Palomo手术治疗儿童精索静脉曲张

目的:对经腹膜后集束结扎精索血管(Palomo手术)治疗儿童精索静脉曲张的可行性及疗效进行评价。方法:对采用Palomo术式治疗的36例精索静脉曲张患儿,于手术前、术后6个月、术后12个月采用彩色Doppler检查睾丸,测量患儿双侧睾丸体积。结果:全部治愈,术后随访12～36个月,所有患儿临床症状消失,所有患儿均未发生睾丸萎缩。术后患侧睾丸血供无明显减少,患侧睾丸体积术后增长明显快于对侧。结论:Palomo手术是一种治疗儿童精索静脉曲张简单易行、疗效可靠的手术方式。     

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.     

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.     

睾丸生殖细胞来源肿瘤分类及其遗传学研究进展

生殖细胞来源的肿瘤包括起源于生长细胞系的各种类型的肿瘤.它们除了发生于性腺,还可见于性腺外的体壁中线上,如骶尾部、腹膜后、纵隔、头颈部、甚至丘脑下部的松果体区,还有极少数可见于体壁中线外的其他器官.这种分布与早期发育时原始生长细胞由卵黄囊向生殖嵴迁移有关.     

小儿肾盂输尿管连接部梗阻术中肾盂穿刺液细菌谱及药敏分析

目的通过了解小儿肾盂输尿管连接部梗阻术中肾盂穿刺液细菌谱和药物敏感特点,以指导围手术期抗菌药物选择。方法回顾性分析医院1995年1月-2010年12月收治的小儿肾盂输尿管连接部梗阻手术患儿262例,术中打开肾盂前常规用无菌注射器穿刺肾盂抽取尿液标本,进行细菌培养及药物敏感试验。结果 31例肾盂穿刺尿液培养阳性,阳性率为11.8%;病原菌中以革兰阴性杆菌为主,位于前3位的革兰阴性杆菌分别为大肠埃希菌、克雷伯菌属和奇异变形菌,分别占33.3%、19.4%、11.1%;大肠埃希菌、克雷伯菌属和奇异变形菌对氨苄西林、哌拉西林和第一、二代头孢菌素均有较高的耐药率,但对亚胺培南、呋喃妥因和阿米卡星高度敏感,对第三代头孢敏感率也较好;肠球菌属对青霉素、庆大霉素、环丙沙星高度耐药,对呋喃妥因和万古霉素高度敏感,对第三代头孢敏感率也较好。结论小儿肾盂输尿管连接部梗阻细菌感染率不高,但对临床常用抗菌药物有较明显的耐药性,术中肾盂穿刺液细菌培养及药物敏感试验对围手术期抗菌药物的选择具有指导作用。     

儿童甲状腺腺瘤的外科治疗

儿童甲状腺腺瘤较为少见,治疗方式多为手术切除,因此采用合理的手术方式非常重要,1983～2003年我院对28例儿童甲状腺腺瘤采用手术切除腺瘤,疗效满意.现予报告并对其外科治疗方面进行讨论.     

新生儿环状胰腺的手术治疗体会

环状胰腺(简称环胰)是新生儿较常见的上消化道畸形及急腹症之一,临床发病率较低,国内报道较少,术前不易确诊,病死率高。我院1997～2003年,收治环状胰腺6例,经手术治愈,现报告如下。一、临床资料1.一般资料:本组6例患儿,男4例,女2例。日龄2～16d,其中早产儿2例。出生时体重2300～3820g,其中小于2500g1例。出生后1～2d内出现频繁呕吐,皆含胆汁,2例呕吐物含咖啡色,伴腹胀者3例。X线平片提示十二指肠或小肠高位近端完全性梗阻,有胃大泡或双泡征,其中1例结肠无充气,5例结肠充气少。皆伴有不同程度的水、电解质、酸碱平衡紊乱。出生时有正常胎粪…     

腹腔镜精索血管高位结扎术治疗小儿精索静脉曲张

2002年5月至2004年12月,我们采用腹腔镜对12例精索静脉曲张患儿行精索血管高位结扎术(Palomo手术),疗效满意。现报告如下。对象与方法本组12例。年龄11～14岁,平均12.6岁。病变均位于左侧。阴囊坠胀不适就诊者8例,无症状偶然发现阴囊肿块者4例。患者术前均采用彩色多普勒测量双侧睾丸容积。睾丸容积计算公式:睾丸容积(ml)=长度(mm)×宽度(mm)×厚度(mm)×0.521。按照Valsalva分度:Ⅱ度3例,Ⅲ度9例。     

婆娑双树基因4干扰载体对儿童睾丸卵黄囊瘤的转染及干扰效果评价

目的 构建婆娑双树基因SALL4的特异性干扰载体并转染儿童睾丸卵黄囊瘤细胞,评价干扰效果,为研究SALL4基因功能提供实验工具.方法 针对不同干扰位点,设计4条特异性siRNA和1条阴性对照序列.构建SALL4干扰质粒载体pGPU6/GFP/Neo-SALL4 shRNA,分空白对照组(Blank组)、阴性对照组(NC组)、单加转染试剂组(MOCK组)、干扰组(SALL4-homo-483、SALL4-homo-1874、SALL4-homo-951、SALL4-homo-3110),分别转染体外培养的儿童睾丸卵黄囊瘤细胞,采用荧光定量PCR和Western bolt技术比较各组细胞SALL4基因mRNA及蛋白表达情况,筛选出最佳干扰序列.结果 荧光定量PCR结果显示,Blank组、NC组、MOCK组、干扰组(SALL4-homo-483、SALL4-homo-951、SALL4-homo-1874、SALL4-homo-3110) SALL4基因mRNA的表达水平分别为100％、(118.0±18.0)％、(128.0±15.0)％、(91.0±21.0)％、(25.0±8.0)％、(59.0±11.0)％、(49.0±13.0)％.SALL4-homo-951、SALL4-homo-3110相比Blank组、NC组、MOCK组mRNA表达下降,差异有统计学意义(F=3.63,P＜0.05).Western bolt的检测结果显示Blank组、NC组、MOCK组、干扰组(SALL4-homo-483、SALL4-homo-951、SALL4-homo-1874、SALL4-homo-3110)SALL4基因蛋白表达水平分别为1、1.1±0.1、1.3±0.3、1.7±0.4、0.2±0.1、0.9±0.2和0.4±0.1.相对Blank组、NC组、MOCK组,SALL4干扰组中SALL4-homo-951、SALL4-homo-3110均能降低蛋白表达,其中SALL4-homo-951降低最多,差异有统计学意义(F=7.96,P＜0.05).结论 SALL4-homo-951、SALL4-homo-3110均能有效干扰SALL4基因在儿童卵黄囊瘤中的表达,其中SALL4-homo-951干扰效果最佳,可用于下一步实验研究.     

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本院1999-09-2005-10手术治疗手法复位失败的婴幼儿嵌顿性腹股沟斜疝330例,其中因手法复位致肠穿孔10例,现报道如下.