野生型p53基因对人卵巢癌细胞株的生长抑制作用

目的：探讨野生型ｐ５３(ｗｔ ｐ５３)基因对人卵巢癌细胞株的生长抑制作用。方法：利用脂质体介导，将含有人全长ｗｔ－ｐ５３基因ｃＤＮＡ的真核表达载体质粒ｐＣＥＰ４／ｐ５３和空载体质粒ｐＣＥＰ４分别转染卵巢癌ＳＫＯＶ３细胞株，分别命名为ＳＫＯＶ３－ｐＣＥＰ４／ｐ５３细胞(C组)、ＳＫＯＶ３－ｐＣＥＰ４细胞(B组)，另设ＳＫＯＶ３细胞为正常对照组（Ａ组）。观察细胞体外生长情况和凋亡变化。结果：外源性ｐ５３基因在Ｃ组细胞中获表达，细胞生长曲线显示ｐ５３的导入使ＳＫＯＶ３细胞生长受到明显抑制，Ｃ组平均细胞集落数（１８．６±１．８个）少于Ａ组（２４．３±２．２个）和Ｂ组（２２．７±２．６），差异有显著性（Ｐ（005）。ＭＴＴ法检测C组细胞活力明显低于Ａ、Ｂ组。Ｃ组细胞Ｇ０～Ｇ１期百分比（５７．７９％）及凋亡细胞百分比（13．９1％）均高于Ｂ组（４６．０２％、２．０８％）和Ａ组（４３．６２％、０）。结论：ｗｔ－ｐ５３基因可介导ＳＫＯＶ３细胞Ｇ１期停滞和细胞凋亡，抑制细胞体外生长。     

生长抑制DNA损伤基因45与p53对卵巢癌细胞凋亡作用的研究

目的：研究生长抑制ＤＮＡ损伤基因（ｇｒｏｗｔｈａｒｅｓｔａｎｄＤＮＡｄａｍａｇｅ,ＧＡＤＤ）对卵巢癌细胞的作用及与ｐ５３蛋白表达的关系。方法：利用ＲＴ－ＰＣＲ鉴定卵巢癌ＳＫＯＶ３和３ＡＯ细胞株ＧＡＤＤ４５的表达;采用ＲＴ－ＰＣＲ和ＷｅｓｔｅｒｎＢｌｏｔ鉴定ＳＫＯＶ３和３ＡＯ细胞株ｐ５３蛋白的表达。通过载体构建将目的基因ＧＡＤＤ４５重组于ｐｃＤＮＡ－Ⅲ真核质粒内。利用脂质体（ＤＯＴＡＰ）法转染ＳＫＯＶ３和３ＡＯ细胞株;Ｇ４１８抗性筛选,ＲＴ－ＰＣＲ进行表达鉴定。采用流式细胞仪分析（ＦＣＭ）以及ＤＮＡ梯度法对细胞凋亡现象进行检测。结果：卵巢癌ＳＫＯＶ３和３ＡＯ细胞株ＧＡＤＤ４５表达阴性。ＳＫＯＶ３细胞株ｐ５３蛋白表达阳性;３ＡＯ细胞株ｐ５３蛋白表达阴性。转染ＧＡＤＤ４５基因的ＳＫＯＶ３细胞株经化学药物足叶已甙（ＶＰ１６）和紫外线诱导后,可以使该细胞产生凋亡现象,而对转染ＧＡＤＤ４５基因的３ＡＯ细胞株经诱导后,未见细胞凋亡现象。结论：转染ＧＡＤＤ４５基因,对ｐ５３蛋白表达阳性的细胞株ＳＫＯＶ３经诱导后,可使细胞进入凋亡状态。对ｐ５３蛋白表达阴性的细胞株３ＡＯ转染ＧＡＤＤ４５基因后,经诱导细胞无凋亡现象,证实ＧＡＤ?     

IL-2基因转移对卵巢癌细胞系SKOV3粘附性的影响

目的：研究ＩＬ－２基因转移对卵巢癌ＳＫＯＶ３细胞系粘附性的影响。方法：细胞对塑料基质、细胞基底膜成分的粘附实验,流式细胞术测定ＣＤ４４Ｈ的表达。结果：ＥＤＴＡ介导的细胞分离实验表明,ＳＫＯＶ３／ＩＬ－２细胞对塑料基质的粘附能力明显下降（Ｐ＜０．０１）;对基质及基底膜成分的粘附实验表明,ＳＫＯＶ３／ＩＬ－２细胞对Ⅳ型胶原和ｍａｔｒｉｇｅｌ的粘附能力下降,但对ＬＮ、ＦＮ的粘附能力无明显改变;粘附分子ＣＤ４４Ｈ的表达：ＳＫＯＶ３、ＳＫＯＶ３／Ｎｅｏ、ＳＫＯＶ３／ＩＬ－２组细胞粘附分子ＣＤ４４Ｈ阳性细胞百分率分别为７６．２％、６６．７％和３７．８％,ＳＫＯＶ３／ＩＬ－２组明显降低（Ｐ＜０．００１）。结论：ＩＬ－２基因修饰可使卵巢癌细胞系ＳＫＯＶ３粘附能力下降。     

粒细胞巨噬细胞集落刺激因子对人卵巢细胞株生长的影响

目的Ｌ：了解粒细胞巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）对人卵巢癌细胞株生长的影响。方法：采用ＸＴＴ比色法及流式细胞仪检测ＧＭ－ＣＳＦ对卵巢癌细胞株ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３细胞生长的影响。结果：用ＸＴＴ法检测ＧＭ－ＣＳＦ对ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３细胞的生长均无明显影响;流式细胞仪检测也未发现ＧＭ－ＣＳＦ对ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３的增殖指数有明显影响。结论：在卵巢癌化疗过程中应用ＧＭ     

多药耐药基因反义寡脱氧核苷酸对卵巢癌细胞株SKOV3多药耐药性逆转的作用

目的 探讨多药耐药基因（ｍｄｒ１）反义寡脱氧苷酸（ＡＳＯＮ）对卵巢癌细胞多药耐药性的逆转作用。方法 以人ｍｄｒ１基因转导产生的耐药卵巢癌ＳＫＯＶ３／ｍｄｒ１细胞为模型,采用２５０μｇ／ｍｌ ｍｄｒ１－ＡＳＯＮ,作用于ＳＫＯＶ３／ｍｄｒ１细胞,采用流式细胞术及罗丹明１２３排出试验,检测ＳＫＯＶ３／ｍｄｒ１细胞ｍｄｒ１基因产物Ｐ－糖蛋白（Ｐ－ｇｐ）的阳性率及其功能。并进行ＳＫＯＶ３／ｍｄｒ１细胞集落培养,观察耐药性。结果 ｍｄｒ１－ＡＳＯＮ作用后,ＳＫＯＶ３／ｍｄｒ１细胞的Ｐ－ｇｐ阳性率由３８．９％减少至２１．３％（Ｐ〈０．０１）,ＳＫＯＶ３／ｍｄｒ１细胞内罗丹明的潴留率,由３２．１％增加至５０．７％（Ｐ〈０．０１）。ＳＫＯＶ３／ｍｄｒ１细胞加ｍｄｒ１－ＡＳＯＮ和空白对照细胞形成抗性细胞集落的相对百分数,在泰素浓度     

腺病毒介导胸苷激酶基因治疗卵巢癌的动物试验研究

目的：探讨腺病毒介导Ｒｏｕｓ肉瘤病毒驱动的单纯疱疹病毒胸苷激酶（ＡＤＶ／ＲＳＶ－ｔｋ）基因人氧鸟苷（ＧＣＶ）治疗卵巢癌的有效性和毒性，方法：首先以卵巢浆液性腺癌细胞系Ｏｖ－ｃａ－２７７４建立荷人卵巢癌裸鼠腹水瘤模型，再分别单用ＡＤＶ／ＲＳＫ－ｔｋ或ＧＣＶ（单药组）或联合应用ＡＤＶ／ＲＳＫ－ｔｋ及ＧＣＶ（联合用药组）进行治疗，观察空白对照组，单药组及联合用药级裸鼠的平均生存时间及药物毒怀，结果：单药     

脂质体—C—erbB2反义脱氧寡核苷酸对人卵癌裸鼠网膜移植瘤 …

目的 探讨脂质体－Ｃ－ｅｒｂＢ２反义脱氧寡苷酸（Ｃ－ｅｒｂＢ２ Ｓ－ＯＤＮｓ２）对人卵巢癌裸鼠网膜移植瘤生长及顺铂（ＤＤＰ）敏感性的影响。方法 对２０只裸鼠建立人卵巢癌裸鼠网膜移植模型,分为对照组（腹腔注射无菌水）、观察组（腹腔注射脂质体－Ｃ－ｅｒｂＢ２ Ｓ－ＯＤＮｓ）、化疗组（腹腔注射无菌水及ＤＤＰ）、观察＋化疗组（腹腔注射脂质体－Ｃ－ｅｒｂＢ２ Ｓ－ＯＤＮｓ及ＤＤＰ）。治疗期间观测裸鼠体重变化     

单纯疱疹病毒胸苷激酶基因转染联合药物对卵巢癌细胞的杀伤效应

目的：探讨单纯疱疹病毒胸苷激酶（ＨＳＶ－ＴＫ）基因转染联合抗病毒药物羟基无环鸟苷（ＧＣＶ）对人卵巢癌细胞系的杀伤效应。方法：采用基因工程技术，将ＨＳＶ－ＴＫ基因的ＤＮＡ序列插入逆转录病毒载体ｐＬＮＳＸ的ＨｉｎｄⅢ位点，构建重组体ｐＬＮＳ／ＨＳＶ－ＴＫ，并采用磷酸钙介导贴壁细胞基因转染法将重组体导入ＰＡ３１７包装细胞，建立重组逆转录病毒载体分泌细胞株ＰＡ３１７／ＴＫ。并采用四甲基偶氮唑蓝比色法检测ＨＳＶ－ＴＫ／ＧＣＶ系统对人类卵巢癌ＡＯ细胞系的生长抑制率。结果：重组逆转录病毒载体ｐＬＮＳ／ＨＳＶ－ＴＫ能有效地将ＨＳＶ－ＴＫ基因导入ＡＯ细胞系内并使其获得对ＧＣＶ的敏感性。当培养液内ＧＣＶ达４０μｍｏｌ时，其对转ＨＳＶ－ＴＫ基因后ＡＯ细胞的生长抑制率为９８．０％。结论：ＡＯ细胞转染ＨＳＶ－ＴＫ基因后，能有效地被ＧＣＶ杀灭。     

c—erbB2反义核酸对卵巢癌细胞系生物学行为及药物敏感性的影响

目的：探讨ｃ－ｅｒｂＢ２反义核酸对卵巢癌细胞生物学行为及其化疗药物敏感性的影响。方法：应用ｐＵＣ－ｅｒｂＢ２和ｐＤＯＲ－ｎｅｏ质粒，构建了含反义ｅｒｂＢ２ｃＤＮＡ３．８ｋｂ的重组逆转录病毒表达载体ｐＤＯＲ－ｅｒｂＢ－ｎｅｏ，经脂质体转染ｅｒｂＢ２高表达的人卵巢癌细胞系ＳＫＯＶ３，再经Ｇ４１８（一种新霉素类似物）筛选得到新霉素抗性细胞，命名为ＳＫＯＶ３－Ａ２。结果：Ｓｏｕｔｈｅｒｎ印迹杂交分析表明，反义ｅｒｂＢ２３．８ｋｂ已完全整合到ＳＫＯＶ３细胞中，与亲本细胞及转染ｐＤＯＲ－ｎｅｏ的对照组细胞相比，ＳＫＯＶ３－Ａ２的生长和ＤＮＡ合成均受到抑制，且对５－氟尿嘧啶（５－ＦＵ）及顺铂（ＤＤＰ）的药物敏感性明显增加。结论：ｃ－ｅｒｂＢ２反义核酸在卵巢癌基因治疗中有一定作用；ｃ－ｅｒｂＢ２癌基因可能与卵巢癌细胞耐药有关。     

肿瘤坏死因子与干扰素协同抗卵巢癌作用的研究

应用肿瘤坏死因子（ＴＮＦ）和γ干扰素（ＩＦＮγ）对靶细胞３ＡＯ和ＮＩＨ：ＯＶＣＡＲ－３卵巢癌细胞株进行抗肿瘤治疗的实验研究。结果：３ＡＯ和ＮＩＨ：ＯＶＣＡＲ－３卵巢癌细胞株对各种浓度的ＴＮＦ和ＩＦＮγ均呈现抵抗作用。ＴＮＦ和ＩＦＮγ协同用药时对靶细胞有杀伤作用，当ＴＮＦ和ＩＦＮγ的浓度均为１×１０￣４Ｕ／Ｌ时，出现对３ＡＯ和ＮＩＨ：ＯＶＣＡＲ－３卵巢癌细胞的明显杀伤作用（Ｐ＜０．０５）；当ＴＮＦ和ＩＦＮγ均为１×１０￣６Ｕ／Ｌ时，杀伤５５％的３ＡＯ和７５％的ＮＩＨ：ＯＶＣＡＲ－３卵巢癌细胞（Ｐ＜０．０１）。提示：ＩＦＮγ可增加３ＡＯ和ＮＩＨ：ＯＶＣＡＲ－３卵巢癌细胞对ＴＮＦ的敏感性，ＴＮＦ与ＩＦＮγ联合用药时有很好的协同抗卵巢癌作用。     

腹主动脉球囊预置术与髂内动脉球囊预置术在植入型凶险性前置胎盘治疗中的比较研究

目的:比较研究腹主动脉球囊预置术与髂内动脉球囊预置术在植入型凶险性前置胎盘治疗中的临床疗效。方法:选择2014年1月至2015年4月住院治疗的植入型凶险性前置胎盘患者64例,随机分为两组,择期剖宫产术前行腹主动脉球囊置管32例(腹主动脉组)和双侧髂内动脉球囊置管32例(髂内动脉组),观察比较两组孕妇术中、术后情况及新生儿出生和婴儿期生长发育情况。结果:两组患者在手术时间、术中出血量、术后最高体温、输血率、子宫切除率、住院时间、球囊预置术后穿刺部位出血率、感觉障碍发生率、新生儿Apgar评分、新生儿身高、出生体质量等方面比较,差异均无统计学意义(P0.05);而腹主动脉组球囊预置时间、透视时间、放射剂量均显著低于髂内动脉组(P0.05)。术后1年内随访,两组婴儿在42天、3个月、6个月、1年的体质量及身高情况比较,差异均无统计学意义(P0.05)。结论:腹主动脉球囊预置术与髂内动脉球囊预置术在植入型凶险性前置胎盘的治疗中均能安全、有效减少术中出血。而腹主动脉球囊预置术X线暴露时间更短、预判效果无需造影剂,对母胎保护性更强,可在临床推广。     

Laparoscopic Morcellator-Related Complications

Morcellation at laparoscopy is a commonly used minimally invasive method to extract bulky tissue from the abdomen without extending abdominal incisions. Despite widespread use of morcellation, complications still remain underreported and poorly understood. We performed a systematic review of surgical centers in the United States to identify, collate and update the morcellator-related injuries and near misses associated with powered tissue removal. We searched articles on morcellator-related injuries published from 1993 through June 2013. In addition, all cases reported to MedSun and the FDA device database (MAUDE) were evaluated for inclusion. We used the search terms “morcellation,” “morcellator,” “parasitic,” and “retained” and model name keywords “Morcellex,” “MOREsolution,” “PlasmaSORD,” “Powerplus,” “Rotocut,” “SAWALHE,” “Steiner,” and “X-Tract.” During the past 15 years, 55 complications were identified. Injuries involved the small and large bowels (n = 31), vascular system (n = 27), kidney (n = 3), ureter (n = 3), bladder (n = 1), and diaphragm (n = 1). Of these injuries, 11 involved more than 1 organ. Complications were identified intraoperatively in most patients (n = 37 [66%]); however, the remainder were not identified until up to 10 days postoperatively. Surgeon inexperience was a contributing factor in most cases in which a cause was ascribed. Six deaths were attributed to morcellator-related complications. Nearly all major complications were identified from the FDA device database and not from the published literature. The laparoscopic morcellator has substantially expanded our ability to complete procedures using minimally invasive techniques. Associated with this opportunity have been increasing reports of major and minor intraoperative complications. These complications are largely unreported, likely because of publication bias associated with catastrophic events. Surgeon experience likely confers some protection against these injuries. Understanding and implementing safe practices associated with the use of the laparoscopic morcellator will reduce these iatrogenic injuries.     

Fístula cutáneo-uterina poscesárea

A fistula is an abnormal communication between two epithelial surfaces. Although fistulas that wrap around the uterus are not infrequent, uterocutaneous fistula is rare. The treatment of choice is abdominal hysterectomy with excision of the fistula up to the skin. We report two cases of uterocutaneous fistula requiring surgical treatment.     

宫腔镜下清宫术治疗剖宫产瘢痕妊娠前不同预处理方式的疗效分析

目的:探讨宫腔镜下清宫术治疗剖宫产瘢痕妊娠(CSP)前不同预处理方式的临床效果。方法:回顾性分析2014年10月至2015年10月本院住院中B超检查提示妊娠囊下缘距子宫切口小于1.0 cm行宫腔镜下清宫术并确诊为CSP的患者170例作为研究对象。其中直接行宫腔镜下清宫术的患者23例(A组);行米非司酮+米索前列醇药物治疗后在宫腔镜下行清宫术的患者29例(B组);行米非司酮+甲氨蝶呤+米索前列醇杀胚治疗后在宫腔镜下行清宫术的患者40例(C组);行子宫动脉灌注+介入栓塞术后在宫腔镜下行清宫术的患者78例(D组),比较4组患者治疗效果。结果:①所有患者手术顺利,无一例发生术中大出血和组织残留,4组患者治疗后血β-HCG较治疗前明显下降,术后血β-HCG、术中出血量、手术时间两两比较,差异无统计学意义(P0.05)。②A组和D组住院时间较短,B组和C组较长,A组与D组住院时间差异无统计学意义(P0.05),但其余两两比较,差异有统计学意义(P0.05)。③A组住院费用最少,D组最多,B组住院费用与C组比较差异无统计学意义(P0.05),但其余两两比较,差异均有统计学意义(P0.05)。④术后不良反应情况:A组术后无一例发生不良反应,B组术后有2例肝功能受损,3例恶心、呕吐等胃肠道反应,C组术后有3例肝功能受损,26例发生胃肠道反应;D组1例发生肝功能受损,16例发热,57例疼痛,对症治疗后好转。结论:CSP的治疗应根据患者病情、经济条件、个人意愿和医院的技术设备条件选择合适的治疗方案。     

The gynecologist and the melanoma: breast and endometrial metastases

Melanoma has an important metastatic potential and its incidence is greatly increasing. Even after many years of negative follow-up, gynecologists should be aware that a gynecological tumor might be a secondary location for a woman with a medical history of melanoma. Because of a poor prognosis and a reduced life expectancy, it is necessary to make a disease staging in order to offer a prompt diagnosis and a personalized strategy of treatment. Considering the increasing incidence of melanoma, gynecologists will face more frequently with this situation.     

Anticoagulación en gestantes portadoras de prótesis mecánicas

Objective

To study distinct anticoagulation regimens in pregnant women with prosthetic heart valves.

Subjects and methods

We performed a systematic review of the literature to determine the required levels of anticoagulation prophylaxis, timing of the introduction of oral anticoagulation and its substitution by heparins, and the maternal and fetal risks associated with different anticoagulation regimens.

Results

A target international normalized ratio (INR) of 2.5-3.5 should be achieved. Although consensus on the heparin of choice is lacking, heparin dose requirements should be based on anti-factor Xa levels (around 1.0 U/mL) or activated partial thromboplastin time (aPTT) (2-3 times control value). The risk of thrombosis in heparin-treated patients is approximately 7%, while the incidence of heparin embryopathy ranges from 1.6-7.4%. The switch from oral anticoagulation to heparin should be made no later than at weeks 35-36 of pregnancy.

Conclusions

The nticoagulation therapy of choice in the first trimester of pregnancy cannot currently be established. Prospective and randomized studies are required to determine the advisability of one treatment over the other