非小细胞肺癌循环肿瘤细胞的定量检测意义

Objective To investigate the sensitivity, specificity and clinical significance of detecting circulating tomor cells (CTCs) in NSCLC. Methods Twenty-five patients who undetwent surgical resection for NSCLC form Jan 2007 to Apr 2007 were in- cluded in this study. Control group included 10 patients with benign pulmonary diseses (2 case of hamartoma and 8 case of pulmonary tu berculosis) and 10 healthy volunteers. The pulmonary veins blood and peripheral vein blood were collected respectively. The CD326 immunomagnetic beads and CK-fluorescein isothiocyanate (CK-FITC) were served as the marker antibodies of CTCs. Firstly mononu- clear cell marked by minibeads conjugated with CD326, the mononuclear cells were enriched and separated though Magnetic-activated cell separation(MACS), then the positive separtion cells were marked by anti-CK-FTTC and anti-leukocyte antibody CD45-phyco- erythrine (anti-CD45-PE), finally those cells detected and analyzed by flow cytometry . Results For stage Ⅰ and stage Ⅱ patients (n= 16), CTCs were detected from peripheral vein blood of in 5 (5/16, 31.25%) and from pulmonary veins blood in 9 (9/16, 56.25%). For stage Ⅲ and stage Ⅳ, CTCs were detected from peripheral vein blood in 5 (5/9, 55.56%) and from pulmonary veins blood in 9(7/9,77.78%).For stage Ⅰ,CTCs were detected from peripheral vein blood in 3(3/13,23%)and from pulmo- nery veins blood in 8(8/13,61.54%).The whole CTCs positive detection rate from pilmonary veins blood was 64%(16/25)which higher than feom peripheral vein blood(40%,10/25)(P<0.05).Conclusion The method was set up by MACS combined with FCM to detect the CTCs of pulmonary veins blood and peripheral vein blood of patients.with NSCLC,MACS combined with FCM may improve detection rate of CTCs.This technique appears to be an efficient to detect circulating tumor cells and may be important for dinical practice in the future.     

非小细胞肺癌循环肿瘤细胞的定量检测意义

Objective To investigate the sensitivity, specificity and clinical significance of detecting circulating tomor cells (CTCs) in NSCLC. Methods Twenty-five patients who undetwent surgical resection for NSCLC form Jan 2007 to Apr 2007 were in- cluded in this study. Control group included 10 patients with benign pulmonary diseses (2 case of hamartoma and 8 case of pulmonary tu berculosis) and 10 healthy volunteers. The pulmonary veins blood and peripheral vein blood were collected respectively. The CD326 immunomagnetic beads and CK-fluorescein isothiocyanate (CK-FITC) were served as the marker antibodies of CTCs. Firstly mononu- clear cell marked by minibeads conjugated with CD326, the mononuclear cells were enriched and separated though Magnetic-activated cell separation(MACS), then the positive separtion cells were marked by anti-CK-FTTC and anti-leukocyte antibody CD45-phyco- erythrine (anti-CD45-PE), finally those cells detected and analyzed by flow cytometry . Results For stage Ⅰ and stage Ⅱ patients (n= 16), CTCs were detected from peripheral vein blood of in 5 (5/16, 31.25%) and from pulmonary veins blood in 9 (9/16, 56.25%). For stage Ⅲ and stage Ⅳ, CTCs were detected from peripheral vein blood in 5 (5/9, 55.56%) and from pulmonary veins blood in 9(7/9,77.78%).For stage Ⅰ,CTCs were detected from peripheral vein blood in 3(3/13,23%)and from pulmo- nery veins blood in 8(8/13,61.54%).The whole CTCs positive detection rate from pilmonary veins blood was 64%(16/25)which higher than feom peripheral vein blood(40%,10/25)(P<0.05).Conclusion The method was set up by MACS combined with FCM to detect the CTCs of pulmonary veins blood and peripheral vein blood of patients.with NSCLC,MACS combined with FCM may improve detection rate of CTCs.This technique appears to be an efficient to detect circulating tumor cells and may be important for dinical practice in the future.     

复合麻醉与硬膜外在经皮肾手术中的应用比较

目的探讨椎管内复合麻醉应用于经皮肾手术治疗的安全性和可靠性。方法选择ASAⅠ～Ⅱ级患者100例，随机分为两组，单纯硬膜外组50例，CSEA组50例。手术治疗均为经皮肾。监测两组患者麻醉效果，对术中镇痛质量进行评价及观察术中患者生命体征变化。结果两组患者麻醉后血压均下降，但两者之间无差别（P〉0．05）。结论CSEA可作为经皮肾手术治疗的麻醉选择，麻醉效果确切而安全。     

非小细胞肺癌循环肿瘤细胞的定量检测意义

Objective To investigate the sensitivity, specificity and clinical significance of detecting circulating tomor cells (CTCs) in NSCLC. Methods Twenty-five patients who undetwent surgical resection for NSCLC form Jan 2007 to Apr 2007 were in- cluded in this study. Control group included 10 patients with benign pulmonary diseses (2 case of hamartoma and 8 case of pulmonary tu berculosis) and 10 healthy volunteers. The pulmonary veins blood and peripheral vein blood were collected respectively. The CD326 immunomagnetic beads and CK-fluorescein isothiocyanate (CK-FITC) were served as the marker antibodies of CTCs. Firstly mononu- clear cell marked by minibeads conjugated with CD326, the mononuclear cells were enriched and separated though Magnetic-activated cell separation(MACS), then the positive separtion cells were marked by anti-CK-FTTC and anti-leukocyte antibody CD45-phyco- erythrine (anti-CD45-PE), finally those cells detected and analyzed by flow cytometry . Results For stage Ⅰ and stage Ⅱ patients (n= 16), CTCs were detected from peripheral vein blood of in 5 (5/16, 31.25%) and from pulmonary veins blood in 9 (9/16, 56.25%). For stage Ⅲ and stage Ⅳ, CTCs were detected from peripheral vein blood in 5 (5/9, 55.56%) and from pulmonary veins blood in 9(7/9,77.78%).For stage Ⅰ,CTCs were detected from peripheral vein blood in 3(3/13,23%)and from pulmo- nery veins blood in 8(8/13,61.54%).The whole CTCs positive detection rate from pilmonary veins blood was 64%(16/25)which higher than feom peripheral vein blood(40%,10/25)(P<0.05).Conclusion The method was set up by MACS combined with FCM to detect the CTCs of pulmonary veins blood and peripheral vein blood of patients.with NSCLC,MACS combined with FCM may improve detection rate of CTCs.This technique appears to be an efficient to detect circulating tumor cells and may be important for dinical practice in the future.     

非小细胞肺癌循环肿瘤细胞检测技术与临床应用的研究进展

转移和复发被认为是影响恶性肿瘤预后的主要因素,对于转移与复发形成的相关基础理论中,"种子和土壤"假说认为,原发肿瘤在生长过程中,有微小肿瘤细胞个体或集团通过循环系统的运送到达远处器官,如同种子播散在土壤中,形成与原发灶特性极其类似的转移灶,这一理论非常形象地说明了恶性肿瘤的转移途径,并点明"种子"肿瘤细胞即循环肿瘤细胞(CTCs)是转移和复发的关键,并有相关可靠的临床研究依据。近年来CTCs又引起肿瘤研究者的热情,将CTCs作为潜在肿瘤生物学标志物及研究肿瘤转移过程成为研究的热点。在本综述中,我们将对CTC生物学和转移相关基因检测、现今最常用的检测技术,及其在临床研究尤其在肺癌领域中的应用进行阐述。     

麦角甾苷眼用固体脂质纳米粒制备及其体外释药的研究

[目的]研究麦角甾苷眼用固体脂质纳米粒的制备方法及其体外释放的情况。[方法]采用乳化蒸发-低温固化法制备麦角甾苷固体脂质纳米粒,超滤离心法测其包封率,并对其粒径、电位、进行进一步考察,用差示扫描量热仪(DSC)表征其性质,采用透析法考察固体脂质纳米粒中麦角甾苷的体外释放行为。[结果]麦角甾苷固体脂质纳米粒的平均粒径为85.56 nm,Zeta电位约为-20.97 m V,药物平均包封率为88.31%,DSC表明其理化性质稳定可靠,体外12 h累计释放率62.46%。[结论]制备的麦角甾苷固体脂质纳米粒包封率较高,粒径小且分布均匀,有良好的缓释作用。     

注射用血栓通（冻干）与3种生物化学注射剂配伍稳定性研究

目的 从制剂分析角度进行中药注射剂与生物化学注射剂配伍稳定性研究。方法 注射用血栓通（冻干）（XST）分别与注射用脑蛋白水解物（Ⅲ）、小牛血清去蛋白注射液以及骨瓜提取物注射液按临床实际条件进行联合使用,采用2,4-二硝基氟苯（DNFB）柱前衍生HPLC法,测定配伍生物化学制剂中16种氨基酸含量。并以2015版《中国药典》为依据,检查配伍7 h内溶液渗透压、不溶性微粒、粒径、pH值、澄明度、颜色等各项制剂分析指标变化情况,综合评价药物配伍联合使用稳定性变化。结果 XST与3种生物化学药物配伍在7 h内各指标（渗透压、pH值及颜色、澄明度）均保持稳定。与小牛血清去蛋白注射液配伍在生理盐水（NS）和5%葡萄糖注射液（5% GS）中粒径波动均较大,但在NS中配伍1 h内粒径值保持稳定;与骨瓜提取物注射液配伍7 h后,在NS输液基质中,甘氨酸含量略有升高、亮氨酸含量略有下降;在5% GS输液基质中,不溶性微粒数值随时间呈增长趋势,但仍处于合格范围内。结论 建议临床XST与小牛血清去蛋白注射液配伍选用NS作为药物溶媒,现用现配,并慢速滴入,观察患者情况;使用XST与骨瓜提取物注射液配伍时建议选用5% GS作为输液剂基质即配即用;为中药注射剂质量再评价和临床合理应用提供理论参考。     

非小细胞肺癌中血管内皮生长因子C表达与淋巴结转移关系的研究

目的 探讨血管内皮生长因子C(VEGF-C)与非小细胞肺癌(NSCLC)淋巴管生成和淋巴结转移的关系.方法 52例NSCLC患者分为淋巴结转移阳性组(25例)和淋巴结转移阴性组(27例),采用半定量反转录PCR及免疫组织化学方法 检测2组患者手术切除癌组织及196枚淋巴结组织(2组各98枚,淋巴结转移阳性组98枚中病理阳性淋巴结72枚,阴性26枚)中VEGF-C mRNA及蛋白的表达.结果 淋巴结转移阳性组患者癌组织VEGF-C mRNA表达水平(0.273±0.179)明显高于淋巴结转移阴性组(0.089±0.087,P<0.01);阳性淋巴结组织VEGF.C mRNA表达水平(0.207±0.174)明显高于淋巴结转移阴性组的淋巴结组织(0.114±0.107,P<0.01).在淋巴结转移阳性组中,阳性与阴性淋巴结组织VEGF-C mRNA表达水平分别为0.207±0.174、0.196±0.186,差异无统计学意义(P>0.05).淋巴结转移阳性组15例患者中14例(93.3%)肺癌组织VEGF-C蛋白表达阳性,46枚阳性淋巴结组织中37枚(80.4%)VEGF-c蛋白表达阳性;而淋巴结转移阴性组15例患者中仅1例(6.7%)肺癌组织VEGF-C蛋白表达阳性,52枚淋巴结组织VEGF-C蛋白表达均为阴性,组间差异均有统计学意义(均P<0.01).结论 VEGF-C mRNA与蛋白的表达水平与NSCLC淋巴结转移关系密切,在淋巴结转移的早期诊断中有潜在的应用价值.     

非小细胞肺癌循环肿瘤细胞的定量检测

目的 探讨非小细胞肺癌循环肿瘤细胞(CTCs)定量检测方法 .方法 CD45免疫磁珠阴性分选组20例,CD326免疫磁珠阳性分选组25例,均为明确诊断的非小细胞肺癌患者.磁性分离富集后肺静脉与外周静脉血标本应用多参数流式细胞仪对CTCs进行定量检测.结果 阴性分选由于只能去除CD45阳性细胞,回收目的 细胞纯度低.阳性分选组中25例术中肺静脉血CTCs定量检测阳性率为64%(16/25),外周静脉血CTCs阳性率40%(10/25,P＜0.05).结论 免疫磁珠富集联合流式细胞分析检测CTCs的敏感性和特异性较高.