AIM To investigate the effects of tetracycline on the expression of adhesive molecules E selectin and intercellular adhesion molecule 1 (ICAM 1) on human umbilical vein endothelial cells (HUVECs) stimulated by tumor necrosis factor alpha (TNF ?) or lipopolysaccharide (LPS). METHODS HUVECs were activated by TNF ? (0 5～50 ?g?L 1 ) or LPS (0 5～50 mg?L -1 ). Tetracycline (10 -6 ～10 -4 mol?L -1 ) was used to incubate HUVECs for 1 h before stimulation with TNF ? ( 1 ?g?L -1 ) or LPS ( 5 mg?L -1 ) and co incubate HUVECs with TNF ? or LPS for 4 h or 18 h. The expression of adhesive molecules on HUVECs was measured by a cellular enzyme linked immunoabsorbent assay (Cell ELISA) method. RESULTS After stimulation of HUVECs with TNF ? or LPS for 4 h induced E selectin expression, and exposure HUVECs to TNF ? or LPS for 18 h increased constitutive expression of ICAM 1 compared with control group ( P

Aim Tostudytheeffectof4-phenlybuty-rate acid(4-PBA) on the heart function of diabetic rats and to explore its underlying mechanism. Methods MaleSprague-Dawley(SD)ratsweredividedintothree groups randomly: ( 1 ) control rats;( 2 ) diabetic rats;(3) 4-PBA treated diabetic rats. Type 1 diabetic mod-el was induced by single intraperitoneal injection of strepotozotocin (60 mg·kg-1 ) . After five weeks dia-betic model was established, diabetic rats were treated with 4-PBA by P. O at the dose of 1 g·kg-1 per day for 20 weeks. At the end of the experiment, hemody-namics of the left ventricle and main artery were detec-ted through right carotid artery cannulation in rats. The level of blood glucose, the activity of superoxide dis-mutase( SOD) and nitric oxide synthetase ( NOS) and the content of malondialdehyde ( MDA ) and nitric ox-ide( NO) of serum and myocardial tissue in rats were measuredbycolorimetry.Results Thetreatmentof4-PBA had no effect on blood glucose, the weight of body and heart, but slightly increased systolic and diastolic pressure of artery, heart rate, systolic pressure of left ventricular, maximum increase rate and minimum de-crease rate of left ventricular pressure( ± dp/dt) in di-abetic rats. It also slightly decreased diastolic pressure of left ventricular, shortened diastolic duration of left ventricular ( P > 0. 05 ) . Moreover, it significantly shortened relaxation time of left ventricular in diabetic rats (P<0. 05). In addition, the treatment of 4-PBA obviously enhanced the activity of SOD and NOS, NO content, and reduced the content of MDA in serum and heartofdiabeticrats(P<0.05).Conclusion 4-PBA has a potential protective effect on heart function in diabetic rats, which is achieved by ameliorating he-modynamics and significantly improving diastolic dys-function in diabetic rats. The underlying mechanism may be related to the inhibition of oxidative stress and promoting the genesis of NO in myocardial tissue.

Objective To investigate the protective effect of angiotensin-converting enzyme inhibitor (ACEI) on kidney function in patients with sepsis.Methods Eighty-seven patients with sepsis were randomly (random number) divided into the routine treatment (A group,n ＝45) and the ACEI intervention group (B group,n =42).Patients were managed by international guidelines for sepsis in A group and were treated with benazepril (ACEI) 20 mg a day in addition in B group.Serum creatinine and cystatin C were detected and APACHE Ⅱ scores and urine output were recorded on the 1st,3rd and 7th day.Those laboratory findings and APACH Ⅱ score were compared between two groups.The incidence of acute kidney injury (AKI) and abnormal cystatin C levels were compared between two groups.Results In B group,serum creatinine and cystatin C of patients were lower compared with A group.The incidence of AKI and abnormal cystatin C and APACHE Ⅱ scores were reduced in B group compared with the A group.Conclusions Benazepril improved sepsis-induced AKI and patient conditions.

Objective To investigate the effect of Bunao capsule on learning,memory and antioxidative abilities of rats with Alzhheimer’s disease(AD)induced by D-galactose combined with amyloid β-protein(Aβ25-35 ),and provide experimental basis for the prevention and treatemtn of AD. Methods A total of 90 SD male rats were randomly divided into model control group,piracetam group,sham operated group,Bunao capsule(0.79,1.58,3.15 g·kg-1 )groups(n= 15 each).The rat models were established by intraperitoneal injection of D-galactose and injection of Aβ25-35 into the bilateral lateral cerebral ventricle.Then rats were given corresponding drugs by gavage in different groups for 8 weeks.The learning and memory abilities were meseured by Morris water maze test.The morphology of brain cells was observed by HE staining.The activities of glutathione peroxidase(GSH-Px)and superoxide dismutase( SOD),and the malondialdehyde( MDA)contents in the brain tissues were measured by spectrophotometry. Results The target quadrant residence time was(20.39±7.75)s and(20.82±5.09)s in Bunao capsule (1.58,3.15 g·kg-1 )groups,which were significantly increased as compared with that in model control group[(12. 35 ± 6.95)s](P<0.01).Brain nerve cell morphology in Bunao capsule(1.58,3.15 g·kg-1 )groups was obviously improved as compared with that in model control group,and was close to that in sham operated group.The activities of GSH-Px and SOD were significantly increased,and MDA contents decreased in Bunao capsule groups as compared with those in model control group (P<0.01). Conclusion Bunao capsule can dose-dependently improve the learning,memory and antioxidative abilities of AD rats.The mechanism may involve upregulation of antioxidative enzyme activities and removal of oxidative products.

Objective To investigate the effects of cold exposure with different time courses on cardiovascular function and skeletal muscle contraction in Sprague-Dawley (SD) rats.Methods The male SD rats were randomly divided into control,12-h cold,24-h cold,and 48-h cold groups.The rats in all cold group were maintained in 4 ℃ environment for indicated time and allowed free access to standard rat chow and water ad libitum.After the animal model was established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Lanendorff heart persusion was used to detect some parameters of cardiac function,such as coronary flow (CF),mean left ventricular systolic pressure (mLVSP),mean left ventricular diastolic pressure (mLVDP) and maximum ascending and declining rate of left ventricular pressure (± dp/dt max) ; and then the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes of above cardiac function indexes were recorded again.Organ bath method was employed to detect contraction response to potassium chloride (KCl) or phenylephrine (PE) and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings.The twitch tension,titanic tension,and fatigue test of soleus and EDL were examined by electrical stimulation to reflect contractive function of skeletal muscle.Results The coronary flow (CF) were significantly reduced in 24-h and 48-h cold groups compared to control group [(9.0 ± 1.7) ml/g and (9.8 ± 1.3) ml/g vs (11.6 ± 1.2) ml/g,P ＜ 0.05] ; 48-h cold exposure also showed significantly increased mLVSP (P ＜0.01) and mLVP (P ＜ 0.05) of rats.After ischemia-reperfusion,the heart re-beat time was shorter in all cold exposure groups (all P ＜ 0.01),mLVP and mLVDP were higher in 24-h and 48-h cold group than control group (all P ＜ 0.01).The contraction of aortic rings responses to 60 mmol/L KCl [(0.63 ±0.13) g and (0.79 ±0.11)g vs (1.28±0.17)g,P ＜0.01] and 1 μmol/L PE [(0.62 ±0.18)g and (0.68 ±0.13)g vs (1.25 ± 0.18)g,P ＜ 0.01] were decreased while the endothelium-dependent relaxation responses to low concentration ACh (3 × 10-8 ～ 3 × 10-7) were increased in 24-h and 48-h cold groups relative to control group (all P ＜ 0.05).The 48-h cold also reduced titanic tension of SOL and EDL in comparison with control [(4.14 ± 0.62) N/cm2 vs (5.50 ± 0.75) N/cm2,P ＜ 0.05 and (3.00 ± 0.57) N/ cm2 vs (4.70 ± 0.85) N/cm2,P ＜ 0.01].Conclusions 24 ～ 48 h cold exposure could reduce CF but enhance the tolerance of heart from rats to ischemia reperfusion injury and also decrease the contraction of aortic rings response to KCl or PE but increase the sensitive responses to ACh of aortic rings from rats.The 48-h cold could decrease the contraction of skeletal muscle elicited by electrical stimulation.

Objective To investigate the effects of fasting with different time courses on the contraction of skeletal muscle and cardiovascular function in Sprague-Dawley (SD) rats.Methods The healthy and male SD rats were randomly divided into (1) control,(2) 12 h fasting,(3) 24 h fasting,and (4) 48 h fasting groups.After animal models of fasting with different time courses were established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Langendorff heart perfusion was used to detect some parameters of cardiac function,including left ventricular systolic pressure (LVSP),left ventricular end-diastolic pressure (LVEDP),and maximum ascending and declining rate of left ventricular pressure (± dP/dt max).Then,the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes in above cardiac function parameters were recorded again.The contraction response to potassium chloride or phenylephrine and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings were measured with organ bath.The changes of twitch tension and titanic tension of soleus and EDL were examined to reflect contractive function of skeletal muscle.Results The + dP/dt max was significantly reduced in 24 h and 48 h fasting groups compared to the control (P ＜ 0.01).After ischemia-reperfusion,the heart re-beat time was significantly longer [(24.3 ± 7.0)min vs (14.0 ± 2.9)min,P ＜0.05],both mLVDP and mLVP were significantly higher in 48 h and 24 h fasting groups than control group (P ＜ 0.05),and those changes did not normalized until re-perfusion 60 min.The 48 h fasting increased significantly endothelium-dependent relaxation of aortic rings response to low concentration acetylcholine (P ＜ 0.05),but did not affect the maximum relaxation of aortic rings response to high concentration acetylcholine and contraction of aortic rings responses to potassium chloride and phenylephrine.The 48 h fasting also enhanced significantly titanic tension of soleus compared to the control [(7.01 ± 1.22) N/cm2 vs (5.5 ± 0.75) N/cm2 for48 h-Fasting vs Control,P ＜0.05].Conclusions The 24 ～48 h fasting may induce inhibition of cardiac function,while fasting 48 h can increase the sensitivity of aortic rings responses to ACh and contraction function of soleus in healthy and male rats.

Objective To study the protection of glutathione (GSH) on renal oxidative damage to mice which caused by mi‐crocystin‐LR(MC‐LR) .Methods Forty healthy KM mice were divided into five groups by randomly sampling ,which were saline control group ,GSH control group ,MC‐LR group ,low dose GSH +MC‐LR group and high dose GSH +MC‐LR group ,and the ex‐periment was lasting 15 days by intraperitoneal injection .Then we took out the kidney for pathological observation and detected the activity of CAT ,SOD ,GSH‐Px and the content of GSH ,MDA .Results Compared with control group ,the MC‐LR increased the content of MDA[(2 .31 ± 0 .22)nmol/mg prot ,P=0 .000] and decreased the content of GSH[(0 .68 ± 0 .02)mg/g prot] .The activi‐ty of CAT[(320 .54 ± 38 .99)nmol/mg prot] ,SOD[(180 .93 ± 15 .30)U/mg prot] ,GSH‐Px[(295 .11 ± 42 .40)U/mg prot](P<0 .05) .However ,after GSH was given ,compared with MC‐LR group ,MDA content[(1 .94 ± 0 .12)nmol/mg prot]of high dose GSH+MC‐LR group significantly decreased (P<0 .05) ,GSH content[(1 .01 ± 0 .08)mg/g prot ,(1 .08 ± 0 .16)mg/g prot]and CAT activity[(383 .46 ± 21 .98)nmol/mg prot ,(428 .50 ± 28 .61)nmol/mg prot] of both GSH groups significantly increased (P<0 .05) ,the activity of SOD[(222 .01 ± 11 .51)U/mg prot] and GSH‐Px[(358 .37 ± 20 .29)U/mg prot] of high dose GSH +MC‐LR group significantly increased (P<0 .05) .Conclusion MC‐LR may cause renal oxidative damage through promoting the lipid perox‐idation on renal cells .The GSH may reach a certain protective effect on kidney by reducing the lipid peroxidation ,improving the an‐tioxidant activity ,and removing oxygen free radicals .

AIM To study the effects of different kinds of chloride channel blockers on KCl and 5 HT induced contractile responses in cerebrovascular smooth muscle. METHOD The tension of rabbit basilar artery rings was measured. RESULTS ① DIDS, furosemide and NPPB inhibited the responses to KCl and 5 HT in a concentration dependent manner. The effects of DIDS, furosemide and NPPB on the response to 5 HT were stronger than that to KCl. ②When SK&F 96365 evoked a maximum inhibitory effect on 5 HT induced response, subsequent additions of DIDS, furosemide and NPPB could further produced vascular relaxation. CONCLUSION The chloride channel participates KCl and 5 HT induced contractile responses in cerebral basilar artery.

AIM To study the effects of protein tyrosine kinase inhibitor and protein tyrosine phosphatase inhibitor on cultured bovine cerebrovascular smooth muscle cells (CSMC) Ca 2+ store operated Ca 2+ influx. METHODS Cell culture and single intracellular free Ca 2+ concentration was measured in fura 2/Am flueorescence probe by MetaFluor Fluorescence ratio imaging system. RESULTS (1) protein tyrosine kinase inhibitor (genistein,2.5,5,10 ?mol?L -1 )decreased Ca 2+ influx significantly induced by endothelin 1(ET 1),ATP,cyclopiazonic acid(CPA) in concentration dependent manner; (2) Protein tyrosine phosphatase inhibitor (vanadate,2,4,8 ?mol?L -1 ) increased Ca 2+ influx significantly induced by ET 1,ATP,CPA in concentration dependent manner. CONCLUSION Protein tyrosine kinase inhibitor and protein tyrosine phosphatase inhibitor have effects on Ca 2+ store operated Ca 2+ influx induced by ET 1, ATP, CPA. Protein tyrosine phosphorylation participats in the signal transduction of Ca 2+ store operat Ca 2+ influx in cerebrovascular smooth muscle cells.

BACKGROUND:L-type calcium channels, as a kind of voltage-dependent calcium channel, is the main way of extracellular calcium ions into the cell, and play an important role in maintaining cellmorphology and physiological activities, characterized by a large single-channel conductance, slow channel attenuation, and longer duration of channel opening. Previous studies showed that basic fibroblast growth factor can promote the proliferation of chondrocytes cultured in vitro.
OBJECTIVE:To explore the effect of the L-type calcium channels on regulating chondrocyte proliferation and differentiation in response to basic fibroblast growth factor with patch-clamp.
METHODS:The chondrocytes were harvested from the joints of 3-day-old New Zealand rabbits. The second passage of chondrocytes was divided into experimental group and control group. Chondrocytes were incubated in media containing 10μg/L basic fibroblast growth factor and media alone separately. The opening of L-type calcium channels under the action of basic fibroblast growth factor was detected by patch-clamp. The intracellular calcium concentration was detected with laser confocal microscopy in the chondrocytes after 2 weeks of culture with basic fibroblast growth factor. Chondrocyte proliferation was analyzed by cellTiter kit after 8 days of culture. Type Ⅱ col agen was assessed quantitatively by immunohistochemistrical staining after 10 days of culture.
RESULTS AND CONCLUSION:Basic fibroblast growth factor has an inhibitory effect on the opening of the L-type calcium channels, resulting in a decrease in intracellular free calcium concentration (P<0.01). cellnumber was higher after culture with basic fibroblast growth factor than that cultured under conventional condition (P<0.01), and staining area of type II col agen significantly increased (P<0.05). Results verified that basic fibroblast growth factor can maintain intracellular Ca2+concentration at a low level by inhibiting the opening of L-type calcium channels, which can promote the proliferation and differentiation of chondrocytes.