肝硬化大鼠肝部分切除术后肝再生的干预研究

目的　以肝硬化大鼠为动物模型 ,研究药物对肝硬化大鼠肝部分切除术后肝再生的影响。方法　取健康的Wistar雄性大鼠 6 4只 ,以 6 0 ?l4油溶液 0 .3ml/ 10 0 g皮下注射 ,同时饮用 5 %酒精溶液 ,4 5d后制成肝硬化动物模型。模型大鼠随机分为 4组 ,16只 /组。全麻下均行左、中叶肝切除术。术后各组按以下方案处理 :A组 (对照组 )注射生理盐水 1mg/ (kg·d) ,B组为泮托拉唑组 ,注射 0 .2mg/ (kg·d) ,C组为重组人生长激素组 ,注射 0 .5U/ (kg·d) ,D组为两药合用组 ,同时给予泮托拉唑注射 0 .2mg/ (kg·d) ,重组人生长激素注射 0 .5U/ (kg·d) ) ,连续给药 1周。抽取静脉血样 ,取肝脏组织 ,检测肝功能、有丝分裂指数 (MI)、增殖细胞核抗原 (PCNA)、细胞核DNA含量。结果　泮托拉唑组、重组人生长激素组、两药合用组MI、PCNA阳性染色细胞量、细胞核DNA含量均高于对照组 (P <0 .0 5 ) ,两药合用组MI、PCNA阳性染色细胞量、细胞核DNA含量均高于泮托拉唑组、重组人生长激素组 (P <0 .0 5 ) ,但各组间肝功能变化无明显差异。结论　泮托拉唑组及重组人生长激素均对肝硬化大鼠肝部分切除术后肝细胞再生有促进作用 ,两药联合应用肝细胞再生更明显 ,其详细机制须待进一步研究。     

重组大鼠肝再生增强因子对急性肾衰竭大鼠肾脏的保护作用

目的 探讨重组大鼠肝再生增强因子(rrALR)对庆大霉素所致急性肾衰竭（ARF）大鼠肾小管上皮细胞及肾功能的保护作用。 方法 雌性Wistar大鼠150只，随机分成5组，每组30只，即健康对照组，ARF模型组，模型+空质粒对照组（空质粒组），模型+rrALR干预组（ALR组）：根据给予rrALR的剂量不同分为ALR1组和ALR2组两个亚组。分别于实验的第4、8、12、16和21天每组随机抽取6只大鼠在留取血、尿标本后，处死大鼠并取肾组织标本。常规生化方法检测各组大鼠BUN、Scr和尿N-乙酰-β-D-葡萄糖苷酶（NAG）酶的变化；PAS染色观察各组大鼠肾组织病理学改变；免疫组化法检测大鼠肾组织中ALR和增殖细胞核抗原（PCNA）的表达；Western印迹法检测肾组织中ALR蛋白的表达量。 结果 ARF大鼠各组BUN、Scr及尿NAG酶水平在第4、8、12、16天时均较对照组显著升高（P < 0.05）。与模型组和空质粒组相比，ALR组BUN、Scr及尿NAG酶水平明显降低（P < 0.05）；肾组织病理损害程度在各时间点明显减轻；而肾组织的ALR蛋白表达增加（P < 0.05）；肾小管上皮细胞增殖活跃；PCNA阳性细胞呈弥漫性分布，增殖指数（PI）明显升高（P < 0.05）。 结论 rrALR对急性损伤的肾小管上皮细胞具有减轻病变和促进再生修复的作用，可明显改善ARF大鼠的肾功能。     

外源性VEGF对自体移植脾组织血管再生的影响

目的：观察外源性VEGF对自体脾组织大网膜内移植后血管再生过程影响。方法：198只昆明种小白鼠随机分为脾切除自体脾组织大网膜内移植组及假手术组,将脾切除自体脾组织大网膜内移植后的小鼠再随机分为实验组（应用外源性VEGF组）和对照组（不用外源性VEGF组）。实验观察组小鼠分别于术后7、15、30d于尾静脉内注射VEGF,最后一次注射后1、7、14、30、60、90d各处死5只,取脾组织标本;对照组于术后观察上述对应时相点后各处死3只,取脾组织标本,进行墨汁灌注血管面密度测定。结果：术后7、15、30d注射VEGF后,较对照组的血管数量明显增多,墨汁灌注显示,实验观察组血管面密度较对照组明显增大。结论：外源性VEGF能够促进移植脾组织内血管生长,改善移植脾组织的血液循环,有利于移植脾组织再生,使其结构恢复更趋完善。     

胶原蛋白-明胶支架材料修复大鼠坐骨神经缺损的研究

目的制备胶原蛋白一明胶神经支架材料（CG材料）并研究其在修复大鼠10mm坐骨神经缺损实验中的疗效。方法以I型胶原蛋白、明胶通过冷冻干燥技术制备具有轴向微管结构的神经支架材料，用其桥接修复sD大鼠坐骨神经10mm缺损。术后16周分别行透射电镜，S-100、β-tubulin class Ⅲ、NFl60免疫荧光染色以及电生理检测，观察支架材料引导神经再生的疗效。结果制备的材料内部为孔径均匀且平行排列的微管结构，术后16周通过透射电镜和免疫荧光染色可见大量再生神经纤维，电生理检测神经传导速度及波幅接近自体神经移植。结论胶原蛋白一明胶支架材料能够有效的促进周围神经再生。     

人发角蛋白桥接周围神经缺损的形态学观察

目的 :了解人发角蛋白在神经再生中的作用 ,为临床桥接周围神经缺损寻求新的替代材料。方法 :将 18只新西兰兔的双侧坐骨神经切断 ,造成 10mm缺损 ,一侧用人发角蛋白桥接 (实验组 ) ,另一侧用空硅胶管桥接 (对照组 ) ,术后 1、 2、 3个月通过肉眼观察 ,光镜、电镜和有髓神经密度测定 ,观察、分析神经再生情况。结果 :实验组再生神经均通过 10mm缺损 ,对照组有 2例无神经生长 ;实验组再生神经排列较紧密、有序 ,髓鞘形成早于对照组 ;神经纤维密度明显高于对照组 (P <0 0 1)。结论 :人发角蛋白可促进周围神经再生 ,是桥接周围神经缺损的理想材料。     

Expression of myosin heavy chain isoforms in Duchenne muscular dystrophy patients and carriers

The expression of MHC isoforms in the skeletal muscles of nine patients with Duchenne muscular dystrophy (DMD) (from 2.5 to 15 yr of age) and three DMD carriers was studied using different specific anti-MHC MAbs. We also analyzed muscle fiber size and fiber reactivity with acridine orange and/or with a surface antigen marker. One-quarter of all fibers of DMD patients, or less with age, were of normal size and contained only adult slow MHC. Half of the muscle fibers contained adult and developmental MHCs. Only half of these fibers were representative of an active regenerative process. MHC co-expression also altered the proportion of normal fast or slow fibers. Adult fast MHCs were expressed as unique MHC only in small and very small fibers in the oldest DMD patients. In DMD carrier muscles, the greatest alterations in MHC expression were observed in patients with the most reduced dystrophin expression. However, MHC changes in dystrophin-positive fibers were similar to those observed in dystrophin-free fibers. In conclusion, disruptions or delays in the switching of all genes coding for adult fast and slow MHC and developmental MHC coincided with dystrophin deletion and with perturbations in its expression.     

Myeloid regeneration after bone-marrow transplantation monitored by serum measurements of myeloperoxidase, lysozyme and lactoferrin

Bone-marrow regeneration after chemo- and radiotherapy-induced aplasia can be monitored by serum levels of myeloperoxidase (MPO), lysozyme (LYS) and lactoferrin (LF). In 10 patients with leukemia, serum measurements were performed before and after bone-marrow transplantation. Bone-marrow regeneration was suggested by increments in serum MPO and LYS 5 and 4 days prior to the increase in mononuclear cells (Mono) and 10 and 9 d before the increase in polymorphonuclear leukocytes (PMN) in the peripheral blood. LF started to rise 4.5 d before detectable circulating PMNs. 2 patients with early relapses of leukemia post transplantation are shown to display atypical patterns of serum MPO and LYS. We conclude that serum measurements of MPO, LYS and LF may be used as early and sensitive means to monitor bone-marrow activity during hematological regeneration. However, the findings also strongly support the earlier proposal that MPO alone may be used to reflect myeloid activity in the bone-marrow in general.     

轴突分叉点位置计算法适用性的分析

某些中枢神经元通过分叉轴突向两个以上核团投射。轴突分叉点的位置通常由计算轴突主干传导时间予以估测。但是,本工作用该法的4个公式计算细胞内或细胞外记录到的数据未能得到一致和可信的数值。结果提示,计算法的原理是不合理的,并至少对在细胞内或细胞外记录条件下估测轴突分叉点是不适用的。     

Chemical traumatization of adult mouse olfactory epithelium in situ stimulates growth and differentiation of olfactory neurons in vitro

This study demonstrates that ZnSO₄ induced chemical trauma results in an in situ regeneration of the olfactory epithelium which, when maintained in vitro, provides an enriched population of olfactory neurons. Therefore, the ability of the olfactory epithelium to respond to chemical trauma with increased mitotic activity can be used to increase growth of neurons in culture. Tissue obtained from normal or vehicle-treated adult mice produced few olfactory neurons, when maintained in culture, compared to cultures established from tissue following an in situ ZnSO₄ trauma. Maximal neuronal yields were obtained in cultures established from tissue that was removed 4–6 days following chemical trauma. The morphological appearance and the presence of cell specific intermediate filament proteins were used to classify the cell types in these olfactory epithelial cultures. Single cells and aggregates of cells which were immunopositive for keratin, but immunonegative for neurofilament protein and GFAP, were identified as epithelioid. Flattened polygonal cells immunopositive for GFAP were identified as glia. A small population of flattened cells was immunonegative for all of the antibodies used in this study. Cells that had processes were immunonegative for GFAP and keratin. Some were immunopositive for 200 kDa and 160 kDa neurofilament proteins but immunonegative for the 68 kDa neurofilament protein. A few of these cells showed positive immunoreactivity with the olfactory marker protein (OMP) antibody and most likely represented the most mature olfactory neurons in the cultures. This trauma-induced culture model using olfactory tissue from adult mice can serve as a source of CNS neurons for comparison with cultured embryonic neurons.     

Research review: DNA polymerases as molecular markers of the regenerating capacity of hepatocytes

Hepatocyte regeneration has been widely investigated, with the mitotic index and the incorporation of [³H]thymidine being used as regeneration markers. We focused on the induction of DNA replication enzymes, particularly DNA polymerases (pol) α, δ, and ε. Using rat models, we have shown that the activity of pol α in crude liver extract well represents the regenerating capacity of hepatocytes. Using pol α as an indicator, we analyzed liver regeneration in rat models under various conditions: obstructive jaundice, external or internal biliary drainage, and the obstruction of portal vein branches. It has been revealed that the ligation of the common bile duct alone induces a certain amount of hepatocyte proliferation. It was striking that external biliary drainage suppressed regeneration capacity in cholestatic rat liver after partial hepatectomy. The strong regeneration in nonligated lobes induced by portal branch ligation was similar to the liver regeneration seen after partial hepatectomy with respect to the induction of DNA polymerases. Taken together, the aspects of DNA replication, particularly the induction of DNA polymerases, may contribute to shedding new light on the regeneration of human liver. This work was supported in part by a Grant-in-Aid for General Scientific Research and for Cancer Research from the Ministry of Education, Science and Culture, Japan, and by grants from the Uehara Memorial Foundation