PB231诱导K562细胞凋亡的初步研究

 目的研究PB231诱导K 562细胞凋亡作用,探讨PB231的抗肿瘤机制.方法采用荧光显微镜观察细胞形态学变化,MTT法测定PB231对K 562细胞生长抑制作用,琼脂糖凝胶电泳检测细胞DNA裂解.结果K562细胞经PB31处理后,在倒置光显微镜下可见细胞变形和凋亡小体形成;荧光显微镜下可见染色质凝集.MTT法检测其IC50值为5×10-5M.琼脂糖凝胶电泳呈典型的"DNA Ladder".结论PB231可诱导K562细胞发生凋亡.这可能是其抑制K562细胞生长的作用机理之一.     

茶多酚诱导肝癌细胞凋亡

为进一步研究茶多酚诱导体外培养的肝癌细胞株HepG-II细胞发生凋亡的作用,采用MTT法、形态学观察、琼脂糖凝胶电泳和末端脱氧核苷酸转移标记法观察被茶多酚处理后的ＨepG－II细胞的形态学和生化等指标的变化。MTT法研究结果显示,当茶多酚浓度为250μg/ml时即时诱导HepG-II细胞凋亡,并与浓度呈正相关;当茶多酚浓度＞2000μg/ml时,抑制率增强的幅度明显减慢。透射电镜下观察到核染色质浓集呈块并可见凋亡小体;荧光染色在荧光显微镜下可见部分细胞核或细胞质内出现致密浓染的黄绿色块状和颗粒状荧光等凋亡细胞的形态学改变。琼脂糖凝胶电泳呈典型的DNA梯形图像。末端脱氧核苷酸转移标记法进一步证实茶多酚可诱导HepG-II细胞的凋亡。提示茶多酚可诱导体外培养的肝癌细胞株HepG-II细胞发生凋亡,具有抗肝癌的作用。     

125I粒子照射诱导人前列腺癌细胞凋亡的实验研究

目的探讨^125I粒子持续低剂量率照射诱导人前列腺癌细胞凋亡的作用机制。方法用DNA琼脂糖凝胶电泳和流式细胞仪分析^125I粒子持续低剂量率照射对PC3细胞的凋亡诱导作用，检测天冬酰胺特异酶切的半胱氨酸蛋白酶Caspase-3活性来观察其对PC3细胞的凋亡诱导途径，并通过间接免疫荧光技术检测其对Bcl-2表达的影响。结果DNA琼脂糖凝胶电泳可观察到清晰的“DNA梯度”，流式细胞仪检测^125I粒子持续低剂量率照射对PC3细胞具有明显的凋亡诱导作用。Caspase-3活性检测表明，Caspase-3活性无明显变化。流式细胞仪分析表明Bcl-2的表达随^125I粒子剂量的增加而下降。结论^125I粒子持续低剂量率照射可诱导PC3细胞凋亡，其诱导凋亡与Bcl-2表达有关，与Caspase-3活性无关。     

纯化后海藻硫酸多糖-蛋白复合物诱导人肝癌细胞SMMC- 7721 凋亡作用的研究

目的 研究海藻硫酸多糖蛋白复合物(Sulfated Polysaccharide-Protein Complex,SPPC)诱导肿瘤细胞凋亡的效率和机制.方法 选用人肝癌细胞株SMMC-7721为实验对象,用MTT法,流式细胞术,DNA凝胶电泳法,细胞形态学检测方法对纯化后不同剂量SPPC处理的SMMC-7721细胞凋亡情况进行观察分析;并用Western blot 法检测凋亡相关蛋白 procaspase-3的表达量变化.结果 纯化后SPPC中、高剂量组(2.0、10.0mg · ml-1)可明显诱导SMMC-7721细胞凋亡, SMMC-7721细胞内凋亡相关蛋白procaspase-3有不同程度的表达降低,DNA凝胶电泳法检测到DNA ladder现象,流式细胞检测显示纯化后SPPC在10.0mg · ml-1浓度下出现了典型的二倍体峰.结论 一定浓度的海藻硫酸多糖蛋白复合物有诱导SMMC-7721细胞凋亡的作用,其生化机制可能是通过caspase-3 的活化来实现的.     

血管内皮生长因子C蛋白在喉鳞癌中的表达与颈淋巴结转移的关系

目的研究血管内皮生长因子C（VEGF-C）蛋白在喉鳞癌和正常组织中的表达情况,探讨其与临床相关因素以及淋巴转移的关系,了解VEGF-C蛋白在喉癌颈淋巴结转移中的作用。方法采用免疫组化法分别检测46例喉鳞癌癌内组织、癌旁组织和正常组织中VEGF-C蛋白的表达,观察并分析其表达与肿瘤病理分化程度、T分期、肿瘤临床分型及颈淋巴结转移等临床病理因素之间的关系。结果喉鳞癌组织不同部位的VEGF-C蛋白表达明显不同。＂~GF-C在癌内、癌旁及正常组织中均有表达,但癌内组织的表达阳性率明显高于癌旁组织和正常组织,其表达与肿瘤病理分化程度、T分期、肿瘤临床分型及颈淋巴结转移密切相关,而VEGF-C在癌旁组织中的表达与这些因素无显著相关。结论喉癌细胞分泌VEGF-C诱导癌内和癌周淋巴管增生扩张是喉癌发生颈淋巴结转移的重要因素之一,VEGF-C检测有望作为临床早期判断和预测颈淋巴结转移的指标之一。     

鹰嘴豆芽提取物对Caco-2细胞凋亡的作用

目的研究鹰嘴豆芽乙醇提取物（CSE）对人结肠腺癌Caco-2细胞增殖及细胞凋亡的影响。方法提取鹰嘴豆芽的活性成分并测定含量;MTT法观察提取物对Caco-2细胞增殖的影响;透射电镜观察细胞超微结构;琼脂糖凝胶电泳法进行细胞凋亡的DNA分析;流式细胞术检测凋亡情况。结果 CSE含23.53%皂苷、14.73%异黄酮;不同浓度的CSE对Caco-2细胞均有一定的增殖抑制作用,且呈量效和时效关系;细胞出现染色质边移和凋亡小体;DNA电泳呈现梯度条带,表明CSE可诱导Caco-2细胞凋亡;3、5、10μg/ml的CSE作用后,细胞凋亡率分别为32.6%、68.8%、73.9%,与对照组（0.9%）比较差异显著（P〈0.05）。结论 CSE可通过诱导细胞凋亡而抑制人结肠腺癌株Caco-2细胞的生长,为鹰嘴豆应用于结肠癌的临床治疗和预防提供了理论依据。     

硫芥诱导大鼠脾脏淋巴细胞凋亡的研究

目的 探讨硫芥诱导脾脏组织淋巴细胞凋亡以及caspase-3在其中的作用.方法 大鼠腹腔注射硫芥,各组动物分别于中毒后1、3、5d后麻醉,开胸取脾.HE染色法观察脾脏组织的病理改变,RT-PCR法检测大鼠脾脏组织caspase-3基因表达,West-em blot法检测caspase-3的蛋白表达.分离脾脏淋巴细胞,制备硫芥染毒细胞模型.DNA琼脂糖凝胶电泳观察caspase-3抑制剂Ac-DEVD-CHO对细胞DNA降解的影响;流式细胞仪检测Ac-DEVD-CHO对染毒细胞凋亡峰(即G1亚G1峰)及线粒体跨膜电位(△Ψm)的影响.结果 硫芥中毒大鼠脾脏组织形态发生病理改变,部分淋巴细胞出现了凋亡的特征;脾脏组织caspase-3的mRNA与蛋白表达均增加:mRNA在中毒后1d的表达量与对照组比较具有统计学意义(P＜0.05);蛋白表达在中毒后3d和5d与对照组比较具有统计学意义(P＜0.05,P＜0.01).caspase-3的特异性抑制剂Ac-DEVD-CHO对体外培养的硫芥染毒淋巴细胞的DNA降解和细胞(G1峰左侧凋亡峰的m现有抑制作用.硫芥可导致细胞线粒体跨膜电位降低,随着中毒时间延长,线粒体膜电位的降低更明显.结论 硫芥中毒可导致大鼠脾脏组织损伤,细胞凋亡是其作用机制之一,caspase-3可能参与了这一过程并在其中发挥重要的作用.     

转化生长因子β1对胃腺癌细胞凋亡诱导的研究

为研究转化生长因子β１能否诱导人胃腺癌ＳＧＣ－７９０１细胞凋亡，运用透射电镜技术了解凋亡细胞超微结构改变，用流式细胞术分析细胞ＤＮＡ，周期变化ＤＮＡ片段改变采用琼脂糖凝胶电泳法，电镜下可见细胞核染色质浓缩，边集，核碎裂及凋亡小体形成，ＤＮＡ梯状电泳带出现ＤＮＡ周期分析显示在整个细胞周期峰群前端存在一个凋亡峰，表明转化生长因子β１可诱导人胃腺癌ＳＧＣ－７９０１细胞凋亡。     

临床颈淋巴转移阴性口腔鳞癌的前哨淋巴结研究

目的探讨前哨淋巴结活检术(SLNB)在口腔鳞癌治疗中的的预测价值。方法口腔鳞癌患者20例,使用亚甲蓝染色法对前哨淋巴结进行染色识别。结果SLNB成功率为70％。SLNB对颈淋巴结微转移的检测准确率为100G。前哨淋巴结(SLN)每例平均2．4枚。14例患者中有6例存在颈部淋巴转移,其中5例仅转移至SLN,1例SLN和非SLN均有转移。结论SLNB能准确预测口腔鳞癌颈淋巴结转移情况,为SLNB阴性的口腔鳞癌患者避免颈淋巴清扫术提供了诊断依据。     

UV-C诱导体外血管平滑肌细胞凋亡模型的建立

应用UV-C垂直照射距离其10cm处的大鼠主动脉SMC,发现经照射后细胞可出现典型的凋亡形态学改变,如细胞变圆、染色质浓缩、细胞膜出泡、凋亡小体等;且提取细胞DNA,其琼脂糖凝胶电泳呈现梯状图谱,从形态学和生化指标方面证明了UV-C可诱导体外血管SMC凋亡。UV-C照射诱导体外SMC凋亡,可作为进一步深入研究SMC凋亡的内在调控机制的一个简便、稳定、可靠的模型。     

Synchronous ipsilateral renal cell and transitional cell carcinoma

The coexistence of multiple and synchronous primary neoplasms in the genitourinary system has only rarely been described in the literature. We present the case of a 78-year-old man with haematuria as the initial presentation, finally proven to be transitional cell carcinoma (TCC) combined with renal cell carcinoma (RCC). Intravenous urography (IVU), CT and arterial angiography studies revealed a space-occupying nodule at the right upper renal pelvicalyces showing mild enhancement with contrast medium. Another strong contrast medium enhancing exophytic tumour was found at the lower pole of kidney; there were hypodense foci and calcified components in this lesion. A right nephroureterectomy was performed. Pathological diagnosis was a papillary TCC and a clear cell type RCC. This is a rare case of combined renal malignancies diagnosed by imaging.     

肾透明细胞癌与肾乳头状癌、嫌色细胞癌MRI表现的对照研究

目的 探讨MR动态增强扫描对肾癌亚型的鉴别诊断价值.方法 搜集77例经病理证实的肾癌患者资料,其中透明细胞癌(CCRCC)55例,乳头状癌(PRCC)14例,嫌色细胞癌(CRCC)8例,回顾性分析各亚型肿瘤患者MR平扫及动态增强扫描表现并与病理对照,根据肿瘤及肾皮质增强前后的皮质期、实质期及延迟期信号变化,分别进行百分比测量、肿瘤-肾皮质增强指数计算,并采用单因素方差分析和LSD法进行比较.结果 CRCC多数信号均匀(7/8);CCRCC及PRCC多数信号不均(分别为51/55和13/14)、常见坏死(36/55和7/14),PRCC最常见出血(9/14)及囊变(9/14).动态增强各期CCRCC强化程度最高,强化模式呈"快进快退",CRCC轻至中度强化,PRCC强化最轻,两者均呈渐进性延迟强化.CCRCC、PRCC及CRCC皮质期信号变化分别为(296.15±60.27)%、(79.70±18.84)%和(119.56±40.76)%,实质期分别为(236.33±58.31)%、(122.81±27.35)%和(163.06±33.91)%,延迟期分别为(216.83±46.72)%、(117.55±20.63)%和(179.72±32.89)%;三者皮质期的肿瘤-皮质增强指数分别为1.26±0.34、0.33±0.12及0.54±0.10,实质期分别为0.92±0.23、0.41±0.23及0.62±0.15,延迟期分别为0.76±0.14、0.35±0.11及0.69±0.12,各亚型增强各期的信号变化(F值分别为940.931、124.515、38.194,P值均＜0.01)、肿瘤-皮质增强指数(F值分别为798.625、78.308、73.699,P值均＜0.01)差异均有统计学意义.3种亚型的MRI表现与病理学所见基本相符.结论 CCRCC、PRCC及CRCC的MRI动态增强有一定特征性的表现,与其病理特点密切相关,在肾癌亚型的鉴别诊断上有着较高的临床应用价值.

Abstract：

Objective To investigate the differential diagnostic features of subtypes of renal cell carcinoma(RCC) using dynamic contrast-enhanced MRI(DCE-MRI).Methods The MRI appearances of 77 RCCs, including 55 clear cell RCCs(CCRCC),14 papillary RCCs(PRCC) and 8 chromophobe RCCs(CRCC), were retrospectively analyzed and compared with findings of pathology. DCE-MRI was conducted in each case after intravenous administration of contrast agent. Region of interest measurements (cortical, nephrographic and delayed Phases) of signals within tumor and uninvolved renal cortex were used to calculate percentage signal intensity change and tumor-to-cortex enhancement index, and the data was analyzed by AVONA and t test. Results On unenhanced and enhanced MRI, most CRCCs showed homogeneous signal(7/8). CCRCC and PRCC often show inhomogenous signal with necrosis(36/55, 7/14). Hemorrhage and cystic degeneration were often found in PRCC (9/14). On the cortical, nephrographic and delayed phase images, CCRCCs showed greater signal intensity change[(296.15±60.27)%, (236.33±58.31)% and (216.83±46.72)%,respectively than PRCCs (79.70±18.84)%, (122.81±27.35)% and (117.55±20.63)%, respectively], and CRCCs showed intermediate change [(119.56±40.76)%, (163.06±33.91)% and (179.72±32.89)%, respectively].A phenomenon of quick staining and quick fainting was observed in CCRCCs. Both of CRCCs and PRCCs showed delayed enhancement. The tumor-to-cortex enhancement index at the cortical, nephrographic and delayed phases was highest for CCRCCs (1.26±0.34, 0.92±0.23 and 0.76±0.14, respectively), lowest for PRCCs (0.33±0.12, 0.41±0.23 and 0.35±0.11, respectively), and intermediate for CRCCs (0.54±0.10, 0.62±0.15 and 0.69±0.12, respectively,P＜0.01). The degree of enhancement was significantly different among the 3 subtypes at the every contrast enhanced phase (F=940.931, 124.515 and 38.194, P＜0.01), so was the tumor-to-cortex enhancement index(F=798.625,78.308 and 73.699, P＜0.01). There was a good consistency between MR appearances of the 3 RCC subtypes and pathological characteristics. Conclusion DCE-MRI could distinctly show imaging features of CCRCC, PRCC and CRCC, which were related to their pathological characteristics, and these features were helpful in predicting a specific subtype of RCC.     

透明细胞乳头状肾细胞癌的影像学表现

目的 探讨透明细胞乳头状肾细胞癌(CCPRCC)的影像学表现.方法 分析15例CCPRCC患者CT及MRI影像特征,采用独立样本t检验比较肿瘤与肾皮质之间平扫CT值、ADC值差异.结果 15例均为单发,边界清晰,大小为(3.1±1.9) cm.13例为实性肿瘤,其中11例伴囊变,2例为囊性肿瘤.4例CT平扫呈等或稍低密...     

Fluorodeoxyglucose cell incorporation as an index of cell proliferation: evaluation of accuracy in cell culture

The use of fluorodeoxyglucose (FDG) and positron emission tomography (PET) is recognized as an accurate tool for the specific diagnosis and staging of cancer. It has also been proposed for the monitoring of anticancer therapy. FDG cell incorporation reflects glycolytic activity whereas inhibition of cell proliferation corresponds to an efficient cancer treatment. The relationship between FDG incorporation and cell proliferation has yet to be demonstrated. Therefore, we aimed to correlate the effects of the toxic agents bleomycin and unlabelled meta-iodobenzylguanidine (mIBG) on cellular metabolism and proliferation. We determined the in vitro metabolic and cytotoxic effects of bleomycin and mIBG by measuring the incorporation of fluorine-18 FDG (%U_FDG) and hydrogen-3 thymidine (%U_THY) in cells of the human premonocytic line U937 in the presence of increasing concentrations of these agents. Proliferation rate of these cells was studied by means of limiting dilution analysis. %U_THY appeared more sensitive to bleomycin or mIBG-mediated cell injury than %U_FDG. After 1 h of exposure to 0.5 M bleomycin, %U_THY was significantly reduced to 62.0% ± 10.4% of control value whereas %U_FDG remained unchanged (91.6% ± 5.3%). Similar results were obtained after 1 h of exposure to increasing concentrations of mIBG (1 M to 1 mM). After 20 h of exposure to bleomycin, %U_THY and %U_FDG were significantly reduced as a function of concentration. After 20 h of exposure to mIBG, a transient increase in %U_FDG up to 149.3% ± 11.2% with 50 M mIBG was further followed by a reduction to 20.1% ± 6.7% with 0.5 mM (P < 0.001). The clonogenic efficiency was reduced as a function of bleomycin (ANOVA, n=255, P) or mIBG concentration (n=80, P) and nearly abolished with 0.1 M bleomycin or 0.1 mM mIBG. In conclusion, %U_THY appears to be a more sensitive index of cytotoxicity in vitro and more accurately relates to cell proliferation than %U_FDG. Correspondence to: D.O. Slosman, Nuclear Medicine Division, Geneva University Hospital, CH-1211 Geneva 14, Switzerland     

一种改进的用于测定细胞周期的细胞制备方法

目的 :减少细胞的聚集数量 ,提高测试效率和结果的准确性。方法 :在用酒精固定细胞时分别加入终浓度为 0 % ,1.5 % ,3% ,6 %和 12 %的小牛血清 ,置 - 2 0℃分别固定细胞 1d ,3d和 7d。比较了不同浓度的血清和保存不同时间粘连细胞的数量及对细胞周期分析结果的影响。结果 :加入血清可明显减轻细胞的粘连 ,减少了细胞的聚集数量 ,尤以 3%小牛血清组最佳。样品可不必过滤 ,在上机测试时 ,进样针不堵塞 ,上样速度快 ,细胞周期分析更准确。随着保存时间的延长 ,聚集细胞的数量有增加的趋势。结论 :在制备用于测定细胞周期的样品时 ,固定细胞的过程中加入终浓度为 3%的小牛血清是一种简单的、能有效地保护细胞膜使细胞不易粘连的技术措施 ,且固定细胞的时间不宜超过 7d。     

Clear cell chondrosarcoma

The clinical, radiologic, and histopathologic features of three cases of clear cell chondrosarcoma are described. On radiographs, this rather benign-appearing tumor resembles a chondroblastoma when it occurs at the end of a long bone, and may occasionally show a calcified matrix. However, it has distinctive tumor cells with a centrally placed vesicular nucleus surrounded by clear cytoplasm. The lesion has a low-grade malignancy and is amenable to en bloc surgical resection, which results in a much better prognosis than that of conventional chondrosarcoma.