豚鼠心室肌细胞的分离及其基本电生理特性的观察

目的 分离豚鼠单个心室肌细胞,并观察其基本电生理特性.方法 Langendorff灌流,胶原酶法分离心室肌细胞;应用膜片钳全细胞模式记录心室肌细胞的动作电位和L型钙通道电流.结果 酶法分离得到存活杆状细胞达50%以上.全细胞电流钳模式下记录,细胞静息电位为(-69.9±3.4)mV,复位到95%时的动作电位时程(APD95)为(313.1±38.9)ms.全细胞电压钳模式下记录到典型L型钙通道电流.结论 该方法能够简单有效地获得性能稳定的心室肌细胞,尤其适用于膜片钳心电生理学的研究.     

豚鼠心房肌细胞的分离及其电生理特性

目的 建立单个心房肌细胞的急性分离方法并记录其电生理特性。方法 采用改进的酶解分离法分离单个细胞并经膜片钳全细胞记录法进行鉴定。结果 采用本方法分离出典型的豚鼠单个心房肌细胞并记录到豚鼠心房细胞毒蕈碱型钾电流及L－型钙电流。结论 此方法简单、快捷，分离出的心房肌细胞具有正常的电生理特性。     

成年小鼠心室肌细胞分离及L型钙电流记录

目的:寻找一种成年小鼠心肌细胞分离的简便、可行且稳定的方法,以获得耐钙心肌细胞并记录心室肌细胞L型钙电流(IL-Ca)。方法:采用Langerdorff逆行主动脉灌流、无钙台氏液+胶原酶Ⅱ消化法分离成年小鼠单个心室肌细胞;利用含钙细胞外液复钙法筛选耐钙心肌细胞;并采用全细胞膜片钳技术记录心室肌细胞L型钙电流。结果:利用该方法分离心肌细胞可获得90%活细胞;复钙后细胞存活率60%-70%;并可记录到典型L型钙电流。结论:无钙台氏液+胶原酶Ⅱ消化法简单可行且稳定,可分离大量活性较佳且耐钙心肌细胞,可用于心肌细胞离子通道电流记录,从而为心脏电生理研究提供有力的技术支持。     

适于膜片钳实验的心肌细胞分离方法及维拉帕米对钙电流的作用

目的：建立适应于膜片钳实验的大鼠单个心室肌细胞的分离方法,探讨L-型钙通道在大鼠心肌电活动中的作用.方法：采用改进Langendorff装置,酶解法分离单个心室肌细胞,应用全细胞膜片钳技术记录心室肌细胞L-型钙电流及维拉帕米的作用.结果：该法分离到的心肌细胞存活率达80％～90％,高倍镜下细胞呈杆状,边缘清楚,表面光滑,纹理清晰;成功记录到典型L-型钙电流（Ica-L）,给予维拉帕米（0.3、3、30 μmol/L）后呈浓度依赖性抑制钙电流,其抑制率分别为（44.35±5.52）％,（61.06±2.99）％,（71.88±4.45）％.结论：改进心肌细胞的分离方法可获得大量耐钙心室肌细胞,这些细胞具有正常的电生理特性.     

鹌鹑心室肌细胞电生理特性的研究

目的　建立鹌鹑单个心室肌细胞的分离方法 ,观察鹌鹑心室肌细胞的电生理特性 ,并探讨不同离子通道在鹌鹑心肌电活动中的作用。方法　采用酶解法分离单个心室肌细胞 ,应用全细胞膜片钳技术记录心室肌细胞动作电位和L型钙电流。结果　鹌鹑单个心室肌细胞的静息电位 (RP)为 ( 6 2 .2 8± 2 .6 4 )mV ,超射值 (OS)为 (5 9.38±3.81 )mV ,5 0 %复极时间 (APD50 )和 90 %复极时间 (APD90 )分别为 (80 .4 0± 1 9.1 0 )ms和 (1 2 1 .95± 38.72 )ms。在实验电压 +1 0mV时 ,L型钙电流的峰值为 ( 1 3.2 7± 3.70 )pA pF(n =4 )。结论　鹌鹑心室肌细胞上存在着L型钙电流 ,此电流为构成鹌鹑心室肌细胞动作电位的重要电流之一     

不同用途的心室肌细胞的分离

目的 :建立适合于不同用途的心室肌细胞的酶解分离方法。方法 :恒流 Langendorff灌流 ,I型胶原酶酶解分离 ,应用膜片箝技术记录离子电流、双波长显微荧光光度术检测细胞内游离钙离子浓度及视频跟踪系统测定心肌单细胞收缩。结果 :采用不同分离技术参数获得的心肌细胞能满足上述三种实验条件下的要求。酶解分离的单个心室肌细胞静息电位为 (- 74 .0 6± 4 .5 4 ) m V,记录到心室肌细胞典型的钾、钠、钙离子电流和稳定的心室肌细胞内游离钙比值 ,心室肌单细胞收缩特性稳定。结论 :通过调整分离技术参数获得的心室肌细胞性能稳定 ,具有正常的电 -机械生理特性和耐钙性。     

急性分离大鼠心室肌细胞的Ca2+电流和细胞内Ca2+浓度

目的 在急性分离SD大鼠心室肌细胞上,检测Ca2 电流(Ica)和Ca2 浓度[Ca2 ].的变化.方法 改良Langendroff法,用含胶原酶Ⅰ(1750 U/mL)和蛋白酶XⅣ(0.28 mg/mL)的无Ca2 台氏液经大鼠主动脉循环灌流法,急性分离大鼠心室肌细胞;采用全细胞膜片钳技术记录心室肌细胞的Ica,动态细胞荧光成像技术检测心室肌细胞[Ca2 ]i的变化.结果 在20 rmin内新鲜分离大鼠心室肌细胞,全细胞膜片钳技术可记录电刺激引起心室肌细胞Ica,1μmol/L异丙肾上腺素可显著增加Ica;并观察到心室肌细胞[Caa2 ]i以及电刺激增强[Ca2 ]i的效应.结论 大鼠心室肌细胞急性分离方法简便实用,所分离的心室肌细胞形态和功能正常,适用全细胞膜片钳和动态细胞荧光成像技术检测实验.     

不同年龄组人心房肌细胞电生理特性鉴定

目的探讨建立稳定的人体心房肌细胞的分离方法,观察分离的正常窦性(normal sinus rhythm,NSR)右心耳心房肌细胞细胞基本电生理特性。方法采用蛋白酶(XXIV型)和胶原酶(V型)两步法对单个人体心房肌细胞进行分离,分为小儿、儿童和成人组成的心房组和小儿、儿童组成的心室组分别进行研究,观察记录L-型钙通道电流(ICa-L)、钠通道电流(INa)、瞬时外向钾通道电流(Ito)、内向整流钾通道电流(IK1)。结果一次取心肌组织约50-70mg,蛋白酶和胶原酶分别到1.1mg/5ml,5.5mg/20ml,可获得300-500个细胞,其中可用于电生理特性记录的细胞数量200-250个。心房室肌细胞均表现某种程度的去极化。结论二步法分离的人体心房肌细胞密度高、细胞膜光滑,数量多且质量好,能记录到多种离子通道电流,满足生理细胞记录使用。     

犬心房肌细胞分离及其L型钙通道电流检测的研究

目的探讨一种稳定可靠的犬心房肌细胞的分离方法,提高膜片钳的实验成功率并检测L型钙通道电流。方法采用改良Langderoff灌流系统行主动脉逆行灌注,采用选择性冠状动脉插管的方法,获得单个心房肌细胞。在显微镜下观察细胞形态,并应用膜片钳全细胞模式记录L型钙通道电流。结果分离细胞存活率为70%～80%,复钙后细胞存活率为30%～40%,耐钙细胞形态呈杆状,折光性强,横纹清晰,并能稳定记录L型钙通道电流。结论改良Langderoff技术有助于稳定分离出存活率高、具有正常电生理特性的犬心房肌细胞,能用于心房肌细胞离子通道的研究。     

乳大鼠心室肌培养细胞代替急性酶分离细胞用于起搏电流电生理研究的可行性探索

目的 探讨体外培养细胞代替急性酶分离细胞用于心室肌电生理研究的可行性.方法 采用膜片钳全细胞记录技术,对比分析心室肌细胞培养的不同时间与急性酶分离的心室肌细胞起搏电流间的电生理学差异.结果 短期的培养就会使细胞膜电容下降,各期培养细胞的膜电容小于急性酶分离细胞的膜电容[(56±7) pF,P＜0.05].4d后膜电容随着培养时间的延长有逐渐增大的趋势,但各期之间的膜电容差异无统计学意义(P＞0.05).各个培养时间段细胞的反转电位、电流密度、活化阈值、半最大激活膜电压(V0.5)等主要电生理指标之间差异无统计学意义(P＞0.05).结论 至少短期培养的乳大鼠心室肌细胞的起搏电流与急性酶分离细胞的起搏电流的主要电生理指标之间无明显差异,可以代替急性分离细胞应用于起搏电流的电生理研究.     

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…     

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.     

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.     

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.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.     

TCM Treatment of Ankylosing Spondylitis by Tonifying the Kidney and Strengthening the Governor Vessel

Ankylosing spondylitis is a chronic and progressive disorder with inflammation mainly involving the central axis joints. It mainly affects the cervical spine and the lumbosacral area, with the pathogenesis closely related to the kidney and the Governor Vessel (GV). TCM holds that the syndrome is deficiency in origin and excess in superficiality, which is due to insufficiency of the kidney, deficiency of GV, and blocking of the channels with the invasion of exogenous evil, leading to poor circulation of qi and blood and malnutrition of the bones, muscles and joints. The TCM method of tonifying the kidney and strengthening GV to regulate circulation of qi and blood and check the arthralgia pain should be adopted, with the Kidney-Tonifying and GV Strengthening Decoction (益肾强督汤) prescribed.     

IMPROVING P-gp EXPRESSION IN HUMAN MONONUCLEAR CELLS IN VITRO TRANSFECTED BY MULTIDRUG RESISTANCE-1 mRNA

CHEMOTHERAPY playsa greatrolein the treat- ment of malignanttumors,especiallyingynecolo- gicalones.But inanticancerchemotherapy,leuko-cytopeniaisfrequentlytheprimarydose-limitingsideeffect factor.Moreover,cancersarefrequentlychemoresistantbe-causeof overexpressionof P-glycoprotein(P-gp), which isencodedby multidrugresistancegene (MDR1 ) and detectableinup to50% ofhuman cancersand renderscellsresistancetoanticancerdrugs.The safetyand potentialtherapeuticbenefitof mdr1 gene transferredto h…