甲基莲心碱增强阿霉素抑制骨肉瘤作用

目的探讨甲基莲心碱增强阿霉素抑制裸鼠体内骨肉瘤生长的作用。方法将人成骨瘤细胞(MG-63)接种于BALB／C裸鼠,建立人成骨肉瘤裸鼠移植模型。随机分4组:生理盐水组 (Ns)、甲基莲心碱组(Nef)、阿霉素组(Adr)、甲基莲心碱+阿霉素组(Nef+Adr)。腹腔内注射以上药物,Nef浓度3．0mg／kg体重、Adr 2．0mg／kg体重。测量瘤重、计算瘤重抑制率、制病理切片,在光镜下观察肿瘤形态与改变。流式细胞术检测肿瘤原发灶细胞凋亡。结果 Ns、Nef、ADR、Nef+ ADR组原发肿瘤重量分别为:(8．03±0．92)、(7．86±0．80)、(4．16±0．54)、(1．86±0．73)g,Nef+ ADR组肿瘤重量较ADR组显著减轻(P<0．05);Nef、ADR合并用药效果α=1．38。4组原发灶细胞凋亡率分别为8．29％、11．80％、25．70％、71．20％,Nef+ADR组较ADR组明显增高(P<0．05)。病理切片发现Nef+ADR组较ADR组肿瘤细胞明显减少,坏死明显增多。结论甲基莲心碱对阿霉素抑制裸鼠体内骨肉瘤生长有增效作用。     

Chemotherapy and surgery in a murine osteosarcoma.

Surgical and chemotherapeutic effects on pulmonary metastatic disease were evaluated in the MGH-OGS murine osteosarcoma. The tumor responded to three sequential injections of doxorubicin with prolonged growth delay but cisplatin administration (although given in doses sufficient to cause weight loss and significant mortality) was not effective in controlling local disease progression. Using a protocol with three injections of doxorubicin (0.006 mg/g of body weight), it was observed that disease-free survival was enhanced when one of the three doses of doxorubicin was given at the time of surgery (perioperatively). By marginally resecting the primary tumor and permitting its regrowth, a model was developed with recurrent primary and metastatic disease present simultaneously. It was observed in this model that amputation or resection of the recurrent primary lesion resulted in pulmonary metastatic growth acceleration. Using this recurrent primary tumor model, doxorubicin's effect on pulmonary metastatic lesions was enhanced when the drug was given at the time of amputation.     

Evaluation of fluorescein diacetate for flow cytometric determination of cell viability in orthopaedic research

Accurate estimation of cellular viability is important both in research and in aspects of orthopaedic clinical practice. We have been interested in the potential for flow cytometric application of fluorescein diacetate (FDA) in evaluating chondrocyte survival following cryopreservation of osteochondral allografts as well as in the assessment of sarcoma necrosis following preoperative chemotherapy. In order to evaluate the suitability of this method for cell viability assays, this study compared FDA with more traditional methodology (trypan blue, clonigenic assay, metabolic activity analysis, measurement of DNA synthesis, and histological assessment of necrosis). Both chondrocytes and sarcoma cells were exposed to various experimental injuries prior to viability analysis. Although it is evident from these experiments that FDA accurately reflects cell survival after physical injury, it underestimates the effect of chemotherapy on cell reproductive potential in vitro. However, FDA is highly correlated with histological assessment of tumor viability after chemotherapy in vivo. It is apparent that the methodology chosen for determination of viability should be appropriate for the type of experimental injury and should analyze the cell function (i.e., metabolic activity or reproductive capacity) that is appropriate for the experimental model.     

40~60岁肢体骨肉瘤病人的临床治疗效果与策略

目的 分析40~60岁肢体骨肉瘤病人的临床治疗疗效,总结预后相关因素并探讨治疗策略。方法 回顾性分析2002年4月至2015年10月国内7家骨肉瘤治疗中心收治的确诊为肢体骨肉瘤的62例病人的资料,其中男34例,女28例;年龄范围限定在40~60岁,平均年龄为50.4岁;收集病人一般情况、发病部位、术前穿刺、化疗情况、手术方式、复发转移及生存情况等。以上述因素为变量指标,应用Kaplan-meier法测算生存率,研究这些因素与3年、5年生存率之间的关系。结果 62例肢体骨肉瘤病人中发病于膝关节周围者50例,约占80.6%。62例病人均得到随访,随访12~115个月,平均随访时间为43个月;3年和5年总生存率为80.6％、56.5%;3年和5年无瘤生存率为41.9%、32.3%;5年总生存率：术前化疗组与未化疗组分别为51.5%、62.1%;术后化疗组与未化疗组分别为52.1%、71.4%;术前术后均化疗组与均未化疗组为51.6%,61.3%;保肢手术组和截肢手术组分别为57.1%、53.8%。术后无复发转移、单纯复发、肺转移、多发转移的病人3年生存率分别为89.7%、78.6%、50.0%、73.3%。结论 40~60岁肢体骨肉瘤病人,膝关节周围发病率高于年轻病人,且预后较差,其预后与术前穿刺明确诊断、术后复发转移情况密切相关,其临床治疗应当建立在充分切除肿瘤的基础之上,严格遵照Enneking外科分期,降低肿瘤的复发转移率,以期改善预后。     

骨膜骨肉瘤与高度恶性表面骨肉瘤

目的报告表面骨肉瘤的两种少见亚型,即骨膜骨肉瘤和高度恶性表面骨肉瘤的诊治经验。方法回顾 5例骨膜骨肉瘤和 4例高度恶性表面骨肉瘤患者的影像学和病理学特点,并对临床治疗结果进行分析。 5例骨膜骨肉瘤患者,男 1例,女 4例;年龄 28～ 42岁,平均 35岁;肿瘤位于胫骨上段者 4例,股骨下段者 1例。 4例高度恶性表面骨肉瘤患者,男 3例,女 1例;年龄 17～ 23岁,平均 19.25岁; 4例患者肿瘤均位于股骨下段。结果 5例骨膜骨肉瘤均予以广泛切除,除 1例因局部复发行截肢术,现无瘤生存 1年 3个月外,余 4例已无瘤生存 3～ 9年,平均 5年 9个月。 4例高度恶性表面骨肉瘤患者,虽经积极综合治疗,但仅 1例无瘤生存 7年; 1例术后 2年 5个月复发而截肢,现无瘤生存 1年 9个月; 1例于术后 2年 4个月死于肺转移;另 1例在确诊后 3个月死亡。结论骨膜骨肉瘤和高度恶性表面骨肉瘤各具鲜明的影像学和病理学特点,骨膜骨肉瘤恶性程度较低,应采用以局部广泛切除为主的手术治疗,预后相对较好;而高度恶性表面骨肉瘤的生物学行为则与经典的髓内骨肉瘤相似,预后较差,必须采用手术与化疗相结合的综合治疗方法。     

肿瘤坏死因子相关凋亡诱导配体联合阿霉素治疗骨肉瘤

目的 探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）与阿霉素（ADM）联用对骨肉瘤细胞株HOS是否具有协同杀伤作用，并对其机制进行初步研究。方法 MTT法检测细胞的抑制率和存活率，流式细胞术（FCM）检测细胞凋亡率，逆转录-聚合酶链反应（RT—PCR）检测TRAIL受体（receptor，R）的表达水平。结果 单用0．5mg／LTRAIL及1．0、10．0、100．0mg／LADM对HOS8603的抑制率分别为13．18％、5．56％、23．02％和76．72％，联合用药的抑制率分别为42．98％、53．12％、81．91％。0．5mg／LTRAIL和1．0mg／LADM有协同作用．TRAIL和ADM联用时TRAIL R2 mRNA表达较TRAIL单用时明显增强。结论 TRAIL与ADM联用对骨肉瘤细胞有明显的协同作用，其机制可能与ADM上调TRAILR2的表达有关。     

Marginal resection for osteosarcoma with effective preoperative chemotherapy

Objective: To present the clinical results of marginal resection with effective preoperative chemotherapy for treatment of osteosarcoma. Methods: Thirty‐eight patients (20 male and 18 female, average age 17 years), underwent marginal resection after confirmation of effective preoperative chemotherapy between 1999 and 2008 and the results were analyzed retrospectively. The distal femur was involved in 22 cases, proximal tibia in 11, proximal humerus in 4, and proximal fibula in 1. Thirty‐seven patients were stage IIB and one IIIB. Twenty‐nine patients were treated with the DIA, and 9 with the MMIA protocol. Twenty‐one patients underwent tumor resection and bone allograft transplantation. The epiphysis was preserved in 9 patients, and not in the other 12. Eleven patients underwent tumor resection and prosthetic replacement, and 4 tumor resection with autograft implantation. One patient underwent tumor resection and allograft with preservation of the epiphysis; another underwent marginal tumor resection only. Results: All patients received effective preoperative chemotherapy. At a median follow‐up of 52 months, local recurrence had developed in one patient (2.6% local recurrence rate). Pulmonary metastases developed in 9 patients (23.7%). Five patients died of metastases, one died of intracranial hemorrhage due to thrombocytopenia caused by postoperative chemotherapy. The overall 2‐year survival rate was 87.3%, and event‐free survival rate 75.5%. The overall 5‐year survival rate was 74.7%, and event‐free survival rate 60.8%. Excellent to good function of affected limbs was achieved in 60.5%. Conclusions: With careful preoperative evaluation and effective preoperative chemotherapy marginal resection of osteosarcoma can produce good results. Marginal resection of osteosarcoma should be performed by an experienced surgeon who is familiar with the limb salvage rules for osteosarcoma.     

Ⅲ期骨肉瘤的治疗

目的探讨Ⅲ期骨肉瘤的诊断、治疗和预后。方法分析1989年12月至2003年12月收治的23例Ⅲ期肢体骨肉瘤患者,男14例,女9例;年龄16～31岁,平均22.4岁。肺转移15例,骨转移7例(跳跃转移5例,骨肉瘤病2例),同时存在肺和骨转移1例。经术前化疗,并行原发肿瘤和转移病灶的切除手术。结果经术前化疗,1例就诊时肺部有1个结节的患者转移瘤消失,1例同时存在肺和骨转移者,切除原发病灶后,出现多处转移,不能行转移灶切除手术;2例骨肉瘤病者仅切除原发病灶;5例跳跃转移者中的2例,切除原发和跳跃转移病灶后,各出现1个肺转移灶。16例行开胸术,其中6例再次出现肺转移灶而行二次开胸术,3例患者二次开胸术后出现肺外转移而放弃治疗。原发病灶与转移病灶的肿瘤坏死率差异无统计学意义。随访5～168个月(平均74.6个月),无瘤生存9例,带瘤生存4例,死亡10例。Cox模型分析提示转移病灶的数目与预后相关(P<0.05)。肺转移与跳跃转移病灶的Kaplan-Meier生存曲线经log-rank检验,差异无统计学意义。结论新辅助化疗及原发病灶和转移灶的手术切除是Ⅲ期骨肉瘤治疗的有效方法,首次肺转移灶切除采用胸骨正中切口,转移瘤的数目与预后相关。     

三氧化二砷抑制骨肉瘤细胞及二倍体细胞增殖的研究

目的　研究三氧化二砷 (As2 O3 )对骨肉瘤细胞及二倍体细胞增殖的影响。方法　应用噻唑蓝 (MTT)法和集落形成能力测定、形态学观察、流式细胞术和电镜观察等方法检测和观察As2 O3 对骨肉瘤细胞株MG 63及二倍体细胞株MRC 5增殖的影响。结果　As2 O3 对骨肉瘤细胞及二倍体细胞株的增殖有抑制作用 ,并具有时间依赖性和一定范围内的剂量依赖性。≥ 1μmol/L浓度的As2 O3 对MG 63骨肉瘤细胞的抑制率达 70 % ,而≥ 8μmol/L浓度的As2 O3 对MRC 5细胞的增殖才达到相近的抑制率 ;0 .5～ 2 .0 μmol/L浓度的As2 O3 对MG 63细胞的生长抑制率高 ,而对MRC 5细胞的生长抑制率低 ,表现为明显的选择性抑制 ( P <0 .0 1)。 1μmol/LAs2 O3 处理的MG 63细胞第 3天呈典型的凋亡形态学改变 ;流式细胞仪分析显示 ,在G1期细胞前出现明显亚二倍体峰 ,凋亡率与As2 O3 处理时间呈正相关 ;MRC 5细胞则未见明显凋亡峰。结论　As2 O3 通过诱导细胞凋亡抑制骨肉瘤细胞和二倍体细胞增殖 ,在一定浓度内As2 O3 可选择性抑制骨肉瘤细胞的生长增殖。     

FDG PET对骨肉瘤新辅助化疗疗效评估的初步研究

目的通过与病理学肿瘤坏死率相比较，评估18氟脱氧葡萄糖正电子发射断层显像（F-18-fluoro-2-deoxy—D—glucose positron emission tomography，FDG PET）对骨肉瘤新辅助化疗效果的判断价值。方法2004年4月至2006年12月，共有15例（女6例，男9例；年龄7～33岁，平均年龄17-3岁）行新辅助化疗的原发性骨肉瘤患者进行全身或局部FDGPET检查。所有患者均行化疗前和化疗后两次FDGPET检查。FDG—PET检查后，分别计算每例患者FDG的最大标准摄取值（SUV）和肿瘤／背景比值（TBR）。化疗结束后行保肢或者截肢手术，术后切除肿瘤按照Salzer-Kuntschik系统评估病理学肿瘤坏死率反应的化疗效果（I～Ⅲ级为效果良好，Ⅳ～Ⅵ级为效果差）。结果肿瘤坏死率评估显示：8例肿瘤坏死率大于90％，化疗效果良好；7例肿瘤坏死率小于90％，化疗效果不佳。统计学分析提示，化疗后与化疗前的最大标准摄取值之比（SUV2／SUVl）、肿瘤／背景比值之比（TBR2／TBRl）以及化疗后肿瘤背景比值（TRB2）均与肿瘤坏死率相关（P〈0．01，P〈0．001，P〈0．001）。所有化疗效果良好的患者TBR2／TBRl值小于0．46，化疗效果不良者均大于0．49。结论FDGPET为评价骨肉瘤患者化疗效果提供了一种可行的无创检查方法。肿瘤／背景比值与最大标准摄取值相比对评价骨肉瘤的化疗效果更有价值。     

Deep venous thrombosis during neoadjuvant chemotherapy in a patient with osteosarcoma: a case report

A 29-year-old man with an osteosarcoma of the left distal femur developed asymptomatic deep venous thrombosis (DVT) during neoadjuvant chemotherapy. The thrombotic event occurred in the left common iliac vein and was revealed by pelvic computed tomography (CT). We successfully performed a limb salvage operation with placement of a permanent inferior vena cava filter. The thrombus spontaneously regressed without thrombolytic therapy after surgery. We should consider DVT in patients with musculoskeletal sarcomas who are under long-term hospitalization and immobilization associated with not only surgery but also chemotherapy. To screen for both metastasis and DVT, we recommend contrast-enhanced whole-body CT, including scans of the pelvis.     

Calcaneal insufficiency fracture in a child with osteosarcoma after chemotherapy and limb-sparing surgery

We report a rare case of a calcaneal insufficiency fracture in an 11-year-old boy with an osteosarcoma of the right proximal tibia. After one course of neoadjuvant chemotherapy for 4 months, we performed limb-sparing surgery. The knee joint was replaced with a custom-made endoprosthesis and the knee extensor mechanism was reconstructed with a medial gastrocnemius muscle flap. Seven months postoperatively, he complained of pain in the right foot and a plain radiograph revealed a calcaneal insufficiency fracture. Osteoporosis induced by chemotherapy and change of forces transmitted to the calcaneus after reconstruction with a gastrocnemius muscle flap might contribute to this fracture. It is important to be aware of the rare clinical entity of insufficiency fracture for making an early diagnosis and for differentiating them from pathological fractures associated with malignancy.     

Prognostic effects of doctor-associated diagnostic delays in osteosarcoma

Introduction  In this retrospective study, we evaluated to what extent diagnostic delays impact prognosis in osteosarcoma. Materials and methods  The authors identified 26 osteosarcoma patients who had undergone inappropriate procedure-associated diagnostic delays of more than 45 days after surgery, calculated overall survival rates, and analyzed clinicopathologic characteristics. Results  Initial clinical impressions were of a benign bone tumor in 15 patients, fracture in 8, and infection in 3. After initial inappropriate procedures, primary surgeons failed to send a tissue sample to a pathologist for definite diagnosis in 12 cases, and pathologists made incorrect diagnoses in the other 14. The average doctor-associated diagnostic delay after inappropriate surgery for these 26 patients was 10.5 months. Following referral to our institute, 22 underwent both surgery and chemotherapy and the remaining 4 patients underwent chemotherapy only. Four of the 26 patients were alive at last follow-ups. Estimated 5- and 10-year overall survival rates were 26 and 10%, respectively. Conclusions  The present study shows that doctor-associated diagnostic delay superimposed on an inappropriate primary procedure has a significant detrimental effect on overall survival in osteosarcoma. This study demonstrates that surgeons and pathologists should spare no effort to minimize diagnostic errors and delays.     

单中心四肢骨肉瘤311例预后分析

 目的 探讨单中心四肢骨肉瘤的治疗、预后及生存状况。方法 回顾性分析1998年至2008年间单中心收治的311例四肢骨肉瘤患者的病历资料,记录患者的基本资料、化疗、手术、并发症、肿瘤转移及预后情况,应用Kaplan-Meier、Cox regression 等统计学方法分析各变量与预后生存的关系。结果 311例四肢骨肉瘤患者,男206例,女105例;年龄5～56岁,平均18.6岁;上肢29例,下肢282例。282例获得广泛或根治性手术,保肢手术149例,截肢手术133例。术后64例患者发生各种并发症,其中17例出现2次以上的术后并发症,25例发生肿瘤局部复发。105例获得规范化疗,206例未获得规范化疗,5年生存率分别为57.4%和36.3%。肺转移患者(76例)和无肺转移患者5年生存率分别为16.8%和50.7%。Enneking 分期ⅡB期和Ⅲ期患者5年生存率分别为44.6%和33.1%。选择保肢手术和截肢手术患者的5年生存率分别为56.6%和31.8%,由于截肢患者中有较大的比例施行了不规范化疗以及其他混杂因素导致5年生存率明显降低。Cox回归分析显示化疗不规范和发生肺转移患者预后较差。结论 新辅助化疗联合肿瘤广泛或根治性手术切除可以挽救约60%青少年四肢骨肉瘤患者的生命,肺转移和不规范化疗严重影响患者的预后。     

Cytotoxic effects of delphinidin in human osteosarcoma cells

Introduction

The aim of this study was to evaluate whether delphinidin is cytoprotective or cytotoxic in osteosarcoma cell lines, and to elucidate the underlying mechanisms.

血管内皮生长因子抗体治疗骨肉瘤的实验研究

目的　探讨血管内皮生长因子 (VEGF)抗体对骨肉瘤OS 732细胞株血管形成及瘤细胞生长状态的影响 ,为从抗血管生成途径治疗骨肉瘤提供依据。　方法　应用鸡胚绒毛尿囊膜模型 ,通过解剖显微镜下血管计数、原位铺片、组织切片HE染色及PCNA免疫组化和原位末端标记技术检测VEGF抗体对肿瘤血管形成、肿瘤生长的抑制作用及其对细胞增殖和凋亡的影响。　结果　VEGF抗体组血管密度 [给予抗体第 8天时为 (30 6± 4 6 ) ]明显低于对照组 [给予抗体第 8天时为 (5 1 0±6 9) ],瘤细胞较对照组减少 ,肿瘤细胞凋亡指数 (16 98± 4 81)显著高于PBS对照组 (3 75± 0 73) ,增殖指数两组差异不明显 ,同时VEGF抗体组凋亡的微血管内皮细胞增多 ,增殖期微血管内皮细胞少见。　结论　VEGF抗体能够显著抑制骨肉瘤OS 732血管形成 ,抑制内皮细胞增殖 ,促进内皮细胞凋亡 ,可能是VEGF抗体发挥抗血管生成作用的途径之一 ,并通过抑制血管形成而促进肿瘤细胞凋亡 ,最终达到抑制瘤细胞生长的作用。     

同源性骨肉瘤新细胞系Zos和Zos-M的生物学比较研究

目的 对来源于同一患者原发灶和跳跃转移灶的两株新建骨肉瘤细胞系-Zos和Zos-M的特性和生物学行为进行鉴定和比较研究.方法 用组织培养块法从1例骨肉瘤患者原发灶和跳跃转移灶骨肉瘤组织,分离并分别建立两株新的骨肉瘤细胞系-Zos和Zos-M;利用形态学观察、核型和细胞周期分析、生长曲线和倍增时间检测、成骨性标志检测和基质胶侵袭试验对两株细胞系进行体外鉴定.通过皮下、原位移植和实验性转移检测细胞系的体内成瘤和转移能力.应用MTT法检测Zos和Zos-M对传统化疗药物的敏感性;RT-PCR方法检测并比较二者转移相关基因的表达.结果 Zos和Zos-M体外连续培养100代后仍保持稳定,形态学检测、RT-PCR检测碱性磷酸酶、骨钙蛋白和骨桥蛋白等成骨系标志提示均符合骨肉瘤特征.Zos和Zos-M倍增时间分别为33.65 h和31.58 h.核型分析表现为非整倍体和多种染色体结构异常.Zos、Zos-M皮下和原位成瘤率均为100%,Zos-M实验性转移率为37.5%(3/8),Zos无实验性转移(0/8).转移相关基因表达检测表明cadherin-11在Zos-M的低表达可能与其高转移能力相关.并且Zos和Zos-M与传统骨肉瘤细胞系相比,对常用化疗药物的敏感性差.结论 人骨肉瘤细胞系Zos和Zos-M及相关动物模型的建立为治疗骨肉瘤的药物筛选提供新的模型,其具有相同的遗传背景和不同的侵袭转移能力,为骨肉瘤转移的研究提供良好的模型.

Abstract：

Objective To characterize and compare the different biological behaviors of two novel human osteosarcoma cell lines, Zos and Zos-M, established respectively from the primary site and the skip metastasis of an osteosarcoma patient. Methods Two novel human osteosarcoma cell lines, Zos and Zos-M,were developed using tissue plant culture method. The vitro examinations included observations of morphology, analysis of karyotype and cell cycle, calculation of doubling time and growth curve, detection of osteoblastic markers and matrigel invasion assay. Subcutaneous, intratibial and intravenous inoculations into nude mice were performed to study the in vivo tumorigenicity and metastatic potentials of both cell lines.MTT were used to detect sensitivity of the cell lines to chemotherapeutic drugs. RT-PCR was performed to assess the expression of and some metastasis-related genes. Results Both cell lines proliferated actively and remained stable for more than 100 passages in vitro without interruption. The morphology and expression of osteoblastic markers of Zos and Zos-M were conformed to the characteristic of osteosarcoma. The karyotype analysis displayed aneuploidy and various structural abnormalities. The population doubling time of Zos and Zos-M were 33.65 h and 31.58 h respectively. Both cell lines were less sensitive to the current chemotherapy protocols compared to U-2OS. Zos and Zos-M were 100% tumorigenic by subcutaneous and othotopic injection. 37.5% of nude mice injected Zos-M and none of nude mice injected Zos developed lung metastasis after intravenous injection. The comparison of the expression patterns of some metastasis-related genes revealed that the decreased expression of cadherin-11 in Zos-M may correlate with a high potential of metastases. Conclusion The two novel established human osteosarcoma cell lines, Zos and Zos-M and related animal models could serve as models for the study of drug resistance and screening of new therapeutics for osteosarcoma. In addition, the study also provide tools for the study of metastasis because the same genetic background and different potential of metastasis of Zos and Zos-M.     

流式细胞术的凋亡检测技术在人胃肠实体瘤药敏试验中的应用

目的化疗药能诱导肿瘤细胞凋亡，据此将流式细胞术的经典凋亡检测技术应用到临床特异性抗肿瘤药物的筛查，实现快速，准确的药物敏感性报告。方法应用流式细胞术的AnnexinV／PI技术，检测67例临床手术切除的胃肠肿瘤在不同化疗药物诱导下发生的凋亡。结果7种化疗药物诱导的胃肠肿瘤凋亡率有显著差异，胃癌的凋亡率以紫杉醇，丝裂霉素，氟尿嘧啶最高；而大肠癌以氟尿嘧啶，丝裂霉素，奥沙利铂诱导的凋亡率最高。联合化疗方案，胃癌以氟尿嘧啶联合紫杉醇的敏感率及凋亡率最高；大肠癌以氟尿嘧啶联合奥沙利铂的敏感率及凋亡率最高。结论不同肿瘤或同一肿瘤的不同个体，对化疗药物的敏感性均不同。临床可应用凋亡检测技术作为快速的药物敏感性检测方法，为肿瘤化疗筛选合适的个体化有效方案，避免无效化疗。     

Apoptotic effects of curcumin on human osteosarcoma U2OS cells

Objective: Curcumin, an active ingredient derived from the rhizome of the plant, Curcuma longa, has antioxidant, anti‐inflammatory and anti‐cancer activities. The aims of this study were to examine whether curcumin can induce apoptosis in an osteosarcoma cell line. Methods: Curcumin‐induced apoptosis in human osteosarcoma U2OS cells was investigated using morphological analysis, marked nuclear condensation and fragmentation of chromatin, which were observed by Hoechst 33258 staining and DNA ladder formation. The U2OS cells were treated with or without curcumin. Cell viability was assessed by the 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐Diphenyltetrazolium (MTT) method. Cell‐cycle, apoptosis and apoptosis‐related proteins in U2OS cells were evaluated by flow cytometry and western blotting. Results: Curcumin showed growth inhibitory effects on U2OS cells in a dose‐and time‐dependent manner, inducing significant G1 arrest and apoptosis in U2OS cells. This curcumin‐induced apoptosis in U2OS cells was accompanied by up‐regulation of Bax, Bak, and p‐Bad and down‐regulation of Bcl‐2, but no effect on the levels of Bcl‐X_L or Bad proteins was noted. Moreover, curcumin treatment resulted in a significant reduction of mitochondrial membrane potential and increase in the concentrations of mitochondrial cytochrome C and caspase‐3. Conclusion: Multiple molecular pathways are involved in curcumin‐induced apoptosis of human U2OS cells. These include pro‐and anti‐apoptotic Bcl‐2 family proteins, mitochondrial membrane potential, mitochondrial cytochrome C and caspase‐3.     

Transmedullary amputation and resection of metastases in combined therapy of osteosarcoma.

The eradication of microfoci of metastatic osteosarcoma by adjuvant chemotherapy permits the surgeon to take a more conservative approach to amputation and to more aggressively resect metastases. In a series of 59 children with osteosarcoma of an extremity, transmedullary amputation carried no greater risk of stump recurrence than either disarticulation or amputation above the joint. We attribute this to careful selection of the level of amputation by use of preoperative bone scans, intraoperative frozen sections of bone stump and intensive postoperative adjuvant chemotherapy. Immediate application of a prosthetic limb has enhanced the physical and emotional rehabilitation of these patients. Resections of pulmonary metastases in 12 patients who were receiving adjuvant chemotherapy has resulted in a median tumor-free survival of 17 mo, with 3 patients still alive without disease for 13, 25, and 72 mo. A more aggressive approach to the resection of pulmonary metastases may substantially improve current rates of tumor-free survival.