热休克反应对卵巢癌细胞凋亡影响的研究

目的 :探讨热休克反应对卵巢癌细胞A2 780凋亡的影响。方法 :将体外培养的卵巢癌细胞A2 780分为非热休克组 (NHS)和热休克组 (HS) ,两组分别由三组浓度放线菌素D(Act-D)诱导。采用脱氧核苷酸末端转移酶介导的dUTP缺口末端标记法 (TUNEL)和3H -TdR掺入试验检测肿瘤细胞凋亡指数 (AI)及DNA合成情况。结果 :HS组比NHS组凋亡指数明显下降 (P <0 .0 1 ) ;cpm值明显升高 (P <0 .0 1 ) ,呈现剂量依赖效应。结论 :热休克反应对卵巢癌细胞A2 780经Act -D诱导凋亡具有抑制作用 ,此作用与Act-D呈剂量依赖效应。     

肺癌细胞凋亡中放线菌素D的作用研究

目的:探讨热休克蛋白在Act-D诱导肺癌GLC-82细胞凋亡中的作用,为进一步探讨HSP抗凋亡机制及肺癌生物治疗提供实验依据。方法:体外培养的肺癌GLC-82细胞于RPMI1640、10%胎牛血清、5%CO2孵育,取指数生长期细胞1×105/ml接种反应板24h,分为非热休克组(NHS)和热休克组(HS),两组分别由不同浓度(0,5,10,20μg/m1)放线菌素D(Act-D)诱导,作用4 h。采用脱氧核苷酸末端转移酶介导的dUTP缺口末端标记法(TUNEL)和3H-胸腺嘧啶脱氧核苷(3H-TdR)掺入试验检测肿瘤细胞凋亡指数(AI)及DNA合成情况。结果:HS组比NHS组凋亡指数明显下降(P<0.01);cpm值明显升高(P<0.01),呈现剂量依赖效应。结论:热休克反应对肺癌GLC-82细胞经Act-D诱导凋亡具有抑制作用,此作用与Act-D呈剂量依赖效应。     

TRAIL与GX15-070联用诱导卵巢癌细胞凋亡的研究

[目的：探讨联合应用肿瘤坏死因子相关凋亡诱导配体（TRAIL）和GX15-070诱导卵巢癌细胞系A2780细胞凋亡的作用及其机制。方法：以联用或未联用GX15-070的TRAIL作用于卵巢癌A2780细胞,采用MTT法检测细胞存活率,Annexin V-FITC/PI双染流式细胞术检测细胞凋亡,JC-1染色流式细胞术检测细胞线粒体膜电位（?ψm）,比色测定试剂盒检测caspase-9、caspase-8和caspase-3活性。结果：TRAIL能在一定程度上降低卵巢癌A2780细胞的存活率、诱导癌细胞凋亡、降低?ψm、提高caspase-9、caspase-8和caspase-3活性,而GX15-070能显著增加TRAIL降低卵巢癌A2780细胞的存活率、诱导癌细胞凋亡、降低?ψm、提高caspase-9和caspase-3活性的效应。结论：GX15-070通过凋亡信号通路的内源性途径,促进卵巢癌A2780细胞对TRAIL诱导凋亡效应的敏感性。     

OSTP增强紫杉醇诱导卵巢癌A2780细胞凋亡的作用

目的 研究卵巢癌特异性结合肽（OSTP）及联合紫杉醇对卵巢癌A2780细胞生长及凋亡的影响.方法 体外培养卵巢癌A2780细胞,随机分为4组：溶剂对照组（1640和0.1％ DMSO组）、OSTP组、紫杉醇组、OSTP联合紫杉醇组.采用四甲基偶氮唑盐（MTT法）检测OSTP对A2780细胞生长的影响;用Annexin-V-FITC和PI双染法标记A2780细胞,通过流式细胞仪检测细胞凋亡情况.结果 MTT法检测OSTP以不同浓度作用于卵巢癌A2780细胞时,可对该细胞的生长起到抑制作用,与对照组相比差异有统计学意义（P＜0.01）.应用流式细胞技术检测细胞凋亡发现：不同浓度的OSTP（20、40、80、160、320μmol/L）对A2780细胞的凋亡率分别7.88％、8.348％、8.727％、9.393％和10.68％,与对照组相比差异有统计学意义（P＜0.01）,提示OSTP可诱导细胞凋亡.OSTP和紫杉醇联合应用,在不同的预处理组（加OSTP80μ mol/L 0、3、6、12h）后再加紫杉醇10μmol/L作用48h,结果发现细胞凋亡率分别是39.40％、53.09％、48.18％和45.62％,均高于OSTP和紫杉醇单独用药及两者的凋亡率相加值,且差异有统计学意义（P ＜0.001）.提示OSTP能增强紫杉醇诱导的卵巢癌A2780细胞凋亡作用.结论 OSTP对卵巢癌A2780细胞有生长抑制及凋亡作用,尤其是OSTP能增强紫杉醇诱导卵巢癌A2780细胞凋亡的作用,对于探讨和开发OSTP作为靶向化疗增敏剂治疗卵巢癌有良好的应用前景.     

卵巢癌紫杉醇耐药细胞耐药性和髓样分化因子88关系的研究

目的：研究卵巢癌亲本A2780细胞与卵巢癌紫杉醇耐药细胞A2780/Taxol生物学特性及与髓样分化因子（Myeloid dif－ferentiation factor 88,MyD88）的关系。方法：应用CCK-8法检测A2780细胞和A2780/Taxol细胞的耐药性和耐药指数,采用流式细胞法检测A2780和A2780/Taxol细胞的细胞周期。应用罗丹明123（Rhodamine 123,Rh123）排出实验测定P-糖蛋白（P-gly－coprotein,P-gp）表达。应用Western blot检测A2780和A2780/Taxol细胞中P-gp的表达情况。应用细胞免疫化学检测A2780和A2780/Taxol细胞中MyD88、P-gp的表达情况。结果：A2780与A2780/Taxol相比,A2780/Taxol生长缓慢,呈G1期阻滞;CCK-8结果显示A2780/Taxol细胞对紫杉醇的IC50为（36.81±2.05） μg/ml,A2780细胞对紫杉醇的IC50为（1.40±0.18） μg/ml;两者的耐药指数（Resistance index,RI）为26.35。Rh123排出实验显示A2780/Taxol平均荧光强度显著降低。Western blot检测发现A2780/Taxol的P-gp表达比A2780明显升高（P<0.05）。细胞免疫化学检测发现A2780/Taxol中的P-gp和MyD88表达明显高于A2780细胞。结论：A2780/Taxol细胞株具有明确的耐药性,紫杉醇耐药与MyD88的表达密切相关,该细胞株可用于卵巢癌紫杉醇耐药的基础研究。     

HSR对TNF-α诱导的肺癌细胞凋亡的拮抗作用

目的 研究热休克反应（HSR）在肺癌细胞GLC-82增殖及凋亡过程中的作用,为肺癌的临床治疗提供实验参考。方法 用体外培养的肺癌细胞GLC-82细胞分为热休克（HS）组和非热休克（NHS）组,每组分为对照组和TNF-α组,TNF-α的浓度分别为25μg／L、50μg／L和100μg／L（T1～T3）,采用脱氧核苷酸末端转移酶介导的dUTP缺口末端标记法（TUNEL）检测肿瘤细胞凋亡指数（AI）;噻唑蓝比色法（MTT、）检测肿瘤细胞的细胞增殖情况;以3H-TdR掺入试验检测肿瘤细胞的DNA合成情况。结果 HS组细胞凋亡百分率明显减少;MTT显示HSR可以使细胞GLC-82抵抗TNF-α的细胞毒作用明显增强,促进细胞增殖;DNA合成明显增加;且与TNF-α呈剂量依赖效应。结论 HSR在肺癌细胞GLC-82生长增殖过程中起促进作用,而对细胞凋亡起拮抗作用。     

稳定过表达XAF1基因对A2780卵巢癌细胞生物学功能的影响

目的 构建稳定过表达XAF1基因A2780卵巢癌细胞株,并观察XAF1基因对卵巢癌细胞增殖、凋亡、细胞周期及对紫杉醇敏感性的影响。方法 分别将质粒pcDNA3.1（+）和pcDNA3.1（+）-XAF1转染至卵巢癌细胞A2780,通过质粒抗性标记（遗传霉素G418）筛选得到阴性对照细胞株（A2780/Negative control, A2780/NC）和稳定表达XAF1的细胞株（A2780/XAF1）;通过行细胞克隆形成实验和CCK8实验检测细胞增殖及对紫杉醇的敏感性,流式细胞仪检测细胞周期及细胞凋亡。结果 成功构建了稳定表达XAF1的卵巢癌细胞A2780/XAF1,细胞形态无明显变化;与A2780/NC相比,A2780/XAF1克隆形成能力能力更低（P= 0.0016）,细胞贴壁后第1天和第3天增殖活性更低（P=0.009,0.0035）,两组细胞的细胞周期分布差异存在统计学意义（P< 0.0001）,两两比较结果显示A2780/XAF1组G2-M期细胞百分比显著增加（P<0.001）。在无凋亡刺激、无血清培养及紫杉醇诱导下,A2780/XAF1的凋亡率均比A2780/NC更高（P<0.001）;在不同紫杉醇浓度作用下,A2780/XAF1的增殖活性均显著低于A2780/NC（P<0.001）,且A2780/XAF1的紫杉醇半数抑制浓度显著低于A2780/NC。结论 成功构建稳定表达XAF1的卵巢癌细胞A2780/XAF1;XAF1调控卵巢癌细胞A2780的增殖、凋亡及细胞周期,并增加了卵巢癌对紫杉醇的敏感性。     

RNA干扰靶向沉默FLIP基因对卵巢癌细胞A2780生物学特性的影响及生长抑制作用

目的探讨靶向FLIP基因的小干扰RNA（siRNA）对人卵巢癌细胞A2780生物学特性的影响及生长抑制作用。方法设计并体外化学合成FLIP序列特异性双链RNA，在脂质体（LipofectamineTM2000）介导下转染人卵巢癌细胞株A2780。采用半定量RT-PCR和Western blot法检测FLIP siRNAs转染前后A2780细胞FLIP基因mRNA和蛋白表达的变化,并筛选出抑制作用最强的FLIP-siRNA。采用四甲基偶氮唑盐（MTT）法、Annexin-V-PI双染法流式细胞术（FCM）分别检测FLIP-siRNA转染对A2780细胞的生长抑制作用和对凋亡的影响。结果特异性FLIP-siRNAs片段能有效降低A2780细胞中FLIP的 mRNA和蛋白水平（P＜0.01）,最大抑制率分别为77.4％和66.1％；转染FLIP-siRNA后，A2780细胞的生长活力明显下降（P＜0.05），RNA干扰组的细胞凋亡也显著增加（P＜0.05）。结论靶向FLIP基因的siRNAs可在转录和翻译水平抑制FLIP基因表达；并可抑制卵巢癌A2780细胞生长，促进其凋亡。     

体外三维培养卵巢癌A2780细胞及其多药耐药机制的实验研究

目的建立卵巢癌A2780细胞形成的多细胞团簇模型，探讨凋亡在卵巢癌细胞群集耐药中的作用。方法以三维培养方法获得人卵巢癌A2780多细胞球体（MCS）模型，单层细胞（MC）为对照；MTT法和liquidoverlay技术检测细胞的药物敏感性；流式细胞术检测细胞周期和凋亡的变化；RT-PCR分析凋亡相关基因caspase-3和bcl-2的表达变化。结果细胞在培养2d后即可形成由多个细胞组成的细胞团，培养5d后，形成有多层细胞结构的细胞球；与A2780／MC相比，A2780／MCS细胞药物敏感性及细胞凋亡率均显著下降，差异有显著性；而与A2780／MC相比，凋亡诱导基因caspase-3在A2780／MCS细胞中明显下降，凋亡抑制基因bcl-2明显增高（P〈0．05）。结论卵巢癌细胞群集耐药可能与肿瘤细胞凋亡相关基因caspase-3和bcl-2的表达相关。     

XIAP基因与耐紫杉醇卵巢癌细胞A2780/Taxol耐药关系的研究

目的 探讨X连锁凋亡抑制蛋白（XIAP）基因与卵巢癌对紫杉醇耐药的相关性。方法 以5、10、20、40、80、160、320 ng/mL的紫杉醇处理卵巢癌紫杉醇敏感细胞A2780和耐药细胞A2780/T,噻唑蓝（MTT）比色法检测各组细胞的抑制率（IR）, 逆转录实时荧光定量PCR（RT-qPCR）、蛋白质印记（Western blot）检测紫杉醇100 ng/mL处理A2780和A2780/T细胞后XIAP基因和蛋白的表达;将A2780/T细胞分为空转染组（转染空载体质粒）、小干扰RNA（siRNA）-XIAP（siRNA-XIAP）组（转染siRNA-XIAP质粒）、非特异性转染组（转染非特异性质粒）、空白对照组（不转染质粒）,RT-qPCR和Western blot检测各组细胞XIAP基因和蛋白的表达,并加入含不同质量浓度的紫杉醇（0、1 000、1 500、2 000、2 500 ng/mL）,流式细胞仪检测细胞凋亡率。结果 不同质量浓度紫杉醇处理后,A2780细胞IR随紫杉醇质量浓度的增加而增高（P<0.05）,A2780/T细胞IR无明显变化。紫杉醇100 ng/mL处理A2780和A2780/T细胞后,A2780细胞XIAP mRNA的表达低于紫杉醇未处理组（P<0.05）,A2780/T细胞中XIAP mRNA的表达与紫杉醇未处理组差异无统计学意义(P>0.05),但A2780/T细胞XIAP mRNA和蛋白表达均高于A2780紫杉醇处理组（P均<0.05）;siRNA-XIAP组XIAP mRNA和蛋白的表达均低于其他各组（P<0.05）,在紫杉醇2 000 ng/mL及2 500 ng/mL作用下,siRNA-XIAP组凋亡率高于其他各组（P<0.05）。结论 卵巢癌对紫杉醇耐药与XIAP基因的高表达有关,特异性的siRNA可通过降低XIAP的表达,促进细胞凋亡,增加耐药癌细胞对紫杉醇的敏感性。     

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…     

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.     

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.     

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.     

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.     

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.