肝癌术后肝再生状态与肝癌复发关系的初步研究

目的:探讨肝癌肝切除术后再生激素的变化，以评估肝再生激素与肝癌复发之间的关系。方法:收集2年内确诊为原发性肝癌行手术治疗的患者129例的临床资料，将小肝癌及大肝癌术后复发患者分别分为术后1，6个月，1年复发组，并以肝癌TAE治疗患者40例作为对照组。分别于术前1d与术后1, 3, 7, 10, 14 d清晨抽取静脉血检测血清肝细胞生长因子(HGF)，观察其在手术前后HGF的变化。手术中切取癌及癌旁组织为标本分别采用半定量RT-PCR及Western Blot法检测HGF受体c-met的mRNA及蛋白表达，比较复发时间早晚与表达量的差别。将所得结果与病理检查发现的脉管癌栓发生率对照。结果:肝切除后，患者血清HGF水平升高，术后约10 d达高峰期，14 d 开始降低。大肝癌组的HGF升高水平明显高于小肝癌组(P<0.05)。大、小两肝癌组的c-met mRNA及蛋白的水平变化均显示肿瘤复发时间越早，c-met水平越高，脉管癌栓发生率越高。结论:肝癌患者肝部分切除术后血清HGF水平明显增高及肿瘤本身c-met 蛋白过度表达可能与术后早期复发有关。     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.     

�˹���֧�ֺ������Ұ�λ������ֲ���Ʊ����Ըι���˥��1������

暴发性肝功能衰竭(fulminant liver failure,FLF)起病急，病情凶险，短期内病死率达80％以上。我院通过人工肝支持和亲属右半肝活体肝移植救治暴发性肝功能衰竭1例。报告如下。     

内镜诊治Ⅲ型先天性胆管扩张症1例

Ⅲ型先天性胆管扩张症为胆总管十二指肠开口部的囊状扩张,是先天性胆管扩张症的其中一型。Ⅲ型先天性胆管扩张症发生率极低。本研究报告了1例成人Ⅲ型先天性胆管扩张症患者,经内镜确诊并实施十二指肠镜胆管造影、乳头括约肌切开、胆总管末端囊肿切开、鼻胆管引流术,患者术后恢复快,效果良好。     

内镜逆行胆胰管造影术处理心脏死亡器官捐献供体肝移植受体术后胆道狭窄临床分析

目的探讨内镜逆行胆胰管造影技术(ERCP)在心脏死亡器官捐献供体(DCD)肝移植术后胆道狭窄治疗中的临床应用价值。方法回顾性分析2011年9月至2014年6月接受DCD肝移植手术治疗的34例肝移植受体的临床资料。结果磁共振胰胆管造影(MRCP)作为诊断肝移植术后胆道狭窄的标准方法,34例DCD肝移植术后发生胆道狭窄8例,发生率为23.53%,其中2例为吻合口狭窄,胆道狭窄组的肝移植等待时间明显长于非胆道狭窄组[(72.5±86.0)d vs(14.4±30.1)d,U=40.0,P=0.01]。1例自行再通,7例接受ERCP胆道内支架置入均获成功,ERCP术后并发胰腺炎4例(57.14%),胆管炎1例(14.29%)和胆管炎合并肝脓肿1例(14.3%)。结论胆道狭窄是DCD肝移植术后常见并发症,MRCP是诊断肝移植术后胆道狭窄的重要非有创性检查手段,ERCP胆道内支架置入是处理肝移植术后胆道狭窄的有效治疗方式,仍需进一步观察ERCP的疗效。     

浆膜下处理胆囊动脉的腹腔镜胆囊切除术

目的 探讨浆膜下处理胆囊动脉的腹腔镜胆囊切除术在临床中的应用价值。 方法 回顾性分析2008年1月～2011年10月169例实施浆膜下处理胆囊动脉的腹腔镜胆囊切除术患者的临床资料,术中离断胆囊管后紧贴胆囊壁肌层平面顺行分离,于胆囊浆膜下处理胆囊动脉进入胆囊壁的分支,游离切除胆囊。胆囊动脉的处理：电钩梯度电凝113例,超声刀处理39例,Ligasure处理8例,钛夹5例,4例胆囊动脉与胆囊管伴行,处理上胆囊动脉与胆囊管一并用Hem-O-Lok夹闭。 结果 全组病例均在腹腔镜下完成手术,其中8例继而行胆总管切开取石,T管引流术,手术时间25～205min,平均43min;术中出血量5～250ml,平均10ml,术中解剖Calot三角内胆囊动脉发自肝右动脉65例,余104例仅解剖其分支,未追求其来源。术后住院时间1～12d,平均4.3d,术后随访2～18个月,平均随访8月,全组无腹腔出血、胆管损伤等并发症。 结论 浆膜下处理胆囊动脉的腹腔镜胆囊切除术更符合安全、微创的理念,是一种合理的手术方式,值得在临床推广。     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.     

腹腔镜技术在肝部分切除术中的临床应用

目的探讨腹腔镜肝部分切除术的方法和应用价值。方法选择经临床筛选的第Ⅱ，Ⅲ，Ⅴ，Ⅵ段或边缘型的肝癌6例、肝海绵状血管瘤4例、左肝外叶结石2例及肝局灶性结节性增生1例。在腹腔镜下应用电凝、超声刀、切割器等方法断肝，采用钛夹夹闭，缝扎，医用胶粘封等多种对肝断面方法进行处理，完成腹腔镜下肝部分切除术。其中3例在全腹腔镜下完成，10例采用手助下腹腔镜肝部分切除术。结果13例患者均在腹腔镜下成功完成手术，包括左肝外叶切除及肝的非规则切除，手术时间54～130min。术后住院6～9d，手术出血40～7ml，无中转开腹手术。术中未出现不能控制的并发症，术后恢复时间较常规开腹的肝切除患者明显缩短。肝癌患者术后随访6～15个月，2例分别于手术后3个月及4个月肝内复发，其余4例随访至今无复发。结论在所经过临床选择的病例中。腹腔镜下肝部分切除术是痛苦小、安全、可行的手术方式。     

原位肝移植术后并发症18例临床分析

目的探讨原位肝移植术后并发症的诊治经验。方法回顾性分析我院18例次原位肝移植(其中2例再次肝移植)病人术后各种并发症的诊治经过。结果对16例患者(5例终末期肝硬化、7例原发性肝癌、1例肝癌术后肝功能衰竭、1例肝豆状核变性、1例慢性。肾功能衰竭合并肝炎后肝硬化、1例急性暴发性肝功能衰竭)进行原位肝移植。手术成功12例．术后最长存活已超过4年。术后并发症包括：腹腔内出血3例、脑血管病变2例、肺部感染6例、急性呼吸窘迫综合征(ARDS)2例、肝上下腔静脉狭窄2例、肝动脉血栓形成1例、胆道结石或胆泥3例、胆漏1例、急性排斥反应2例、慢性排斥反应2例、急性肾功能衰竭2例。围手术期死亡6例，直接死亡原因：脑出血1例、ARDS1例、急性。肾功能衰竭1例、肝动脉血栓1例、急性排斥反应1例、肝功能衰竭1例。结论肝移植围手术期采取合理的防治措施能有效减少肝移植术后并发症的发生，对肝移植术后并发症的及时诊断和正确处理是提高肝移植成功率的关键。