大鼠心肌缺血再灌流损伤的免疫组化研究

用16只 SD 大白鼠建立缺血再灌流心肌损伤模型,其中8只预先静脉注射吗啡以抗心律失常。此外,再用8只大鼠开胸作正常对照。其心脏标本进行 HE 染色及抗肌动蛋白单克隆抗体(HHF_(35))免疫组化观察:对照组心肌呈均匀一致的棕褐色染色;注射吗啡后再灌流组心肌呈小灶性分布的 HHF_(35)染色缺染区;未注射吗啡组心肌缺染呈广泛大面积分布。结果表明,HHF_(35)免疫组化法对显示实验性心肌缺血再灌流损伤有重要价值,且心肌缺血再灌流损伤程度与心律失常有关。     

Lawyers,CABx and the Community Legal Service: A new dawn for social welfare law provision?

This article explores the prognosis for social welfare law provision in the light of the government's plans for the Community Legal Service. It considers whether the involvement of both the advice sector and the traditional legal profession can provide the basis for stable and comprehensive provision of social welfare law. I conducted in-depth interviews with CAB workers and local solicitors in the large conurbation served by two CABx who participated in the non-solicitor franchise pilot in 1997. The research focused on the relationships between the legal service providers and the impact of the franchise operation on the CAB. Although harmonious relationships were found to be largely intact at a local level, the franchise raised considerable tension within the CABx that must serve as a cautionary note to any optimism about 'establishing the Community Legal Service'.     

Immunocytochemical diagnosis of early myocardial ischaemic/hypoxic damage

The sensitive and reliable dinitrophenyl (DNP) hapten sandwich staining (DHSS) procedure (B. Jasani et al., Virchows Arch (Pathol. Anat.), 406 (1985) 441-448) was used to study the distribution of immunoperoxidase staining seen with antibodies to seven protein markers in post-mortem heart tissue. This was obtained from 12 cases with macroscopic myocardial infarction and 17 cases without myocardial infarction (10 with and 7 without significant coronary artery atherosclerosis). The immunostaining patterns were compared with the appearances seen in adjacent sections stained by the routine haematoxylin and eosin (H & E) and phosphotungstic acid haematoxylin (PTAH) methods and a method previously recommended for the detection of early myocardial infarction, the haematoxylin basic fuchsin picric acid (HBFP) stain. Loss of immunostaining with an antibody to myoglobin was found to be a reliable and more objective marker of both early and established myocardial infarction compared with the histological stains. Antibodies to myosin, caeruloplasmin, C-reactive protein and pre-albumin gave similar but less reliable results, whilst those to complement factor C3b and alpha-1 anti-trypsin gave the least reliable results for early myocardial ischaemic/hypoxic damage. The immunocytochemical results are considered sufficiently encouraging to extend the work to a large number of sudden death cases in order to establish a new, more reliable approach to the detection of histologically latent ischaemic/hypoxic damage in the myocardium.     

Expression of TGF-beta 1 and proliferation of collagen in myocardium in viral myocarditis

To investigate the tissue repair of acute viral myocarditis, and to explore the diagnostic method of slight viral myocarditis in forensic pathology. Slight viral myocarditis model was induced in Balb/c murine by CVB3. Collagen proliferation in myocardium of mice with myocarditis was observed by special staining. The hearts of mice and human(9 cases) with myocarditis were studied LSAB-immunohistochemically with anti-TGF-beta 1 antibody. In the study, the proliferation of collagen was seen in myocardium in acute viral myocarditis. Generous expression of TGF-beta 1 was found in the myocardium of mice and human with myocarditis. The quantity of collagen proliferation and expression of TGF-beta 1 was positive correlation. It is concluded that the tissue repair exists in acute viral myocarditis and that positive staining of myocardium for TGF-beta 1 is a sensitive index of myocardial damage and tissue repair.     

Further evaluation of immunocytochemical staining in the diagnosis of early myocardial ischaemic/hypoxic damage

The pattern of immunocytochemical staining with antibodies to caeruloplasmin, myosin, myoglobin and C-reactive protein seen in myocardium taken from deaths with macroscopic evidence of myocardial infarction and/or significant coronary artery atherosclerosis and from deaths with neither of these lesions has been correlated with H&E, PTAH and HBFP staining of myocardium and circumstances of each death indicative of antemortem hypoxia and/or ischaemia. Loss of staining with these antibodies correlated well with fuchsinorrhagia and both techniques are more sensitive than H&E and PTAH staining in the detection of early ischaemic/hypoxic damage to myocardium. However, their sensitivity is such that they appear to detect agonal changes and, therefore, cannot be used for specific diagnosis of early myocardial infarction.     

高氧环境下大鼠肺白细胞毒素的免疫组化研究

目的研究白细胞毒素在高氧环境下大鼠肺的组织学分布及与肺部疾病之间的关系。方法将大鼠分组后分别置于高氧环境和正常空气中24h、48h、72h和96h,制作石蜡标本切片,用特异的抗白细胞毒素抗体进行免疫组化染色。结果 置于高氧环境下大鼠肺中的中性粒细胞对白细胞毒素呈阳性反应,且随置入高氧环境时间的延长而明显增强。此外,巨噬细胞也呈现阳性反应。结论 用免疫组化方法证明了中性粒细胞产生白细胞毒素,提示白细胞毒素是导致高氧环境下肺损伤的重要化学介质之一。     

A New Method for the Characterization of the Degree of Fire Damage to Gypsum Wallboard for Use in Fire Investigations

A new method to characterize the degree of fire damage to gypsum wallboard is introduced, implemented, and tested to determine the efficacy of its application among novices. The method was evaluated by comparing degree of fire damage assessments of novices with and without the method. Thirty-nine “novice” raters assessed damage to a gypsum wallboard surface, completing 66 ratings, first without the method, and then again using the method. The inter-rater reliability was evaluated for ratings of damage without and with the method. For novice fire investigators rating degree of damage without the aid of the method, ICC(1,2) = 0.277 with 95% CI (0.211, 0.365), and with the method, ICC(2,1) = 0.593 with 95% CI (0.509, 0.684). Results indicate that the raters were more reliable in their analysis of the degree of fire damage when using the method, which support the use of standardized processes to decrease the variability in data collection and interpretation.     

Isolated viable rat cardiac myocytes as a tool for evaluating histochemical methods for diagnosis of myocardial ischemia. I. Mitochondria-bound oxidoreductases — A preliminary report

A pilot study has been performed to evaluate the usefulness of isolated viable rat cardiac myocytes in the evaluation of histochemical oxidoreductase methods for estimating cellular anoxic damage. Data are presented in support of the use of succinate dehydrogenase and β-hydroxybutyrate dehydrogenase for evaluating different degrees of cellular damage. It appears that upon anoxia biochemical signs of membrane leakage do occur earlier than positive histochemical reactions. However, trypan blue staining occurs at a later time point. Probably the histochemical reaction-rates are limited by the penetration of both the substrate and the electron acceptor, in this case nitro blue tetrazolium, into the cells and the mitochondria.     

实验性脑挫伤后 Fos蛋白和 NGFR变化与损伤时间关系的研究

目的 检测分析不同损伤时间组实验大鼠脑挫伤灶周围神经细胞中 NGFR阳性染色强度变化与损伤后经过时间之间的关系,对其在脑挫伤时间推断中的作用作出评价。方法 运用免疫组织化学技术（ SABC法）,结合计算机彩色图像分析技术观察大鼠实验性脑挫伤后原癌基因 c－ fos的表达产物 Fos蛋白和神经生长因子受体（ NGFR）的染色变化。结果 （ 1）伤后 0.5h组在挫伤灶周围可见 Fos阳性染色细胞出现。伤后 3h组阳性表达数量和强度达峰值,且分布广泛。后表达逐渐降低。 1d后各组未见 Fos阳性染色强度增加的细胞。对照组染色为阴性。（ 2）伤后 1d组在挫伤灶周围可见少量 NGFR阳性染色细胞,以神经细胞表达为主。伤后 3d组,阳性细胞表达与 1d组相比有显著性增强 (P< 0.01), 数 量 和 着 色 强 度 达 到 高 峰 , 量 多 且 染 色 深 。 之 后 逐 渐 缓 慢 减 弱 。 对 照 组 染 色 为 阴 性 。 可 见 脑 挫 伤 后 Fos和 NGFR在 损 伤 局 部 的 阳 性 染 色 分 别 在 伤 后 早 期 及 随 后 较 晚 时 段 里 出 现 。 前 者 表 达 时 间 短 暂 , 而 后 者 则 持 续 时 间 较 长 , 但 都 有 一 定 的 规 律 性 。 结 论 Fos蛋 白 和 NGFR变 化 可 用 于 脑 挫 伤 后 不 同 时 段 的 损 伤 时 间 推 断 。     

大鼠心肌早期缺血内皮素的表达

为探讨心肌缺血后内皮素在心肌组织内表达的变化。用免疫组化SABC法研究内皮素的表达情况。结果发现缺血60min后 ,缺血区心肌细胞呈局灶性阳性 ;缺血120min后 ,血管平滑肌、内皮细胞及心肌细胞浆均呈阳性着色。对照和缺血30min组心肌组织未见有阳性反应。上述各组HE染色未见明显病理改变。     

广泛软组织损伤肾肌红蛋白免疫组化研究

目的探讨肌红蛋白对挤压综合征肾功能损害的病理形态学改变及法医病理学诊断价值。方法应用EnVision(两步法)免疫组化方法和LSAB免疫组化方法,对22例生前有较广泛肌肉软组织损伤的人尸体肾脏组织(损伤组)进行肌红蛋白检测,12例无上述病史的人尸体肾脏组织作对照(对照组)。结果损伤组有8例肾远曲小管及集合管管腔管型染色呈阳性或强阳性,均见于伤后36h至10d死亡者;有7例近曲小管、远曲小管和集合管上皮腔缘附近的胞浆及腔面肌红蛋白阳性或强阳性,最早见于伤后8h死亡者,伤后24h至48h死亡者最为显著;余9例未见上述改变。对照组12例上述部位均为阴性。结论肌红蛋白管型堵塞肾小管和肌红蛋白对肾小管上皮的直接损害是挤压综合征肾功能衰竭的重要病理形态学基础,两者对挤压综合征的法医病理诊断均具有重要价值。     

显示和判定心肌早期病变的染色方法研究

作者用探讨的变色酸2R亮绿法和其他10余种染色方法,对心血管病猝死的尸检心肌和经心电、Mb、CPK、LDH、SOD诊断有急性心肌损伤的多种大量动物实验材料观察结果,证明心肌早期病变有染色性质异常,对显示和判定心肌早期病变有实用价值。提出并讨论了心肌早期病变及其显示、判定方法。     

大鼠急性心肌缺血后心肌NF-kB的检测及其法医学意义

目的检测大鼠急性心肌缺血后核因子-kB(NF-kB)在心肌组织内的变化,探讨其在心肌早期缺血死后诊断中的法医学意义。方法30只大鼠随机分为正常对照,缺血0.5h,1h,2h,4h,8h共6组,每组5只建立大鼠急性心肌缺血模型,用免疫组化染色技术(S-P法),观察不同时间缺血心肌中NF-kB阳性着色,并应用图像分析系统进行定量分析。结果大鼠心肌缺血30m in,心肌细胞核出现散在NF-kB阳性染色;缺血60m in,心肌阳性染色细胞核增多;4h达高峰;8h组开始下降;免疫组化染色定量检则的阳性单位:缺血2h、4h、8h分别为20.042±1.084、22.028±3.452和20.524±1.595;正常对照组未见核阳性反应。结论大鼠心肌缺血8h内,心肌细胞核NF-kB阳性染色呈现一定规律性变化,对早期急性心肌缺血的死后诊断具有法医学意义。     

Vapor-phase staining of cyanoacrylate-fumed latent fingerprints using p-dimethylaminobenzaldehyde

Contrasting or enhancing of cyanoacrylate ester-fumed latent fingerprints deposited on solvent-sensitive materials such as oil marker writings and rough surface materials such as unglazed earthenware is not easy by conventional dye solutions dipping or dye powder dusting. In this study, a new vapor-phase staining method using p-dimethylaminobenzaldehyde (DMAB) is proposed for staining such materials. DMAB has high volatility and selective absorbability to cyanoacrylate-fumed fingerprints, so that cyanoacrylate-treated samples can be easily stained by leaving them simply in a closed container along with DMAB crystals for 48-96 h at room temperature or in conjunction with the use of mild heating. The stained fingerprint could be excited by UV irradiation (365 nm), and the fluorescent fingerprint was photographed through a UV cut-off filter (420 nm). The new method achieved minimally destructive fluorescent staining for the solvent-sensitive samples and the rough surfaced samples.     

大鼠急性心肌缺血再灌流损伤c-fos mRNA原位杂交实验研究

目的 探讨急性心肌缺血和/或再灌流早期不同时间不同区域心肌细胞内原癌基因c-fos mRNA表达变化,为心肌早期缺血死后诊断提供依据。方法 复制心肌早期缺血模型,大鼠分为正常、缺血及缺血再灌流组。采用原位杂交方法结合图像分析与统计学处理方法研究心肌细胞内c-fos mRNA表达情况。结果 缺血10min再灌流30min,部分大鼠再灌流区心肌细胞胞浆内c-fos mRNA弱阳性反应,散在分布。缺血60min再灌流30min时达高峰,120min开始减弱。正常和单纯缺血组中缺血少于60min组未见有阳性表达。HE染色无明显病理改变。结论 原位杂交法检测c-fos mRNA方法灵敏特异,有望成为一项急性心肌缺血/再灌流早期损伤的诊断指标。     

Immunostaining by complement C9: a tool for early diagnosis of myocardial infarction and application in forensic medicine

Before the first 12 hours, diagnosis of early myocardial infarctions is always difficult for forensic pathologists. We tested complement C9 expression in 121 formalin-fixed and paraffin-embedded heart samples by an immunohistochemical procedure. The heart specimens were separated into four groups: 33 cases in group 1 with typical ischemic damages histologically located, 20 cases in group 2 with death related to myocardial infarction on the basis of ischemic presentation on electrocardiogram but no obvious histological ischemic damage, 35 cases in group 3 with severe coronary disease without cause of death found at the autopsy, and 33 cases in group 4 without sign of myocardial infarction and without coronary disease. In the first group, all 33 heart samples showed a well-defined C9 expression in the necrotic areas. The second group in 17 of 20 cases showed positive areas for C9 expression. In the other three heart specimens, only few stained cells were observed whereas the painful symptoms had begun less than 1 h before death. The third group showed C9 immunopositive areas in six of 35 cases, few stained cells in 8 cases, and no C9 deposition in the 21 other cases. The last group showed no staining area. To avoid nonspecific C9 staining due to tissue autolysis, we studied C9 expression during a controlled putrefactive process in four cases included in group 1; staining was found only in infarcted myocardial areas, and was observed up to ten days. Specificity of C9 expression was evaluated to be 100% [89.4 to 100%] and sensitivity to be 85% [62.11 to 96.79%]. In conclusion, evaluation of immunohistochemical expression of C9 appears to be a highly sensitive and specific marker of early myocardial infarction, useful in forensic medicine if survival is more than 1 h after the beginning of myocyte damage.     

苏木素碱性品红苦味酸（HBFP）染色法对诊断心肌早期缺血的评价

法医病理学检验中,常需一种诊断心肌早期缺血性损害的准确方法,以便解决疑为心肌早期缺血,迅速死亡的案例.这些案例往往因为HE染色病变不典型,难以作出急性心肌缺血的诊断.为此,我们采用Lie等介绍的苏木素碱性苦味酸(HBFP)染色法,对26例法医尸体解剖的心肌组织切片进行研究,并同HE染色进行了对比观察.期望在诊断早期心肌缺血方面作出一恰当的评价.     

Crossbow suicide: mechanisms of injury and neuropathologic findings

Crossbow injuries are rarely reported events in modern times. Two cases of death due to self-inflicted crossbow injuries to the head are reported in 2 men aged 18 and 27 years, respectively. Despite relatively low velocity and concussive force, the sharpness and propulsion force of crossbow bolts may be sufficient to enable penetration of the skull at short range. Due to the relatively low concussive force of the crossbow bolt, however, death may not be instantaneous but may occur from intraparenchymal cerebral damage sometime thereafter. Detailed neuropathologic evaluation of such cases may therefore demonstrate "red cell" hypoxic injury, as well as axonal injury, not limited to the region of the missile tract, but widely distributed, even to the point of extensive brain stem involvement. These changes may result from primary mechanical deformation at the time of injury, from secondary hypoxic damage, or from a combination of both factors. Immunohistochemical staining of brains for amyloid precursor protein to delineate more clearly the pattern of axonal damage may assist in determining the extent of injury in such cases.     

大鼠急性心肌缺血zif/268蛋白表达及其意义

目的 探讨急性心肌缺血早期不同时间不同区域心肌细胞内原癌基因蛋白zif/268的表达变化,为心肌早期缺血死后诊断提供新指标。方法 建立心肌早期缺血模型,大鼠分为正常、缺血组。采用免疫组化SABC法研究心肌细胞核蛋白zif/268的累积情况。结果 缺血 60min后,缺血区大鼠心肌细胞有部分心肌细胞核呈弱阳性着色,以后随缺血时间延长核阳性增强。正常和缺血30min组及未缺血区心肌细胞核未见有阳性反应。HE染色无明显病理改变。结论 免疫组化染色法检测心肌细胞核zif/268的表达对急性心肌缺血的死后诊断是一种有价值有意义的手段。     

A silver staining method for the detection of polymorphic proteins in minute bloodstains after isoelectric focusing

A silver staining method has been developed to study polymorphic proteins in bloodstains after isoelectric focusing. This method is highly sensitive and permits the detection of polymorphic proteins (i.e. alpha 1-antitrypsin, Gc and Tf C subtypes) in bloodstains as small as 0.2 microliter or less. The method is simple and reproducible and can be used after immunofixation. Blood stains can be identified after longer storage periods than is possible by using conventional staining methods.