吗啡对大鼠培养海马神经元钙离子作用机制的研究

目的 研究吗啡对海马神经元游离子浓度([Ca^2 ]i)影响的机制，探索吗啡成瘾的神经生物学机制及对吗啡成瘾可能的治疗途径。方法 运用新型荧光探针Fluo-4 , 利用激光共聚焦显微镜研究吗啡对大鼠培养海马神经元[Ca^2 ]i作用机制。结果 10μmol/L吗啡急性刺激引起海马神经元[Ca^2 ]i升高，μ阿片受体选择性拮抗剂CTOP（1μmol/L）不能阻断吗啡引起的细胞内[Ca^2 ]i增加，δ2阿片受体选择性拮抗剂Naltrindole(1μmol/L）阻断了吗啡引起的细胞内[Ca^2 ]i反应，特异性的内质网钙泵抑制剂Thapsigargin(TG,1μmol/L）预处理海马神经元组断吗啡引起的细胞内[Ca^2 ]i增加，L－钙通道阻断剂Verapamil(20μmol/L）预处理海马神经元不能完全抑制吗啡引起的细胞内[Ca^2 ]i增加；100μmol/L吗啡长时程（24h）作用于海马神经元，细胞内[Ca^2 ]i升高，加入10μmol/L纳络酮急性戒断后，不能阻断细胞内[Ca^2 ]i升高，反而引起[Ca^2 ]i异常升高。结论 吗啡急性刺激引起的海马神经元内钙增加主要来源于δ2阿片受体介导的IP3敏感的钙库释放。     

2,3,7,8-TCDD对培养乳鼠心肌细胞内游离钙离子浓度影响的研究

目的 测定培养SD大鼠乳鼠心肌细胞内游离钙离子浓度[Ca^2＋]i,观察2,3,7,8-四氯二苯并二噁英（2,3,7,8-tetrachlorodibenzo-p-dioxin,以下略为TCDD）对心肌细胞[Ca^2＋]i的影响。方法将实验动物分为0．05、0．5、5、10μg／kg4个染毒组,采用Fluo-3／AM同时加入Pluronic F-127作为细胞内游离钙离子的荧光探针,负载培养的心肌细胞,应用共聚焦激光扫描显微镜（CLSM）技术检测空白组及染毒组乳鼠心肌细胞[Ca^2＋]i。结果静息时,心肌细胞[Ca^2＋]i为（77．68-4-12．00）nmol／L。染毒组心肌细胞的[Ca^2＋]i显示不同程度的升高,且呈剂量依赖性。结论应用Fluo-3／AM和Pluronic F-127结合CLSM技术可以精确的反映细胞内游离钙离子浓度,TCDD可能通过某种机制增加心肌细胞[Ca^2＋]i从而对乳鼠心脏功能产生一定损伤。     

肾上腺素对视网膜色素上皮细胞ATP酶活性及细胞内cAMP和钙离子浓度的影响

目的 研究。肾上腺素对培养的成人视网膜色素上皮细胞（retinal pigment epithelium，RPE）钠-钾三磷酸腺苷酶（Na^＋-K^＋ATP酶）、钙离子三磷酸腺苷酶（Ca^2＋-ATP酶）和细胞内环腺嘌呤核苷酸（CAMP）、[Ca^2＋]i的影响与超微结构的变化，以探讨视网膜下液转运和吸收的机制。方法 采用不同浓度。肾上腺素诱导RPE细胞24h后，采用ATP酶试剂盒检测细胞Na^＋-K^＋ATP酶、Ca^2＋-ATP酶活力，^125I-cAMP放免试剂盒检测胞内cAMP浓度，Fura-2／AM荧光探针检测细胞内[Ca^2＋]i，并采用透射电镜技术观察细胞超微结构的改变。结果 0．1～100nmol／L。肾上腺素能显著降低RPE细胞Na^＋-K^＋ATP酶活性，并提升Ca^2＋-ATP酶活性（均P〈0．01）。10nmol／L。肾上腺素可明显升高REP细胞内cAMP水平，加入10nmol／L。肾上腺素30min可明显引起RPE细胞内[Ca^2＋]i降低，透射电镜观察发现。肾上腺素诱导后的RPE细胞内水肿样改变。结论 肾上腺素可能通过β-肾上腺素能受体-cAMP途径影响RPE细胞离子与液体转运。     

大麻素WIN55，212-2对体外培养牛眼小梁细胞cAMP表达的影响

目的通过研究大麻素WIN55，212-2对体外培养牛眼小梁细胞（BTMC）环腺苷酸（cAMP）表达的影响，探讨其增加房水排出而降低眼压的机制。方法对BTMC进行体外原代培养及传代培养，应用免疫组化的方法鉴定细胞。然后用不同浓度的WIN55，212-2作用于BTMC，放射免疫法检测细胞上清液中cAMP的水平。结果WIN55，212-2浓度在0．1-20μmol／L范围内，培养液中cAMP水平随浓度增加而显著性升高，若WIN55，212-2浓度继续增高（40-60btmol／L），cAMP水平不再升高。结论WIN55，212-2可能通过提高BTMC内cAMP水平而起到降低眼压的作用。     

嘌呤受体P2X7激活介导大鼠视网膜神经节细胞死亡的实验研究

 【目的】研究嘌呤受体P2X7激动剂、拮抗剂对大鼠视网膜神经节细胞（RGCs）的影响。【方法】（1）对新生Long-Evan大鼠进行上丘注射荧光标记物Aminostilbamidine以标记RGCs。检测P2b激动剂BzATP（0、50、100、500μmol／L）和特异性拮抗剂OxATP（100μmol／L）对体外培养RGCs存活率的影响；（2）体外培养未经Aminostilbamidine标记的新生大鼠RGCs，以10μmol／L钙离子（Ca^2＋）荧光染料Fura-2标记后。利用Ca^2＋影像测定仪分别测定P2X7激动剂BzATP（50μmol／L）和拮抗剂OxATP（100μmol／L）对RGCs胞内Ca^2＋浓度的影响。【结果】P2X7受体激动剂BzATP可引起体外培养的RGCs死亡，反应呈剂量依赖性，EC50=35μmol／L。在50μmol／L浓度下，BzATP约杀死30％的RGCs；而100μmol／L OxATP则可明显减轻BzATP对RGCs的毒性作用，使RGCs存活率从77％±4％提高至96％±3％（P〈0．001）。BzATP可引起RGCs胞内Ca^2＋持续升高，在50μmol／L浓度下可使胞内Ca^2＋升高（1022±113）nmol／L。在OxATP作用下，BzATP介导的Ca^2＋升高幅度显著降低，仅升高（63±13）nmol／L（P〈0．001）。【结论】嘌呤能P2X7受体激活可导致大鼠视网膜神经节细胞死亡和胞内钙离子浓度升高。     

大麻素对大鼠三叉神经节神经元γ-氨基丁酸激活电流的抑制作用

目的探讨人工合成大麻素WIN55,212-2对大鼠三叉神经节神经元γ-氨基丁酸(GABA)激活电流(IGABA)的调制作用。方法采用全细胞膜片钳技术。结果①实验中大部分受检细胞(91.84%,99/108)对胞外给予GABA(10～1000μmol/L)敏感,可记录到具有浓度依赖性的内向电流,该电流可被GABAA受体特异性拮抗剂荷包牡丹碱(Bicuculline)阻断。②预加WIN55,212-2(0.03～10μmol/L)对IGABA产生抑制作用,该抑制作用呈可逆性、浓度依赖性和非电压依赖性。WIN55,212-2使IGABA的量效曲线明显下移,而两者的阈值基本不变;最大反应浓度时IGABA幅值减少了(48.83±4.78)%;两条曲线的半数有效浓度(EC50)值比较接近(36.85μmol/Lvs25.76μmol/L)。③WIN55,212-2对IGABA的抑制作用可被大麻素CB1受体选择性拮抗剂AM281阻断,不能被大麻素CB2受体选择性拮抗剂AM630阻断。细胞外灌流蛋白激酶C(PKC)的抑制剂BIM可部分逆转WIN55,212-2对IGABA的抑制作用。结论大麻素WIN55,212-2作用于CB1受体,部分通过激活PKC途径来减少GABAA受体介导的电流,加强突触前抑制作用,这可能是大麻素的外周镇痛机制之一。     

生长抑素对人胰腺癌细胞株ASPC—1的生长调控

目的：观察生长抑素（SS）对人胰腺癌细胞ASPC－1的生长作用；测定神经鞘磷脂（SM）、环磷酸腺苷（cAMP）和细胞内[Ca^2 ]i水平。方法：细胞培养，分别加入不同浓度的的SS。应用MTT法来观察细胞增殖程度；薄层层析法测定细胞内SM，放射免疫法测定细胞内cAMP含量；荧光法测定[Ca^2 ]i。结果：不同浓度的SS（10^-6～10^-12mol/L）均能有效地抑制ASPC－1的生长，能抑制ASPC－1细胞内SM，cAMP生成和细胞内[Ca^2 ]i水平（P＜0．01），cAMP生成量和细胞内[Ca^2 ]i水平与SS浓度呈负相关。结论：生长抑素能抑制ASPC－1细胞的生长，经过特异性受体调节。     

吗啡对大鼠海马星形胶质细胞[Ca2+]i的影响

目的 研究吗啡对大鼠海马星形胶持细胞[Ca^2 ]i的影响，探索吗啡对星形胶质细胞的作用效应。方法 运用荧光探针Fluo-4，利用激光共聚焦显微镜研究培养纯化大鼠海马星形胶质细胞[Ca^2 ]i的变化。结果 经0.5,1,2,10μmol/L浓的吗啡短期处理3d后，细胞内[Ca^2 ]i浓度与正常细胞比较差异无显著性（P＞0．05）。吗啡急性作用于正常星形胶质细胞后，[Ca^2 ]i呈单峰样升高；而作用于吗啡处理后的细胞，作用浓度需高于处理浓度出现[Ca^2 ]i呈单峰样升高的现象，纳络酮可阻断上述效应，NMDA受体阻断剂MK－801则不能。结论 星形胶质细胞[Ca^2 ]i对吗啡的反应性变化可能参与吗啡耐受的形成。     

生长抑素抑制胰腺癌细胞株生长的作用

目的：观察生长抑素对人胰腺癌细胞株HS766T生长的作用；测定神经鞘磷脂（sphingomyolin,SM）、cAMP、细胞内Ca^2 水平，探讨生长抑素的受体后信息传递作用。方法：细胞培养，分别加入不同浓度的生长抑素，PACAP1－38。应用MTT法来观察细胞增殖程度；薄层层析法测定细胞内SM，放免法测定细胞内cAMP含量；Fura-2/AM测定[Ca^2 ]i。结果：不同浓度的生长抑素（10^-6-10^-12mol/L）均能有效地抑制S766T的生长。能抑制HS766T细胞内SM、cAMP生成和细胞内Ca^2 水平，cAMP生成量和细胞内Ca^2 水平与生长抑素浓度呈负相关。结论：生长抑素能抑制HS766T细胞的生长，它的抑制作用是通过受体介导的。     

氯胺酮、咪达唑仑及丙泊酚对发育期神经元细胞内钙的影响

目的：探讨氯胺酮、咪达唑仑及丙泊酚对大鼠发育期海马神经元细胞内钙浓度的影响．方法：将原代培养第5日的大鼠海马神经元用10μmol／L的Ca2＋指示剂Fluo-4共孵育30min洗涤后,分别加入150μmol／L氯胺酮,咪达唑仑3μmol／L,丙泊酚10μmol／L,采用激光共聚焦显微镜选定多个细胞分别测定荧光强度的变化．结果：氯胺酮使体外培养第5日的海马神经元代表钙浓度的荧光强度明显下降[（987±307）vs（766±226）,P〈0．05],咪达唑仑,丙泊酚均使体外培养5d的海马神经元荧光强度明显升高[（1707±514）vs（2663±572）,（1057±353）vs（1749±708）,P〈0．05]．结论：阻滞NMDA受体的氯胺酮降低发育期海马神经元细胞内钙浓度,而兴奋GABA。受体的咪达唑仑,丙泊酚则会升高细胞内钙浓度．     

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…     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.