面部原发性皮肤隐球菌病1例

报道1例自身免疫功能正常的原发性皮肤隐球菌病。患者因"面部红斑进行性加重半年,破溃伴结痂2个月余"来我科就诊,皮肤科检查可见左侧面颊部3 cm×5 cm大小红斑,边界清楚,中央可见数片浅溃疡,覆盖灰褐色痂皮。病理检查示真皮浅层及深层可见大量炎性细胞浸润,PAS染色见大量紫红色孢子聚集在多核巨细胞内外;阿申蓝染色阳性;粘蛋白卡红染色阳性;免疫组化染色(抗体为兔抗隐球菌抗体)可见棕色圆形孢子。根据其临床症状、病理检查、特殊染色及免疫组化染色,确诊为原发性皮肤隐球菌病。经伊曲康唑治疗痊愈。     

卵巢囊性颗粒细胞瘤的临床病理学特征、诊断及鉴别诊断

目的:探讨卵巢囊性颗粒细胞瘤的临床病理学特征、诊断和鉴别诊断要点。方法:复习1例卵巢囊性颗粒细胞瘤患者的临床资料、肿物的大体以及镜下病理组织学特征、免疫组化染色特征。结果:患者左侧单房囊性颗粒细胞瘤,大小为14.6 cm×18.9 cm×9.8 cm,囊壁厚薄不均一;镜下可见内壁由颗粒细胞组成,细胞层数不一,可见典型的Call-Exner小体;免疫组化染色可见α-inhibin,Vimentin,CD99均阳性。结论:卵巢囊性颗粒细胞瘤可依据镜下发现不典型增生的颗粒细胞、典型的Call-Exner小体和核沟等特征性的组织形态学作出诊断。     

新鲜组织肥大细胞快速染色方法的探讨与应用

目的探讨如何在组织内更好地显示肥大细胞。方法取大鼠后肢皮肤组织,经恒冷箱切片机切片,然后将组织切片直接入1%甲苯胺蓝染液进行染色。结果在新鲜组织切片内可见到较多充满蓝紫色颗粒的肥大细胞和它们分泌的蓝紫色颗粒。结论新鲜组织直接进行甲苯胺蓝染色可显示较多的肥大细胞。     

小鼠骨髓来源肥大细胞的培养及鉴定

目的:探讨小鼠骨髓来源肥大细胞体外大量培养及鉴定方法。方法:采用IL-3(10 ng/mL)和不同浓度的SCF联合刺激小鼠骨髓细胞,培养四周,采用普通光学显微镜观察细胞形态,牛鲍氏血细胞计数板进行细胞计数,甲苯胺蓝染色观察其胞浆颗粒物质,流式细胞仪检测细胞表面CD117和FcεRI的表达情况,β-己糖苷酶释放试验评估肥大细胞活化能力。结果:培养条件为10ng/mL IL-3和20 ng/mL SCF时,悬浮细胞数目能达到1×10~8细胞,其中CD117和FcεRI双阳性细胞比例达90%以上;甲苯胺蓝染色可见细胞胞浆含有大量紫红色颗粒;细胞经IgE-DNP致敏及DNP-HSA激发后,β-己糖苷酶释放率明显增加,且SCF能促进肥大细胞释放β-己糖苷酶。结论:采用10 ng/mL IL-3和20 ng/mL SCF诱导骨髓细胞能够获得大量高纯度的小鼠骨髓来源肥大细胞,为肥大细胞特性及功能学研究提供了基础。     

神经皮肤黑变病中黑色素的来源初探

目的探讨食蟹猴神经皮肤黑变病（NCM）中黑色素的来源。方法收集北京协和医学院比较医学中心病理室食蟹猴神经皮肤黑变病的病理学标本一例,复习其病理切片并辅助免疫组化及电镜技术对黑色素的来源做进一步探讨。结果皮肤病理检查显示真皮内黑色素主要由黑色素细胞产生,黑色素细胞CD31免疫组化染色阴性。脑膜的组织病理学显示脑膜下黑色素主要由黑色素细胞产生,黑色素细胞CD31免疫组化染色阴性。脑实质的组织病理学显示其黑色素由黑色素细胞和血管内皮细胞产生,CD31免疫组化染色显示含有黑色素的血管内皮细胞呈阳性着色。电镜检查结果同样显示血管内皮细胞内可见黑色素小体沉积。结论我们在世界上首次揭示了食蟹猴NCM大脑中血管内皮细胞可产生黑色素,这给内皮细胞增加了一项新的功能。     

甲状腺浆细胞瘤1 例及文献复习

目的:探讨甲状腺浆细胞瘤的病理学特点及临床表现。方法:对1例甲状腺浆细胞瘤进行组织学表现、免疫组化染色观察及文献复习。结果:组织学特点:肿瘤细胞成分单一,大多数为分化较为成熟的肿瘤性浆细胞,胞核圆形或卵圆形,大小较一致,常偏位,染色质呈车辐状或钟面状。免疫组化特点:肿瘤细胞表达CD20(++)、CD79a(+)、CD38(++)、CD138(+)、κ链(++)。结论:甲状腺浆细胞瘤是甲状腺一种少见的肿瘤,应同炎症反应的浆细胞增生及低分化癌、淋巴瘤、髓样癌等相鉴别,可以通过免疫组化染色及形态学观察进行鉴别诊断。     

特殊染色及免疫组化双重染色在恒河猴糖尿病模型中的应用

目的在链脲佐菌素（STZ）诱导恒河猴糖尿病动物模型基础上,用特殊染色、免疫组化双染方法显示胰腺等组织中的特征性病变。方法健康恒河猴5只,小剂量（30mg／kg）多次静脉注射STZ,濒死状态时将动物安乐死。取胰腺、心脏、肾脏、脾脏、肝脏、眼球、脑等器官制成石蜡切片,用HE、PAS、Masson、天狼星红和甲苯胺蓝等方法进行特殊染色,用免疫组化双重染色法同时显示胰岛A、B细胞。结果模型动物的胰岛萎缩,数量减少。Masson染色见外分泌部间质内纤维增生。局灶性慢性肾炎,病变区肾间质纤维组织增生,部分肾小球及肾小管萎缩。天狼星红染色见脾脏中央动脉管壁增厚。免疫组化双染见胰岛A胰高血糖素表达增多,胞浆呈棕褐色。B胰岛素表达减少,胞浆粉红色。结论HE染色结合特殊染色和免疫组化双重染色可较好地对STZ诱导糖尿病动物模型进行组织学评价。     

从甲状腺肥大细胞探讨胎儿器官肥大细胞的差异

研究肥大细胞在人胎儿甲状腺发育中数量、分布及组化性质的改变,以探讨胎儿器官发育中肥大细胞的差异。取45例不同胎龄的人胎甲状腺石蜡切片做甲苯胺蓝染色和阿尔辛蓝－－藏红染色,并测定肥大细胞的临界电解质浓度值及进行硫酸小蘖硷荧光染色。结果显示：3月龄胎儿甲状腺内开始出现肥大细胞,数量极少,主要分布在被膜及小叶间结缔组织内,甲苯胺蓝染色肥大细胞颗粒呈淡紫蓝色,阿尔辛蓝－－藏红染色呈蓝色,临界电解质浓度值较低,硫酸小蘖硷染色未见显黄色荧乐的肥大细胞,从3月龄到足月随着胎龄增长,肥大细胞数量缓慢增多,8月龄时肥大细胞经甲苯胺蓝染色,其颗粒呈紫红色,阿尔辛蓝－－藏红染色出现少量含红色和红蓝混合染色颗粒的肥大细胞,临界电解质浓度值偏高,可见少量显黄色荧光的肥大细胞,结果表明：在人胎儿3月龄时甲状腺发育中开始出现肥大细胞,但随胎儿发育肥大细胞的组化性质改变不明显。     

人毛乳头细胞组织化学研究

毛乳头细胞是一种高度特殊化的成纤维细胞。本文通过对体外培养的毛乳头细胞进行组织化学染色研究发现,它对阿新蓝、甲苯胺蓝和PAS染色均呈阳性,并对甲苯胺蓝显异染性．与原位时的细胞染色结果相同,表明在体外培养下．毛乳头细胞合成和分泌酸性、中性粘多糖的能力仍能维持较长时间;在细胞聚集区和多层化细胞团中有丰富的细胞外基质,阿新蓝和PAS染色呈强阳性,说明细胞外基质的存在与毛乳头细胞的聚集有很大关系;另外毛囊真皮鞘细胞对阿新蓝、甲苯胺蓝染色呈阳性反应．无甲苯胺蓝的异染性,PAS染色阴性,而真皮成纤维细胞这些染色均阴性,说明它与毛乳头细胞关系密切。     

树脂包埋半薄切片在视神经组织学和免疫组织化学研究中的应用

目的探讨半薄切片在视神经组织学和免疫组织化学研究中的应用,寻找一种视神经形态学研究和疾病诊断的有效方法。方法健康Sprague-Dawley大鼠视神经分别行树脂包埋制成半薄切片与石蜡包埋制成石蜡切片,分别采用HE、甲苯胺蓝染色及髓鞘碱性蛋白(myelin basic protein,MBP)免疫组织化学方法染色,光学显微镜下观察并比较半薄切片与石蜡切片染色结果的差异。结果石蜡切片HE和甲苯胺蓝染色均显示视神经髓鞘不着色,MBP免疫组织化学染色示MBP阳性着色较重,着色较紊乱,髓鞘着色不清晰。半薄切片HE及甲苯胺蓝染色均显示髓鞘呈环形,着色清晰,颜色对比鲜明;MBP免疫组化染色结果可见髓鞘呈环形,非特异性染色不明显,阳性对比显著,可清晰显示髓鞘结构。结论半薄切片在视神经组织学和免疫组织化学研究中优于石蜡切片,可用于视神经疾病的诊断和研究。     

Utility of toluidine blue staining and brush biopsy in precancerous and cancerous oral lesions

OBJECTIVE: To evaluate the usefulness of toluidine blue and brush biopsy in precancerous oral lesions and squamous cell carcinoma. STUDY DESIGN: The study was conducted at Moti Lal Nehru Medical College, Allahabad, India. Ninety-six patients with suspicious oral lesions who attended the outpatient clinics of otorhinolaryngology were screened with in vivo toluidine blue staining and oral brush biopsy. RESULTS: Oral brush biopsy showed high specificity and sensitivity. Toluidine blue staining was highly sensitive and moderately specific for malignant lesions but less sensitive for premalignant lesions. CONCLUSIONS: Early detection of oral carcinoma is possible even at the precancerous stages by using noninvasive, painless and outpatient procedures, such as in vivo toluidine blue staining and brush biopsy.     

KM突变小鼠慢性炎症性皮肤病的免疫学改变

目的 观察KM突变小鼠皮肤慢性炎症的病理变化,探讨该小鼠皮肤免疫学改变.方法 通过外部特征、常规HE病理组织学、免疫组织化学、特染方法对3月龄、6月龄KM突变小鼠皮肤炎症细胞及炎症因子进行检测并与KM野生小鼠皮肤炎症细胞及炎症因子浸润进行比较.结果 KM突变小鼠皮肤毛稀、皮屑、皮皱等;组织病理表皮细胞坏死,上皮角化过度或不全,颗粒层增厚,基底细胞层水肿,真皮浅层血管扩张,结缔组织炎细胞浸润等,皮肤CD3+、CD4+T细胞、巨噬细胞、肥大细胞等增多,同时炎症因子IL-6、IL-22、TNF-α、IFN-γ等增多;且这些炎症细胞及炎症因子浸润3月龄较6月龄增多.结论 KM自发突变小鼠皮肤组织出现自发的慢性炎症病变,与人类慢性炎症性皮肤病变有相类似的病理改变和细胞分子改变,有望培育成为一种新的慢性炎症性皮肤病的动物模型.     

Comparison of histochemical staining techniques for detecting mast cells in oral lesions

ABSTRACT

Mast cells are large cells with granular cytoplasm that participate in wound healing, angiogenesis and defense against pathogens. They also contribute to inflammation by initiating innate and acquired immunity. The granules of these cells exhibit characteristic staining properties. We investigated toluidine blue, astra blue, Alcian blue-pyronin Y and May-Grunwald Giemsa stains for mast cells in various oral lesions and assessed the efficacy of each for identifying mast cells. Sections were obtained from 10 each of diagnosed cases of inflammatory fibrous hyperplasia, periapical cyst, mild dysplasia, oral submucous fibrosis and oral squamous cell carcinoma and stained using the stains listed above. Mast cells were assessed for their presence, contrast of the mast cell in the connective tissue background and number. We found that May-Grunwald Giemsa stain was the best for identification of mast cells, although toluidine blue staining is less time-consuming. Overall we obtained better results using May-Grunwald Giemsa and toluidine blue for staining mast cells.     

Sclerotic fibroma-like dermatofibroma: an uncommon distinctive variant of dermatofibroma

Dermatofibroma (DF) is a common benign cutaneous tumor with many variants based on alterations in the morphology and composition of its diverse elements. One very infrequent type is sclerotic fibroma-like DF (SF-DF). We report 7 new cases of SF-DF. In addition, their main clinicopathological and immunohistochemical features were compared with 14 unselected common DFs and with 3 sclerotic fibromas (SFs). Microscopically, the 7 cases of SF-DFs showed an unencapsulated, well-circumscribed, hypocellular central nodule with thick collagen bundles arranged in a storiform pattern with prominent clefts. The overlying epidermis was attenuated. The periphery of this nodule was more cellular with histopathologic features of common DF. The 7 SF-DFs patients were 4 women and 3 men with a mean (+/-SD) age of 44.8 (+/-15.5) years. These 7 patients were younger than those suffering from SFs [71.0 (+/-17.3) years; (p=0.04)] and older than those presenting common DFs [30.5 (+/-12.3) years; (p=0.03)]. Immunohistochemically, spindle cells in all 7 SF-DFs were negative for CD34 and CD99. On the contrary, the 3 cases of SF were positive for CD34 and CD99. All of the common DFs were negative for CD34 and only 4 (28.6%) of them were positive for CD99. In conclusion, SF-DF is an uncommon variant of DF with similar clinicopathological and immunohistochemical features. SF-DF shares certain histopathologic features with SF but they are immunophenotypically different. Therefore, both entities should be differentiated.     

胶原蛋白/BMP复合材料的制备和成骨性能研究

以胶原膜(含87.5 mg I型胶原蛋白)为载体, 复合3.5 mg rhBMP-2(人基因重组骨形成蛋白-2), 制备胶原蛋白/BMP复合材料。复合材料首先在兔背阔肌中埋置, 预构新生骨组织, 并采用ALP染色、Von Kossa染色和HE染色等观察复合材料的成骨过程和组织形态。然后将形成的新骨组织游离移植修复自体下颌骨体部洞穿性缺损; 并设以胶原为载体的rhBMP-2复合骨修复材料直接修复为对照组, 骨缺损不修复组为空白组。采用X线、抗压强度、硬组织切片、四环素荧光染色、骨形态计量检查, 观察复合材料修复骨缺损的质量和效果。结果表明, 胶原蛋白/BMP复合材料在兔背阔肌中4～6周成骨, 胶原材料于3～5周降解; 成骨过程为是以软骨成骨为主的方式, 新骨形态为编织骨, 可见明显的微血管分布; 游离移植修复自体下颌骨缺损, 6周缺损区为骨性愈合, 与对照组在抗压强度(P = 0.041)、新骨量(P = 0.034)均有显著性差异。胶原蛋白/BMP复合材料在骨骼肌中形成的新生骨组织可作为供骨修复一定范围的骨缺损。     

胶原蛋白/BMP复合材料的制备和成骨性能研究

以胶原膜(含87.5 mg I型胶原蛋白)为载体, 复合3.5 mg rhBMP-2(人基因重组骨形成蛋白-2), 制备胶原蛋白/BMP复合材料。复合材料首先在兔背阔肌中埋置, 预构新生骨组织, 并采用ALP染色、Von Kossa染色和HE染色等观察复合材料的成骨过程和组织形态。然后将形成的新骨组织游离移植修复自体下颌骨体部洞穿性缺损; 并设以胶原为载体的rhBMP-2复合骨修复材料直接修复为对照组, 骨缺损不修复组为空白组。采用X线、抗压强度、硬组织切片、四环素荧光染色、骨形态计量检查, 观察复合材料修复骨缺损的质量和效果。结果表明, 胶原蛋白/BMP复合材料在兔背阔肌中4～6周成骨, 胶原材料于3～5周降解; 成骨过程为是以软骨成骨为主的方式, 新骨形态为编织骨, 可见明显的微血管分布; 游离移植修复自体下颌骨缺损, 6周缺损区为骨性愈合, 与对照组在抗压强度(P = 0.041)、新骨量(P = 0.034)均有显著性差异。胶原蛋白/BMP复合材料在骨骼肌中形成的新生骨组织可作为供骨修复一定范围的骨缺损。     

孤立性纤维性肿瘤15例CT影像病理特征分析

对比研究孤立性纤维性肿瘤(SFTs)的CT影像学特征及其与病理学的关系,评估CT在SFTs诊断中的应用价值。方法:收集15例孤立性纤维性肿瘤,回顾分析CT表现以及病理形态学和免疫表型变化。结果:15例中6例发生于胸腔内(3例发生于肺,3例位于壁层胸膜);此外鼻腔2例,上颌窦、腹腔、咽旁、腮腺、肾脏、眼眶、髂窝各1例。临床主要表现为局部肿块及其引起的压迫症状,CT扫描显示肿块边界清楚,增强扫描早期呈明显均质性强化。病理学改变显示瘤细胞呈梭形细胞为主,具有细胞密集区与细胞疏松区特征,间质可见瘢痕样的胶原和鹿角样厚壁血管,瘤细胞排列方式多样,免疫组化染色CD34阳性(91．67%), CD99(88．88%),Bcl-2(87．5%)和Vim(100%)。结论:SFT因有完整的包膜,故多为境界清楚的软组织密度肿瘤,如境界不清多提示侵袭性生长。因病理学上肿瘤细胞分布以及有无变性坏死而呈现不同的密度特征,因富血管表现而在增强CT扫描时有较为明显的强化;最终诊断需要免疫组织化学染色证实。     

Expression of lumican related to CD34 and VEGF in the articular disc of the human temporomandibular joint

Lumican belongs to the small leucine-rich repeat proteoglycan (SLRP) gene family and has been reported to exist in the cornea, intervertebral disc and tendon. Lumican plays a significant role in the assembly and regulation of collagen fibres. The human temporomandibular joint (TMJ) disc is made up of fibrocartilage with an extracellular matrix (ECM) composed of collagen and proteoglycans. The existence and behaviour of lumican have not been studied in the human TMJ disc. Therefore, we used immunohistochemical methods to detect lumican, CD34 and vascular endothelial growth factor (VEGF) and histochemical staining with toluidine blue in 13 human TMJ specimens (10 surgically removed and 3 obtained from autopsy). In both normal and deformed discs we observed staining with toluidine blue. We found that the area of metachromasia inside the deformed disc was uneven and expression of lumican was strong in the areas negative for metachromasia. Staining of VEGF and CD34 inside the deformed disc was seen. We confirmed the expression of lumican in the human TMJ disc and showed that a large number of fibroblast-like cells existed in the area of strong lumican expression. These new findings about the behaviour of lumican suggest that it may play a key role in the generation of a new collagen network by fibroblast-like cells.Key words: TMJ disc, lumican, CD34, VEGF, immunohistochemistry, metachromasia.     

Age-related changes of immunophenotypically immature lymphocytes in normal human peripheral blood.

The presence of phenotypically immature lymphocytes in umbilical cord blood has been a controversial topic. Moreover, their changes with age have not been systematically evaluated. In the present study, relative and absolute numbers of CD34+, CD10+CD19+, and CD4+CD8+ cell subsets were determined in umbilical cord blood from 12 full-term normal newborns, 43 children aged 1 month to 6 years, and 10 young adults. The samples were processed by whole-blood lysis and monoclonal antibody staining, and cells were analyzed by flow cytometry. Immature cells were present in cord blood and progressively declined in both absolute and percentage numbers with age, each according to a particular curve, reaching youth values roughly at age 2-4 years. These results demonstrate that phenotypically immature cells normally circulate at low levels in peripheral blood, mostly at birth and during infancy, but also during youth.     

Hereditary melanoblastoma in miniature pigs and its successful therapy by devitalization technique.

Applying selective breeding for 10 years we established the MeLiM (Melanoblastoma-bearing Libechov Minipigs) strain. Melanoblastoma (MB) in this strain shows a hereditary occurrence. Cutaneous tumours are usually nodular, multiple and distributed on various parts of body. They appear in darkly pigmented animals already at the birth or during two months thereafter (57% of all animals). Numerous organ metastases mainly into the spleen, lymph nodes and lungs are regularly ascertained in animals with cutaneous MB. Tumour cells were surprisingly found also in the inner organs of phenotypically healthy minipigs in which no cutaneous MBs were observed visually (27% of all animals). About 34% of all affected piglets die during the first 2 months of age. These features document a malignancy of this tumour in the MeLiM strain. Original surgical technique was applied in more than 40 affected minipigs at 1-2 months of age. It consists in a devitalization (ischemization) of one of cutaneous tumours by the mattress sutures conducted around the tumour base without any excision of tumour tissue. This simple procedure causes a total destruction of MB cells in all cutaneous tumours as well as in all organ metastases during 4-6 months. Animals treated by this technique were fully healed of tumour cells and no relapses were observed. This technique could bring similar positive results also in therapy of human MB.