叶酸靶向长春新碱纳米微球对腺样囊性癌细胞活性的影响

目的 制备连接有叶酸(folic acid,FA)的长春新碱(vinscristine,VCR)靶向缓释纳米微球(nanopaticles,NP)FA-PLGA(VCR)-NP,观察其表征及对体外培养的腺样囊性癌低转移株(adenoid cystic carcinoma,ACC-2)细胞增殖的抑制作用.方法 采用改良的复乳法制备FA-PLGA(VCR)-NP,测定NP的粒径分布、载药率及体外释放曲线等;MTT比色法检测高浓度下的PLGA-NP对肿瘤细胞的毒性作用,比较药物组VCR、PLGA(VCR)-NP、FA-PLGA(VCR)-NP 3种药物在不同浓度和不同作用时间对ACC-2细胞的抑制作用;异硫氰荧光素标记PLGA-NP和FA-PLGA-NP,荧光显微镜观察FA-PLGA-NP的靶向性及游离FA对其影响.结果 FA-PLGA(VCR)-NP形态圆整,大小均匀,平均粒径249.2nm,载药率4.5%,体外释放时间达14 d;PLGA-NP与ACC-2细胞共培养5 d,细胞存活率达80.0%以上;给药后第4～5天FA-PLGA(VCR)-NP对ACC-2细胞的抑制率显著高于VCR,3种药物对细胞抑制作用均呈时间和浓度依赖性;FA-PLGA-NP可靶向附着于ACC-2细胞表面,这种识别可以被FA竞争性抑制.结论 FA-PLGA(VCR)-NP具有稳定的栽药率和体外释放行为,具有良好的靶向识别力,体外实验证实具有优于VCR原药的抗肿瘤能力.     

多药耐药相关基因在眼眶腺样囊性癌中的表达

目的 了解P53、GST-π、P170和鼠抗人MDR相关蛋白（MRP）在眼眶腺样囊性癌（ACC）组织中的表达情况.方法 免疫组织化学法检测51例眼眶ACC标本和10例正常眼眶组织中P53、GST-π、P170和MRP的表达状况.结果 P53、GST-π和P170在眼眶ACC组织中表达阳性率分别为52.94%、100%和82.35%,与眼眶正常组织相比差异有统计学意义.P53和P170在实体型ACC中的表达率明显高于其在腺样-管状型中的表达率;P53在复发组中的表达率明显高于其在初发组中的表达率;各指标之间在表达上无相关性.结论 眼眶ACC的多药耐药性与P53、GST-π和P170的过表达有关,但作用机制不尽相同,各指标的检测对于判断预后有参考价值.     

长春新碱靶向缓释微球治疗鼠腺样囊性癌的研究

目的 制备连接有叶酸(FA)的长春新碱(VCR)纳米微球(NP),简称为FA-PLGA(VCR)-NP,观察其对体外培养的腺样囊性癌(ACC-2)细胞及裸鼠眼眶移植瘤的抑制作用.方法 实验研究.采用改良的复乳法制备FA-PLGA(VCR)-NP;四甲基偶氮唑盐比色法比较细胞生长抑制率,设VCR、长春新碱微球PLGA(VCR)-NP及FA-PLGA(VCR)-NP组,药物浓度分为0.05、0.25、0.50、1.00、5.00、10.00、30.00 mg/L,检测加药后第1、2、3、4、5天吸光度值;瘤细胞悬液接种法建立裸鼠腺样囊性癌眼眶移植瘤动物模型,分VCR、PLGA(VCR)-NP、FA-PLGA(VCR)-NP及对照组,观察给药后第1、7、14天,各组肿瘤体积抑制率、高效液相色谱法测定肿瘤内残余药物浓度,以及不同时间组织电镜和组织病理学观察.不同时间正常对照与空白微球A值比较采用t检验.考虑到药物、浓度、时间以及三者之间的交互作用,采用多因素方差分析法对比药物对ACC-2细胞增殖的影响.体积抑制率及药物残余浓度比较均采用单因素方差分析,多重比较采用LSD法.结果 FA-PLGA(VCR)-NP呈规则球形,平均粒径249.2 nm,载药率4.53%,体外释放时间达14 d;PLGA-NP与ACC-2细胞共培养5 d,细胞存活率达80%以上(t=1.952～3.285,P=0.081～0.190);FA-PLGA(VCR)-NP对ACC-2细胞的抑制率显著高于VCR,并呈时间和浓度依赖性(F=4.798～563.479,P=0.000～0.006);靶向微球附着于肿瘤细胞表面,这种识别可以被FA竞争性抑制;PLGA(VCR)-NP组和FA-PLGA(VCR)-NP组对裸鼠移植瘤的体积抑制率显著高于VCR组(P=0.029,0.016);FA-PLGA(VCR)-NP组高于PLGA(VCR)-NP组,但差异无统计学意义(P=0.376);给药后第1、7、14天肿瘤残留药物浓度存在差异,时间对浓度的影响具有统计学意义(P=0.000);透射电镜观察14 d肿瘤细胞内仍有高电子密度的纳米颗粒聚集,肿瘤细胞坏死明显,注射周围组织结构形态正常.结论 VCR靶向缓释微球具有稳定的载药率和体外释放行为,具有良好的靶向识别力,体外及体内实验证实具有优于原药的抗肿瘤能力.     

泪腺腺样囊性癌的研究现状

泪腺腺样囊性癌是原发性泪腺上皮性肿瘤中最常见且恶性程度最高的肿瘤.近年来国内外对该病的组织学分类及病理学改变、主要临床表现、治疗手段有了新的认识;在细胞与分子生物学方面,对与泪腺腺样囊性癌的发生、侵袭性生长及转移、嗜神经性生长等特性相关的生物学指标的研究也取得了一些进展.就泪腺腺样囊性癌的研究现状做一综述.     

爱先蓝和细胞角蛋白双重染色技术在泪腺腺样囊性癌诊断中的应用

目的:探讨爱先蓝(Alcian Blue,AB)和细胞角蛋白(cytokeratin,CK)双重染色技术在泪腺腺样囊性癌组织中的应用,提高泪腺腺样囊性癌染色效率.方法:选取中山大学中山眼科中心临床病理科2015年1月至2017年1月期间诊断为泪腺腺样囊性癌病例标本23例,在同一张切片上先进行AB染色,再进行CK染色,观察染色效果.结果:23例泪腺腺样囊性癌标本组织中黏液物质全部呈蓝色;癌细胞胞质CK阳性,呈棕黄色.结论:AB和CK双重染色方法稳定,颜色对比鲜明,能够良好显示癌细胞及黏液的关系,并且比分开的两次单种染色省时、经济.     

人泪腺腺样囊性癌细胞的原代培养及其形态学特征

目的 探讨人泪腺腺样囊性癌细胞的原代培养方法.方法 实验研究.取2011年5月16日至6月1日在天津医科大学眼科医院接受手术治疗的1位泪腺腺样囊性癌患者的术中标本,采用组织块培养法获得原代细胞.运用连续传代、机械刮除、反复贴壁和消化排除等方法去除杂细胞.通过相差显微镜、扫描电镜、透射电镜等观察细胞形态.免疫细胞化学法检测细胞波形蛋白(vimentin)、结蛋白(desmin)、S-100蛋白、细胞角蛋白(cytokeratin,pan)、CD34等蛋白的表达情况.免疫组织化学法检测肿瘤组织vimentin、desmin、S-100蛋白、cytokeratin(pan)等蛋白的表达情况.消化法绘制细胞生长曲线,计算细胞倍增时间.结果 组织块原代培养第5天见周围开始有细胞爬出,此后细胞缓慢增殖,至第32天进行首次传代,第69天发现典型的上皮样细胞克隆.运用多种方法去除杂细胞后,至第8代细胞基本纯化.纯化后细胞现已稳定传至100代以上.相差显微镜下观察第25代细胞呈典型的上皮样细胞形态,可出现接触抑制消失.扫描电镜和透射电镜下第25代细胞呈现低分化恶性肿瘤细胞的典型特点,细胞间信号沟通丰富.细胞免疫化学法检测第25代癌细胞vimentin、cytokeratin (pan)和S-100蛋白呈阳性,desmin、CD34蛋白呈阴性,与源肿瘤组织染色结果一致.细胞生长曲线类似“S”形,细胞倍增时间37.1 h.结论 采用组织块培养法可以获得基本纯化的人泪腺腺样囊性癌细胞,有可能通过此法获得稳定的人泪腺腺样囊性癌细胞系.     

眼眶腺样囊性癌的影像学研究

目的分析眼眶腺样囊性癌(ACC)的超声、CT及MRI等影像学特征。设计回顾性病例系列。研究对象经病理诊断证实的泪腺腺样囊性癌38例和副鼻窦来源的腺样囊性癌4例。方法分析眼眶腺样囊性癌超声、CT和MRI等各种影像学检查结果,记录图像特征。主要指标标准化A/B超、CT和MRI影像。结果ACC标准化A超表现为前部高波峰,后部低波峰和不规则高低混杂波峰两种类型。B超可显示肿瘤的轮廓,对于瘤内钙化及液化坏死等病变显示优良。CT扫描显示ACC有梭形、扁平形、团块状和不规则形等四种形状,对于肿瘤的边界、空间位置,骨破坏和骨吸收等继发改变显示清晰。ACC在T_1加权像为中信号,T_2加权像有均一的等信号,等、低信号混杂和等、高信号混杂三种,多数不均匀强化。强化MRI有利于显示肿瘤的蔓延途径,泪腺ACC有多种蔓延途径。副鼻窦ACC易累及筛窦和上颌窦。结论各种影像学检查可显示眼眶ACC的不同特性。综合运用和分析各种影像学检查结果可提高诊断水平。(眼科,2007,16:391-394)     

泪腺腺样囊性癌CT表现的研究

目的 分析泪腺腺样囊性癌常见的CT图像特征,从而探讨CT(computed tomography)影像对于泪腺腺样囊性癌的诊断价值.方法 病历来源为天津医科大学眼科中心及天津医科大学第二医院,经病理确诊的眼眶腺样囊腺癌39例.其中原发肿瘤20例,复发肿瘤19例,均为单眼发病,所有患者均行眼眶CT检查,获取水平及冠状扫描图像.结果 泪腺腺样囊性癌中原发肿瘤形状多为厚扁平状,贴附于眶壁增长,在体层像上呈锥形者10例、不规则形5例;复发肿瘤形状为不规则型8例、厚扁平状,贴附于眶外上壁向后增长者8例.原发与复发泪腺腺样囊腺癌的好发位置位于眶外上方分别为18例与11例.结论 CT可清晰显示肿瘤的位置,形状,范围、眶骨壁改变,对于泪腺腺样囊性癌的诊断和制定治疗方案具有重要指导意义.

Abstract：

Objective To analyze imaging characteristics of adenoid cystic carcinoma(ACC)of lachrymal gland on CT for the diagnosis of ACC in clinic. Methods Thirty-nine patients with ACC consisting of 20 cases of primary tumor and 19 cases of recurred tumors confirmed by pathology were collected from The Eye Centre of Tianjin Medical University and The Second Hospital of Tianjin Medical University. All the patients were unilateral, and examined by CT. The imaging characteristics of ACC on CT were analyzed. Results The most common shape of primary ACC included 10 cases of conical-shape, 5 cases of irregular-shape. The recurred cases contained 8 cases of irregular-shape, 8 cases of conical-shape. The primary and recurred ACC usually existed on superior-external side, with which there were 18 cases and 11 cases respectively. Conclusions CT examination can exactly reveal the shape, size, position, and ossa orbitale destruction of ACC and played an important role in the diagnosis of ACC.     

凋亡相关基因在眼眶腺样囊性癌组织中的表达意义

目的:分析凋亡相关基因p53和bcl-2在眼眶腺样囊性癌组织中的表达,分析其关系及临床意义。方法:免疫组化法检测51例眼眶腺样囊性癌临床标本和10例正常眼眶组织中P53和Bcl-2蛋白的表达状况。结果:P53和Bcl-2在眼眶腺样囊性癌组织中的表达阳性率分别为53%和65%,显著高于眼眶正常组织(53%,65%vs10%,P<0.01)。实性型腺样囊性癌中P53的表达率显著高于其在腺样-管状型中的表达率(86%vs30%,P<0.01);P53和Bcl-2在复发组中的表达率显著高于其在初发组中的表达率(85%vs42%,P<0.01),(100%vs53%,P<0.01);P53和Bcl-2二者在表达上具有相关性(χ2=1.461,P>0.05)。结论:P53和Bcl-2蛋白的检测对于判断肿瘤对化疗的敏感性和预后有参考价值。     

泪腺腺样囊性癌的放射治疗

泪腺腺样囊性癌(adenoidcysticcarcinoma,ACC)是泪腺恶性上皮性肿瘤中最常见的,也是恶性度最高的肿瘤。其发病率占泪腺上皮性肿瘤的25%～30%[1],占泪腺恶性上皮性肿瘤的41.7%[1],复发率达77.14%[26],远处转移率达37%[26],本病病程短,可经周围神经浸润性生长、血行及淋巴系统转移,预后不佳。目前多采用包括放化疗在内的综合治疗的方案。本文着重从放疗的角度对ACC治疗的具体方法加以阐述。     

耐长春新碱视网膜母细胞瘤细胞株的建立

目的：研究视网膜母细胞瘤获得性多药耐药现象，探讨其产生机制。方法：利用长春新碱(vincristree，VCR)对人视网膜母细胞瘤细胞系HXO-RB₄₄细胞在体外进行浓度递增法逐步诱导实验，建立一株耐长春新碱视网膜母细胞瘤细胞亚株HXO—RB/VCR。并对其细胞生长，药物含量，药物敏感性与放射性敏感性进行检测。 结果：该耐药细胞株HXO—RB/VCR细胞对长春酰胺、丝裂霉素C、阿霉素、足叶乙甙、顺铂、卡铂等有交叉耐受，对卡氮芥、氟尿嘧啶无明显抗药性，而对氨甲喋呤的敏感性反而增加。对⁶⁰ Coγ射线有轻度耐受性。细胞内药物含量测定显示：耐药细胞内长春新碱浓度明显低于敏感细胞，钙通道阻断剂异博定可使细胞内VCR浓度增加，部分逆转其耐药性。 结论：VCR可诱导视网膜母细胞瘤产生多药耐药和轻度放射耐受，获得性多药耐药作用可被异博定逆转。 （中华眼底病杂志，1997，13：6-9）     

眼眶腺样囊性癌临床病理分析

目的：探讨眼眶腺样囊性癌（adenoid cystic carcinoma ACC)的临床特征及病理组织学特点。方法：回顾性分析20例ACC患者的临床及病理资料。结果：ACC主要表现为眶内包块（20/20），眼球突出移位（14/20），眼球运动障碍（14/20），眶周疼痛（13/20）等；病理组织学分5型；筛状型，管状型，实体型，粉刺型和硬化型。结论：ACC的临床特征有助于其诊断，最终确诊有赖于病理组织学检查。     

microRNA-125b对人视网膜母细胞瘤多药耐药性影响的实验研究

目的 探讨microRNA-125b（miR-125b）在人视网膜母细胞瘤细胞中多药耐药的作用及其机制。设计 实验研究。 研究对象 人视网膜母细胞瘤SO-RB50细胞。方法 用RT-PCR方法检测miR-125b视网膜母细胞瘤细胞株SO-RB50和耐药细胞株SO-Rb50/VCR中的表达变化;化学合成的miR-125b过表达（miR-125mimic 组）和抑制载体（miR-125inhibitor组）转染SO-Rb50细胞株,用MTT法和Annexin V-FITC法检测在药物长春新碱、依托泊苷和卡铂,依次作用上述转染细胞后,细胞增生力和细胞凋亡的变化;用蛋白印迹法检测miR-125b过表达和抑制表达后细胞株SO-RB50中 MAGE-A/P53蛋白的表达变化。主要指标 细胞存活率和细胞凋亡率。结果 SO-Rb50/VCR组与SO-RB50组相比,miR-125b的表达显著增高（P=0.002）;长春新碱、依托泊苷和卡铂依次作用于转染后的SO-RB50细胞株后,miR-125mimic组与miR-125inhibitor组相比,细胞存活率显著增高（P=0.000）,细胞凋亡率显著下降（P=0.000）,P53蛋白表达水平显著下降（P=0.001）,MAGE-A蛋白表达水平显著增高（P=0.004）。结论 在SO-RB50细胞中,下调miR-125b后提高肿瘤细胞对化疗药物敏感性,且miR-125b可能是通过MAGE-A/P53通路调控视网膜母细胞瘤多药耐药性。     

Clinical analysis of the effect of intraarterial cytoreductive chemotherapy in the treatment of lacrimal gland adenoid cystic carcinoma

PURPOSE: To determine the effect of intraarterial cytoreductive chemotherapy (IACC) as an adjunct to conventional surgery and radiation therapy for lacrimal gland adenoid cystic carcinoma (ACC). DESIGN: A retrospective, comparative, interventional case series. METHODS: setting: Institutional. patient population: Nine consecutive patients with lacrimal gland ACC were treated with IACC, followed by orbital exenteration and chemoradiotherapy. This case series was compared with a series of seven patients treated by conventional local therapies in the same institution. intervention procedure: Clinical records, imaging studies, histologic sections, and archival specimens from all 16 patients were reviewed. Information analyzed included site of disease, histologic characteristics, extent of disease, incidence of locoregional recurrence or distant metastases, and disease-free survival and overall survival time. main outcome measure: The effect of IACC was assessed by the radiographic and histologic response and survival outcome in comparison to a historical cohort of patients managed by conventional local therapies. RESULTS: The difference between the carcinoma cause-specific death rate of the study group versus conventional treatment was significant (P = .029, log rank test). The cumulative 5-year carcinoma cause-specific death rate in the IACC treated group was 16.7% compared with 57.1% in the conventional treatment group. The cumulative 5-year recurrence rate in the IACC treated group was 23.8% compared with 71.4% in the conventional treatment group. CONCLUSIONS: The preliminary data suggest that IACC as an integral component of a multimodal treatment strategy is potentially effective in improving local disease control and overall disease-free survival in lacrimal gland adenoid cystic carcinoma.     

Choroidal metastasis from adenoid cystic carcinoma of the lung

PURPOSE: We report a case of a choroidal metastasis from an adenoid cystic carcinoma of the lung.Interventional case report. DESIGN: A 40-year old man, 9 months' status postresection of a bronchial adenoid cystic carcinoma, was diagnosed by clinical evaluation and fine-needle aspiration biopsy and treated with palladium-103 ophthalmic plaque brachytherapy. RESULTS: This unusual patient with uveal metastasis from adenoid cystic carcinoma presented with decreased vision in the right eye and a diaphanous amelanotic choroidal tumor in the superotemporal macula. Ultrasound revealed a dome-shaped tumor that measured 4.0 mm in apical height and 11 x 10 mm in basal diameter. Fluorescein angiography revealed a double circulation and late intense subretinal fluorescence. Plaque brachytherapy provided local control and preservation of the eye. CONCLUSION: Bronchogenic adenoid cystic carcinoma can metastasize to the choroid.     

Space-occupying lesions of the lacrimal gland at one tertiary eye center in China: a retrospective clinical study of 95 patients

AIM: To investigate the treatment status and prognosis of space-occupying lacrimal gland lesions at one tertiary eye center in China. METHODS: A retrospective clinical study was performed on 95 patients with space-occupying lesions of the lacrimal gland surgically treated at the Eye & ENT Hospital of Fudan University from 2003 to 2007. The reviewed clinical data included age, gender, side of the lesion, duration of signs and symptoms, histopathological diagnosis, treatment modality, recurrence (local, regional, and distant metastasis) and survival. RESULTS: Of the 95 cases (99 eyes), pleomorphic adenomas were the most common lesions (43 cases), followed by lymphoid disorders (14), inflammatory pseudotumors (11), carcinoma ex-pleomorphic adenomas (11), and adenoid cystic carcinomas (ACC, 6). There were 8 patients with relapsed pleomorphic adenomas. Five of these 8 cases had malignant pathological changes. All patients with ACC had metastasis and three of them died during their follow-up. CONCLUSION: Our study indicated that the most common lacrimal gland lesions were pleomorphic adenomas. Multiple recurrence and surgical procedures may increase the risk of tumor progression. ACC had a high incidence of tumor metastasis and a poor prognosis.     

泪腺腺样囊性癌CT表现的研究

目的 分析泪腺腺样囊性癌常见的CT图像特征,从而探讨CT(computed tomography)影像对于泪腺腺样囊性癌的诊断价值.方法 病历来源为天津医科大学眼科中心及天津医科大学第二医院,经病理确诊的眼眶腺样囊腺癌39例.其中原发肿瘤20例,复发肿瘤19例,均为单眼发病,所有患者均行眼眶CT检查,获取水平及冠状扫描图像.结果 泪腺腺样囊性癌中原发肿瘤形状多为厚扁平状,贴附于眶壁增长,在体层像上呈锥形者10例、不规则形5例;复发肿瘤形状为不规则型8例、厚扁平状,贴附于眶外上壁向后增长者8例.原发与复发泪腺腺样囊腺癌的好发位置位于眶外上方分别为18例与11例.结论 CT可清晰显示肿瘤的位置,形状,范围、眶骨壁改变,对于泪腺腺样囊性癌的诊断和制定治疗方案具有重要指导意义.     

【In Press】 Expression of multidrug resistance proteins in retinoblastoma

AIM: To elucidate the mechanism of multidrug resistance in retinoblastoma (RB), and to acquire more insights into in vivo drug resistance. METHODS: Three anticancer drug resistant Y79 human RBcells were generated against vincristine, etoposide or carboplatin, which are used for conventional chemotherapy in RB. Primary cultures from enucleated eyes after chemotherapy (PCNC) were also prepared. Their chemosensitivity to chemotherapeutic agents (vincristine, etoposide and carboplatin) were measured using MTT assay. Western blot analysis was performed to evaluate the expression of p53, Bcl-2 and various multidrug resistant proteins in RB cells. RESULTS: Following exposure to chemotherapeutic drugs, PCNC showed less sensitivity to drugs. No significant changes observed in the p53 expression, whereas Bcl-2 expression was found to be increased in the drug resistant cells as well as in PCNC. Increased expression of P-glycoprotein (P-gp) was observed in drug resistant Y79 cells; however there was no significant change in the expression of P-gp found between primary cultures of primarily enucleated eyes and PCNC. Multidrug resistance protein 1 (Mrp-1) expression was found to be elevated in the drug resistant Y79 cells as well as in PCNC. No significant change in the expression of lung resistance associated protein (Lrp) was observed in the drug resistant Y79 cells as well as in PCNC. CONCLUSION: Our results suggest that multidrug resistant proteins are intrinsically present in RB which causes treatment failure in managingRB with chemotherapy.