体外不同比例再生肝细胞与结肠癌细胞的共培养

Objective To investigate the proliferation potential of colon cancer cells co-cultured with different proportions regenerating hepatocytes and role of the cell growth factors related to liver regeneration in vitro.Methods Active regenerating hepatocytes were obtained by collagenase perfusion in situ of rats models underwent 70% liver resection after 24 hours.Co-cultures with different proportions of hepatocytes to human colon cell line SW480 were performed respectively.The proportion of hepatocytes to human colon cell line SW480 Was 10:1 in group A,1:1 in group B.There was only human colon cell line SW480 in group C(control group).Proliferation capacity was assessed with the percentage by 3H-thy incorporation.Concentration of epidermal growth factor(EGF),insulin-like growth factor-1(IGF-1)and hepatocyte growth factor(HGF)was analyzed by ELISA.Results There was no significant difference among the 3 groups in 3H-thy incorporation percentage,and concentration of EGF,IGF-land HGF after 24 hours' culture(P>0.05).3H-thy incorporation percentage(15.9±1.4,13.2±1.5),and concerntration of EGF [(722.9±55.4)ng/L,(498.2+41.5)ng/L]and IGF-1[(755.2±35.7)ng/L,(538.1±37.5)ng/L]in Group A and B were higher than that in group C after 72 hours(P<0.05).Especially the indexes above in group A were higher than that in group B(P<0.05).there was no significant difference in concerntration of HGF among the 3 groups after 72 hours(P>O.05).Conclusions These results imply that the regenerating hepatocytes contribute to the hyperproliferative state of co-culturing colon cancer cells.The mechanism maybe have relationship with EGF and IGF-1 high expression in colon cancer cells caused by growth signals coming from hepatocytes.     

小肠间质瘤的临床分析

目的:探讨小肠间质瘤(small intestine stromal tumors ,SISTs)的临床表现、影像学特点和治疗措施.方法:收集2002年8月至2007年8月本院普外科治疗的14例SISTs的患者的临床资料进行回顾性分析.结果:14例患者中男6例,女8例,平均年龄(56.8±11.4)岁.SISTs常见的临床表现为:腹痛、腹部包块、肠梗阻、胃肠道出血及体重下降;血清肿瘤学指标均正常.术前9例行消化道气钡双对比造影检查,4例检出病变,均未提示间质瘤可能.14例行CT检查,全部检出肿瘤,定性诊断准确8例;14例患者均行手术治疗,无围手术期死亡.术后病理结果为SISTs,其中良性间质瘤2例,潜在恶性3例,恶性9例.结论:SISTs无特异性临床症状,消化道气钡双对比造影诊断率低,CT扫描具有一定的特征,两种检查互相配合,结合临床具有较高的定位定性诊断价值.早期诊断对改善SISTs的预后十分重要,积极手术切除和甲磺酸伊马替尼辅助治疗可以获得良好效果     

加速康复外科在胆道外科中的临床应用

目的:探讨加速康复外科(ERAS)在胆道外科围手术期应用的临床效果及安全性。方法:选取我院2017年1~12月收治的胆总管结石患者198例,随机分为观察组和对照组各99例。对照组予以传统围手术期处理,观察组实施ERAS措施。观察两组术后恢复以及不良反应发生情况。结果:观察组下床时间、术后进食时间、白细胞恢复时间、术后补液量、术后住院天数、治疗费用、术后5 d体重下降量以及术后并发症发生率均明显低于对照组,术后5 d血红蛋白水平显著高于对照组(P0.05)。结论:胆道外科手术围手术期应用ERAS理念和措施,有助于减轻患者术后疼痛,加快胃肠功能恢复,促进术后早期下床活动,从而减少术后并发症发生率,缩短了住院时间,降低住院费用,加速患者的康复速度,值得临床推广应用。     

缺氧对人胃癌细胞的细胞周期调控及己糖激酶Ⅱ表达的影响研究

目的 探讨缺氧对人胃癌细胞株SGC-7901的细胞周期调控机制及对乏氧诱导因子-1α(HIF-1α)、己糖激酶Ⅱ(HK Ⅱ)表达的影响.方法 人胃癌细胞株SGC-7901分成3组:缺氧12 h组、缺氧24 h组及对照组.缺氧组分别在缺氧条件下(37℃、5%CO2、2.0%O2饱和度)培养12 h和24 h,对照组置于正常氧浓度条件下(37℃、5%CO2、21%O2饱和度)培养24 h.流式细胞仪检测细胞周期分布,免疫组织化学S2P法检测HIF-1α表达,荧光定量PCR法检测HK Ⅱ mRNA表达量.结果 缺氧情况下细胞周期分析处于G0/G1期细胞百分比明显增多,处于G2/M期的细胞减少,并且可见到较明显的凋亡峰的形成,缺氧12 h组、缺氧24 h组的G0/G1期比例显著高于对照组(P＜0.05);HIF-1α和HK Ⅱ在两组缺氧组与对照组比较,其表达量明显增加(P＜0.05),缺氧24 h组与缺氧12 h组比较差异有统计学意义(P＜0.05).结论 缺氧导致细胞阻滞在G0/G1期,HIF-1α可刺激人胃癌细胞HK Ⅱ表达的增加,以HIF-1α为药物靶点可望成为治疗胃癌的重要新方法.     

肿瘤坏死因子相关细胞凋亡诱导配体表达载体的制备

目的构建一种携带肿瘤坏死因子相关细胞凋亡诱导配体（TRAIL）基因的重组腺相关病毒（rAAV）载体。方法首先构建携带可溶性肿瘤坏死因子相关细胞凋亡诱导配体（sTRAIL）基因的穿梭质粒腺相关病毒穿梭质粒-可溶性肿瘤坏死因子相关细胞凋亡诱导配体（pAAV-sTRAIL），将穿梭质粒转染入HEK293细胞（人胚肾细胞系）中，采用细胞内质粒DNA同源重组法构建重组腺相关病毒载体rAAV-sTRAIL。以噬斑分析法筛选单克隆rAAV-sTRAIL；PCR法鉴定阳性rAAV-sTRAIL；氯化铯密度梯度离心法纯化rAAV-sTRAIL；紫外分光光度仪测定rAAV-sTRAIL颗粒数及纯度，噬斑分析法测定rAAV-sTRAIL感染滴度；Western免疫印迹检测rAAV-sTRAIL在HEK293细胞中的表达情况。结果成功构建了rAAV-sTRAIL，制备的病毒纯度好、滴度高，且在HEK293转导细胞中能有效表达目的基因sTRAIL。结论构建的重组腺相关病毒载体rAAV-sTRAIL，为研究TRAIL抗肿瘤细胞效应及临床应用提供先进的载体系统。     

大鼠肝移植自发性免疫耐受的研究

目的建立大鼠肝移植自发性免疫耐受模型，初步了解其移植后的免疫学变化。方法按Kamada的双管套法，选用近交系Lewis（LEW）、Wistar Furth（WF）大鼠进行原位全肝移植，术后动态观察宿主的肝功能和肝脏病理变化。另外，观察移植后不同时期，宿主植入供体同源的心脏后的免疫耐受情况。结果LEW→WF肝移植后血清谷丙转氨酶（ALT）及总胆红素（TB）的峰值在术后第4天，ALT在2周内、TB在4周内基本恢复到正常范围内；但肝脏炎症细胞的浸润则在第2周最为明显。通过再移植供体心脏的方法提示宿主对供体的特异性免疫耐受是移植2周后建立的。结论该组合早期虽存在急性排斥反应，但术后不需给予免疫抑制剂，可以自然克服这种损害而长期生存，属自发性免疫耐受模型。     

胰腺实性假乳头状肿瘤病例报告并文献复习

目的 报道1例中年女性胰腺实性假乳头状肿瘤病例,并通过文献回顾和复习,提高临床医生对此类罕见病的认识,减少该疾病的误诊、漏诊.方法 回顾性分析1例中年女性胰腺实性假乳头状肿瘤的诊治经过,结合以往的文献报道,总结该疾病的临床表现、诊治方法.结果 回顾文献显示:胰腺实性假乳头状肿瘤比较罕见,好发于年轻女性,属于低度恶性肿瘤...     

体外不同比例再生肝细胞与结肠癌细胞的共培养

Objective To investigate the proliferation potential of colon cancer cells co-cultured with different proportions regenerating hepatocytes and role of the cell growth factors related to liver regeneration in vitro.Methods Active regenerating hepatocytes were obtained by collagenase perfusion in situ of rats models underwent 70% liver resection after 24 hours.Co-cultures with different proportions of hepatocytes to human colon cell line SW480 were performed respectively.The proportion of hepatocytes to human colon cell line SW480 Was 10:1 in group A,1:1 in group B.There was only human colon cell line SW480 in group C(control group).Proliferation capacity was assessed with the percentage by 3H-thy incorporation.Concentration of epidermal growth factor(EGF),insulin-like growth factor-1(IGF-1)and hepatocyte growth factor(HGF)was analyzed by ELISA.Results There was no significant difference among the 3 groups in 3H-thy incorporation percentage,and concentration of EGF,IGF-land HGF after 24 hours' culture(P>0.05).3H-thy incorporation percentage(15.9±1.4,13.2±1.5),and concerntration of EGF [(722.9±55.4)ng/L,(498.2+41.5)ng/L]and IGF-1[(755.2±35.7)ng/L,(538.1±37.5)ng/L]in Group A and B were higher than that in group C after 72 hours(P<0.05).Especially the indexes above in group A were higher than that in group B(P<0.05).there was no significant difference in concerntration of HGF among the 3 groups after 72 hours(P>O.05).Conclusions These results imply that the regenerating hepatocytes contribute to the hyperproliferative state of co-culturing colon cancer cells.The mechanism maybe have relationship with EGF and IGF-1 high expression in colon cancer cells caused by growth signals coming from hepatocytes.     

不同灌注方式对供肝质量的影响

目的探讨不同的灌注方式对供肝质量的影响并阐明其作用机制。方法通过体外、体内两种实验方法检验。54只大鼠用于体外实验,采用不同的灌注液(HCA液 UW液)冷灌注供肝,按8,16,32 h时间冷保存肝脏,然后检测相应冷保存时限后,经肝脏灌注的灌洗液中丙氨酸转氨酶(ALT)的变化,并动态观察肝脏保存前后重量差、组织学的改变;48只大鼠用于体内实验,采用kamada的双套管法,设单纯UW液灌注作为对照组,实验组是先用HCA液再用UW液灌注的方法灌注冷保存后(12,24 h)进行原位肝移植,移植12 h后抽血检测反映肝功能变化的ALT、以及肝脏病理改变。结果①体外实验显示冷保存8 h,灌注液中ALT值,3组间差异无显著性(P>0.05),但冷保存16,32 h后的ALT值对照组均明显低于实验组1、实验组2,差异有显著性(P<0.05);16 h冷保存后肝重量差显示,实验组1、实验组2肝重量差均小于对照组,差异有显著性(P<0.05),但3组冷保存32 h后的肝脏均较保存前加重,差异无显著性(P>0.05)。②体内实验提示肝脏冷保存12,24 h后移植,实验组的ALT值高于对照组,组间差异有显著性(P<0.05);肝脏病理支持前述的改变。结论体外、体内的实验均显示:先予HCA液、再用UW液的灌注模式,和单纯UW液灌注方法相比较,前者的方法在抑制肝脏水肿、维持肝细胞机能稳定等方面作用较差,且随冷保存时间的延长,这种供肝灌注模式的负面影响更加明显。     

急诊手术治疗原发性肝癌破裂出血的临床分析

目的：分析总结原发性肝癌破裂出血时急诊行肝切除治疗的经验.方法：回顾性分析18例急诊行肝切除手术治疗的原发性肝癌破裂出血病例的临床资料。结果：18例原发性肝癌破裂出血者均顺利完成手术．无手术死亡。16例获得随访，生存时间10～58个月。1、3年生存率分别为75％（12／16）、50％（8／16）：结论：在经过临床选择的病例中，急诊肝部分切除术治疗原发性肝癌破裂出血是安全可行的，疗效显著，如患者的病情允许可作为首选