利用电压钳技术初步研究抗菌肽抗菌机制

目的为了证实电压钳技术测得的电流变化能说明诱导的昆虫离体细胞Tn-5B1产生的抗菌活性肽类物质的抗菌机制。方法将制备的抗菌肽作用于去掉滤泡膜的爪蟾卵母细胞,利用电压钳技术记录的电流研究抗菌肽抗菌机制。结果去掉滤泡膜的非洲爪蟾卵母细胞可作为模式细胞研究离子通道,抗菌肽作用于爪蟾卵母细胞14 min后,检测到了电流的变化,而且表现为外向电流持续升高。结论细胞经抗菌肽作用后产生了离子通道,此离子通道使得阳离子外流,初步证实该抗菌肽通过破坏坏细菌细胞来达到杀菌目的 。     

非洲爪蟾卵母细胞内源性及表达的P2型嘌呤受体介导的膜电流记录方法

目的 记录和区分卵母细胞内源性P2型嘌呤受体及表达的外源性P2型嘌呤受体介导的膜电流。方法 应用双电极电压钳技术分别在带滤泡膜及去滤泡膜后注入RNA并经孵育的卵母细胞记录膜电流。结果 根据所用卵母细胞标本的制备（是否去滤泡膜及是否注入RNA并孵育）以及电流的形式及幅值，可将内源性P2型嘌呤受体和表达的外源性P2型嘌呤受体介导的膜电流区分开来。结论 应用以上方法可区分两种不同的三磷酸腺苷（adenosine triphosphate,ATP)激活电流。     

NMDA受体亚基NR1的体外表达及功能检测

目的：体外表达ＮＭＤＡ受体亚基ＮＲ１并了解其电生理学特性,探讨体外表达受体亚基的意义。方法：将克隆的ＮＭＤＡ受体亚基ＮＲ１质粒ＤＮＡ经体外转录成ＲＮＡ,显微注射到爪蟾卵母细胞后进行受体表达,采用双电极电压钳位技术记录表达的ＮＲ１亚基的激活电流。结果：注射ＮＲ１亚基ＲＮＡ的爪蟾卵母细胞能表达出具有电生理学功能的受体,其激活电流能被Ｄ－ＡＰ５阻断等特性,说明该表达的ＮＲ１亚基具有ＮＭＤＡ受体的功能。结     

表达的辣椒素受体对甲醛刺激的反应

目的：验证Nielsen假设的正确性。方法：体外转录辣椒素受体(VRI)cDNA，获得CRmRNA，注入非洲爪蟾卵母细胞，异源性表达VRI。用甲醛刺激异源性表达的VRI。用双电极电压钳记录。结果：实验组有3个细胞对激动剂辣椒素(capsaicin 10^-7mol／L)有反应，并且VRI拮抗剂(capsazepine 10^-5mol／L)能完全阻断此内向电流。结论：表达的VRI对甲醛刺激发生反应。验证了Nielsen假设的正确性。     

大鼠心室肌细胞肿胀激活氯电流ICl,swell的研究

目的：探讨大鼠心室肌细胞肿胀激活氯电流ICl,swell在等渗(1T)及低渗(0．6T)溶液中的变化。方法：①采用图文采集及分析系统测量并比较大鼠心室肌细胞在等渗(1T)及低渗(0．6T)溶液中体积的变化。②采用全细胞电压钳方式记录大鼠心室肌细胞在1T及0．6T溶液中ICl,swell的变化。结果：①大鼠心室肌细胞在0．6T溶液中比1T中体积增大37．4％。②心室肌细胞在1T溶液中未见电流激活，在0．6T溶液中可见到内向及外向电流增大，其外向电流可被1mmoL／L9AC抑制。结论：大鼠心室肌细胞低渗性肿胀可激活ICl,swell，且能被9AC抑制，后者可能在充血性心力衰竭心律失常的发生起重要作用。     

脑片膜片钳法——改进的盲法和离子孔道法

脑片技术和膜片钳技术的结合是目前离体神经系统神经元网络研究的前沿技术。作者对其中的盲法和离子孔道法进行了改进，用解剖显微镜代替倒置显微镜，对记录电极的拉制进行改良，使之不必进行抛光处理，在离子孔道记录时，去极化电压钳位可稳定记录达０～４０ｍＶ。作者并于实验中筛选出典型的电反应波形以便于判断获得良好的电极膜封闭。上述改进降低了仪器费用，同时也利于获得稳定的记录，因而使此技术更易于推广应用。     

脑片膜片钳法———改进的盲法和离子孔道法

脑片技术和脑片钳技术的结合是目前离体神经系统神经元网络研究的前沿技术。作者对其中的盲法和离子孔道法进行了改进，用剖解是微镜代替倒置显微镜，对记录电极的拉制进行改良，使之不必进行抛光处理，在离子孔道记录时，去极化电压钳位可稳定记录达０．４０ｍＶ。作者并于实验中筛选出典型的电反应波形便于判断获得良好的电极膜封闭。上述改进降低了仪器费用，同时也利用获得稳定的记录，因而使此技术更易于推广应用。     

鼠脑神经递质受体在卵母细胞上的表达

把提取的鼠脑mRNA注射到卵母细胞内，在卵母细胞上以双电极电压钳技术研究某些递质的诱导电流，实验发现除卵母细胞本身所具有的muscarine受体外，还有多种受体明显地在卵母细胞上表达，卵母细胞膜上的表达受体的反应可分平滑型，振荡型，复合型三类，其反应类型与所开启的通道及通透离子有关，平滑型反应的受体与离了通道为一复合体，激动剂开启其自身离子通道，而振汇型反应是递质通过第二信使的介导，引起胞内Ca^2 库中Ca^2 释放，开启卵母细胞膜本身的Cl^-通道，产生Ca^2 依赖的Cl^-电流，构成振荡型电流反应的主要成分，复合型二者兼有。     

牛蛙球囊毛细胞电压依赖性钾通道电流的研究

目的 :研究牛蛙球囊毛细胞膜上钾通道的类型。方法 :全细胞膜片钳技术。结果 :1当钳制电位为 - 1 0 0 m V,以 1 0 m V的步距阶跃 ,阶跃从 - 70 m V至 + 2 0 m V,随着膜电位的去极化 ,可记录到一系列快速、瞬时的外向电流 ,4- Ap对其有特异性阻断作用。激活电压为 - 53.85± 7.8m V。2 A型钾通道的数量在牛蛙毛细胞间存在差异。 3当钳制电位为 - 70 m V,以 1 0 m V的步距阶跃去极化 ( - 50 m V～ + 4 0 m V) ,可产生一延迟整流性钾离子流 ,四乙基氯化氨 ( TEA)能使该电流幅度下降 59%± 1 1 %。结论 :牛蛙毛细胞膜上含有 A型钾通道和延迟整流性钾通道。     

离体新生大鼠运动神经元对5-羟色胺的反应

<正> 在离体新生大鼠脊髓切片对218个经逆行激活鉴定的运动神经元(MN)进行电流或电位记录,发现灌流(10～100μmol/L)或压力注射5-羟色胺(5-HT),使81%的 MN 发生去极化或内向电流,9%发生超极化或外向电流,在57%的 MN 诱发了包含 EPSP 和(或) IPSP 的突触活动。5-HT 诱发的突触活动不仅可被低     

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.     

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     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

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.     

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…     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.     

Possible Spread of adenovirus type 3 from poultry to humans ：indirect evidence from an outbreak in China

Objective:To explore the epidemiology and etiology for an outbreak of acute respiratory tract infection that occurred in one county of Jiangsu Province, China 2004. Methods: Only cases meeting the case definition were included in the study. We reviewed the medical records of the cases who were admitted to the local hospitals, interviewed cases by a standard questionnaire, and then described the epidemiotogic features and analyzed risk factors by means of a case-control study. We collected pharyngeal swab specimens and sent them to different laboratories for isolation and culture. The laboratory used different detection methods such as DIP, PCR, electron microscope examination and microneutralization assay, to identify and then type the positive specimens. Results:A total of 871 cases were reported during the period from April 18 to July 4,2004. The distribution of onset times presented two peaks, one in late May and another in middle June. The epidemic occurred mainly in the elementary and junior high schools in ten townships of one county, and the mean age of the cases was 12 years (range 7 months to 18 years). The course of the disease was acute, and was characterized by fever accompanied with sore throat and tonsillitis. The WBC count of cases was normal or elevated. The mean duration of illness was 5 days (range 2 to 12 days). No fatalities from illness were reported. A case-control study indicated that the possible risk factors were close contact with a case and/or poultry before onset and sharing of towels among members of the family. The typical CPE was observed through inoculating pharyngeal swab specimens into the HEP-2 cell cultures in different laboratories. An infection of adenovirus type 3 was verified by detecting positive specimens in different methods. Conclusion:This investigation demonstrated that the acute respiratory infection in cases was caused by adenovirus type 3. Cases occurred in over 70 schools in ten townships in 2004, and the route of transmission was possibly close contact with cases or droplet transmission.