肾母细胞瘤裸鼠移植瘤模型的建立及其生物学特性

目的 采取手术标本,在裸鼠皮下连续传代建立人肾母细胞瘤裸鼠移植瘤模型。方法 切取肾母细胞瘤患儿肿瘤组织植入裸小鼠右前肢皮下,连续传代。观测肿瘤生长,对各代移植瘤进行光镜和电镜观察,波形蛋白（ｖｉｍｅｎｔｉｎ）及上皮膜抗原（ＥＭＡ）免疫组化染色,ＷＴ１基因检测。结果 １０个月中在裸鼠皮下传４代,总成瘤率为４３％。移植瘤生长曲线与Ｇｏｍｐｅｎｔｚ函数符合。     

神经母细胞瘤裸鼠移植瘤模型的建立及相关应用研究进展

自 196 9年Ｒｙｇａｒｒｄ和Ｐｏｖｌｓｅｎ(Ｃｏｎ ｃｅｒＲｅｓ,1975 )首次将人结肠癌成功移植于裸鼠 ,建立裸鼠移植瘤模型以后。人们已将这一方法应用于许多肿瘤的研究 ,神经母细胞瘤也不例外。 1975年ＬａｗｒｅｎｃｅＨｅｌｓｏｎ第一次成功的将人神经母细胞瘤ＳＫ Ｎ ＳＨ、ＳＫ Ｎ ＭＣ细胞移植于裸鼠皮下建立起移植瘤模型。裸鼠移植瘤模型可以直接模拟人体内肿瘤的自然生长过程 ,与体外细胞培养相比 ,移植瘤模型具有以下优点 :①模型的建立和长期传代相对方便 ,不受成纤维细胞污染 ;②移植瘤的增生受到宿主的营养状况和血供的影响 ,…     

小儿睾丸卵黄囊瘤细胞的体外培养及生物学特性研究

目的 采用组织培养技术在体外培养取自人体的睾丸卵黄囊瘤组织,探讨小儿睾丸卵黄囊瘤组织的体外培养方法及生物学特性.方法 采用组织块培养法培养1例睾丸卵黄囊瘤患儿手术的标本,从形态学、细胞生长动力学、肿瘤内分泌、染色体分析、细胞DNA分析等方面初步研究了小儿睾丸卵黄囊瘤细胞的生物学特性.结果 小儿睾丸卵黄囊瘤细胞无论形态学观察,还是功能学测定,均符合卵黄囊瘤细胞的特征,染色体众数39～97条,并具有体外分泌AFP功能.其特有表现为:细胞贴壁生长,呈短梭形或多角形,细胞密集时可多层重叠生长,肿瘤细胞表面有明显的微绒毛,细胞核形态怪异,细胞倍增时间较长.结论 小儿睾丸卵黄囊瘤细胞在体外生长稳定,增殖活跃,肿瘤细胞的纯度较高,可以用于后续的实验研究.

Abstract：

Objective To establish a method for the culture of testicular yolk sac tumor in childhood and then investigate biological characteristics in vitro. Methods One specimen from testicular yolk sac tumor was cultured in vitro. Testicular yolk sac tumor cell lines were studied morphologically and subjected to karyotype analysis, DNA analysis, and tumor formation evaluation. Results Morphological observation and functional analysis show that cell lines have characteristics of testicular yolk sac tumor. The number of chromosomes varied from 39 to 97. Turours were immunostained positively for AFP in vitro and found to form multiple layers with microvilli. The nuclei were variable and bizarre in size and shape. Conclusions Testicular yolk sac tumor cell lines were cultured proliferated in a stable manner in vitro, which provides a convenient and economical object for basic researches on yolk sac tumor in future.     

儿童毛细血管瘤裸鼠移植模型的制作研究

目的　建立人毛细血管瘤裸鼠移植瘤模型。同时观察肿瘤生长情况 ,探讨血管瘤裸鼠模型建立的条件。方法　将手术切除的 1例雌激素受体阳性的儿童增生期草莓状血管瘤组织制成组织块 ,植入 16只幼裸小鼠 (BALB/cnudemice)皮下 ,每只 4处。将 16只裸小鼠分为 2个实验组。实验 1组在移植后给予普通鼠食喂养 ;实验 2组在 1组基础上每周肌注雌二醇 0 .1mg。喂养 90d ,观察2个实验组中移植瘤的生长情况。 90d后对成活的移植瘤标本进行病理学光镜检查 ,血管内皮细胞单克隆抗体CD3 4 、细胞增殖标记抗体Ki 6 7免疫组化染色。证实成活的移植瘤来源于原人体血管瘤 ,并了解其生物学特点。结果　 90d后 ,单纯喂养的实验 1组所有移植瘤均未成活 ,被吸收或形成脓肿。而实验 2组移植瘤均成活。光镜下成活的移植瘤与原血管瘤组织生物学特点相似。CD3 4 阳性指数达 4 8.4 8± 3.90。Ki 6 7阳性指数 15 .0 4± 2 .4 4 ,与正常人体皮肤的Ki 6 7阳性指数 1.14± 0 .4 0相比 ,差异具有显著性意义 (P <0 .0 1)。结论　通过移植人体血管瘤在裸鼠体内可建立人类血管瘤的动物模型。该模型保留了人类血管瘤的主要生物学特点 ,是血管瘤基础和临床研究的良好模型。     

小儿卵黄囊瘤临床病理及免疫组化分析

目的：探讨小儿卵黄囊瘤临床病理特点。方法：对61例小儿卵黄囊瘤临床病理资料回顾分析,部分进行免疫组织化学染色。结果：男36例,女25例,发病年龄出生～14岁,平均 22.3月;<3岁的占 78.7 %。性腺内39例,性腺外22例。镜下病理形态多样,以内胚窦结构(SD小体)最具特征性,但并非每例都能见到,网状结构、玻璃样小体、微囊结构及腺样结构在所有病例均可见到 ,常伴有出血坏死。免疫组织化学染色显示AFP+,AAT+,CK+,HCG-,PCNA+,P53+。结论：小儿卵黄囊瘤多为单一成分的卵黄囊瘤,常见于睾丸(占54.1%),预后较成人好。性腺外卵黄囊瘤主要位于骶尾部(20/22例),且多为女性(18/20例),预后较差 。诊断卵黄囊瘤时不必过分强调S D小体的存在,免疫组织化学染色有助于鉴别诊断。     

Avastin联合环磷酰胺治疗神经母细胞瘤裸鼠移植瘤的研究

目的 探讨Avastin抗神经母细胞瘤(NB)裸鼠移植瘤生长的作用,以及对血清血管内皮生长因子(VEGF)水平的影响.方法 培养NB细胞并建立NB裸鼠移植瘤.用Avastin单药或与环磷酰胺(CPM)联合治疗NB裸鼠移植瘤.Avastin剂量为5 rag/kg,由尾静脉注入,每周1次;CPM75 mg/kg,仅1次.于第22天处死裸鼠,计算肿瘤体积及抑瘤率.应用抗CD34免疫组化法检测肿瘤微血管密度,ELISA法测定各组治疗结束后血清VEGF(sVEGF)水平.结果 Avastin单药治疗、CPM单药化疗、Avastin联合CPM治疗NB裸鼠移植瘤的抑瘤率分别是44.0%、38.1%、62.8%.Avastin联合CPM化疗与Avastin单药治疗和CPM单药化疗比较,差异均有统计学意义(P＜0.05).Avastin对NB血管生成的抑制率为64%.Avastin治疗后尽管裸鼠肿瘤已明显缩小,但sVEGF升高至对照组的1.9～2.4倍.结论 Avastin能抑制NB裸鼠移植瘤的生长,而且与CPM有协同抗NB作用.sVEGF不能用于监测Avastin抗瘤效果.     

FASN在小儿卵黄囊瘤中的表达及临床意义

目的分析小儿卵黄囊瘤的临床特点,探讨脂肪酸合酶(Fatty acid synthase,FASN)在小儿卵黄囊瘤中的表达及意义。方法回顾性分析26例卵黄囊瘤患儿临床资料,运用免疫组化检测26例卵黄囊瘤、17例未成熟畸胎瘤、1例胚胎性癌及12例相关部位正常组织标本中FASN的表达情况。结果26例小儿卵黄囊瘤发病平均年龄2岁5个月,16例单纯以肿块就诊,5例单纯以局部继发症状就诊,5例同时具有肿块及局部症状。FASN在卵黄囊瘤的表达程度(73%)显著高于未成熟畸胎瘤(28%)、胚胎性癌(0%)、正常组织(16%)。FASN在性腺内和性腺外小儿卵黄囊瘤的阳性表达率分别为56%和100%。伴淋巴结或远处转移组和无转移组阳性表达率分别为100%和61%。FASN的阳性表达率与临床分期呈正相关。以上差别均有统计学意义。化疗前后FASN表达率分别为72%和75%,差别无统计学意义。结论小儿卵黄囊瘤起病隐匿,进展迅速,手术治疗联合化疗有效,FASN的表达可为诊治提供参考。     

儿童盆腔卵黄囊瘤的64层螺旋CT诊断与临床病理对照分析

目的分析儿童盆腔卵黄囊瘤的临床、病理及64层螺旋CT(MSCT)特点,以提高对本病的诊断水平。方法回顾性分析34例经手术病理检查证实的盆腔卵黄囊瘤儿童的临床、病理、MSCT及多平面重建资料。术前均行盆腔MSCT平扫及增强扫描。结果本组34例中,女性21例,男性13例;患儿年龄3个月至7岁3个月,平均(18±15)个月龄。临床表现主要为腹部或骶尾部肿块以及腹痛。33例甲胎蛋白(AFP)明显升高,1例(占2.9%)AFP值为2.2 ng/mL。34例患儿中,18例来源于盆腔腹膜腔内,16例位于腹膜腔外向骶尾部生长,其中9例主体部分位于盆腔,7例主体部分位于骶尾部。MSCT均表现为卵圆形或不规则形囊实性肿块,增强后肿瘤的实性部分明显强化,肿块最大径为2.8~10.7 cm。AFP值与肿块大小及部位无显著相关性。结论盆腔卵黄囊瘤的临床及MSCT表现有一定特征,CT能较准确描述肿瘤内部结构与血供,结合血清AFP检查,有利于盆腔卵黄囊瘤的正确诊断、制订术前手术方案及判断术后复发情况。     

三氧化二砷抑制人胃癌裸鼠皮下移植瘤生长及机制的研究

目的探讨三氧化二砷(As2O3)抗人胃癌裸鼠皮下移植瘤的作用及其机制。方法实验用30只裸鼠建立人胃癌裸鼠移植瘤模型,随机分为3组,即生理盐人组、低剂量As2O3组和高剂量As2O3组,比较各组的抑瘤作用,并对标本分别进行光镜和电镜观察及原位末端标记(TUNEL)检测。结果As2O3能显著抑制胃癌裸鼠移植瘤的增长,并能诱导肿瘤细胞凋亡,未见As2O3引起肝、肾系统损害。结论As2O3对人胃癌细胞裸鼠移植瘤具有显著的抗癌作用,此作用与诱导癌细胞凋亡密切相关。     

CT对儿童原发肝脏卵黄囊瘤和肝母细胞瘤的鉴别诊断

目的 探讨儿童原发肝脏卵黄囊瘤的影像学表现,并寻找可能与肝母细胞瘤相鉴别的影像学特征。方法 回顾性分析北京大学第一医院收治的1例原发肝脏卵黄囊瘤患儿的临床表现,尤其是其影像学资料,与22例肝母细胞瘤资料进行对比分析,并复习相关文献。结果 原发肝脏卵黄囊瘤患儿为1岁11个月男童,甲胎蛋白(AFP)明显升高,腹部增强CT及B超均提示肝脏右叶巨大单发占位,未见明显钙化,肿物内可见出血、坏死,有包膜下积液等肿瘤破裂迹象。肝母细胞瘤患儿AFP亦明显升高,腹部增强CT提示多为单发肝脏巨大肿物,结节状多见,无包膜,多见点状或片状钙化,肿物周边有明显血流信号,未见有肿瘤破裂迹象。复习文献8例原发肝脏卵黄囊瘤患儿的影像学资料显示多有出血和坏死表现,1例有明显肿瘤破裂表现,无钙化。结论 原发肝脏卵黄囊瘤和肝母细胞瘤都表现为肝脏巨大占位伴AFP明显升高,前者影像学特征是肿物更容易出现出血、坏死和肿瘤破裂迹象,有包膜,无钙化,对二者有一定的临床鉴别价值。     

人肾母细胞瘤裸小鼠肾原位移植模型的建立和生物学特性的研究

目的 建立人肾母细胞瘤的裸小鼠肾原位移植模型。方法 直接将未经处理的患儿肾母细胞瘤组织块接种在裸小鼠肾包膜下作原代以及系列传代移植；原、2、3、4代接种裸小鼠数目分别为2、5、5、18只，每代间隔时间均为8周。原、2、3代裸小鼠分别于接种8周后全部处死，4代裸小鼠从接种后3周至8周，每周处死3只至全部，取出移植瘤进行称重，以反映肿瘤生长动力学。采用流式细胞仪检测原代和系列传代肿瘤样本中细胞周期/凋亡分布以及倍体数。光学显微镜下观察肿瘤标本形态学以及角质蛋白18和结蛋白的免疫组化染色特征，聚合酶链反应扩增基因组微卫星序列D14S68、D18S69and D20S199，以确定其人源性。结果 人肾母细胞瘤在原代裸小鼠中形成肾原位移植瘤，随后系列传四代，共30只裸小鼠，其中27只在肾原位可以检测到移植瘤生长；移植瘤生长曲线符合指数模型，其倍体数、细胞周期分布在各代之间相似。所有移植瘤都保留了最初的组织形态学和免疫组化染色特征。而且，移植瘤基因组微卫量DNA序列与人肾母细胞瘤完全一致。结论 该异种原位移植瘤模型，准确地重现所移植的人肾母细胞瘤的形态和恶性生物学特点，具有重要的基础和临床研究应用价值。     

SALL4在儿童卵黄囊瘤中表达的研究

目的 研究分析一种新的肿瘤标志物SALL4在儿童卵黄囊瘤(yolk sac tumor,YST)的表达情况及临床意义.方法 收集儿童卵黄囊瘤病理标本21例(含有穿刺标本8例).选取成熟畸胎瘤(mature teratoma,MT)10例、未成熟畸胎瘤(immature teratoma,IMT)5例、无性细胞瘤1例进行对照研究.应用免疫组织化学方法检测SALL4在上述肿瘤中的表达情况,并就SALL4与甲胎蛋白(alpha fetoprotein,AFP)在YST的表达情况进行比较.结果 ①SALL4在21例儿童YST全部表达,且每例都有较高的阳性表达强度;②在儿童YST中,SALL4的阳性表达强度强,为6例(++)、15例(+++),而AFP在YST的表达相对较弱,分别为2例(-)、5例(+)、9例(++)、5例(+++),SALL4表达强度比AFP更强(P＜0.01);在儿童YST中,SALL4与AFP的表达强度之间进行相关性分析,发现两者无明显相关性(P＞0.05);③8例穿刺标本中,SALL4染色3例(++)、5例(+++),AFP染色1例(-)、2例(+)、4例(++)、1例(+++),两者比较差异有统计学意义(P＜0.05).结论 ①在儿童常见生殖细胞肿瘤(GCT)中,SALL4可能作为YST相对敏感的新一代瘤标;②在免疫组化染色分析中,SALL4比AFP在儿童YST中阳性表达强度更高;但SALL4与AFP的表达强度之间无明显相关性;③SALL4免疫组化染色可帮助儿童YST的诊断,尤其针对组织量少的穿刺标本,更敏感的瘤标SALL4有助于标本的病理分子学诊断.

Abstract：

Objective To study the expression of SALL4 in yolk sac tumors (YST) in children.Methods Twenty one patients were diagnosed with yolk sac tumor and treated in the children's hospital of Chongqing medical university. Their tumor tissue specimens including 8 puncture biopsy specimens were collected for this study. In addition, 10 cases of mature teratoma (MT), 5 cases of immature teratoma (IMT). 1 dysgerminoma were collected as control The expressions of SALL4 in YST,MT, IMT and dysgerminoma were studied by SP immunohistochemical method. Alpha fetoprotein 21 patients. Among them, the expression of SALL4 in YST was strong ( + + ) in 6 patients, and very It is negative in 2 cases, detectable in 5, strong in 9, and very strong in 5. The expression of SALL4 expression of SALL4 was strong ( + + ) in 3 cases, and very strong ( + + + ) in 5 cases. The staining of AFP was negative in 1 case ( - ), detectable in 2 ( + ), strong in 4 ( + + ) and very strong in 1 ( ++ + ). A significant difference was found between SALL4 and AFP expression in these specimens (P＜0. 05). Conclusions SALL4 could be a new relatively sensitive tumor marker for YST. The expression of SALL4 is stronger than AFP in YST. But their expression doesn't correlate with each other.Immunohistochemical staining of SALL4 could be helpful to diagnose YST in children, especially in the small amount specimens.     

RNA-binding protein LIN28 is a sensitive marker of pediatric yolk sac tumors

Background

RNA-binding protein LIN28 is involved in maintaining the pluripotency of embryonic stem cells. It has been detected in different types of testicular and ovarian germ cell tumors (GCTs), but its status in pediatric YSTs (yolk sac tumors) is still unknown. The aim of this study was to determine the immunohistochemical profile of LIN28 in pediatric YSTs.

Methods and results

Immunohistochemistry detection of LIN28 was performed in 22 cases of pediatric YSTs and 10 mature teratomas. The percentage of tumor cells stained was scored as 0, 1+ (1–30 % cells), 2+ (31–60 %), 3+ (61–90 %), and 4+ (>90 %). To compare its sensitive and specificity with alpha-fetoprotein (AFP), we also stained AFP in 22 cases of pediatric YSTs and 10 mature teratomas in children. LIN28 staining was high in all 22 pediatric yolk sac tumor (2+ in 1, 3+ in 1, and 4+ in 20), and weak staining of LIN28 was seen in 1 of 10 mature teratomas (1+), 9 of 10 mature teratomas were negative expression. However, the expression of AFP in pediatric YST was lower compared with Lin28 (- in 1, 1+ in 8, 2+ in 12, and 3+ in 1), and weak expression of AFP was seen in 2 of 10 mature teratomas (1+), 8 of 10 mature teratomas were negative. LIN28 had higher intensity expression than AFP in pediatric YSTs (P < 0.001).

Conclusions

LIN28 is a sensitive marker for pediatric YSTs and it can be used to distinguish them from mature teratomas. LIN28 is likely to become a new and valuable biomarker for diagnosing of pediatric YST.

     

Yolk sac tumor with a unique uniform hepatoid pattern histology

Abstract Background: Yolk sac tumors (YST) exhibit several different histological subtypes and the mechanisms of cellular differentiation and prognosis of each subtype remain unknown.
Results: We report two infants with sacrococcygeal YST; one represented a typical histological subtype and the other showed a hepatoid subtype with uniform histology. The isoform of alpha-fetoprotein (AFP) in the patient with the hepatoid pattern was examined by lectin-affinity Immunoelectrophoresis and represented as a YST. but not hepatocellular, subtype. The patient with typical YST responded well to VAB-6 combination chemotherapy. However, this regimen was only partially effective to the patient with the pure hepatoid histological subtype, and an etoposide with ifosfamide and cisplatin (VIP) regimen as a salvage chemotherapy combined with complete tumor resection was useful to achieve complete remission (CR). Both of the patients have been in CR for more than four years.     

舒尼替尼及联合顺铂对小儿睾丸卵黄囊瘤荷瘤鼠模型的抗肿瘤作用

目的 探讨舒尼替尼(Sunitinib)、顺铂(CDDP)及两者联合用药对小儿睾丸卵黄囊瘤(TYST)异种移植荷瘤鼠模型的抗肿瘤作用及相关作用机制.方法 肿瘤标本来自本实验室的小儿睾丸卵黄囊瘤裸鼠第17代模型,并接种在雄性裸鼠单侧腹股沟皮下区,成瘤后随机分成4组(n=5):对照组、CDDP组、Sunitinib组和Sunitinib+CDDP组.绘制肿瘤体积和裸鼠体重变化曲线图,计算肿瘤消退率;HE染色观察肿瘤组织形态学变化;免疫组织化学法检测AFP、Ki-67、Glypican-3、CD105在各组肿瘤中的表达:CD105测定微血管密度(MVD),Ki-6表示细胞增殖率(PI);TUNEL法检测肿瘤细胞凋亡率(AI);实时荧光定量PCR(RT-qPCR)验证靶向因子的mRNA表达变化.结果 各治疗组均能显著抑制肿瘤生长,并能消退肿瘤体积.在治疗后肿瘤体积上,除顺铂组与舒尼替尼组间无统计学差异外(41.61±7.61比67.15±5.39,P＞0.05),其余各组间都有统计学差异:对照组与顺铂(651.72±121.16比41.61±7.61,P＜0.05),对照组与舒尼替尼组(651.72±121.16比67.15±5.39,P＜0.05),对照组联合舒尼替尼±顺铂组(651.72±121.16比23.03±2.37,P＜0.05),舒尼替尼组与联合舒尼替尼+顺铂组(67.15±5.39比23.03±2.37,P＜0.05),顺铂组与联合舒尼替尼+顺铂组(41.61±7.61比23.03±2.37,P＜0.05);在裸鼠体重上,相比对照组,除舒尼替尼组无统计学差异外(25.90±0.75比26.66±0.65,P＞0.05),其余各组间差异均有统计学意义:对照组与顺铂组(25.90±0.75比18.90±0.63,P＜0.05),对照组与联合舒尼替尼+顺铂组(25.90±0.75比18.26±1.20,P＜0.05);AFP、Glypican-3在各治疗组阳性表达面积(Pixels)均少于对照组(AFP:对照组与顺铂组,1.26×106土1.48×105比5.54×105±8.14×104,P＜0.05;对照组与舒尼替尼组,1.26×106±1.48×105比7.09×105±6.64×104,P＜0.05;对照组与联合舒尼替尼+顺铂组,1.26×106±1.48×105比3.62×105±4.83×104,P＜0.05.Glypican-3:对照组与顺铂组,9.68×105±7.63×104比4.04×105±5.04×104,P＜0.05;对照组与舒尼替尼组,9.68×105±7.63×104比4.59×105±2.32×104,P＜0.05;对照组与联合舒尼替尼+顺铂组,9.68×105±7.63×104比1.89×105±2.55×104,P＜0.05).两者在顺铂组与舒尼替尼组的比较中差异无统计学意义(P＞0.05);PI和AI在各治疗组中相比对照组,差异都具有统计学意义(PI和AI:对照组与顺铂组,58.97土1.38比42.36±1.28和1.69±0.20比54.62±2.49,P＜0.01;对照组与舒尼替尼组,58.97±1.38比43.48±1.00和1.69±0.20比47.32±2.00,P＜0.01;对照组与联合舒尼替尼+顺铂组,58.97±1.38比33.34±1.19和1.69±0.20比63.41土2.23,P＜0.01).顺铂组相比联合舒尼替尼+顺铂组,PI:P=0.001,AI:P=0.002;舒尼替尼组相比联合舒尼替尼+顺铂组,PI和AI:P＜0.001;顺铂组相比舒尼替尼组,PI.P=0.597,AI:P=0.059;RT-qPCR证实M-CSFR、PDGFR-β、RET、VEGFR-2的mRNA表达受到抑制.结论 Sunitinib能显著抑制小儿睾丸卵黄囊瘤的生长,并消退肿瘤体积:主要通过直接抑制肿瘤细胞的生长和肿瘤血管的新生,从而诱导肿瘤细胞凋亡,同时伴有直接细胞毒作用引起坏死;Sunitinib相比CDDP具有更轻的毒副作用,且联合CDDP具有增强抗肿瘤作用.

Abstract：

Objective To study the antitumor effects of Sunitinib or Sunitinib combind with cis - diamminedichloroplatinum (CDDP) on an athymic mouse human testicular yolk sac tumor xenograft model. Methods The athymic mouse human testicular yolk sac tumor xenograft model was established by subcutaneous injection of 17th passage pediatric testicular yolk sac tumor cells in the unilateral inguinal region of the male nude mice. The mice of control group didn't receive any treatment. The tumor bearing mice were treated with either CDDP, or Sunitinib group, or Sunitinib combined with CDDP. The tumor-bearing nude mice were divided into 4 groups (5 in each) according the treatment they underwent. The tumor volumes and mice weight were measured to calculate the regression rate of tumor. The tumor was collected for H&E staining and immunohistochemical staining of AFP, Ki-67,Glypican-3 and CD105. Microvessel density (MVD) was measured by analyzing the CD105 expression. The tumors' proliferation index (PI) was studied by analyzing Ki-67 expression. The apoptosis of the tumor was quantitated using TUNEL staining. The mRNA expressions of cytokines were determined by quantitative real-time PCR (RT-qPCR). Results The tumor volumes were significantly decreased after chemotherapy. No difference of tumor volume was found between CDDP group and Sunitinib group (41.61 ± 7. 61 vs. 67. 15 ± 5. 39, P＞0. 05). Significant differences of tumor volumes were found between CDDP group and CDDP+ Sunitinib group (41.61 ± 7. 61 vs. 23. 03 ± 2. 37, P＜0. 05), and between Sunitinib group and CDDP+ Sunitinib group (67. 15 ± 5. 39 vs. 23.03 ± 2. 37, P＜0. 05). Of the body weight of tumor-bearing mice, no difference was found between controls and Sunitinib group (25. 90 ± 0. 75 vs. 26. 66 ± 0. 65, P＞0. 05). And significant differences of the body weight were noted between controls and CDDP group (25.90 ± 0. 75 vs. 18. 90 ± 0. 63, P＜0. 05),controls and CDDP+ Sunitinib group (25. 90 ± 0. 75 vs. 18. 26 ± 1.20,P＜0. 05). The positive areas (pixels) of AFP in the mice with chemotherapy were less than those of control mice (Control vs. CDDP: 1.26 × 106±1.48 × 105 vs. 5. 54 × 105 ± 8. 14 × 104 , P＜0. 05. Control vs. Sunitinib: 1.26 × 106 ± 1.48 × 105 vs. 7. 09 × 105 ± 6. 64 × 104, P＜0. 05. Control vs. CDDP + Sunitinib: 1.26 × 106 ± 1.48 × 105 vs. 3. 62 × 105 ± 4. 83 × 104, P＜0. 05). The positive areas (pixels) of Glypican-3 in the mice with chemotherapy were less than those of control mice (Control vs. CDDP: 9. 68 × 105 ± 7. 63 × 104 vs. 4. 04 × 105 ± 5. 04 × 104 , P＜0. 05. Control vs. Sunitinib: 9. 68 × 105 ± 7. 63 × 104 vs. 4. 59 × 105 ± 2. 32 × 104 , P＜0. 05. Control vs. CDDP + Sunitinib: 9. 68 × 105 ± 7. 63 × 104 vs. 1.89 × 105 ± 2. 55 × 104, P＜0. 05). However, there were no statistical differences of the positive areas (pixels) of AFP and Glypican-3 between CDDP and Sunitinib groups (P＞0. 05). The PI was significantly decreased after chemotherapy (Control vs. CDDP: 58. 97 ± 1.38 vs. 42. 36 ± 1.28, P＜ 0. 01. Control vs.Sunitinib: 58. 97 ± 1.38 vs. 43. 48 ± 1.00, P＜0. 01. Control vs. CDDP+ Sunitinib: 58. 97 ± 1.38 vs.33. 34 ± 1.19, P＜0. 01 ). The apoptosis index (AI) was also significantly increased after chemotherapy (Control vs. CDDP: 1.69 ± 0. 20 vs. 54. 62 ± 2. 49, P＜0. 01. Control vs. Sunitinib: 1.69 ± 0. 20vs. 47. 32 ± 2. 00, P＜0. 01. Control vs. CDDP + Sunitinib: 1.69 ± 0. 20 vs. 63. 41 ± 2. 23, P＜0. 01). Significantly differences of PI and AI were found between CDDP and CDDP + Sunitinib groups (P＜0. 01 ), and Sunitinib and CDDP + Sunitinib (P＜0. 01 ). RT-qPCR study confirmed that the mRNA expressions of M-CSFR, PDGFR-β, RET and VEGFR-2 were decreased in the mice underwent chemotherapy. Conclusions Sunitinib is effective to suppress the pediatric testicular yolk sac tumor growth, and reduce the tumor volume. Sunitinib can inhibit the angiogenesis in tumor, induce apoptosis of tumor cells, and kill the tumor cells directly. Sunitinib is less toxic than CDDP, and synergistic with the antitumor effect of CDDP.     

The nude mouse model for human retinoblastoma: A system for evaluation of retinoblastoma therapy

We have critically examined the nude mouse model for human retinoblastoma to determine whether or not characteristics found in the parent tumor are retained in the mouse. We have demonstrated that most tumors grown in the anterior chamber of the nude mouse maintain a similar karyotype, show the same degree of differentiation and develop an adequate tumor blood supply when compared to the primary tumor from which they were obtained. Because of these findings, we suggest that this model may be particularly useful for evaluating new methods or combinations of treatment for human retinobastoma.     

Risk of germ cell malignancy in children with XY intersex versus isolated cryptorchidism by immunohistochemistry

The risk of subsequent development of testicular germ cell neoplasia is related to presence of underlying developmental defects such as cryptorchidism, in which the risk is around 0.5%, and XY intersex with abdominal testes, in which the risk may be as high as 20-25%. We examined the hypothesis that the increased risk of germ cell malignancy in intersex testes with Y chromosome was a direct consequence of an abnormal increase in number of PLAP/CD117+ immature germ cells into postnatal life. Archival cases of uncomplicated cryptorchidism (CO) and XY intersex (INT) were identified and anonymized, and a subgroup of aged-matched cases had sections immunostained with placental alkaline phosphatase (PLAP) and CD117. From a total of 89 intersex and 105 cryptorchid cases identified, a power calculation to detect a 20% difference in expression between groups (alpha = 0.05, power = 80%) determined that 18 intersex and 36 cryptorchid cases were required. Thus, 58 cases were examined, median age 3 (range birth-11) years, including 39 CO and 19 INT. The prevalence of any PLAP+ germ cells was 2/39 (5.1%) versus 3/19 (15.7%), respectively. (Z = 1.4, p = 0.17). In contrast, 94% of cases showed presence of any CD117+ germ cells, but the frequency of CD117+ cells was not significantly different between groups (t = 0.56, p = 0.58). CD117 and PLAP identify different populations of germ cells in pediatric testes. The extent of increased risk of malignancy in XY INT is not simply related to increased numbers of immature PLAP+/CD117+ germ cells present; additional factors play a pathogenic role.