顺铂热化疗对骨肉瘤细胞株细胞毒性的实验研究

目的：研究顺铂热化疗对骨肉瘤细胞株(OS-732)细胞毒性的影响，探讨热疗在骨肉瘤顺铂化疗中增效作用的机理。方法：OS-732细胞株分别经不同温度的热疗、不同浓度顺铂化疗、43℃热疗 顺铂联合作用各1h，采用体外细胞毒性试验(MTT法)、流式细胞仪(FCM)分析及电子显微镜观察其对细胞毒性和细胞周期的影响，以及诱导细胞凋亡的情况。结果：41℃、43℃、45℃热疗对OS-732细胞株有明显的细胞毒性作用。在37℃恒温条件下，细胞株单独与不同浓度的顺铂作用1h，随着顺铂浓度的升高，其细胞毒性作用明显增加。43℃热疗 顺铂共同作用1h，其细胞毒性作用进一步增加，细胞毒性指数最高达72．37％；FCM分析显示对细胞周期有明显的影响，S期细胞比例明显增加，G2与M期细胞比例明显减少。43℃热疗、43℃热疗 顺铂联合作用各1h均出现凋亡细胞峰(亚G1峰)，细胞凋亡比例最高达56．47％；透射电镜观察有典型的细胞凋亡特征性变化。结论：顺铂热化疗有明显的增强OS-732细胞株的细胞毒性作用。43℃热疗、43℃热疗 顺铂均可诱导骨肉瘤细胞凋亡。43℃热疗诱导骨肉瘤细胞凋亡可能是热疗在骨肉瘤顺铂化疗中增效作用的潜在机制之一。     

浅蓝菌素在诱导骨肉瘤细胞株U2-OS凋亡过程中对Bcl-2和Bax的影响

目的研究凋亡蛋白Bcl-2、Bax是否参与浅蓝菌素（Cerulenin）诱发骨肉瘤细胞株U2-OS凋亡信号传导。方法采用Western—blotting检测Cerulenin作用前、后骨肉瘤细胞株U2—OS中凋亡蛋白Bcl-2和Bax表达变化情况,了解不同的Cerulenin剂量和作用时间骨肉瘤细胞株U2-OS中凋亡蛋白Bcl-2和Bax表达变化情况。结果骨肉瘤细胞株U2-OS中Bcl-2和Bax呈阳性表达,Cerulenin作用细胞株后Bcl-2表达量随着浓度增加和作用时间延长而递减,Bax表达量随着浓度增加和作用时间延长而递增。结论Cerulenin可能通过下调Bcl-2的表达和上调Bax的表达而诱导骨肉瘤细胞凋亡。     

热疗对纤维肉瘤S180株的抑瘤作用及其生物学机制

目的 探讨热疗对纤维肉瘤S180株的抑瘤作用及其生物学机制.方法 40只昆明小鼠,接种S180肿瘤细胞株后用加热进行实验干预,标本于实验后6、2、54h取材,原位末端标记法(TUNEL)检测凋亡细胞,流式细胞仪测定细胞周期分布,RT-PCR检测bcl-2基因的mRNA表达.结果 热疗对纤维肉瘤有显著的治疗作用,可以诱导纤维肉瘤S180株细胞凋亡,改变细胞周期分布,使发生G1期阻滞,下调Bcl-2基因.结论 热疗可以诱导纤维肉瘤S180株细胞凋亡,改变细胞周期分布,使发生G1期阻滞,下调bcl-2基因,为进一步的基因配合热放疗治疗肿瘤和选择肿瘤放化疗结合时机和化疗药物提供了理论基础.     

加热对骨肉瘤细胞系OS-9901和SOSP-M3的体外灭活作用

目的 探讨两株不同转移性成骨肉瘤细胞系的体外灭活最佳温度与时间。方法 不同温度不同时间对OS-9901和SOSP-M3细胞系加热作用后，用MTT法测定琥珀酸脱氢酶（SDH）活性以及用肿瘤干细胞集落形成法测定集落形成能力。结果 （1）两株不同转移性的成骨肉瘤细胞系对热的敏感性是一致的。（2）SDH的活性随温度升高和时间延长而降低。（3）随着加热温度升高和时间延长肿瘤干细胞的集落形成能力不断下降，45℃60min或46℃30min以上时细胞基本无集落形成能力。结论 50℃30min可以作为成骨肉瘤的加疗温度，时间。     

紫杉醇联合热疗对鼻咽癌CNE-1细胞株增殖及凋亡的影响

目的 探讨紫杉醇(Paclitaxel)联合热疗对鼻咽癌细胞株CNE-1增殖及凋亡的影响,为临床紫杉醇与热疗联合应用提供实验依据.方法 将不同温度(39℃、41℃和43℃)及不同浓度(0.01、0.10、1.00、10.00和100.00 nmol/L)紫杉醇采用分组设计(共15个组),各浓度组设37℃为对照组(共5个组).在紫杉醇对CNE-1细胞作用48h后加热30min.使用MTT法、克隆形成能力试验、细胞形态学、超微结构观察及琼脂糖凝胶电泳等方法 测定紫杉醇联合热疗对CNE-1细胞体外增殖抑制及细胞凋亡的影响.结果 加热41℃组细胞增殖率与其他不同温度同剂量组比较均降低,差异有显著性.与其他各组比较均降低,差异均有显著性(P<0.05);39℃和43℃组细胞增殖率与37℃对照组比较均升高,差异有显著性(P<0.05).琼脂糖凝胶电泳显示紫杉醇作用48h后便可出现DNA梯带,并呈时间剂量性增强.最低0.10nmol/L浓度可见DNA梯带.倒置显微镜和电镜下可见细胞损伤及凋亡细胞.结论 加热41℃时不同浓度紫杉醇对CNE-1细胞均有增殖抑制及诱导细胞凋亡作用;39℃或43℃热疗联合0.01 nmol/L紫衫醇化疗时CNE-1细胞增殖能力最强.     

放疗对人肝癌细胞株Hep-G2中Bax和Bcl-2表达的影响

目的:探讨放疗对人肝癌细胞株Hep-G2中Bax和Bcl-2蛋白表达水平的影响.方法:6MV-X射线照射细胞,采用免疫组织化学染色观察人肝癌细胞株中Bax和Bcl-2的表达.结果:Bax和Bcl-2的表达在照射时间和照射剂量之间存在交互作用(F=1.733,P=0.042;F=1.700,P=0.048).4Gy照射剂量作用细胞48h后,Hep-G2细胞中Bax的表达达高峰、Bcl-2的表达至低谷.结论:放射线可能通过促进Bax表达和抑制Bcl-2表达诱导肝癌细胞凋亡.     

羟基喜树碱对胃癌细胞的诱导凋亡及凋亡相关基因表达的影响

[目的]探讨羟基喜树碱(HCPT)对胃癌MGC-803细胞株的诱导凋亡和抗增殖作用及凋亡相关基因表达的影响.[方法]通过MTT比色法、HE染色、吖啶橙(AO)/溴化乙锭(EB)染色及各种显微镜观察,并且采用原位细胞凋亡检测法(TUNEL法)及免疫细胞化学法,观察HCPT对胃癌MGC-803细胞株的诱导凋亡和抗增殖作用及凋亡相关基因的表达情况.[结果]HCPT在质量浓度0.5~48.0 mg/L范围内对胃癌MGC-803细胞株均有抑制生长作用,并表现出显效关系;在倒置显微镜、HE染色光学显微镜、AO/EB染色荧光显微镜和透射电子显微镜下,均可见MGC-803细胞的凋亡形态学改变;不同质量浓度HCPT均可诱导胃癌MGC-803细胞凋亡,细胞凋亡率随药物质量浓度的增高而上升,当药物作用48 h时细胞凋亡率达高峰;增殖细胞核抗原Ki-67的阳性表达率随HCPT质量浓度的增高和作用时间的延长而降低;当12 mg/L HCPT作用于胃癌MGC-803细胞株48 h后,明显下调抑制凋亡基因Bcl-2的表达和上调促进凋亡基因Bax的表达.[结论]HCPT对胃癌MGC-803细胞株有诱导凋亡和抗增殖作用,其机制可能与下调抑制凋亡基因Bcl-2的表达和上调促进凋亡基因Bax的表达有关.     

加热诱导结肠癌细胞Lovo凋亡的研究

目的：观察加热对人结肠癌细胞株Lovo的凋亡作用。方法：用Hochest33258荧光染色、凝胶电泳和流式细胞术观察不同时间一温度作用下Lovo细胞的变化。结果：不同温度一时间处理的细胞均发生凋亡，出现明显的形态学改变及凋亡特有的亚二倍体峰和DNA梯度。其中43℃处理3、4h和44℃处理1、2h时细胞凋亡最明显，细胞周期也发生变化。结论：加热可诱导人结肠癌细胞株Lovo发生凋亡，并具有一定的周期特异性。     

Bim对热疗诱导的人宫颈癌Hela细胞凋亡的作用

目的研究Bim在热疗诱导的人宫颈癌Hela细胞凋亡中的作用。方法用不同温度(41、42.5、43.5℃)水浴对Hela细胞加热刺激,处理后继续培养24 h。用Hoechst 33258染色细胞核,计数细胞凋亡率;采用Western blot方法检测Bim及Caspase-3蛋白的表达水平;用lipofectamine 2000转染干扰片段后观察干扰效率及对凋亡的保护作用。结果人宫颈癌Hela细胞受到不同温度(41、42.5、43.5℃)刺激后,分别引起约(16.81±0.23)%、(29.52±0.56)%和(31.22±0.67)%细胞凋亡。Bim mRNA及蛋白质呈温度依赖地表达上调。小分子干扰Bim的表达后,细胞凋亡率从(30.51±0.54)%下降到si-Bim-1(12.52±0.67)%、si-Bim-2(15.23±0.43)%。结论 Bim介导了热疗诱导的人宫颈癌Hela细胞凋亡,可能在热疗治疗宫颈癌中发挥重要作用。     

聚焦超声诱导卵巢癌细胞株COC1凋亡及其机制的初步研究

目的:研究低频脉冲聚焦超声波对人卵巢癌细胞株COC1的抗肿瘤作用,并探讨其诱导细胞凋亡的分子机制.方法:以低频脉冲聚焦超声(超声频率0.8MHz,声强83.6W/cm2,脉肿持续时间:5μs)辐照卵巢癌COC1细胞10s,然后采用MTT法观察超声对COC1细胞增殖的抑制作用;流式细胞仪(FCM)、TUNEL法、荧光染色和光、电镜检测细胞凋亡;应用FCM免疫荧光技术观察Bcl-2、bax基因蛋白表达的变化.结果:超声可明显抑制存活卯巢癌COC1细胞的增殖,辐照后12h、24h、36h、48h,COC1细胞MTT光吸收值与对照组相比显著降低;FCM检测超声辐照后6h的COC1细胞凋亡率达(23.57±4.62)%;Bcl-2蛋白的荧光指数(FI)降低,而bax蛋白FI升高.同时TUNEL法观察到棕褐色着染的凋亡细胞;荧光显微镜和电镜下观察到凋亡小体形成等典型的形态学改变.结论:低频脉冲聚焦超声辐照对人卵巢癌细胞株COC1具有细胞毒作用,其机制为诱导辐照后存活的人卵巢癌细胞株COC1凋亡,bcl-2和bax蛋白可能参与了COC1细胞凋亡过程.     

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…     

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.     

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.