinant plasmids were identified by using restriction enzymes and then sequenced.The results indicated that the cloned DNA sequences and the Genbandk Sequences were inentical.The cRNA probes made from them can be used in the in situ hybridization immunohistochemistry to detect the mRNA expression level in the future study.

Objective To evaluate orthotopic splenectomy and pericardial devascularization for the treatment of portal hypertension.Methods The modified anterior approach splenectomy includes dissection of the peri-splenic vessels and ligments before division of short gastric vessels.During pericardial devascularization,the dessection panel was close to the esophagus and the stomach,leaving intact both the anterior and posterior vagus trunks.Results 63 patients underwent this modified operation.The free portal pressure decreased from (38 ±4) cmH2O to(28 ±4)cmH2O.The average blood loss was (530 ± 37)ml.There was no mortality,nor perioperative gastric paralysis and portal venous thrombosis.By 12-36 months follow-up,there was no pancreatic leakage,hepatic coma and recurrence of bleeding.Intrahepatic portal venous thromboses were detected in 4 cases at the sixth postoperative months.Conclusions This modified splenectomy plus selective pericardial devascularization carries less bleeding and is safe and effective for the treatment of portal hypertension.

Objective To investigate the clinical significance of detecting mammaglobin in sentinel lymph nodes(SLN) of breast cancer. Methods In 32 breast cancer patients, methylene blue was injected around parechyma of breast cancer to locate the SLNs, and nested RT PCR(reverse transcriptase polgmecane chain reaction) was used to examine the expression of mammaglobin mRNA. Results The SLN was successfully identified in 30 of the 32 cases(93.8%).The micrometastases detection between group SLNs and non SLNs had significant statistical difference(P

AIM: To select optimum purification of Qingluotongbi decoction (Caulis Sinomenii etc.). METHODS: Qingluotongbi decoction was refined respectively by ceramic membrane microfiltration, ultrafiltration, alcohol sedimentation, high speed centrifugalization, flocculation and macroporous resin absorption. Their refining effect was compared according to indexes of color, clarity, loss rate of sinomenine, removal ratio of impurity. RESULTS: All kinds of purification made decoction light and clear. Removal ratio of impurity by macroporous resin absorption was the highest and reached 80% or more. Removal ratio of impurity by ceramic membrane microfiltration was 21.17% , less than that by alcohol sedimentation or ultrafiltration, but more than that by flocculation and centrifugalization. Loss rate of sinomenine by AB 8 type macroporous resin absorption was the lowest and reached 6.39% , and that by ceramic membrane microfiltration was 15.31% , less than that by ultrafiltration, alcohol sedimentation, centrifugalization and flocculation. The chromatogram of decoction disposed by macroporous resin was different from that of decoction unsettled, but that disposed by ceramic membrane was similar to that of decoction unsettled. CONCLUSION: The technique of ceramic membrane microfiltration is optimum purification of Qingluotongbi decoction with the virtues of moderate removal ratio of impurity, small amount lose rate of sinomenine and simple process.

Objective To explore the changes of elasticity in the large arteries and its clinical significance of sublingual administration of nitroglycerin in patients with elevated PWV,and mild and moderate essential hypertension.Methods Totally,779 patients with mild and moderate hypertension and PWV more than 1400 cm/s were selected and divided into four groups,one with simple hypertension,the second one with only one risk factor for it,the third one with two risk factors,and the fourth one with three risk factors or damage in their target organs.Sublingual administration of 0.5 mg nitroglycerin was administered and blood pressure and PWV were measured for all of them.and data were statistically compared and analyzed by groups and before and after nitroglycerin administration.Results Systolic blood pressure decreased to(135±20)mm Hg from(149±15)mm Hg,pulse pressure decreased to(50±11)mm Hg from(59±12)mm Hg,PwV decreased to(1329±288)cm/s from(1688±378)cm/s,respectively,after nitroglycerin treatment for hypertensive patients(t=3.036,2.473,3.885,all P＜0.05).Improvement of elasticity of the large arteries in them after nitroglycerin treatment inversely correlated with the number of risk factors for hypertension they had,with 89.1 percent,74.3 percent,72.6percent and 44.0 percent of improvement in PWV,respectively for those with simple hypertension,only one risk factor.two risk factors and three risk factors or with damage in target organs.Conclusions Systolic blood pressure and pulse pressure significantly correlated with elasticity of the large arteries in hypertensive patients,and their stifiness improved after sublingual administration of nitroglycerin due to decrease in systolic blood pressure and pulse pressure.The more risk factors the hypertensive patients had,the poorer elasticity of their large arteries and less sensitive to nitroglycerin treatment were.

This article was aimed to explore the effect of acupuncture on HDAC2 activity in peripheral blood of chronic obstructive pulmonary disease (COPD) rats. The COPD rat model was established by cigarette smoking. Acupuncture was given for 3 consecutive weeks. The lung ventilation of rat was detected by equipment. Peripheral blood of rat was taken after 3 weeks. The HDAC2 enzyme activity was detected by fluorescence technology. ELISA was used in the detection of IL-10, IL-8 and TNF-αexpression in peripheral blood. The results showed that after acupuncture treatment, the lung ventilation was obviously increased compared with the model group (P < 0.05). Meanwhile, the HDAC2 activity in the peripheral blood of rat was obviously increased (P < 0.05). However, the expression of inflammatory cytokines IL-10, IL-8 and TNF-α was obviously reduced (P < 0.05). It was concluded that acupuncture treatment had a good efficacy on COPD rat. It indicated that acupuncture treatment might regulate HDAC2 activity to reduce inflammation.

BACKGROUND:Vascular endothelial growth factor is a potent angiogenesis and permeability inducible factor. Vascular endothelial growth factor 165 and vascular endothelial growth factor 121 are mainly expressed in vivo, with a strong role of angiogenesis.
OBJECTIVE:To observe the feasibility of vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cel s to differentiate into vascular endothelial cel s.
METHODS:Bone marrow derived mesenchymal stem cel s were isolated and col ected from 50 g Sprague-Dawley rats,and identified by flow cytometry. The plasmid pGLV-EF1a carrying a vascular endothelial growth factor 165 gene was transfected to the mesenchymal stem cel s using lentiviral. Expression of green fluorescent protein was observed under a fluorescence microscope.
RESULTS AND CONCLUSION:After 12 hours of transfection, expression of green fluorescent protein was observed, increased at 48 hours, peaked at 72 hours and gradual y declined thereafter. Results prove that vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cel s have the expression of green fluorescent protein, indicating successful transfection. It is feasible to induce bone marrow mesenchymal stem cel s to differentiate into vascular endothelial cel s.

Objective To investigate the effects of dexamethasone on expressions of epithelial neutrophil activating protein-78 (ENA-78) and transforming growth factor- beta 1 (TGF-β1) in neutrophil of asthma rats.Methods Thirty male SD rats were randomly divided into three groups on average, including asthma group, control group and dexamethasone treated group. In this experiment, the rat model of asthma were established by sentization and challenge with ovalbumin. Blood neutrophil were isolated and purified. The expression of ENA-78 was detected by flow cytometry. The expression of TGF-β1 was detected by immunohistochemical method in blood neutrophil and bronchial wall. Results Expression of ENA-78 in blood neutrophil in dexamethasone treated group(71.82 ±8. 87 mean fluorescence intensity)was lower than that in asthma group, but higher than that in control group(all P ＜0. 01). And expressions of TGF-β1 protein in dexamethasone treated group(0. 173 ± 0. 014,0. 202 ± 0. 019 optical density, respectively) was lower than that in asthma group(all P ＜0. 01) ,but higher than that in control group(all P ＜0. 01). There were significant positive correlation between ENA-78 expression at blood neutrophil and numbers of total inflammation cells in bronchoalveolar lavage fluid (n = 29, γ = 0. 762, P ＜ 0. 01). Conclusion The beneficial effect of glucocorticoid(dexamethasone) on airway inflammation in asthma rats could be at least in part due to their direct inhibitory effect on ENA-78 and TGF-β1 protein generation by neutrophil.

Aim To investigate if anti-depressive effect of venlafaxine was associated with improving oxi-dative stress and expression of hippocampus NET and 5-HTT in rat model induced by CUS. MethodsEighty SD male rats were randomly divided into four groups:model group(MG),normal group(NG), ven-lafaxine-treated normal group ( VNG ) , and venlafax-ine-treated model group ( VMG) . VNG and VMG were given venlafaxine (23. 4 mg·kg-1 ·d-1 ) once daily;NG and MG were given the same volume solvent. Soli-tary condition with chronic unpredicted stress ( CUS ) was taken to establish rat depression model. The force swimming test was used to evaluate the behavior chan-ges of experimental rats. The malondialdehyde ( MDA) level and activity of superoxide dismutase ( SOD ) in serum were determined by biochemical methods. The mRNA and protein expressions of NET and 5-HTT in
hippocampus were determined by Real-Time Reverse transcription polymerase chain reaction ( real-time RT-PCR) and Western blot ( WB) , respectively. Results Compared with NG rats, obviously increasing immo-bile time of rats in force swimming test and serum MDA level, as well as significantly decreasing SOD activity in serum was observed with clearly decreasing 5-HTT expression and elevating NET expression in hippocam-pus of MG rats. The treatment of venlafaxine distinctly suppressed changes above from CUS-induced rats. However, significant changes failed to be found in VNG rats. Conclusion The anti-depressive effect of venlafaxine may at least partly involve in improving ox-idative stress/anti-oxidative stress balance and revers-ing abnormal expression of NET and 5-HTT.