不同实验方法在人胎脑海马神经干细胞检测中的应用

目的探讨精确可靠的检测人胎脑神经干细胞(NSCs)的方法。方法收集胎龄32周的水囊引产胎儿10例,灌注固定后分别予石蜡包埋和冷冻切片,采用HE检测Nestin蛋白表达免疫组织化学染色和光镜技术对人胎脑海马部位两种切片组织结构的改变和NSCs进行检测。结果石蜡切片海马组织Nestin蛋白表达阳性NSCs数较少,每个NSCs中Nestin蛋白表达的量也较少:而冷冻切片Nestin蛋白表达阳性NSCs数较多,且每个NSCs中Nestin蛋白表达的量也较多,石蜡切片中组织及细胞结构完整性优于冷冻切片。结论水囊引产、灌注固定及冷冻切片三结合研究人胎脑组织NSCs,是一种可靠、精确而敏感的实验方法。     

肾组织石蜡切片Ⅳ型胶原α链免疫组织化学染色法诊断Alport综合征

目的 探讨肾组织石蜡切片Ⅳ型胶原α链免疫组织化学染色方法 在Alport综合征(AS)患者诊断中的应用,从而为AS的诊断提供新的技术手段.方法 将对照组及已确诊的X连锁型、常染色体隐性遗传型AS及血尿息儿各2例肾组织分别进行石蜡切片Ⅳ型胶原α3和α5链免疫组织化学染色和冰冻切片的间接免疫荧光染色,比较分析染色结果 .具体方法 根据PV-9000试剂盒提供的实验方法 ,进行染色.采用高压锅加热、胃蛋白酶消化和蛋白酶消化3种抗原修复方法 分别对正常肾组织切片进行抗原修复,并比较其修复效果.免疫荧光染色采用间接免疫荧光染色法.结果 肾小球基底膜Ⅳ型胶原α3、α5链免疫组织化学染色在对照和血尿组为连续的线状阳性;在x连锁显性遗传型女性患儿为间断线状阳性,在男性患儿为阴性;在常染色体隐性遗传患者均为阴性,在肾小囊及部分肾小管基底膜上Ⅳ型胶原α5链为阳性.结果 均与免疫荧光染色法结果 一致.结论 肾脏组织石蜡切片Ⅳ型胶原α链免疫组织化学染色法可用于诊断As,为诊断As提供了新的手段.     

肠三叶因子对小鼠炎症性肠病肠道病变的保护作用

目的 探讨肠三叶因子(ITF)对小鼠炎症性肠病(IBD)肠道病变的影响,分析ITF对小鼠IBD肠道先天性免疫功能的调节作用.方法 48只小鼠随机分为3组,每组16只.三硝基苯磺酸(TNBS)组:用TNBS建立IBD模型后每只小鼠给予9g.L-1盐水0.1 mL腹腔注射;基因重组肠三叶因子(rITF)组:建立IBD模型后每只小鼠给予rITF0.1 mL腹腔注射;乙醇对照组:未造模型小鼠每只小鼠给予9g.L-1盐水0.1 mL腹腔注射.各组均连续5d注射,第14天评估小鼠临床表现,麻醉处死小鼠后取其结肠组织作组织学评分,HE染色行肠组织病理学检查,并采用免疫组织化学法及实时定量PCR法检测其肠道TNF-α及Toll样受体(TLR4)和核因子( NF-κBp65)基因、蛋白表达水平.结果 与TNBS组小鼠比较,r-ITF组小鼠死亡只数、大便性状、血便情况、体质量下降情况等临床表现及肠道局部炎症均明显减轻,肠道病理大体观及组织学评分均明显下降(Pa＜0.05),TNF-α蛋白表达水平明显下降(Pa＜0.05),TLR4、NF-κBp65基因及蛋白表达明显下调(Pa＜0.01).乙醇对照组无死亡,临床症状及肠道局部炎症轻微,TNBS组TNF-α、TLR4、NF-κBp65基因及蛋白表达明显高于乙醇对照组(Pa＜0.01).结论 ITF对IBD小鼠肠道病变具有保护作用,可能与调节肠道先天免疫有关.     

纳米银薇乔线在小鼠皮内连续缝合线道周围近期抗炎疗效的观察研究

目的 探索纳米银薇乔线用于小鼠背部皮内连续缝合线道周围炎性细胞和炎性蛋白的表达情况,分析其近期抗炎疗效。方法 对4-0普通薇乔线采用化学涂层技术制备纳米银薇乔线。18只10～12周Balb/C小鼠随机分为3组(每组各6只),剃除小鼠背部毛发,纵行切开15 mm皮肤全层切口。分别采用4-0带针普通薇乔线、抗生素薇乔线、纳米银薇乔线行皮内连续缝合。术后5 d,以纵形切口为中心,取材全层皮肤组织。采用免疫组织化学染色方法比较线道周围中性粒细胞、巨噬细胞、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达水平,并计算平均光密度(mean optical density,MOD)。原位末端转移酶标记(mediated dUTP-biotin nick end labeling,TUNEL)染色了解线道周围细胞凋亡情况。Masson染色了解线道周围胶原蛋白沉积情况。统计学方法采用单因素方差分析和student’s t检验。结果 免疫组织化学染色显示,普通薇乔线、抗生素薇乔线和纳米银薇乔线的中性粒细胞MOD值为...     

肥胖相关性肾小球病小鼠肿瘤坏死因子α表达的变化

目的通过检测肥胖相关性肾小球病(ORG)小鼠血清中TNF-α水平及肾小球中TNF-αmRNA及蛋白表达,探讨ORG的发病机制。方法选取40只清洁级健康35日龄C57BL/6雄性小鼠,按体质量随机分为肥胖组和对照组各20只。肥胖组予高脂高能量饲料喂养,对照组予普通饲料喂养。2组分别于8周末留取尿液,ELISA法检测其尿微量清蛋白(Alb)、转铁蛋白(TRF)、尿视黄醇结合蛋白(RBP)、β2-微球蛋白(β2-MG)等尿微量系列蛋白;留取静脉血,ELISA法检测其血清TNF-α水平;游离其肾脏组织,固定、切片、染色,分别于光镜、电镜下观察肾脏组织病理学改变;提取肾组织RNA,实时定量(RT)-PCR检测TNF-αmRNA表达;提取肾组织,采用Western blot检测TNF-α蛋白表达。结果采用SPSS 13.0软件进行统计学处理。结果与对照组比较,肥胖组尿Alb、TRF、RBP、β2-MG及血清TNF-α水平均明显增高,差异均有统计学意义(Pa<0.01)。肥胖组肾组织TNF-αmRNA及蛋白表达明显增高(Pa<0.01)。肾组织病理学检查发现肥胖组均出现肾小球肥大;电镜下见上皮细胞足突融合,部分肾小管上皮细胞胞质内见较多脂滴,基膜增厚,三层结构消失,节段系膜插入。结论 ORG小鼠早期可出现尿微量蛋白异常;血清及肾组织中TNF-α表达异常可能在ORG的发生发展中扮演重要角色。     

布地奈德对慢性支气管哮喘小鼠肺组织HIF-1α和VEGF表达及气道重塑的影响

目的：研究布地奈德对慢性哮喘小鼠缺氧诱导因子-1α（HIF-1α）、血管内皮生长因子(VEGF)表达的调控和对血管生成、气道重塑的影响。方法：30只雌性BALB/c小鼠随机分为对照组、模型组和治疗组,每组10只。卵清蛋白（OVA）激发试验建立哮喘小鼠模型,治疗组从第28天开始每日在OVA激发前1 h雾化吸入布地奈德100 μg/kg,对照组以PBS代替OVA。苏木精-伊红染色分析气管管壁厚度变化,Masson染色分析肺组织胶原沉积,免疫组织化学染色法检测肺组织血管生成情况,免疫组织化学染色法和Western blot检测HIF-1α、VEGF表达水平。分析气管管壁厚度、血管面积百分比与VEGF、HIF-1α表达的相关性。结果：模型组较对照组气道管壁增厚,肺组织胶原沉积、血管面积、HIF-1α和VEGF表达显著增加;治疗组较模型组气道管壁厚度、肺组织胶原沉积及血管面积减少,HIF-1α和VEGF表达显著下降。气道管壁厚度、血管面积分别与HIF-1α及VEGF表达呈正相关,HIF-1α与VEGF表达亦呈正相关。结论：布地奈德通过抑制HIF-1α、VEGF表达,明显减轻哮喘小鼠肺组织血管生成和气道重塑。     

瘤体内注射糖皮质激素对婴幼儿血管瘤组织中血管内皮生长因子表达的影响

目的探讨糖皮质激素瘤体内注射对婴幼儿增生期血管瘤组织中血管内皮生长因子(VEGF)表达的影响及意义。方法选取拟手术治疗的增生期血管瘤患儿20例,分别于术前3d、6d、9d、2周注射曲安奈德1次(1mg/cm2),各5例;对照组为消退期血管瘤5例和增生期血管瘤5例,均不予用药。手术切除后瘤体标本,立即予40g/L甲醛固定,常规石蜡包埋,切片,HE染色,光镜下观察组织学变化,采用兔抗人VEGF多克隆抗体,第2代通用型二步免疫组织化学法染色,测定每张图片的平均吸光度(A)值,每张切片随机选取5个高倍镜视野。结果HE染色切片光镜下观察,各用药组与未用药组组织学比较均无显著性差异。免疫组织化学染色切片见VEGF表达在增生期血管瘤组织已出现血管腔的内皮细胞胞质中和尚未出现血管腔的内皮细胞胞质或胞膜中,在增生期未用药组中可见棕褐色颗粒,呈强阳性表达,随着激素影响时间的延长,1周后其表达强度渐下降,染色深度渐变浅,阳性细胞渐减少。测定其免疫反应阳性颗粒积分A值,未用药组与术前6d、9d、2周和消退期血管瘤组行样本均数的两两比较有统计学差异(F=17.9191P<0.01),糖皮质激素影响时间和血管瘤VEGF的表达呈显著负相关(r=-0.5922P<0.01)。结论应用曲安奈德瘤体内注射治疗婴幼儿增生期血管瘤,能明显下调VEGF表达,促进血管瘤消退。     

GATA-3在支气管哮喘小鼠肺组织表达及地塞米松对其抑制作用

目的探讨转录因子GATA-3在支气管哮喘小鼠肺组织中表达及地塞米松（Dex）对其干预作用。方法采用卵清蛋白（OVA）致敏方法建立支气管哮喘小鼠模型;Balb/c小鼠24只随机分为对照组、哮喘组、Dex治疗组各8只。采用免疫组织化学方法检测肺组织GATA-3蛋白表达;反转录聚合酶链反应（RT-PCR）方法检测肺组织GATA-3 mRNA相对水平;流式细胞技术检测小鼠脾CD4 T细胞白细胞介素-4（IL-4）水平;HE染色评价各组小鼠气管炎性反应变化。结果哮喘组小鼠肺组织GATA-3及mRNA水平、IL-4水平均明显高于对照组（P〈0.05）,Dex治疗组GATA-3阳性细胞数及mRNA水平、IL-4水平均明显低于哮喘组,具有显著性差异（P〈0.05）。HE染色结果表明Dex治疗组气管炎性反应明显缓解。结论GATA-3在支气管哮喘小鼠肺组织呈高表达。Dex可阻断哮喘鼠肺组织GATA-3表达,抑制Th2型细胞因子释放,减轻气管炎性反应。     

胆总管结扎胆汁淤积性肝硬化动物模型的建立与评价

目的探讨胆总管结扎手术致胆汁淤积性肝硬化小鼠模型的建立价值,并对模型进行评价。方法将84只balb/c小鼠随机分为实验组(采用胆总管结扎术)和对照组(采用假手术),术后观察小鼠一般情况和肝胆大体结构;采用全自动生化分析仪检测术后1 d、3 d、5 d、7 d、14 d、21 d、28 d总胆红素(total bilirubin,TBIL)、直接胆红素(direct bilirubin,DBIL)、谷氨酸转氨酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST);采用H&E染色评估肝脏组织形态学变化;采用masson染色和免疫组织化学检测术后28 d小鼠肝脏组织切片,评估两组胶原纤维增生和a-SMA、CK-19蛋白表达情况。结果实验组与对照组小鼠相比,术后一般情况相对更差,胆汁淤积,胆囊、胆总管体积明显增大,各项血清生化指标明显增高(P<0.05);HE染色可见肝小叶结构改变,胆管增生;术后28 d实验组小鼠masson染色可见胶原纤维形成,免疫组织化学染色提示a-SMA、CK-19表达量显著增多(P<0.05)。结论经腹中线小切口行胆总管结扎手术可成功构建小鼠胆汁淤积性肝硬化模型。     

增殖核抗原在胆管上皮细胞的表达

目的通过先天性胆总管囊肿，胆管癌和流产胎儿胆管标本的免疫组织化学染色，探讨增殖核抗原（PCNA）在胆管黏膜上皮细胞的表达，了解胆管上皮细胞的增殖分化情况。方法取先天性胆总管囊肿标本20例，胆管癌标本8例，流产胎儿胆管标本20例，用福尔马林固定24h。标本常规石蜡包埋，切片，切片厚度3μm。行PCNA免疫组织化学染色。结果PCNA阳性染色时细胞核为棕色。正常胎儿胆管的表达为（5．45±1．90）％，先天性胆总管囊肿的表达为（47．75±5．48）％，胆管癌的表达为（83．88±7．24）％。3组之间比较，差异有统计学意义。结论PCNA在3种细胞的表达不同，先天性胆总管囊肿的上皮细胞处于增殖状态。     

The use of rapid assessment of enteric ICC and neuronal morphology may improve patient management in pediatric surgery: a new clinical pathological protocol

Alterations in interstitial cells of Cajal (ICC) distribution and density may seriously influence gut motility. We and others have documented a disturbance of ICC and neurons in Hirschsprung’s disease (HD), intestinal ischemia, and inflammation. The ability to remove intestine with permanent dysmotility improves significantly the prognosis. This may be important in the developing intestine especially in HD. More complete pathological information increases the accuracy of surgical decisions. Therefore we sought to develop a rapid, intraoperative immunohistochemical protocol for ICC and neurons in surgical specimens for routine diagnostic and therapeutic assessment of pediatric patients. To date, cKit is the only reliable immunohistochemistry for ICC identification. There are multiple antibodies in use for neuronal identification. A comparison of fixation methods and immunostaining using cKit and multiple intestinal neuronal antibodies was done. Fresh segments of surgically resected intestine were sectioned and stained using antibodies for ICC cell identification (anti-cKit) and for neuronal characterization. By carefully changing tissue fixation methods, different neuronal antibodies were tested to determine an optimal rapid protocol. Each suggested protocol was tested on normal and pathological intestinal tissue and compared to the previous overnight immunostaining of the same tissue. A new rapid tissue fixation and immunostaining protocol using cKit for ICC identification and NF 68 was developed. By employing this protocol, we could obtain ICC and neuro-immunohistochemistry in unfixed frozen sections within 1 h with tissue vibration to diminish the time for immunostaining. Without vibration the protocol takes 3 h. ICC and enteric neuronal changes could be readily observed and the quality of staining was comparable to standard immunohistochemistry. Each gut pathology displayed characteristic changes in ICC and neuronal density/distribution in the affected bowel. The time-scale of the 1-h immunoprocessing is still longer than the standard clinical pathological “quick” sections using H&E staining; however, the protocol duration is within the surgery timescale. Standard H&E stain used in combination with our rapid neuronal and ICC immunohistochemistry protocol enables a fast, comprehensive, and accurate assessment of the pathophysiology of signaling networks controlling gut motility while the patient is still in the operating room. We propose that the addition of this simple and rapid immunohistochemical assessment in the pathologist evaluation of surgical specimens would result in a more complete characterization for diagnosis and prognosis of the pediatric patient. Specifically, we propose that this test will differentiate good versus poor prognosis HD patients based on their neuron/ICC ratio and present a rapid, standardized method for use in general pathology.     

Idiopathic Hyperammonemia following an Unrelated Cord Blood Transplant for Mucopolysaccharidosis I

Bone marrow transplantation (BMT) has been shown to reverse or stabilize some manifestations of mucopolysaccharidosis I (Hurler syndrome). Idiopathic hyperammonemia (IHA) is a rare complication of solid organ and BMT that is characterized by elevated serum ammonia, normal liver enzymes, and abrupt onset of neurologic deterioration. We present the case of a 14-month-old male patient with Hurler syndrome who developed fatal IHA (ammonia = 2297 mmol/L) 31 days after a cord blood transplant. A complete autopsy was performed, with examination of both frozen and formalin-fixed paraffin-embedded (FFPE) tissues using a variety of special stains and electron microscopy. Hyperammonemia was documented by analysis of antemortem serum and postmortem cerebrospinal and vitreous fluid. Other causes of hyperammonemia, including Reye syndrome, were excluded. Histologic changes included centrilobular microvesicular steatosis of the liver and storage product present in multiple organs. The highly water-soluble mucopolysaccharide (MPS) storage product was best identified by colloidal iron staining of FFPE and unfixed air-dried fresh frozen liver sections. Alcian blue stains failed to convincingly demonstrate MPS in any of the liver sections. This is the first published report, to our knowledge, of IHA in a posttransplant patient younger than 18 years old or following transplantation for Hurler syndrome. Demonstration of the hepatic centrilobular microvesicular steatosis characteristic of IHA was complicated by the diffuse storage of MPS within the liver. MPS storage can be best detected in the liver using colloidal iron staining. Oil-red-O staining may be useful to document microvesicular steatosis in cases with a clinical history of hyperammonemia following solid organ or BMT. Determining if certain subsets of children are at increased risk for IHA requires further study.     

A variant pattern of Calretinin immunohistochemistry on rectal suction-biopsies is fully specific of short-segment Hirschsprung’s disease

Background/purpose

Calretinin immunohistochemistry is now widely used to diagnose Hirschsprung’s disease (HD), since loss of calretinin expression within the mucosa and muscularis mucosae of rectal suction-biopsy is pathognomonic of HD. However, a stippled staining may be observed within hypertrophic nerves in the submucosae in some HD patients. The aim of the study was to test the hypothesis that such findings may announce the beginning of the transitional zone.

Methods

We retrieved 44 consecutive patients (10 girls and 34 boys; median age 6.5 days), diagnosed with aganglionosis on rectal suction biopsies, followed by surgery. According to calretinin immunohistochemistry performed on all paraffin-embedded rectal biopsies, we defined two HD groups: P? showing an absence of any staining within mucosa, muscularis mucosae and submucosa et P+ showing an absence of staining within the mucosa and muscularis mucosae, but a positivity of some submucosal hypertrophic nerves. These data were correlated to the length of total pathological segment (aganglionic and transitional zones) obtained from the original surgery reports.

Results

18/44 patients (40.9 %) belonged to the P+ group and 26/44 (59 %) patients were within the P? group. In the P+ group, the maximal length of the aganglionic zone was 9 cm [mean 4 (1–9)] and the total pathological zone never exceeded 14 cm [mean 8 (3.8–14)]. In the P? group, the maximal length of aganglionic zone was 55.5 cm [mean 11.3 (2.5; 55.5)] and the total pathological zone extended to 59.5 cm [mean 17.75 (4.5; 59.5)]. Aganglionic segment was significantly shorter in the P+ group (p < 0.0001).

Conclusion

Staining of some hypertrophic nerves in the submucosa in suction rectalbiopsy of HD patients using calretinin immunohistochemistry is only encountered in short-segment aganglionosis with a pathological zone always restricted to rectal and sigmoid colon. This information could be crucial for the surgeons in the decision to choose a transanal procedure.     

Safety and efficacy of a new extensively hydrolyzed formula for infants with cow's milk protein allergy

Cow's milk protein allergy (CMPA) is best treated by complete elimination of cow's milk from the diet. For infants with CMPA who cannot be breast-fed, formulas based on extensively hydrolyzed proteins or on amino acids are the preferred substitutes for cow's milk-based formulas. In this study, we compared the tolerance and growth of infants with CMPA who were fed a new extensively hydrolyzed formula containing lactose (eHF) with those who were fed an amino acid formula (AAF). This was a prospective, multi-center, randomized, reference-controlled study. Seventy-seven infants <12 months old with suspected CMPA were enrolled. In 66 of these, CMPA was confirmed by oral challenge in a double-blind, placebo-controlled food challenge (DBPCFC) or by a medical history of severe allergic reaction to cow's milk and a positive skin prick test. These infants were then tested for their reaction to eHF and AAF in a DBPCFC. All infants tolerated both formulas and were randomized to receive either eHF (n = 34) or AAF (n = 32) for 180 days. Growth (weight, length, and head circumference) and tolerance [skin, gastro-intestinal, and respiratory tract symptoms of allergy] were evaluated after 30, 60, 90, and 180 days. There were no significant differences between the two groups in any of the growth measurements. Length and head circumference were similar to Euro-growth standards, but weight was slightly lower. Gastro-intestinal and respiratory tract symptoms of allergy were also similar in the two groups. However, whereas SCORAD scores for atopic dermatitis remained constant throughout the study in infants-fed eHF, there was a slight decrease in those fed AAF. Infants-fed eHF had significantly fewer incidents of vomiting than infants-fed AAF and a significantly higher frequency of soft stools. The new eHF is safe and well tolerated in infants diagnosed with CMPA.     

Safety and efficacy of a new extensively hydrolyzed formula for infants with cow's milk protein allergy.

Cow's milk protein allergy (CMPA) is best treated by complete elimination of cow's milk from the diet. For infants with CMPA who cannot be breast-fed, formulas based on extensively hydrolyzed proteins or on amino acids are the preferred substitutes for cow's milk-based formulas. In this study, we compared the tolerance and growth of infants with CMPA who were fed a new extensively hydrolyzed formula containing lactose (eHF) with those who were fed an amino acid formula (AAF). This was a prospective, multi-center, randomized, reference-controlled study. Seventy-seven infants <12 months old with suspected CMPA were enrolled. In 66 of these, CMPA was confirmed by oral challenge in a double-blind, placebo-controlled food challenge (DBPCFC) or by a medical history of severe allergic reaction to cow's milk and a positive skin prick test. These infants were then tested for their reaction to eHF and AAF in a DBPCFC. All infants tolerated both formulas and were randomized to receive either eHF (n = 34) or AAF (n = 32) for 180 days. Growth (weight, length, and head circumference) and tolerance [skin, gastro-intestinal, and respiratory tract symptoms of allergy] were evaluated after 30, 60, 90, and 180 days. There were no significant differences between the two groups in any of the growth measurements. Length and head circumference were similar to Euro-growth standards, but weight was slightly lower. Gastro-intestinal and respiratory tract symptoms of allergy were also similar in the two groups. However, whereas SCORAD scores for atopic dermatitis remained constant throughout the study in infants-fed eHF, there was a slight decrease in those fed AAF. Infants-fed eHF had significantly fewer incidents of vomiting than infants-fed AAF and a significantly higher frequency of soft stools. The new eHF is safe and well tolerated in infants diagnosed with CMPA.     

表皮生长因子在血管瘤和血管畸形中的表达

目的 了解表皮生长因子(epidermal growth factor,EGF)与血管瘤及血管畸形的关系.方法 标本选自1998年3月至2005年10月经手术切除的先天性皮肤血管病变92例患儿,女52例,男40例,年龄为40 d至13岁,平均年龄为1.6岁.组织标本取出后立即浸入10%的中性甲醛溶液中固定,常规石蜡包埋,5μm连续切片,HE染色,同时采用含有正常血管的皮肤标本10例作为对照.采用免疫组织化学方法,对72例血管瘤和20例血管畸形组织切片进行EGF表达的测定.结果 EGF在血管瘤组织中高表达,而在血管畸形组织中低表达(P<0.01).在血管瘤增殖期、退化早期和退化完成期三者之间,EGF的表达存在着差异.在血管瘤中EGF的表达与其内皮细胞的增生程度有关.内皮细胞增生越强,EGF的表达越高(P<0.05).结论 提示EGF与血管瘤的增生及退化有关,而与血管畸形无明显关系.     

先天性巨结肠的一种术中快速诊断方法

目的 探讨先天性巨结肠的术中快速诊断方法。方法 17例术前诊断为巨结肠的患儿，术中分别在狭窄段、移行段和扩张段取三份全层标本，以直肠肛门畸形患儿结肠造瘘术后关闭瘘口时的结肠标本为正常对照，利用一种快速AchE染色方法，将标本孵育时间分别确定为1、3、5、10、15、30min，同时进行冰冻切片HE染色并和正常组比较。结果 快速AchE染色法标准孵育时间5min后出现深褐色的沉淀，我们可以识别出增生的胆碱能神经纤维，而正常组神经纤维不显色，如果结合冰冻切片HE染色观察神经节细胞使结果更准确。结论 这种简单而可靠的快速AChE染色方法可以和冰冻切片HE染色联合应用在术中诊断HD和IND，并判断异常肠段的范围。     

Preliminary Report: Immunoperoxidase-Linked Human Placental Lactogen as a Histopathologic Index of Perinatal Morbidity and Mortality

Anatomic study of placental dysfunction may benefit from the applications of new routine techniques. Immunohistochemical staining for human placental lactogen (HPL) was used in 3 cases illustrating different perinatal disorders. The amount of HPL labeling ranged from high in an acute anoxic death due to abruptio placentae, through decreased in a case of maternal hypertension, to low in severe intrauterine growth retardation. Such information complements standard clinical and pathologic studies. Ten percent buffered Formalin was superior to Bouin's fixative and alcoholic Formalin for the demonstration of HPL. Even after 4 days of refrigeration at 4°C, all of the syncytiotrophoblastic tissue was labeled in sections of paraffin-embedded, Formalin-fixed normal placentas.     

表皮生长因子在血管瘤和血管畸形中的表达

目的 了解表皮生长因子(epidermal growth factor,EGF)与血管瘤及血管畸形的关系.方法 标本选自1998年3月至2005年10月经手术切除的先天性皮肤血管病变92例患儿,女52例,男40例,年龄为40 d至13岁,平均年龄为1.6岁.组织标本取出后立即浸入10%的中性甲醛溶液中固定,常规石蜡包埋,5μm连续切片,HE染色,同时采用含有正常血管的皮肤标本10例作为对照.采用免疫组织化学方法,对72例血管瘤和20例血管畸形组织切片进行EGF表达的测定.结果 EGF在血管瘤组织中高表达,而在血管畸形组织中低表达(P<0.01).在血管瘤增殖期、退化早期和退化完成期三者之间,EGF的表达存在着差异.在血管瘤中EGF的表达与其内皮细胞的增生程度有关.内皮细胞增生越强,EGF的表达越高(P<0.05).结论 提示EGF与血管瘤的增生及退化有关,而与血管畸形无明显关系.