代森锰锌对小鼠生精细胞T型Ca2+通道的作用

目的:研究农药代森锰锌(mancozeb)对小鼠生精细胞T型钙电流(ICaT)的影响?方法:采用膜片钳全细胞记录技术观察代森锰锌对急性机械分离的小鼠生精细胞ICaT 的影响?结果:Mancozeb(10?20?40?80?120 μmol/L)呈浓度?电压依赖性抑制小鼠生精细胞钙电流,抑制率分别为(9.93±0.90)%?(20.93±2.50)%?(25.92±2.80)%?(42.64±3.10)%?(56.80±3.20)% (n=6,P<0.05)?Mancozeb 对T型钙通道的半数最大抑制浓度(K50)为35.60 μmol/L?120 μmol/L mancozeb显著改变T型Ca2+通道的激活和失活特性:半数激活电压(V1/2a)和激活斜率因子(κa)分别从(-47.09±1.05)mV和(8.67±0.78) mV变为(-51.46±0.84)mV和(10.56±0.619)mV (n=5, P < 0.05);而半数失活电压(V1/2i)和失活斜率因子(κi)分别从(-62.96±3.36)mV和(9.93±1.41)mV变为 (-64.11±5.55)mV和(13.64±1.95)mV (n=5,P < 0.05)?结论:Mancozeb对生精细胞T型钙通道有抑制作用,且呈浓度依赖性?     

丹参酮ⅡA磺酸钠对原代培养猪冠脉平滑肌钙激活钾通道的影响

目的 :研究丹参酮ⅡA磺酸钠 (DS 2 0 1)对培养猪冠脉平滑肌细胞上钙激活性钾通道的作用。方法 :采用膜片钳单通道记录技术 ,观察给药前后通道特性变化。结果 :证实 :①猪冠脉平滑肌钙激活钾通道的单通道电导值高 ,为 2 91 74± 17 6 3pS。对膜电位变化及胞内钙离子浓度变化敏感。能被 5～ 2 0mmol/LTEA所阻断。②在细胞贴附式膜片方式下 ,5～ 30 μmol/L的DS 2 0 1对通道均无明显作用 ;而在内面向外式膜片方式下 ,5～ 2 0 μmol/L的DS 2 0 1对通道表现出明显的激活效应。结论 :DS 2 0 1对通道具有明显的直接激活作用     

二十二碳六烯酸对大鼠心肌细胞通道的影响

目的:探讨二十二碳六烯酸(DHA)对大鼠心室肌细胞动作电位(AP)及钠通道电流(INa)的影响。阐述DHA抗心律失常的机制。方法:采用酶消化法获得大鼠心室肌细胞,用膜片钳技术分别记录0、20、40、60、80、100和120μmol/L DHA对大鼠心室肌细胞AP复极25%、50%和90%时程(APD25、APD50和APD90),对AP最大上升速率(Vmax)、幅值(APA)、超射值(OS)及INa的影响。结果:①不同浓度DHA对APD25、APD50和APD90呈浓度依赖性延长(P<0.05,n=20),对Vmax、APA和OS的影响无显著性差异(P>0.05,n=20)。②不同浓度DHA对INa呈浓度依赖性阻滞、I-V曲线上移、稳态失活曲线左移、失活后恢复时间延长,对稳态激活曲线无影响。在指令电压-30 mV时,上述浓度DHA对INa阻滞分别为(1.51±1.32)%、(21.13±4.62)%、(51.61±5.73)%、(67.62±6.52)%、(73.49±7.59)%和(79.95±7.62)%(P<0.05,n=20),DHA对INa的半效作用浓度(EC50)为(47.91±1.57)mol/L。结论:DHA对APD的延长及对钠通道的抑制作用可能是其抗心律失常机制之一。     

巴曲酶对原代培养的大鼠皮层神经元钙激活钾通道的作用研究

目的观察巴曲酶对原代培养新生SD大鼠皮层神经元上钙激活钾通道(Kca)作用。方法采用细胞贴附和内面向外两种膜片钳单通道记录技术检测巴曲酶对Kca作用。结果在钙浴液内,细胞贴附式膜片上,钳制膜电位+30mV,加入不同浓度巴曲酶0.15mmol/L、0.25mmol/L、0.50mmol/L、0.75mmol/L、1.0mmol/L,通道开放概率由0.005分别增加为0.013±0.002、0.082±0.011、0.131±0.012、0.211±0.010、0.062±0.009(P<0.01),在0.75mmol/L以内表现出浓度依赖性。在无钙浴液内,细胞贴附式膜片上,钳制膜电位+50mV,随巴曲酶浓度增加为0.15mmol/L、0.40mmol/L、0.60mmol/L、1.0mmol/L时,通道开放概率由0.005分别增加为0.013±0.001、0.112±0.007、0.193±0.010、0.301±0.009(P<0.05)。6例内面向外式膜片上,钳制膜电位+40mV,分别加入巴曲酶0mmol/L、0.25mmol/L、0.50mmol/L3min后,通道开放概率分别为0.012±0.007、0.011±0.009、0.013±0.008(P>0.05),巴曲酶在胞内Kca开放概率,平均开放/关闭时间,电流幅值均无明显变化。结论巴曲酶通过影响胞内游离钙水平间接调节Kca,可能对神经元有直接保护作用。     

多巴胺对大鼠前额叶皮层锥体神经元NMDA受体通道的影响

[目的]探讨多巴胺(DA)对前额叶皮层锥体神经元谷氨酸/N-甲基-D-天冬氨酸(NMDA)受体的调节作用.[方法]使用细胞贴附式膜片钳技术进行单通道记录,观察不同浓度DA对急性分离的前额叶皮层锥体神经元NMDA受体通道电流的影响.[结果]结果表明:当DA浓度为0逐渐升高到60 μmol/L时,NMDA受体通道的电流幅度由(2.78±0.68)pA升高到(5.42±1.26)pA;短开放时间常数变化不明显,但长开放时间常数由(13.59±1.26)ms升高到(36.19±1.17)ms;长关闭时间常数由(19.73±1.46)ms降低到(2.67±0.75)ms;通道开放概率由0.25±0.09升高到0.56±0.25;当DA浓度继续由60 μmol/L升高到100 μmol/L时,NMDA受体通道的电流幅度回落到(3.56±0.79)pA,长开放时间常数回落到(14.40±1.37)ms;长关闭时间常数恢复为(17.74±1.79)ms,与对照组比较无显著性变化;通道开放概率由0.56±0.25降低到0.17±0.08.[结论]DA可以影响前额叶皮层锥体神经元NMDA受体通道的活动,而且具有浓度依赖性,低浓度有明显促进作用,高浓度则促进作用减弱,甚至有抑制作用.     

心肌细胞肿胀激活氯离子通道的单通道特性

目的研究急性分离小鼠心肌细胞肿胀激活氯离子通道的单通道特征.方法采用膜片钳单通道(细胞贴附式和内面向外式)方法,记录急性分离的小鼠心肌细胞暴露于低渗溶液时出现的肿胀激活氯通道电流(ICl.swell).结果等渗状态下,用细胞贴附式记录,多数心肌细胞膜片无通道活动,用低渗溶液灌流细胞,在细胞容积增大的同时,出现单通道电流的激活.该肿胀激活单通道氯电压在-100mV~+100mV均有激活.在电压+100mV时,其单通道电导为(38.1±2.5)pS,开放概率为0.76±0.08,电流-电压曲线显示其反转电位(-34.5±2.8)mV,接近氯离子平衡电位的理论值(ECl=-38.6mV),且反转电位随氯离子浓度的改变而改变.氯通道阻断剂DIDS可逆性阻断该单通道电流.结论小鼠心肌细胞肿胀激活氯通道的单通道电导为(38.1±2.5)pS,电流呈外向整流特征并对DIDS敏感.     

甘氨脱氧胆酸对肝细胞钙激活性钾通道的影响

目的观察甘氨脱氧胆酸(GDC)对大鼠肝细胞钙激活性钾通道(KCa)开放概率的影响.方法体外培养大鼠肝细胞,以GDC为处理因素,应用膜片钳技术通过贴附式和内面向外式膜片测定大鼠KCa开放概率.结果①细胞贴附式膜片下,浴液中Ca2+浓度10-7mol/L,GDC终浓度为50,100,250μmol/L时,概率由0.038±0.09分别增加到0.620±0.338,0.591±0.163,0.895±0.176(n=4,P均＜0.01).浴液中不加GDC,改变Ca2+浓度(10-7、10-6、10-5mol/L),KCa的开放概率并不发生明显改变,而在此液中加入GDC(终浓度100μmol/L),KCa开放概率由0.042±0.015增加到0.385±0.330和0.551±0.163(n=4,P均＜0.05).浴液中不含Ca2+,并加入EGTA,GDC浓度为50,100,250μmol/L,KCa开放概率显著增加(n=4,P均＜0.05),但比浴液中有Ca2+时增加程度小.②内面向外式膜片下,浴液中无Ca2+,加入GDC(10,50,100,250μmol/L),KCa开放概率无明显改变(n=2,P＞0.05).结论①GDC对KCa有明显的激活作用,并有浓度依赖性.②GDC既有引起胞外Ca2+内流,也能诱导内贮Ca2+的释放.③GDC对KCa的激活是通过Ca2+实现的.     

丹参酮ⅡA磺酸钠对原代培养猪冠脉平滑肌ATP敏感钾通道、钙激活钾通道的影响

目的 :探讨丹参酮ⅡA磺酸钠 (DS 2 0 1)对原代培养猪冠脉平滑肌上ATP敏感的的钾通道 (KATP)、钙激活钾通道 (KCa)的作用。方法 :采用膜片钳单通道电流记录技术进行定量研究。结果 :KATP(33 15pS)可被 30 μmol/L优降糖特异性阻断 ,而KCa(2 46 .5 3pS)则表现出明显的钙离子依赖性和TEA阻断作用等特点。内面向外式膜片下 ,浴液中加入DS 2 0 1可激活KATP及Ca。膜电位为 5 0mV时 ,5、10、15 μmol/LDS 2 0 1分别使KATP开放概率 (NPo)从0 0 0 4± 0 0 0 1增加到 0 44 9± 0 0 0 4(n =5 ,P <0 0 1)、0 6 0 2± 0 0 32 (n =5 ,P <0 0 1)、0 90 4± 0 10 5 (n =5 ,P <0 0 1) ,2 0 μmol/LDS 2 0 1使KCa的NPo从 0 0 0 0增加到 0 0 11± 0 0 0 2 (n =3,P <0 0 1)。结论 :DS 2 0 1对KATP和KCa均有直接激活作用。     

紫杉醇对卵巢癌细胞作用的浓度依赖性及其机制探讨

目的 探讨不同浓度紫杉醇对卵巢癌细胞发挥抗癌效应的内在机制.方法 以0.01、0.1、1、10、50 μmol/L紫杉醇作用于人卵巢癌细胞系A2780,采用碘化丙啶(PI)单染法流式细胞术检测不同浓度紫杉醇作用下细胞周期的变化情况.进而对细胞进行同步化处理,采用PI单染法流式细胞术分析低浓度(0.01 μmol/L)紫杉醇对细胞周期的影响;Annexin Ⅴ/PI双染法流式细胞术分析高浓度(50 μmol/L)紫杉醇对A2780细胞坏死的影响.结果 ①紫杉醇浓度为0.1、1、10 μmol/L时,G2/M期细胞比例明显高于0.01 μmol/L组(P<0.01)和50 μmol/L组(P<0.01).②A2780细胞经同步化处理后给予0.01 μmol/L紫杉醇,经过一个细胞倍增时间未观察到G0/G1期阻滞.③0.01 μmol/L组,1 μmol/L组坏死细胞比例明显低于50 μmol/L组(P<0.01).结论 在0.1～10 μmol/L 的浓度范围内,紫杉醇可能通过诱导G2/M期阻滞使卵巢癌细胞发生凋亡,浓度为0.01 μmol/L时紫杉醇诱导凋亡与G0/G1期或G2/M期阻滞均无关,浓度为50 μmol/L时紫杉醇可能主要通过诱导细胞坏死发挥抗癌效应.     

双氢青蒿素对人宫颈癌细胞Hela生物学特性的影响

目的 探讨双氢青蒿素(dihydroartemisinin,DHA)对人宫颈癌细胞Hela细胞周期、增殖、凋亡和侵袭的影响及机制。方法以Hela细胞为研究对象,根据DHA的不同浓度,分组：空白对照组（等体积生理盐水）、低浓度DHA组 （50 μmol/L）及高浓度DHA 组（400 μmol/L）。RT-PCR、Western blot检测各组细胞Caspase-3、Bax基因和蛋白的表达变化;流式细胞仪对细胞周期及凋亡检测;平板克隆实验检测细胞增殖;Transwell侵袭实验检测细胞侵袭力。结果Caspase-3、Bax蛋白表达空白对照组、低浓度DHA组分别为0.48±0.07、0.51±0.08,0.95±0.11、1.03±0.12,高浓度DHA 组为2.28±0.21、2.35±0.23,明显高于上述两组（P<0.05）;细胞周期结果显示G0/G1期与S 期细胞空白对照组、低浓度DHA组分别为53.52±3.98、32.84±3.18,61.64±3.92、22.47±2.31,高浓度DHA 组为76.48±5.26、13.25±1.22,细胞周期阻止于G0/G1期（P<0.05),S期细胞数减少（P<0.05）;高浓度DHA 组凋亡率为（20.41±0.92）%,明显高于空白对照组和低浓度DHA组［（2.84± 0.61）%、（10.36±0.63）%］（P<0.05）;平板克隆实验显示高浓度DHA 组细胞克隆形成数为（62.54±12.46）个,明显低于空白对照组和低浓度DHA组［（188.56±14.84）个,（119.56±14.62）个］（P<0.05）;穿膜细胞数高浓度DHA 组为（92.75±13.54）个,明显低于空白对照组和低浓度DHA组［（232.58±26.42）个,（128.75±22.16）个］（P<0.05）。结论DHA干预调高Bax及Caspase-3蛋白表达,抑制Hela细胞的增殖,促进细胞凋亡,减少S期细胞,降低细胞侵袭能力,高浓度时效果更显著。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.