Sulfur mustard (SM), a bifunctional alkylating agent that causes severe lung damage, is a significant threat to both military and civilian populations. The mechanisms mediating the cytotoxic effects of SM are unknown and were investigated in this study. The purpose of this study was to establish a rat model of SM-induced lung injury to observe the resulting changes in the lungs. Male rats (Sprague Dawley) were anesthetized, intratracheally intubated, and exposed to 2 mg/kg of SM by intratracheal instillation. Animals were euthanized 6, 24, 48, and 72 h post-exposure, and bronchoalveolar lavage fluid (BALF) and lung tissues were collected. Exposure of rats to SM resulted in rapid pulmonary toxicity, including partial bronchiolar epithelium cell shedding, focal ulceration, and an increased amount of inflammatory exudate and number of cells in the alveoli. There was also evidence that the protein content and cell count of BALF peaked at 48 h, and the alveolar septum was widened and filled with lymphocytes. SM exposure also resulted in partial loss of type I alveolar epithelial cell membranes, fuzzy mitochondrial cristae, detachment and dissociation of ribosomes attached to the surface of rough endoplasmic reticulum, cracked, missing, and disorganized microvilli of type II alveolar epithelial cells, and increased apoptotic cells in the alveolar septum. The propylene glycol control group, however, was the same as the normal group. These data demonstrate that the mechanism of a high concentration of SM (2 mg/kg) induced acute lung injury include histologic changes, inflammatory reactions, apoptosis, oxidative stress, and nuclear DNA damage; the degree of injury is time dependent.

目的:观察门静脉高压巨脾大部切除后残脾红髓笔毛微动脉(PA)超微结构的变化,探讨高血流动力对脾小血管内皮细胞超微结构的影响.方法:选取门静脉高压脾肿大患者13例,术后切取脾组织为巨脾组,术后8年穿刺获取脾组织为残脾组,另设外伤性脾组织13例为正常组.采用透射电镜观察脾脏红髓PA内皮细胞的超微结构改变.结果:巨脾组PA内皮细胞细胞核结构完整,染色质分布正常,一部分线粒体肿胀,变大变圆,基质变浅,线粒体嵴变短变少,甚至消失;一部分线粒体扁平嵴变成烧瓶状,甚至空泡状,基质致密,出现髓样变.残脾组PA内皮细胞核结构完整,染色质分布正常,线粒体基质均匀,嵴断裂减少,未见空泡样变和髓样变.正常组PA内皮细胞细胞核结构完整,染色质分布正常,线粒体嵴片状排列,外膜连续,基质均匀,连续内膜中可见包裹.结论:巨脾大部切除术可减少残脾血管PA增生,逆转PA内皮细胞线粒体的变化.

Objective To establish an animal model for rat respiratory tract injury due to sulfur mustard (SM),and observe the morphologic changes of tracheal tissues and epithelial cells in SM-induced injury.Methods Male SD rats (n =72)were randomly divided into three groups (SM group,propylene glycol group,and normal group).The rats in the SM (2 mg/kg)group were injected intratracheally with diluted SM (0.1 ml).Histomorphologic and cytomorphogic changes of the specimen were observed under light and electron microscopy.Results In the SM group,histomorphologic changes included shed tracheal epithelial cells,focal ulcer formation,deranged cilia,increased glands in the lamia propria mucosae,and inflammatory cells invading the submucosa.The cytomorphologic changes were as follows:the cellular membranes lacking,medullary changes in the mitochondria with difficult to discern mitochondrial cristae appeared in goblet cells,and difficult to discern karyotheca,karyopyknosis,and margination of the nuclear chromatin in cilia,basal cells,fibroblasts.The structure of tracheal tissue and epithelial cells of the propylene glycol group was the same as the control group.Conclusions SM (2 mg/kg)can cause acute injury of tissues and multi-epithelial cells of the rat trachea.The degree of injury is positively correlated to the duration of time.SM mainly affects the cellular membranes and organelles of goblet cells,as well as nuclear chromatin of cilia,basal cells,and fibroblasts.

Sulfur mustard (SM) is believed to be a major threat to civilian populations because of the persistent asymmetric threat by nonstate actors, such as terrorist groups, the ease of synthesis and handling, and the risk of theft from stockpiles. The purpose of this study was to establish mechanisms of acute tracheal injury in rats induced by SM using histopathologic, immunohistochemical, and biochemical parameters. Male rats (Sprague-Dawley) were anesthetized, intratracheally intubated, and exposed to 2 mg/kg of SM. Animals were euthanized 6-, 24-, 48-, and 72-hour postexposure, and intracavitary blood samples from the heart and tracheal tissues were collected. Exposure of rats to SM resulted in rapid tracheal injury, including tracheal epithelial cell shedding, focal ulceration, and abundant lymphocyte invasion of the submucosa. There was also evidence of a large number of apoptotic cells in the epithelium and submucosa, the serum levels of tumor necrosis factor α, interleukin 1β (IL) 1β, IL-6, and γ-glutamyl transferase peaked at 24 hours, and the serum levels of lactate dehydrogenase, glutathione peroxidase, and thiobarbituric acid reactive substance peaked at 6 hours. The SM exposure also resulted in a loss of the cellular membrane, leakage of cytoplasm, fuzzy mitochondrial cristae, medullary changes in ciliated and goblet cells, and the nuclear chromatin appeared marginated in basal cells and fibroblasts. The results in the propylene glycol group were the same as the control group. These data demonstrated the histologic changes, inflammatory reactions, apoptosis, oxidative stress, and DNA damage following SM (2 mg/kg)-induced acute tracheal injury; the severity of changes was time dependent.

Objective:To explore the relationship between endogenous hydrogen sulfide(H2S)of the blood plasma and the contraction of myometrium.Methods:The sixty cord plasma samples were collected and divided into two groups according to the delivery has not launched the contractions.The concentration of H2S was assayed by sensitive sulphur electrode.The expressions of cystathionine beta-synthase(CBS) and cystathionine gamma-lyase(CSE) in placenta were measured by using immunohistochemistry.Results:H2S contents in the group which mothers did not start uterine contraction were higher than that mother started uterine contraction(P=0.01);The expressions of CSE were detected in cytoplasm of placental villi trophoblast cells and at higher levels in the group with mothers did not start uterine contraction.The expressions of CBS had not seen in both two groups.Conclusion:Cytoplasm of placental villi tropho-blast cells are included CSE and no CBS.The production of H2S in cord plasma is regulated by trophoblast cells.H2S content in cord placenta is related to initiation of parturition.

Objective: To investigate the ultrastructural changes of long-time existing the residual spleen after subtotal splenectomy on endothelial cells,lymphocyte,Macrophage,and reticular cells,in order to evaluate immune cells remodeling of the residual spleen.Methods: 8 cases splenomegoly of portal hypertension were Collected.Specimens were obtained by operating and puncturing biopsy,and the splenomegoly groups and residual spleen groups were divided.The other control group was 8 cases of trauma-induced splenic rupture.The thinest slices was made.The ultrastructural change of cells were observed under electrical microscope.Results: 1)The endothelial cells were the form of "the long rod",the basement membrane and the fenestra were clear,the mitochondrial structure roughly normal.2)The ultrastructure of small and media lymphocytes was roughly normal.3)The partal macrophage apoptosis which the nuclear membrane have disappeared and the chromatin of cell nucleus have spilled over were appeared.4)The tonofilament was clear,and the structure of organelles such as mitochondria normal in the reticular cell.Conclusion: Under the residual spleen was departed hypertensive state,the ultrastructure of cells can be occurred adaptive change,it conduce to educe the function of immune cells of the residual spleen.

To observe the ultrapathological changes of mice kidney after toxoflavin(TXF) intoxication.TXF(1.5mg/kg) was injected into the peritoneal cavity of mice.The cortex of kidney was taken after 2 h of intoxication for ultrathin section and the ultrastructural changes were observed by using transmission electron microscopy.TXF can injure the epithelial cells of proximal tubules as well as the endothelium,podocyte and intraglomerular mesangial cells of glomerulus of kidney of experimental mice.The mitochondria of the epithelial cells of proximal tubules and the basement membrane of endothelial cells of glomerulus are the main injured organelles.The toxic effect of TXF on experimental mice is probably to injure the inner membrane system of mitochondria and capillary basement membrane.The results provided ultramorphological evidence on mechanism of TXF intoxication.

目的研究灭活A组链球菌诱发红细胞破裂的机制,为A组链球菌导致的溶血性疾病治疗提供新思路。方法将杂种家兔随机分为对照组和实验组,每组2只并分别称重,实验组从耳缘静脉注射2.5mL灭活的A组链球菌全菌体,对照组从耳缘静脉注射2.5mL生理盐水。分别于注射后第6h、48h、96h、7d、14d时从耳缘静脉取血2mL,常规分离红细胞并提取膜蛋白,十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)分析各组红细胞膜蛋白的百分含量。每次取材均做涂片,瑞氏染色,光镜下观察红细胞形态和计数异常率。结果实验组家兔的体重在相同时间点较对照组明显降低。注射A组链球菌第96h、7d、和第14d,实验组家兔光镜下均见红细胞形态异常,大小不等,部分红细胞中央淡染区消失,部分红细胞中央淡染区扩大。计数红细胞畸形,实验组高于对照组(P<0.05),差异有显著性。SDS-PAGE电泳分析各组红细胞膜蛋白的百分含量,发现有多条带实验组红细胞膜蛋白含量显著低于生理盐水对照组。结论灭活A组链球菌从耳缘静脉注射第96h始能引起红细胞形态异常,其机制可能是红细胞膜蛋白的含量变化所致。

目的 研究脐血浆中硫化氢(H2S)和羊水污染的相关性.方法 收集孕产妇脐血浆60份,根据分娩时羊水是否污染分为2组.用敏感硫电极检测脐血浆中硫离子的浓度,并换算出H2S的含量;免疫组织化学方法检测胎盘中胱硫醚-γ-裂解酶(CSE)和胱硫醚-β-合成酶(CBS)的表达程度.结果 羊水污染组中H2S含量高于羊水清组,差异有显著性.免疫组化显示:CSE表达于胎盘绒毛的细胞滋养层胞浆中,羊水污染组CSE呈现高表达.CBS在任何组中均未见有表达.结论 胎盘绒毛的细胞滋养层胞浆中含有CSE,不舍CBS;脐血浆中H2S由细胞滋养层细胞参与调控生成;孕产妇脐血浆中H2S与羊水污染有密切关系.

超薄切片质量是电镜技术的基础,理想的超薄切片应该是厚度适中,厚薄均匀,结构清晰,没有颤痕、皱褶、重叠、刀痕、空洞及污染.优质超薄切片的获得,与样品的取材、脱水、浸透、包埋和切片的好坏诸因素息息相关.笔者根据多年的工作经验,就样品制备的几个关键环节在实践中不断研究和探讨,摸索出一些技巧,供同行们参考.

目的 观察钙离子螯合剂对白血病患者骨髓单个核细胞(BMMNC)生成硫化氢(H2S)的影响.方法 采用BAPTA-AM和EGTA螯合细胞内外的钙离子,用敏感硫电极检测细胞外硫离子,计算出H2S含量的变化;免疫组化方法和图像分析系统检测bcl-2,bax的表达变化.结果 BAPTA-AM和EGTA各实验组与对照组比较,H2S生成量均明显降低(P<0.05).免疫组化结果显示,bcl-2和bax均表达于细胞浆和胞膜上.图像分析光密度值(OD值),BAPTA-AM和EGTA各实验组bcl-2蛋白表达与对照组比较,12h,24h和48h均高于对照组(P<0.05).bax蛋白的表达在BAPTA-AM和EGTA各实验组与对照组相比,12h,24h和48h均低于对照组(P<0.05).结论 细胞内外钙离子螯合剂均能减少白血病患者BMMNC生成H2S,H2S生成降低在较短时间内可能抑制细胞凋亡,促进细胞增殖.

传统的组织胚胎学实验课以观察玻璃切片为主,玻璃切片在实验教学使用中存在易褪色、不易永久保存和使用时受时间和空间限制等缺陷,利用全自动显微镜扫描系统将玻璃切片制作为数字切片,结合显微数码互动系统和校园网络系统应用于组织胚胎学实验教学,不仅能有效解决优质典型玻璃切片片源不足带来的教学问题,也有利于教师的备课讲解、学生的复习与讨论以及师生间的互动,推动了组织胚胎学实验教学和考试模式的改革.该文主要介绍组织胚胎学数字切片的特点、应用及发展前景.

Hepatic cirrhosis is the end-stage of chronic liver diseases. The majority of patients with hepatic cirrhosis die from life-threatening complications at early age. Liver transplantation has been the most effective treatment for patients with hepatic cirrhosis. Since liver transplantation is critically limited by the shortage of available donor livers, searching for an effective alternative therapy has attracted great interest in preclinical studies. The encouraging advances in stem cell research have paved the way towards the treatment of the end-stage of chronic liver diseases. In view of the pathogenic fundamentals of hepatic cirrhosis, cell-based treatment should be aimed to complement or replace damaged liver cells and to correct the imbalanced extracellular matrix regeneration/degradation. Understanding the transition of hepatocyte regeneration to hepatic fibrogenesis during chronic liver injury could guide the appropriate utilization of cell-based therapy. This chapter is intended to describe the characteristics and therapeutic potential of various stem cells, including hepatocytes, liver progenitor cells, hematopoietic stem cells, mesenchymal stem cells, embryonic stem cells and induced pluripotent stem cells. Since autologous adult stem cells have the least obstacles for clinical application, their potential interventions on cirrhosis are especially illustrated in terms of the cellular and molecular mechanisms of hepatic fibrogenesis.

硫化氢(H2S)作为第三种气体信号分子在生命活动中具有特殊的作用。在心血管系统中H2S主要由胱硫醚-γ-裂解酶(CSE)催化生成,CSE在心肌和主动脉上高表达。H2S可直接作用于KATP通道实现对血管的调节作用。现已证明H2S与多种心血管疾病关系密切。

组织胚胎学是主要的形态学课程之一,是从微观上来研究人体结构及相关功能的一门基础学科,实验教学占该课程总学时50%左右,是教学的重要环节之一,在人才培养中有着重要的地位和作用.提高组织胚胎学实验课教学效果,有助于学生更好地掌握相关基础知识,为今后学习其他医学课程奠定扎实的基础.就如何搞好组织胚胎学实验教学的探讨,旨在总结经验,更有效地提高组织胚胎学实验课教学质量.

Objective To explore the effect of chloratum kalium( KCl) on the production of H_2S and ultra- microstructure of pulmonary artery smooth muscle cells( PASMCs). Methods Rat's cultured PASMCs were randomly di- vided into control group and KCl groups at different time poits. Each group contained 4 samples ( V =5ml,D = 1～2× 10~5/L). The KCl was added into PASMCs at the concentration of 50mol/L. 5ml of each sample was sucked at the 5th, 15th and 30th minute. The concentration of sulfion was measured by sensitive sulfion electrode and the cultured PASMCs were observed by electron microscope. Results Compared with the control group, the concentration of sulfion was re- spectively increased by 162. 72% ( P 0. 05 ) ,253. 68% ( P 0. 01 ) ,63. 23% ( P 0. 05 ) for PASMCs in each KCl group;the number of rough endoplasmic reticulum( RER) in KCl group is much more than that in control group. Conclu- sion KCl can sublimate the concentration of sulfion in rat's PASMCs.

Objective To study the survival and infiltration condition of cord blood derived-T lymphocytes in mice and its therapeutic action for erythroleukemic(EL) mice;and to provide the experimental and theoretical basis for studying the biological characteristics and anti-leukemia effects of cord blood-derived T cells.Methods Fourty-eight BALB/c mice were randomly divided into three groups,the control group(CG),the experimental group 1(EG 1)and the experimental group 2(EG 2).Cord blood derived-T lymphocytes were separated and prepared,and transplanted it into EL mice via the tail vein or peritoneal injection after being signed to their group.This was done in order to observe the survival time and infiltrate condition of the cord blood-derived T cells in mice and its effect on the morphosis of EL mice spleen by immunofluorescence(IMF),immunohistochemistry(IMC) and transmission electron microscope(TEM).Results Most activated cells were CD3~+ T lymphocytes by PHA,and the positive rate was(83.42±1.26)%;human CD3~+ cells and Brdu~+ cells were detected in peripheral blood,spleens and liver of all mice at the 3rd and 7th day after cord blood derived-T lymphocytes were transplanting;CD25~+ cells could be detected in the liver and spleen of EL mice in both experimental groups;and the average survival time of EL mice in the two experimental groups was(27.25±7.06) days and(24.74±2.93) days respectively(this was statistical significance compared to control group,P0.05).The results of Hoechst 33258 staining showed: 1.Nuclear fluorescence was uniform in the control group and there were few apoptotic cells;2.In the experimental group,some of the leukemia cells appeared to have morphological changes consistent with apoptosis; chromatin had a high degree of cohesion;dense concentration,or splited into massives,edge was smooth and clear;Under TEM,leukemic cells infiltrate was generally observed in the spleens of the control group mice.Parts of the leukemic cells showed typical apoptosis morphological changes in the experimental groups: nuclear chromatin concentration and edge accumulation was seen;nuclei fragmentation and formation of apoptotic bodies;the expansion of RER,and the vacuolar or medullar degeneration of mitochondria could be found.Conclusion The cord blood-derived T lymphocytes could be activated in vivo,and transferred to the site of leukemia,and educed relevant function,thus having some therapeutic action for leukemia.

Objective: To explore the effects of mixed cord blood plasma(CBP) on phytohemagglutin(PHA) induction of cord blood-derived T cells.Methods: Cord blood plasma and umbilical cord blood mononuclear cells(CBMCs) were collected;CBMCs were divided two groups: CBP+PHA group and PHA group,and the proliferation index and immune phenotypes of each group were detected.Results: The proliferation index of CBP+PHA group was lower than that in the PHA group without significant difference(P>0.05);the immune phenotypes of two groups were both mainly CD3+ and CD4+ without significant difference(P>0.05).Conclusion: The effects of CBP on proliferation and differentiation of cord blood-derived T cells were similar to those of 20% RPMI1640.