猪胆总管脱细胞支架的制备及组织学评价

目的 用不同的脱细胞方法对猪胆总管进行处理,比较脱细胞前后的组织学变化,筛选适宜的脱细胞方法,为组织工程胆管支架材料的应用提供理论依据。 方法 30例猪胆总管随机分为5组：对照组（A组）：0.05% 胰蛋白酶+核酸酶（B组）：0.1%十二烷基硫酸钠（SDS）+核酸酶（C组）：1.0% Triton X-100+核酸酶（D组）：1.0% Triton X-100+0.1% SDS +核酸酶（E组）。通过HE染色光学显微镜下观察脱细胞基质的组织结构和细胞残留情况;用紫外分光光度法检测脱细胞基质的DNA含量,计算脱细胞率。结果 脱细胞B组有少量细胞残留,纤维有损伤;脱细胞C组、D组、E组的细胞均被去除,纤维无明显损伤。A组的DNA含量为（71.24 ± 2.56）μg /100 mg。B、C、Ｄ、Ｅ4个脱细胞组的DNA含量与A组的差异有统计学意义（F =15.29, P<0.01）,均有明显的脱细胞效果（P<0.01）。Ｂ组的脱细胞率为77.03%,比C组、Ｄ组、Ｅ组的脱细胞效果稍差（P<0.05）,Ｅ组的脱细胞率高达99.03%。 结论 应用1.0% Triton X-100+0.1% SDS +核酸酶的脱细胞效果好,能更好地降低胆总管的免疫原性,是一种比较理想的猪胆总管脱细胞方法。     

两种方法制备脱细胞血管材料在动物体内免疫反应的比较

文题释义：去垢剂：又称表面活性剂，是一类既具有亲水基又具有疏水基的物质，一般具有乳化、分散和增溶作用，可分阴离子、阳离子和中性去垢剂等多种类型，中性去垢剂在蛋白提取中应用较多。  巨噬细胞的极化：巨噬细胞可分化为包括M1、M2a、M2b、M2c在内的极化表型，其中的M2表型巨噬细胞在组织修复和伤口愈合中发挥了重要作用。M1型巨噬细胞可促进单核巨噬细胞的聚集，分泌大量促症因子包括白细胞介素12、白细胞介素6、肿瘤坏死因子等，然而M2型可以分泌大量抗炎症因子，诱导巨噬细胞清除凋亡细胞及组织修复的作用。背景：去垢剂作为目前较成熟的脱细胞方法，具有操作简便、破坏小、实验条件容易控制等优点，有研究将不同的去垢剂联合应用于脱细胞中取得了不错的效果。 目的：对比两种去垢剂联合方案脱细胞的效果，以及两种脱细胞血管材料植入动物皮下后的免疫反应。 方法：采用两种方案对猪颈动脉进行脱细胞处理，一组置于1%十二烷基硫酸钠与1%脱氧胆酸钠混合溶液内处理72 h(SDS+SDC组)，另一组置于1%十二烷基硫酸钠与1%Triton X-100混合溶液内处理72 h(SDS+Triton X-100组)，脱细胞后分别进行组织学分析、扫描电镜分析、力学分析与DNA含量检测。将两组脱细胞血管材料分别植入SD大鼠背部皮下，术后1，2，4，8周取出移植血管材料，进行苏木精-伊红染色与免疫荧光染色。实验已通过江南大学实验动物伦理委员会批准。 结果与结论：①组织学分析显示，SDS+SDC组方案有效去除了细胞成分，较完整的保留了细胞外基质结构，脱细胞效果优于SDS+Triton X-100组；②SDS+SDC组DNA含量明显低于SDS+Triton X-100组(P < 0.05)，两组爆裂强度与缝合耐受强度比较差异无显著性意义(P > 0.05)；③扫描电镜显示，SDS+Triton X-100组细胞外基质破坏程度大于较SDS+SDC组；④皮下植入实验中，术后1周时两组颈动脉材料周围均可见大量炎性细胞浸润，术后8周时SDS+SDC组脱细胞材料周围已无炎性细胞浸润，SDS+Triton X-100组脱细胞材料周围仍可见明显的淋巴细胞浸润；SDS+Triton X-100组脱细胞材料主要诱导巨噬细胞向M1型分化，SDS+SDC组脱细胞材料主要诱导巨噬细胞向M2型分化；⑤结果表明相对比1%十二烷基硫酸钠联合1%Triton X-100脱细胞处理方案，1%十二烷基硫酸钠联合1%脱氧胆酸钠脱是较有前景的脱细胞细胞处理方案。ORCID: 0000-0002-3120-0203(尹晶) 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程     

不同脱细胞方法对猪尾纤维环生物力学特性及组织结构的影响

目的探讨不同脱细胞处理方法对猪尾椎间盘纤维环生物力学特性及组织结构的影响,为构建组织工程纤维环提供实验依据。方法取新鲜猪尾纤维环60个,随机分为4组,每组15个。Triton X-100组(A组):先将纤维环放入Tris-HCl低渗缓冲液中振荡48 h,再用TritonX-100、DNaseⅠ和RNase A对纤维环脱细胞处理;SDS组(B组):将纤维环冻融3次,接着用SDS、DNaseⅠ和RNase A对纤维环脱细胞处理;胰蛋白酶组(C组):用含胰蛋白酶、DNaseⅠ和RNase A的Tris缓冲液对纤维环脱细胞处理;对照组(D组):纤维环不做任何处理。采用HE染色与扫描电镜方法,观察各组脱细胞情况以及纤维环超微结构的变化;采用生物化学和生物力学方法检测各组纤维环的胶原含量、氨基葡聚糖(GAG)含量及力学参数。结果与对照组比较,HE染色及扫描电镜可见A、B、C 3组均无细胞残留;A组纤维环超微结构未见破坏,C组纤维环超微结构可见轻度破坏,B组纤维环超微结构可见明显破坏。A、B、C 3组纤维环胶原含量与对照组比较无统计学差异(P>0.05),但GAG含量均明显低于对照组(P<0.05)。A、C两组的力学参数(极限载荷、极限应力、韧度、弹性模量、断裂功耗)与对照组无统计学差异(P>0.05),B组上述力学参数均低于对照组(P<0.05)。结论 Triton X-100组处理后的猪尾椎间盘纤维环无细胞残留,结构无破坏,基质成分及力学性能保存良好,适合于构建组织工程纤维环。     

IL-1β通过加重脂质诱导的内质网应激导致人系膜细胞损伤

 目的： 探讨白细胞介素1β（IL-1β）加重脂质诱导的内质网应激（ERS）而导致人肾小球系膜细胞（HMCs）损伤的分子机制。方法：将HMCs分为对照组、高脂组（LDL）、IL-1β+高脂组（IL-1β+LDL）及4-苯基丁酸(4-PBA)干预组（4-PBA+IL-1β+LDL）。油红O染色检测HMCs胞内脂质沉积;real-time PCR检测葡萄糖调节蛋白78(GRP78)、蛋白激酶R样内质网激酶（PERK）和α-平滑肌肌动蛋白（α-SMA） mRNA水平;免疫细胞化学分析GRP78蛋白水平; Western blotting检测NF-κB p65水平;ELISA法测定细胞培养上清IL-6和TGF-β1含量。结果：与LDL组相比,IL-1β+LDL组胞内脂质沉积、GRP78 mRNA与蛋白水平、PERK mRNA水平、NF-κB p65水平及IL-6分泌量均显著增高（均P＜0.05）;4-PBA可减少胞内脂质沉积,降低GRP78 mRNA与蛋白及PERK mRNA水平,抑制NF-κB p65的表达,减少IL-6的分泌（均P＜0.05 vs IL-1β+LDL组）。与LDL组相比,IL-1β+LDL组α-SMA mRNA表达增高（P＜0.05）,4-PBA可显著降低其表达（P＜0.05 vs IL-1β+LDL组）。结论：IL-1β可加重脂质诱导的ERS,从而导致细胞损伤。     

TGFBR1基因调控Wnt/β-catenin信号通路对结缔组织病相关的肺间质病变中抗纤维化的机制研究

目的：探讨TGF-β受体Ⅰ（TGFBR1）基因调控Wnt/β-catenin信号通路对结缔组织病相关的肺间质病变（CTD-ILD）的抗纤维化作用。方法：以肺癌的癌旁组织作为正常对照组,Western blot检测CTD-ILD患者肺组织TGFBR1的蛋白表达;人胚肺成纤维细胞系MRC-5分为空白对照组、TGF-β1组、阴性对照组和TGFBR1-siRNA组,各组细胞培养48 h,Western blot检测TGFBR1、Ⅰ型胶原、结缔组织生长因子（CTGF）、α-平滑肌肌动蛋白（α-SMA）、E-钙黏蛋白（E-cad）及Wnt/β-catenin信号通路β 连环蛋白（β-catenin）和c-Myc的蛋白表达;CCK8法检测各组细胞的活力。结果：TGFBR1在CTD-ILD肺组织中的表达显著高于正常肺组织（P<0.05）;与空白对照组比较,TGF-β1组细胞活力、TGFBR1、Ⅰ型胶原、CTGF、α-SMA、β-catenin和c-Myc的蛋白表达均显著升高,E-cad蛋白表达显著降低（P<0.05）;TGF-β1组和阴性对照组细胞活力、TGFBR1、Ⅰ型胶原、CTGF、α-SMA、E-cad、β-catenin和c-Myc的蛋白表达差异均无统计学意义（P＞0.05）;与TGF-β1组比较,TGFBR1-siRNA组细胞活力、TGFBR1、Ⅰ型胶原、CTGF、α-SMA、β-catenin和c-Myc的蛋白表达均显著降低,E-cad蛋白表达显著升高（P<0.05）。结论：TGFBR1基因在CTD-ILD肺组织中表达升高,抑制TGFBR1的表达可通过下调Wnt/β-catenin信号通路降低CTD-ILD纤维化的发生。     

脱细胞真皮基质补片修复滇南小耳猪胆管损伤☆

背景：目前胆管缺损的修复效果不佳,脱细胞真皮基质已广泛用于烧伤、疝修复、口腔黏膜及软组织修复等。 目的：观察脱细胞真皮基质修复滇南小耳猪胆管损伤的效果。 方法：将滇南小耳猪建立胆管损伤动物模型,建模成功后按修复方式分为胆肠吻合组、膨体聚四氟乙烯组及脱细胞真皮基质组,胆肠吻合组进行胆肠吻合修补,膨体聚四氟乙烯组及脱细胞真皮基质组分别用膨体聚四氟乙烯补片和脱细胞真皮基质补片制成直径稍宽管状进行修补。 结果与结论：治疗后2~4个月,脱细胞真皮基质组的总胆红素低于其他2组(P < 0.05);免疫组织化学染色显示,治疗后1~4个月,其转化生长因子β1表达均高于其他2组(P < 0.05),表达随时间增加而降低(P < 0.05)。脱细胞真皮基质组胆总管吻合口及周围组织出现胆道上皮、腺体、血管及平滑肌等再生。脱细胞真皮基质组治疗后未出现胆道狭窄、感染及排斥反应发生。结果证实,脱细胞真皮基质可生理性修复胆道并重建其功能,并发症少,生物相容性较好。 关健词：脱细胞真皮基质;补片;胆管损伤;转化生长因子β1;猪;生物相容性;细胞外基质材料 doi:10.3969/j.issn.1673-8225.2012.12.012     

ILK siRNA对高糖刺激的人肾小管上皮细胞GSK-3β及β-catenin表达的影响*

 目的：探讨高糖诱导肾小管上皮细胞转分化中整合素连接激酶小干扰RNA（ILK siRNA）对糖原合成酶激酶3β（GSK-3β）磷酸化和β-连环蛋白（β-catenin）核内表达的影响及意义 。方法：体外培养人近端肾小管上皮细胞系HKC,分为正常对照组（NG）、高糖组（HG）、高糖＋阴性转染对照组 (HG+HK)和高糖＋ILK siRNA组(HG+ILK siRNA)。倒置荧光显微镜下观察绿色荧光蛋白表达。RT-PCR及Western blotting检测ILK  mRNA及蛋白表达水平;免疫细胞化学检测磷酸化GSK-3β（p-GSK-3β）和β-catenin表达。Western blotting检测总GSK-3β、p-GSK-3β、核β-catenin、总β-catenin、E-钙黏蛋白（E-cadherin）和α-平滑肌肌动蛋白（α-SMA）的表达水平。结果：（1）倒置荧光显微镜下可见绿色荧光蛋白表达,证实构建的siRNA重组质粒成功转染HKC细胞;（2）与HG和 HG+HK组相比,HG+ILK siRNA组ILK mRNA及蛋白水平下降,但较NG组表达仍高;（3）HG+ILK siRNA组ILK基因沉默后,p-GSK-3β与核β-catenin蛋白表达较HG及HG+HK组均下降,但较NG组表达仍高。而总GSK-3β与总β-catenin 在各组表达无明显差异。结论:ILK、GSK-3β和β-catenin可能参与了高糖介导的肾小管上皮细胞转分化过程。ILK可能通过调节Wnt/β-catenin途径下游效应蛋白GSK-3β和β-catenin的表达而促使肾小管上皮细胞转分化。     

氧化苦参碱抑制高糖诱导的大鼠肾小管上皮-间充质转化及其机制研究

 目的：探讨氧化苦参碱(OM)对高糖诱导的大鼠肾小管上皮-间充质转化（EMT）的抑制作用及其可能机制。方法：体外培养大鼠近端肾小管上皮NRK52E细胞,随机分为：对照组、高糖组、高糖+OM不同浓度组和高糖+0.50 g/L OM动态观察组。采用real-time PCR和Western blotting方法检测转化生长因子β1（TGF-β1）、Smad7、α-平滑肌肌动蛋白（α-SMA）、E-钙黏素（E-cadherin）mRNA和蛋白的表达。结果：（1）与对照组相比,高糖组TGF-β1、α-SMA mRNA和蛋白表达水平均进行性增高,Smad7蛋白表达进行性降低,E-cadherin mRNA和蛋白表达进行性降低,呈时间依赖性（P＜0.05）,而Smad7 mRNA表达进行性增高（P＜0.05）;（2）与高糖组相比,高糖+ OM不同浓度组随OM剂量增加,TGF-β1、α-SMA mRNA和蛋白表达均逐渐降低,Smad7蛋白表达水平逐渐增高,E-cadherin mRNA和蛋白表达水平逐渐增高,且呈剂量依赖性（P＜0.05）,而Smad7 mRNA表达无明显差异;（3）与高糖组相比,高糖+0.50 g/L OM动态观察组TGF-β1、α-SMA mRNA和蛋白表达持续降低,Smad7蛋白表达持续增高,E-cadherin mRNA和蛋白表达持续增高（P＜0.05）,而Smad7 mRNA表达无明显差异。结论：OM可抑制高糖诱导的NRK52E细胞发生EMT,其机制可能与OM下调TGF-β1表达及上调Smad7蛋白表达,进而抑制TGF-β1/Smads信号通路的致纤维化效应有关。     

比较两种方法制备脱细胞小血管支架

目的:比较0.5%Triton X-100 0.05%NH4OH和酶法制备脱细胞小血管支架的效果.方法:分别用含0.5%Triton X-100 0.05%NH4OH处理3 d或1.0H Triton X-100 0.125%胰蛋白酶 Dnase Rnase孵育48 h,脱去犬股静脉中的细胞成分,50Co辐照消毒,血清浸泡24 h,将平滑肌细胞和内皮细胞接种到脱细胞支架中;进行H-E染色、胶原纤维和弹力纤维染色,扫描电镜观察及力学检测.结果:0.5%Triton X-100 0.05%NH4OH法完全地脱去了血管细胞;细胞外基质较完整地保留下来,其形态结构与脱细胞前无明显改变;见种植细胞在支架内生长良好,连成片,支架具有良好的生物相容性;力学结果显示其弹性回复率和最大断裂强度好于酶组.结论:两种方法比较,0.5%Triton X-100 0.05%NH4OH法简便易行,成本低,脱细胞效果好,组织相容性佳,对力学性状影响小,是比较理想的制备脱细胞小血管支架的方法.     

硫化氢对高糖诱导的小鼠足突细胞损伤的影响*

 目的： 观察硫化氢对高糖诱导的小鼠足突细胞损伤的影响并探讨其作用机制。方法: 将体外培养的小鼠足突细胞系MPC5分为高糖组、正常糖组、正常糖+DL-炔丙基甘氨酸(PPG)组和高糖+NaHS组。处理后,用Western blotting检测各组细胞胱硫醚γ-裂解酶（CSE)、紧密连接蛋白2（ZO-2）、裂孔蛋白（nephrin）及β-连环蛋白（β-catenin）蛋白水平表达差异。结果: （1）高糖显著降低CSE、nephrin和ZO-2表达（P＜0.05）,而β-catenin的水平明显增高（P＜0.05）,4种蛋白变化均表现出时间依赖性;（2）正常糖培养加不同浓度PPG可显著抑制ZO-2和nephrin的表达（P＜0.05）,显著提高β-catenin的水平（P＜0.05）,3种蛋白变化呈浓度依赖性;（3）高糖诱导的足突细胞加NaHS培养后,ZO-2和nephrin表达可部分恢复（P＜0.01）,β-catenin表达可被部分抑制（P＜0.01）。结论: 本研究结果提示高糖诱导的足突细胞CSE表达降低可能是足突细胞损伤的重要机制之一。而外源性硫化氢对高糖诱导的足突细胞损伤具有一定保护作用,其机制可能与增加ZO-2表达、抑制Wnt/β-catenin通路有关。     

乙酸消融胆道的实验研究

目的探讨乙酸消融家兔胆道后病理及肝功改变,为临床应用消融栓塞胆道治疗肝内胆管结石提供实验依据。方法观察家兔右外叶胆道于25%至5%浓度乙酸消融后肝脏功能状态、肝组织病理改变和术后存活情况。结果25%乙酸胆道消融造成右外叶肝脏大块坏死,仅肝叶边缘有少量存活肝组织。20%乙酸胆道消融后近肝门部肝实质以汇管区为中心的坏死,肝叶中部见汇管区及周围少量肝细胞坏死,肝叶边缘见汇管区点状坏死。15%乙酸胆道消融后近肝门部见汇管区及周围少量组织坏死,肝叶中部及边缘组织见胆管无明显改变,周围组织点状坏死。10%乙酸胆道消融后近肝门部汇管区胆管坏死,周围充血,肝叶中部胆管上皮完整,周围组织充血,肝叶边缘胆管完整,周围少量肝细胞肿胀。5%乙酸胆道消融后近肝门部胆管上皮坏死,肝叶中部胆管上皮完整,仅充血或少量肝细胞肿胀。结论20%为乙酸消融胆道较理想的浓度。     

Abnormal bile duct epithelium accompanying septicaemia

Summary A 56-year-old female without previous hepatobiliary disease developed a severe obstructive cholestasis followingE. coli urinary tract infection with septicaemia. Liver biopsy showed cholangiolitis and a unique abnormality of almost all the interlobular bile ducts; the epithelium was irregular with polymorphic, angular, and hyperchromatic or pyknotic nuclei. Some ducts were ectatic, others narrowed due to protrusion of proliferating epithelium. In some areas the ducts were blurred or completely destroyed. Cholangitis or granulomas were, however, not present.Abnormal interlobular bile ducts have to our knowledge not previously been described in septicaemia. The lesion is morphologically distinguishable from other types of abnormal bile ducts. It is considered to be caused by endotoxaemia and seems to be reversible. The cholestasis may be due to endotoxic alteration of biliary secretion, bacterially induced inspissation of bile, and/or mechanical obstruction due to the bile duct lesions.     

Muscularis mucosae versus muscularis propria in gallbladder, cystic duct, and common bile duct: smoothelin and desmin immunohistochemical study

The muscle layer in the cystic duct and common bile duct is not well defined, and it is unresolved whether it represents muscularis mucosae or muscularis propria. Smoothelin is a novel smooth muscle–specific contractile protein expressed only in fully differentiated smooth muscle cells of the muscularis propria and not in proliferative or noncontractile smooth muscle cells of the muscularis mucosae. In this study, we characterize the histologic aspects of the muscle layer in gallbladder, cystic duct, and common bile duct by evaluation of routine histologic sections and the utilization of immunohistochemistry using desmin and smoothelin. Formalin-fixed, paraffin-embedded sections of the gallbladder (15 cases), cystic duct (11 cases), and common bile duct (10 cases) were stained for smoothelin and desmin. Staining intensity was evaluated as weak or strong. The staining pattern score was evaluated as follows: 0 or negative = less than or equal to 5% positivity, +1 or focal = 6% to 10% positivity, +2 or moderate = 11% to 50% positivity, and +3 = greater than 50% muscle cells positivity. With desmin, strong and diffuse (+3) staining was observed in all gallbladder cases (15/15, 100%), highlighting one continuous muscle layer. The muscle layer was discontinuous and interrupted in all cystic duct cases and in most common bile ducts, highlighted by the desmin stain. Smoothelin intensely stained (at least +2) muscle fibers in the gallbladder in 11 (73%) of 15 cases similar to that observed with desmin staining. In contrast, common bile ducts predominantly had absent or weak and focal immunostaining (0 or +1 staining) with smoothelin (7/10, 70%), with only a few cases (3/10, 30%) having +2 staining (no cases with +3). Cystic ducts also showed absent or weak and focal immunostaining with smoothelin, with 5 (44%) of 11 cases showing 2+ immunostaining with smoothelin (no cases with 3+). Based on our findings, we conclude that, in the gallbladder wall, the muscle layer is muscularis propria and there is no muscularis mucosae present. In the cystic duct and common bile duct, only an attenuated and incomplete muscle layer of muscularis mucosae is present; because there is no muscularis propria, there probably is limited contractile function. Differentiating these anatomical muscle structures may be important for the pathologic staging of carcinoma in these organs.     

纳米银－猪脱细胞真皮基质敷料治疗浅Ⅱ度烧伤创面的临床观察

目的观察纳米银－猪脱细胞真皮基质敷料在临床上治疗浅Ⅱ度烧伤创面的临床疗效。 方法2014年1月到2015年12月,选取北京军区总医院烧伤整形科收治的浅Ⅱ度烧伤患者90例,按入院顺序依次编码,采用随机排列数字表法将90例患者分为纳米银敷料组、猪脱细胞真皮基质敷料组及纳米银－猪脱细胞真皮基质敷料组,每组30例。患者入院当天,拍照计算创面面积,并用咽拭子取创面分泌物作细菌培养,行创面清创术,分别在创面上敷以纳米银敷料、猪脱细胞真皮基质敷料以及纳米银－猪脱细胞真皮基质敷料。于治疗后第5天,用咽拭子取创面分泌物作细菌培养;采用痛觉评分标准,通过询问与观察患者换药时的痛觉情况,评估患者换药时痛觉评分。于治疗后第7天,拍照计算创面面积,计算创面愈合率。记录创面最终愈合时间。数据比较采用单因素方差分析、χ²检验及SNK－q检验。 结果纳米银敷料组、猪脱细胞真皮基质敷料组及纳米银－猪脱细胞真皮基质敷料组在治疗后第5天创面细菌培养阳性数结果分别为2例(6.6%)、9例(30.0%)、1例(3.3%),3组结果比较,差异有统计学意义(χ²＝10.962,P＝0.004);纳米银－猪脱细胞真皮基质敷料组的细菌培养阳性数结果显著优于猪脱细胞真皮基质敷料组,差异有统计学意义(χ²＝7.680,P＝0.006)。纳米银敷料组、猪脱细胞真皮基质敷料组及纳米银－猪脱细胞真皮基质敷料组治疗后第5天的痛觉评分[(8.6±0.5)、(6.6±0.8)、(0.6±1.3)分],治疗后第7天的创面愈合率[(61.67±18.22)%、(86.77±15.32)%、(99.80±0.56)%],创面愈合时间[(11.5±1.3)、(10.3±0.7)、(7.3±0.7)d],组间差异均有统计学意义差异(F＝201.7、19.9、55.7,P值均小于0.05);纳米银－猪脱细胞真皮基质敷料组治疗后第5天的痛觉评分显著低于其余两组,差异均有统计学意义(P值均小于0.05);治疗后第7天的创面愈合率明显优于其余两组,差异均有统计学意义(P值均小于0.05);创面愈合时间显著短于其余两组,比较差异均有统计学意义(P值均小于0.05)。 结论纳米银－猪脱细胞真皮基质敷料具有抗感染、促进创面愈合及减轻换药痛觉的作用。     

新型生物可降解支架材料生物特性及在损伤胆道修复中的应用

BACKGROUND:A variety of factors contribute to biliary injury that is difficult to be repaired. Stent implantation is extensively used for bile duct injury, but either scaffolds made by metal or plastics can lead to certain adverse reactions. OBJECTIVE: To explore the biological characteristics of a novel biodegradable scaffold and its repair effects on bile duct injury. METHODS: The biological characteristics of the novel biodegradable scaffold were detected by fresh bile, and its degradation was observed at different time points. Thirty Bama mini pigs were included and were randomly divided into observation group (n=15) and control group (n=15). After bile duct injury models were prepared, the control group was subjected to the bile duct interrupted suture, while the observation group was subjected to the novel biodegradable scaffold combined with omentum majus. The biological properties of the scaffolds were observed. Hepatic enzymes and serum total bilirubin levels were detected, as well as hematoxylin-eosin staining, Masson staining and immunohistochemistry detection of α-smooth muscle actin were performed. RESULTS AND CONCLUSION: Before and 1, 3 and 6 months after surgery, hepatic enzymes and total bilirubin of two groups were detected, and neither intra-group nor intergroup comparisons had significant differences (P > 0.05). Hematoxylin-eosin staining and Masson staining revealed that inflammatory reactions and fiber hyperplasia at the anastomotic site in the observation group were lighter than those in the control group at different time points after surgery. The α-smooth muscle actin-positive scores in both two groups were in a rise at 1 and 3 months after surgery, and peaked at the 3rd month, and then began to decline. Moreover, the α-smooth muscle actin-positive scores in the observation group were significantly lower than those in the control group at 3 and 6 months after surgery (P < 0.05). These results show that the novel biodegradable scaffold has good biological characteristics and can obtain ideal repair effects in the bile duct injury.     

自体微粒皮与异体脱细胞微粒真皮混合移植对大鼠创面细胞骨架蛋白与新生血管化的影响

目的探讨自体微粒皮与异体脱细胞微粒真皮混合移植对创面愈合的影响,并对有关机理做进一步研究。方法用雄性Wistar大鼠制做异体脱细胞真皮,90只雌性SD大鼠背部建立全层皮肤损伤模型,分为A、B、C、D、E组,每组18只,A组为自体微粒皮组,B组为异体脱细胞微粒真皮组,C、D、E组为自体微粒皮与异体脱细胞微粒真皮按不同的比例（C组为1∶1,D组为1∶0.5,E组为1∶0.25）混合移植（统称为混合移植组）,比较各组的创面愈合率、微血管计数及细胞骨架蛋白（desmin）的表达。结果 （1）移植后2、3周,混合移植组创面愈合率均高于A组和B组,E组移植后2、3周创面愈合率分别为（88.00±5.71）%和（96.18±6.62）%,高于C组（79.81±7.07）%和（86.76±3.35）%（P〈0.05）;（2）移植后2、3周混合移植组微血管数均明显高于A组、B组（P〈0.05）,移植后2周,E组、D组的血管数（35.67±1.53）、（34.33±1.53）明显多于C组（P〈0.05）。（3）细胞骨架蛋白在创面形成后表达上调,混合移植组细胞骨架蛋白的表达高于A组（P〈0.05）,尤以D组、E组明显。结论自体微粒皮与脱细胞微粒真皮混合移植创面愈合率高于自体微粒皮移植,且自体微粒皮与脱细胞微粒真皮混合移植的面积比例按1∶0.25混合移植效果最佳,这可能与创面血管化进程加快,及细胞骨架蛋白的适度表达有关。     

Factors producing bile infarction and bile duct proliferation in biliary obstruction

To clarify the factors producing bile infarction and bile duct proliferation in obstructive jaundice, the incidence of the hepatic lesions and the serum levels of the bile constituents were examined in three rat models. (1) Ligation of the common bile duct induced bile infarction, bile duct proliferation, retention of bile in the liver, and elevation of the serum levels of total bilirubin and total bile acids. (2) The rats treated by choledochotomy had bile in the abdominal cavity, but there was no retention of bile in the liver. The degree of development of bile infarction was similar to that of the common bile duct ligation group, but bile duct proliferation was not found: the serum levels of total bilirubin and total bile acids were elevated. (3) In the rats subjected to partial bile duct ligation, bile infarction and bile duct proliferation were seen only in the lobes with ligation of the hepatic ducts: only slight or no elevation of the serum levels of total bilirubin and total bile acids was found. These data suggest that bile infarction is caused by the toxic action of bile constituents other than bilirubin and bile acids, absorbed into the blood from the obstructed biliary system, and that bile duct proliferation is due to mechanical factors following bile retention or direct actions of retained bile in the liver.     

抑制素α、βA、βB亚基单克隆抗体制备及其初步应用

目的:制备INHα、βA、βB亚基的McAbs,并进行肿瘤免疫组化研究。方法:抗原为猪卵泡液INH提取物及人工合成的INHα、βA、βB亚基肽段(6个)。以上述抗原免疫BALB/c小鼠,经细胞融合、筛选、克隆化,获得杂交瘤细胞14株,对其免疫学特性进行了分析。用纯化的McAbs进行了肿瘤免疫组化研究(285例次)。结果:获得分别分泌抗INHα、βA、βB亚基的McAbs细胞株14个,效价1×10^-4-1×10^-6。免疫组化除生殖系统肿瘤阳性外,还发现乳腺癌的阳性率超过70%。结论:成功制备了分泌抗INH、、B亚基肽的McAbs,纯化后进行了肿瘤免疫组化研究。