三苯氧胺治疗乳腺癌分子机制及其与CDK4、PCNA的关系

目的 探讨三苯氧胺(TAM)治疗乳腺癌分子作用机制及其与细胞周期的关系.方法 TAM作用于ER(-)小鼠乳腺癌MA782细胞,用流式细胞仪检测细胞凋亡率和细胞周期分布;MTT法检测细胞增殖活性;免疫法检测CDK4、PCNA蛋白表达量变化值,同时用病理图像分析软件进行半定量分析.结果 TAM(6、10 μmol/L)在体外能明显抑制MA782细胞生长,诱导细胞凋亡, TAM(6、10 μmol/L)作用于细胞24 h后,细胞凋亡率分别为7.04%﹑19.04%,与对照组比较结果有明显差异, TAM(10 μmol/L)作用于细胞24、48 h后细胞凋亡率从19.04%上升到51.27%,两时间段之间差异显著.免疫组织化学染色结果显示 TAM (2 μmol/L)作用于细胞不同时间后,CDK4、PCNA蛋白表达量无明显变化,而6、10 μmol/L TAM作用于细胞不同时间后,CDK4、PCNA蛋白表达量有不同程度的下降,与对照组相比差异显著(P＜0.05或P＜0.01).结论 TAM诱导MA782细胞凋亡的分子作用机制,可能与TAM下调细胞周期正性调节因子CDK4、PCNA蛋白表达量有关.     

三苯氧胺诱导MA782细胞凋亡与Cyclin D1的关系

目的　探讨三苯氧胺Tamoxifen(TAM)诱导ER( - )小鼠乳腺癌细胞MA782细胞凋亡及其分子作用机制。方法　TAM体外作用于MA782细胞 ,用MTT法检测细胞增殖活性 ,流式细胞仪检测细胞凋亡和细胞周期分布变化 ,免疫组化检测MA782细胞cyclinD1蛋白表达(并用病理图像分析软件进行半定量分析 ) ,RT -PCR检测MA782细胞cyclinD1mRNA表达情况。结果　TAM在体外能明显抑制MA782细胞生长 ,诱导细胞凋亡 ,下调cyclinD1mRNA及蛋白表达 ,免疫组化半定量分析显示 6μmol/LTAM作用MA782细胞 72小时后 ,cyclinD1蛋白平均灰度值从 1 32 .5± 0 .0 2下降至 1 2 6.1 8± 0 .0 3,1 0 μmol/LTAM作用 72小时后下降为 73.5 6± 0 .0 2 ,与对照组相比差异极其显著 (P <0 .0 1 )。RT -PCR检测结果显示 :2 μmol/LTAM组无明显改变 ,而 6、1 0 μmol/LTAM作用于细胞 2 4、48小时后 ,细胞cyclinD1mRNA表达呈现出不同程度的下降 ,与对照组比较有统计学意义 (P <0 .0 5或P <0 .0 1 )。结论　TAM诱导MA782细胞凋亡其分子作用机制 ,可能与下调细胞cyclinD1mRNA及蛋白表达有关     

Tamoxifen诱导MA782细胞凋亡与P21/CIP1、PCNA的关系

目的　探讨三苯氧胺 (TamoxifenTAM )诱导ER(- )小鼠乳腺癌细胞MA782细胞凋亡及其分子作用机制。方法　TAM体外作用于MA782细胞 ,用MTT法检测细胞增殖活性、流式细胞仪检测细胞凋亡和细胞周期分布 ,透射电镜观察细胞凋亡超微结构改变 ,免疫组化检测MA782细胞p2 1/CIP1、PCNA蛋白表达 ,并用病理图像分析软件进行半定量分析。 结果　 6 μmol/L、10 μmol/LTAM在体外能明显抑制MA782细胞生长 ,诱导细胞凋亡 ,作用 2 4小时细胞凋亡率分别为 7.0 4 %、19.0 4 % ,10 μmol/LTAM作用 4 8小时后细胞凋亡率从 19.0 4 %上升到 5 1.2 7% ,与对照组比较有显著差异 (P <0 .0 1)。 2 μmol/LTAM作用不同时间后 ,细胞上述蛋白无明显变化 ,而 6 μmol/L、10 μmol/LTAM作用不同时间后 ,PCNA蛋白表达有不同程度的下降 ,p2 1/CIP1蛋白表达有不同程度的上升与对照组相比差异显著 (P <0 .0 5或P <0 .0 1)。结论　TAM诱导MA782细胞凋亡其分子作用机制可能与下调细胞PCNA蛋白表达及上调 p2 1/CIP1蛋白表达有关。     

TGF-β1与TAM诱导MA782细胞凋亡的关系

目的探讨三苯氧胺Tamoxifen(TAM)诱导ER(-)小鼠乳腺癌细胞MA782细胞凋亡及其分子作用机制。方法TAM体外作用于MA782细胞。流式细胞仪检测细胞凋亡和细胞周期分布情况。MTT法检测细胞增殖活性。免疫组化法检测MA782细胞TGF-β1蛋白表达量,并用病理图像分析软件对蛋白表达进行半定量分析。RT-PCR法检测MA782细胞TGF-β1mRNA表达情况。结果TAM在体外能明显抑制MA782细胞生长,诱导细胞凋亡,上调TGF-β1mRNA及蛋白表达量,免疫组化半定量分析显示TGF-β1平均灰度值为(59.72±0.02),2μmol/L TAM作用48h后无明显变化,72h后上升到(97.1±0.03),6、10μmol/LTAM作用48h后分别上升为(83.2±0.04)、(96.83±0.02),72别上升到(121.75±0.03)、(139.01±0.05),与对照组相比差异极其显著(P<0.01)。RT-PCR检测结果显示2μmol/L TAM组无明显改变,而6、10μmol/LTAM作用于细胞24、48h后,细胞TGF-β1mRNA水平表现出不同程度的上升,与对照组比较有统计学意义(P<0.05或P<0.01)。结论TAM诱导MA782细胞凋亡,其分子作用机制可能与上调细胞TGF-β1mRNA及蛋白表达有关。     

姜黄素诱导髓系白血病细胞凋亡的分子途径

目的研究姜黄素诱导白血病细胞系NB4、K562、THP1凋亡的分子途径。方法培养上述3种细胞系，用不同浓度的姜黄素（25，12．5，6．25，3．125，0μmol／L）作用24h和25umol／L姜黄素作用不同时间（0，12，24，48h），流式细胞仪测定细胞凋亡率及Fas抗原（CD95）表达，Western blot测定凋亡蛋白Caspase-8、Caspase-9的表达变化。结果①姜黄素以时间和浓度依赖的方式诱导白血病细胞系NB4、K562、THP-1凋亡，25μmol／L姜黄素作用48h凋亡细胞超过50％。②25μmol／L姜黄素作用24h后Fas抗原表达明显升高（P〈0．01）。③25μmol／L姜黄素作用24h后凋亡蛋白Caspase-8、Caspase-9的表达明显增高（P〈0．01）。结论姜黄素诱导白血病细胞凋亡涉及死亡受体参与的外源性途径和线粒体参与的内源性凋亡途径。     

高浓度葡萄糖损伤人腹膜间皮细胞分子机制的研究

目的：探讨高浓度葡萄糖损伤人腹膜间皮细胞的机制。方法：（1）人腹膜间皮细胞分别经含1．5％、2．5％、4．25％葡萄糖的M199培养基培养48小时后，检测间皮细胞线粒体膜电位（Ψ）、凋亡率、caspase-3活性、bax和bcl-2基因mRNA表达；（2）分别用0．1μmol／L和0．01μmol／L环孢素A（CsA）与4．25％葡萄糖共同作用人腹膜间皮细胞，检测细胞凋亡率和caspase-3活性。结果：（1）随着葡萄糖浓度增加，Ψ丧失的细胞百分率、bax mRNA表达量、caspase-3活性、凋亡率增加（P〈0．05），而bcl-2 mRNA表达量减少；（2）随着葡萄糖浓度增加，bax mRNA的表达量与皿丧失的间皮细胞百分率呈显著正相关（P〈0．01）；bcl-2 mRNA的表达量与Ψ丧失的间皮细胞百分率呈显著负相关（P〈0．01），Ψ丧失的间皮细胞百分率与间皮细胞凋亡率、caspase-3活性呈显著正相关（P〈0．01）：（3）0．1μmol／L CsA组间皮细胞凋亡率和caspase-3活性显著低于高糖对照组（P〈0．05）。结论：（1）高糖可能通过上调bax基因表达和下调bcl-2基因表达，诱导人腹膜间皮细胞线粒体活化、caspase-3活化和凋亡。（2）高糖诱导人腹膜间皮细胞caspase-3活化和凋亡可能依赖于线粒体活化。     

紫杉醇诱导黑色素瘤细胞凋亡的研究

目的：探讨紫杉醇体外抑制人黑色素瘤细胞（A375细胞）增殖及诱导凋亡作用机制。方法：四甲基偶氮唑蓝（MTT）还原法测定细胞活力；光镜和电镜观察细胞形态变化；流式细胞术（FCM）检测细胞周期和凋亡率。结果：紫杉醇通过诱导细胞凋亡抑制A375细胞生长。并呈时间和剂量依赖性（P〈0．01）；当0．01-1μmol／L紫杉醇作用于培养细胞24h后即可引起猾亡小体等典型细胞凋亡形态学改变；FCM分析显示：紫杉醇在0．001μmol／L即可诱导细胞凋亡，但不影响细胞周期，在0．01μmol／L时使G0／G1期细胞明显减少，在0．1μmol／L以上时使细胞发生G2／M期阻滞。结论：紫杉醇具有抑制A375细胞生长和诱导细胞凋亡作用。不同浓度紫杉醇诱导细胞凋亡的作用机制不同。     

巨幼细胞贫血所致黄疸分析

目的分析巨幼细胞贫血（MA）患者的血清胆红素浓度改变。方法选择47例经骨髓象检查确诊的初诊MA患者,另选取41例无贫血健康人员作为对照组。结果MA组、对照组直接胆红素（DBil）均数分别为（9．07±4．43）μmol／L及（3．614±1．29）μmol／L,P〈0．0001;两组间接胆红素（IBil）中位数分别为18．950μmol／L、7．340μmol／L,P=0．0000。MA患者中,37例（78．72％）总胆红素（TBil）〉17．μmol／L,13例（27．66％）TBil〉34．2μmol／L,30例（63．83％）DBil〉6．8μmoL／L,最大值为19．42μmoL／L,36例（76．60％）IBil〉14．0μmol／L,最大值为98．07μmol／L。37例伴黄疸MA患者中,4例（10．81％）DBiL／TBil〈0．20,33例（89．19％）DBil／TBil为0．20～0．50。结论MA患者中,近3／4者TBil升高,近3／10呈显性黄疸,近9／10者MA黄疸类似肝细胞性黄疸,需与肝病相鉴别。     

激光共聚焦显微镜观测蛋白酶抑制剂MG-132诱导人脑胶质瘤细胞凋亡的研究

目的探讨MG-132诱导人脑胶质瘤细胞SHG-44损伤中钙超载的作用。方法通过传代方法培养人脑胶质瘤细胞。选取MG-132处理24小时,采用处理前（对照组）和不同浓度（5、10、15、50μmol／L）处理后的胶质瘤细胞,采用MTT比色法检测细胞活力,流式细胞仪检测细胞凋亡情况,激光共聚焦检测细胞内钙离子荧光强度。结果MTT比色法检测发现,MG-132处理24h后,SHG-44细胞活力明显下降43％（P〈0．05）,且随着MG-132浓度的加大,继续呈现显著下降的趋势（P〈0．05）。流式细胞仪检测结果发现,与对照组比较,MG-132作用24h后,各组细胞凋亡率显著升高（P〈0．05）,5、10、15μmol／L组细胞调往情况相似（P〉0．05）,但50μmol／L细胞凋亡率升高较为明显（P〈0．05）。激光共聚焦测钙离子荧光强度结果显示,与空白对照组相比钙离子荧光强度明显升高（P〈0．05）,5、10、15μmol／L组钙离子荧光强度相似（P〉0．05）,但50μmol／L细胞钙离子荧光强度明显升高（P〈0．05）。结论MG-132处理可以通过激发细胞内钙超载诱导胶质瘤细胞凋亡。     

环境毒素导致多巴胺能神经元蛋白水解应激的实验研究

目的探讨环境毒素诱发蛋白水解应激在多巴胺能神经元变性死亡中的作用。方法6-羟多巴（6-OHDA）、1-甲基-4-苯基吡啶离子（MPP^＋）、鱼藤酮作用于神经生长因子诱导的PC12细胞,MTT法检测各种药物的神经毒性作用,应用共聚焦免疫荧光双标染色观察药物处理后细胞内仪．突触核蛋白及泛素化蛋白的表达及聚集。荧光分光光度计测量各组细胞蛋白酶体中各蛋白水解酶活性的变化。结果3种神经诱变剂处理PC12细胞后,细胞活力呈浓度依赖性下降,其中6-OHDA 100μmol／L、MPP^＋ 75μmol／L、鱼藤酮20nmol／L作用24h使细胞活力分别下降52％、44％、40％（P〈0．01）。共聚焦显微镜观察显示药物处理后细胞胞质内α-突触核蛋白及泛素表达上调,并形成蛋白颗粒物。酶活性测定提示MPP^＋75μmol／L、鱼藤酮20nmol／L处理使胰蛋白酶,糜蛋白酶及PgH水解酶活性明显下降（P〈0．05）,而6-OHDA 100μmol／L处理使细胞蛋白酶水解能力轻度降低,差异无统计学意义（P〉0．05）,6-OHDA 200μmol／L则诱导3种蛋白酶活性进一步下降,荧光指数分别为7．23±0．58,79．60±2．73和4．23±0．49（正常组为13．90±1．75,99．30±5．23和6．90±0．60,P〈0．01）。结论神经诱变剂可导致多巴胺能神经元泛素蛋白酶体功能失调及仪．突触核蛋白和泛素化蛋白的积聚,诱发蛋白水解应激异常状态。     

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…     

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.     

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.