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程序性死亡因子配体1在脂肪间充质干细胞抑制大鼠肝移植术后排斥反应中的作用

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中华实验外科杂志 2014年第8期31卷 1731-1734,F0003页

作者：张宇万赤丹华中科技大学同济医学院附属协和医院肝胆外科武汉430022

目的探讨脂肪间充质干细胞（ADMSCs）对大鼠肝移植术后排斥反应的影响及其机制.方法分离培养SD大鼠ADMSCs,构建RNA干扰慢病毒载体转染ADMSCs阻断程序性死亡因子配体1（PD-L1）.以SD为供体、Wistar大鼠为受体建立肝移植模型,随机分为A... 详细信息

目的探讨脂肪间充质干细胞（ADMSCs）对大鼠肝移植术后排斥反应的影响及其机制.方法分离培养SD大鼠ADMSCs,构建RNA干扰慢病毒载体转染ADMSCs阻断程序性死亡因子配体1（PD-L1）.以SD为供体、Wistar大鼠为受体建立肝移植模型,随机分为A（空白组）、B（对照组）、C（实验组）3组,分别以生理盐水2 ml/ADMSCs（1×107个）/慢病毒转染后ADMSCs（1×10 7个）处理,比较3组大鼠肝移植术后7d肝功能、白细胞介素（IL）-2和IL-10、肝脏病理改变及干细胞凋亡、肝脏组织B细胞淋巴瘤/白血病-2（bcl-2）和bcl相关X蛋白（bax）表达差异.结果 A组血清谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TBIL）、IL-2和IL-10水平分别为（188.9±30.8）U/L、（177.1 ±27.9） U/L、（53.8±14.1）μmol/L、（296.32±56.25） ng/L、（74.92±10.98） ng/L;B组为（114.2±22.7） U/L、（134.8±18.1） U/L、（27.7±5.2）μmol/L、（186.36±17.84） ng/L、（194.56±36.87） ng/L;C组为（160.5 ±28.3） U/L、（155.6±25.6） U/L、（41.2±10.3） μmol/L、（250.95±42.68） ng/L、（125.52 ±26.34） ng/L,3组间差异有统计学意义（P＜0.05）.原位缺口末端标记法（TUNEL）检测A、B、C组的凋亡指数分别为35.86 ±5.92、12.39±2.23和28.47 ±4.32,3组间差异有统计学意义（P＜0.05）.B组大鼠肝移植模型术后bcl-2表达显著高于A组和C组,C组bcl-2表达低于A组;A组bax表达高于A组和C组,C组bax表达高于A组.结论 ADMSCs可显著降低肝移植术后排斥反应,PD-L1通路可能在其抑制排除反应中起重要作用.

关键词：脂肪间充质干细胞肝移植术排斥反应程序性死亡因子配体1

Experimental Study on the Cryopreservation of LLC-PK1 Epithelial Cells with Hypoxic UW Solution

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第4期27卷 426-428页

作者：万赤丹王春友刘涛王宏博杨智勇 Department of General Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology

The effects of oxygen partial pressure on cryopreservation of the cells with organ preservation solution were explored. Hypoxic UW solution was made by purging the UW solution with argon. The pig proximal tubule epithelial cells （LLC-PK1 cells） were cryopreserved in hypoxic UW solution （Ar-UW group） or standard UW solution （UW group） at 4℃ for 48 h. Trypan blue staining and LDH detection were performed to evaluate the injury of the cells. The results showed that the oxygen partial pressure in Ar-UW group was significantly declined from 242±6 mmHg to 83±10 mmHg. After cryopreservation at 4℃ for 48 h, LDH leakage rate and Trypan blue-stained rate in Ar-UW group were （11.3±3.4）% and （10.5±4.7）%, respectively, which were significantly lower than in UW group [（49.5±6.9）% and （47.6±9.3）% respectively, both P〈0.01]. It was concluded that lower oxygen partial pressure of UW solution was more beneficial to the cryopreservation of LLC.

关键词： organ preservation solution oxygen partial pressure cryopreservation

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荧光显像技术在胆道外科中的应用

上海医学 2022年第4期45卷 236-238页

作者：万赤丹华中科技大学同济医学院附属协和医院肝胆外科湖北武汉430022

有效识别胆道解剖结构是保证胆道手术安全的关键因素。应用荧光染料吲哚菁绿进行术中荧光胆道显像,有助于辅助胆道解剖结构的识别,实现实时手术导航。通过对荧光胆道显像技术的有效应用,可以提高胆道手术安全性和手术效率,降低胆道手术... 详细信息

有效识别胆道解剖结构是保证胆道手术安全的关键因素。应用荧光染料吲哚菁绿进行术中荧光胆道显像,有助于辅助胆道解剖结构的识别,实现实时手术导航。通过对荧光胆道显像技术的有效应用,可以提高胆道手术安全性和手术效率,降低胆道手术的风险。合理的给药方案及规范的手术流程是提高荧光胆道显像应用效果的关键。

关键词： ICG 荧光胆道显像胆囊切除

PDX-1 Expression in Pancreatic Ductal Cells after Partial Pancreatectomy in Adult Rats

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第5期24卷 464-466页

作者：刘涛王春友万赤丹熊炯忻许逸卿周峰 Pancreatic Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

To investigate the protein and mRNA expression of pancreas/duodenal homeobox-1 (PDX-1), a transcription factor as a marker for pancreatic stem cells, in pancreatic ductal cells of rats after partial (90 %) pancreatectomy and evaluated the significance of the PDX-1 expression. Western blot and Reverse transcriptase-polymerase chain reaction (RT-PCR) were used to detect the expression of PDX-1 protein and mRNA respectively. PDX-1 protein was only faintly detected in pancreatic ductal cells on the day 1 after partial pancreatectomy. On the day 2 and 3 after operation in operation group, a 2—3 fold increased PDX-1 protein was observed, corresponding to the characteristic 42—kD protein in Western blot. There was significant difference between operation group and sham-operation group (P0.05). RT-PCR revealed the PDX-1 mRNA expression showed no significant difference between operation group at various time points and sham-operation group (P>0.05). These results indicate that there was overexpression of PDX-1 in the cells of pancreatic epithelium during the regeneration of remnant pancreas after partial pancreatectomy in adult rats, suggesting the pancreatic stem cells in pancreatic ductal epithelial cells are involved in the regeneration of remnant pancreas and the expression of PDX-1 in ductal cells was regulated posttranscription.

关键词： pancreas/duodenal homeobox-1 pancreatic epithelium ductal cells rats

Experience in Diagnosis and Treatment of Bleeding Complications in Severe Acute Pancreatitis by TAE

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2005年第2期25卷 182-184页

作者：周峰王春友熊炯炘万赤丹郑传胜 Pancreas Surgery Center Union Hospital Tongji Medical College of Huazhong University ofScience and Technology Wuhan 430022 China

Summary: The experience in diagnosis and treatment of bleeding complications in severe acute pancreatitis (SAP) by transcatheter arterial embolization was summarized. The clinical data of 19 SAP patients complicated with intra-abdominal bleeding in our hospital from Jan. 2000 to Jan. 2003 were analyzed retrospectively and the therapeutic outcome of TAE was evaluated statistically. The results showed that the short-term successful rate of hemostasis by TAE was 89.5 % (17/19), the incidence of re-bleeding after TAE was 36.8 % (7/19) and the successful rate of hemostatis by second TAE was 71.4 % (5/7). It was concluded that the intra-abdominal bleeding in SAP was mainly caused by the rupture of erosive/infected pseudoaneurysm. Mostly, the broken vessels were splenic artery and gastroduodenal artery. In terms of emergence hemostatis, TAE is the most effective method. Surgical hemostasis is necessary if hemostasis by TAE is failed or re-bleeding occurs after TAE.

关键词： severe acute pancreatitis intra-abdominal bleeding erosive/infected pseudoaneurysm transcatheter arterial embolization

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脾切断流术后门静脉血栓形成防治的研究进展

中华肝胆外科杂志 2021年第10期27卷 797-800页

作者：辜嘉宇唐勇孙释然万赤丹华中科技大学同济医学院附属协和医院肝胆外科武汉430022

门静脉血栓形成(PVT)是脾切断流术后的常见并发症,具有隐匿性和潜在危险性,削弱了手术的疗效。且行脾切断流术患者常伴有严重肝硬化,其异常的止血功能使术后血栓防治成为临床医师需要面对的难题。本文就脾切断流术后PVT发生的危险因素... 详细信息

门静脉血栓形成(PVT)是脾切断流术后的常见并发症,具有隐匿性和潜在危险性,削弱了手术的疗效。且行脾切断流术患者常伴有严重肝硬化,其异常的止血功能使术后血栓防治成为临床医师需要面对的难题。本文就脾切断流术后PVT发生的危险因素、机制及防治方法的研究进行了综述。

关键词：门静脉血栓形成肝硬化脾切断流术门静脉高压

Expression of NALP3 in the Spleen of Mice with Portal Hypertension

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2010年第2期30卷 170-172页

作者：夏泽锋王国斌万赤丹刘涛王帅王博程锐 Department of General Surgery Union HospitalTongji Medical CollegeHuazhong University of Science and Technology

This study examined the mRNA expression of NALP3 in the spleen of the mice with hypersplenism due to portal hypertension （PH）. The mouse hypersplenism models were established by oral administration of tetrachloromethane （2 mL/kg/week for 12 weeks by oral gavage）. All the mice were randomly divided into a control group and an experimental group. The blood routine test was conducted, spleen index was calculated and spleen was histologically examined. Portal vein sera were taken for detection of the level of uric acid. The mRNA expressions of NALP3 and IL-1β in the spleen were detected by reverse transcriptase-polymerase chain reaction （RT-PCR）. The results showed that the platelet count was significantly lower in the experimental group [（674±102）× 10^9/L] than in the control group [（1307±181） × 10^9/L] （P〈0.05）, while the spleen index was significantly higher [（9.83 ±1.36） μg/g] in the experimental group than in the control group [（4.11 ±0.47） μg/g] （P〈0.05）. The histopathological changes of spleen followed the pattern of congestive splenomegaly. No significant difference was found in the uric acid level in the portal vein between the control group and the experiment group. The mRNA expressions of NALP3 and IL-1β were up-regulated significantly in the spleen in the experimental group as compared with those in the control group （P〈0.05）. It was concluded that NALP3 and IL-1β may play important roles in the pathogenesis of hypersplenism.

关键词： NALP3 interleukin-1β hypersplenism hypertension, portal

腹腔镜下吲哚菁绿荧光显像结合超声引导消融治疗肝脏肿瘤

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中华肝胆外科杂志 2019年第2期25卷 98-101页

作者：熊俊唐勇李民蔡雄万赤丹华中科技大学同济医学院附属协和医院肝胆外科武汉430022

目的探讨腹腔镜下吲哚菁绿(ICG)荧光显像结合术中超声引导消融治疗肝脏肿瘤的临床应用价值。方法采用回顾性研究方法,收集2016年5月至2017年12月华中科技大学同济医学院附属协和医院收治的81例采用荧光染色结合超声引导下完成的腹腔镜... 详细信息

目的探讨腹腔镜下吲哚菁绿(ICG)荧光显像结合术中超声引导消融治疗肝脏肿瘤的临床应用价值。方法采用回顾性研究方法,收集2016年5月至2017年12月华中科技大学同济医学院附属协和医院收治的81例采用荧光染色结合超声引导下完成的腹腔镜消融治疗原发性肝癌、转移性肝癌和肝血管瘤患者的临床资料。ICG荧光染色采用术前静脉注射和术中正染法。结果原发性肝癌36例。其中20例为肝内病灶,荧光染色不显影,在术中超声引导下完成消融;16例为包膜下病灶,荧光染色均显影,但15例肉眼不易分辨边界。转移性肝癌18例,病灶总数54枚,肝内及肝包膜下病灶荧光染色均未显影,包膜下病灶与周边染色肝组织对比明显。肝血管瘤27例,病灶均处于肝表面,荧光染色不显影,与周围染色肝组织对比明显。81例患者均手术成功,无中转开腹,平均单枚肿瘤消融时间(8.5±3.5)min,术中出血0~20ml,术中皮下气肿2例,血红蛋白尿1例,术后出血0例,术后肾功能不全0例,术后胆漏、周边组织损伤0例。结论腹腔镜超声引导消融术中及术后并发症发生率极低,是安全有效的消融方式,结合荧光显像可更加清晰地分辨正常肝组织及肿瘤,并对肝血管瘤消融的即时评估有帮助,具有良好的临床应用价值。

关键词：肝肿瘤导管消融术腹腔镜荧光显像超声引导

Proliferation and Differentiation of Duct Epithelial Cells after Partial Pancreatectomy in Rats

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第5期26卷 567-569页

作者：刘涛王春友万赤丹熊炯炘周峰 Department of Pancreatic Surgery Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

The proliferation and differentiation of pancreatic duct epithelial cells in remnant pancreas during regeneration after partial pancreatectomy in rats were studied, and the source of pancreatic stem cells was characterized. Partial （90 %） pancreatectomy was performed on 4- to 5-week-old Sprague-Dawley rats, and different duct epithelial cells and acinar cells were detected by immunohistrochemical stain method and scored using 5-bromo-2＇-deoxyuridine （BrdU） labeling index （LI） at various time points after partial pancreatectomy. It was found that at 24 h after partial pancreatectomy proliferation started in the main, large and small duct cells, and persisted in small duct cells to day 5. There was significant difference between the experimental group and the control group （P〈0.001）. Acinar cells positive for BrdU were greatly increased and reached the peak LI on day 5. The destroyed lobular architecture almost totally recovered on day 7, and the newly islet cells appeared around the pancreatic ducts. These results suggest that regeneration after partial pancreatectomy is involved in proliferation and differentiation of pancreatic stem cells, and pancreatic stem cells may locate in the pancreatic ductules.

关键词： pancreas stem cells regeneration 5-bromo-2'-deoxyuridine rats