Expression of CD44V6 and PCNA in squamous cell carcinomas

Objective To investigate the expression of cluster of differentiation 44 variant 6 (CD(44)V6) and proliferating cell nuclear antigen (PCNA) in ocular squamous cell carcinomas. Methods Streptavidin-biotin complex (SABC) immunohistochemistry was used to explore the expression of CD(44V6)and PCNA in 35 cases of ocular squamous cell carcinomas, 20 cases of papillomas, and 11 cases of normal eyelid tissue.Results The CD(44V6)positive rate was 62.9% (22/35) in ocular squamous cell carcinomas, 15.0% (3/20) in papillomas, but not detectable in the 11 cases of normal eyelid tissue. The positive expression rates of CD(44V6)in ocular squamous cell carcinomas were significantly higher than in benign tumors （χ[2=11.57, P<0.01）or control tissue (P=0.001), and the positive expression rates of CD(44V6)in metastasis were significantly higher than without metastasis （P=0.049）. PCNA labeling indexes (PI) in tumors with CD(44V6)expression were significantly higher than those without (t=20.21, P<0.01). Conclusions Overexpression of CD(44V6)is correlated with the progress and metastasis of ocular squamous cell carcinomas. CD(44V6)protein positive staining is associated with high PI. CD(44V6)and PCNA are useful for evaluating prognosis.     

Primary cutaneous CD30-positive anaplastic large cell lymphoma analysis

Objective To examine 10 cases with primary cutaneous CD30-positive anaplastic large cell lymphoma (ALCL), analyze their clinical manifestations and pathological and immunohistochemical features, and improve early diagnosis of this disease. Methods We studied the morphological characteristics of primary cutaneous CD30-positive ALCL using histopathological methods.Leukocyte common antigen (LCA), CD20, CD30, CD45RO, CD68, epithelial membrane antigen (EMA), cytokeratin (CK) and HMB45 antibodies were used to determine the expression of their respective antigens from routine paraffin samples of the patients.Results Ten patients (7 men and 3 women, aged 31 to 84 years) complained of subcutaneous masses or papular eruptions over their lower trunks and extremities.Histopathologically, the lesions were composed of numerous large round or oval pleomorphic cells.The cytoplasm was usually abundant, amphophilic or basophilic, and finely vacuolated.Nuclei were commonly eccentrically localized and lobated or horseshoed in shape, and multinucleated giant cells and Reed-Sternberg-like cells were seen.Nucleoli were generally multiple and large.Of the 10 patients, tumor cells displayed positive antigen expression of CD30 in all cases, positive CD45RO in 6 cases, positive CD20 in only 1 case, but negative CD45RO and CD20 expressions in 3 cases.Two patients died at 7 weeks and 34 years of follow-up, respectivelyConclusion Our study highlights the importance of histopathologic features and positive CD30 staining for differentiation of this disease from other malignant skin tumors.     

THE EXPRESSION OF C-erbB-1 AND C-erbB-2 ONCOGENES IN BASAL CELL CARCINOMA AND SQUAMOUS CELL CARCINOMA OF SKIN

The expression of c-erbB-1 and c-erbB-2 oncogenes were investigated by immunohistochemistry using monoclonal antibodies to c-erbB-1 and c-erbB-2 protein in 43 cases of basal cell carcinoma (BCC) and 26 cases of squamous cell carcinoma (SCC). We found that the expression of c-erbB-1 oncogene in all BCC increased by different degrees and the expression of c-erbB-2 oncogene in BCC was significantly reduced or lost when compared to that in normal epidermal cells.Furthermore,apparent negative and positive relationships were observed respectively between the tumor differentiation and the expression of c-erbB-1 and c-erbB-2 oncogenes in SCC.It is suggested that the abnormal expression of c-erbB-1 and c-erbB-2 oncogenes in BCC and SCC may play a role in the development of skin tumors.The pattern of the c-erbB-1 and c-erbB-2 oncogenes expression in SCC may assist in distinguishing the biological behavior and prognosis of SCC.     

The distribution and significance of renal infiltrating cells in patients with diffuse crescentic glomerulonephritis

Objective To study the significance and distribution of renal infiltrating cells, includin g monocytes (CD68(+) cells), proliferative cell nuclear antigen positive cells ( PCNA(+) cells), CD4(+) and CD8(+) cells in diffuse crescent glomerulonephritis (DCGN).Methods Fifty-six patients with DCGN were studied, including 10 cases of anti-glomerul ar basement membrane (GBM)-type Ⅰ DCGN, 26 immune complex-type Ⅱ DCGN, and 2 0 cases of pauci-immune-type Ⅲ DCGN. Glomerular and interstitial infiltrates of CD68(+) and PCNA(+) cells, and interstitial infiltrates of CD4(+) and CD8(+) cells were detected by using four-layer PAP methods. Results There was a significant increase of renal infiltrating CD68(+), PCNA(+), CD4(+) and CD8(+) cells in patients with DCGN compared with that in normal controls. I n patients with type Ⅰ DCGN, there was a higher number of renal infiltrating C D68(+) and PCNA(+) cells than that in patients with type Ⅱ and Ⅲ DCGN. A glom erular infiltrates of CD68(+) and PCNA(+) cells correlated with the interstitial infiltrates of CD4(+) cells in type Ⅰ or Ⅲ DCGN patients. In lupus DCGN pati ents, the numbers of renal infiltrating CD68(+) and PCNA(+) cells were similar t o vasculitis or type Ⅲ DCGN patients.Conclusion These findings demonstrate that the renal infiltrates of CD68(+) and PCNA(+) cel ls play an important role in patients with DCGN, that the infiltrates of CD4(+) cells correlate with the infiltrates of CD68(+) and PCNA(+) cells may be an acti ve marker of DCGN, and that cell-mediated immunity may contribute to crescent f ormation in lupus DCGN patients.     

Primary signet ring cell carcinoma of prostate (report of 2 cases)

Objective To study the clinical features of primary signet ring cell carcinoma of prostate. Methods 2 cases of primary signet ring cell carcinoma of the prostate were studied and reviewed. Results The age of the 2 patients was 64 and 73. The clinical symptoms were dysuria, vesical irritability and perineum discomfort. Histologically, signet ring cell carcinoma was composed of round cells with abundant clear cytoplasm and crescent-shaped nuclei on one side. Mitosis were frequently observed. Immunohistochemical testing showed the cancer cell was positive for prostate specific antigen (PSA.), prostate acid phosphatase ( PAP ), AR, cytokeratin and negative for caicinoernbryonic antigen (CEA), alcian blue/ periodic arid-schiff (AB/PAS). One case (stage D) died 6 months after bilateral orchiectomy and flutamide therapy because of wide-spead metastasis; the other (stage B2) has been surviving 25 months after radical prostatectomy, bilateral orchiectomy, endocrine therapy and local irradiation ministration. Conc     

Transplantation of human limbal cells cultivated on amniotic membrane for reconstruction of rat corneal epithelium after alkaline burn

Background The transplantation of limbal epithelial cells cultivated on amniotic membrane is a newly developed treatment for limbal stem cell deficiency. The purpose of our study was to investigate the biological characteristics of limbal epithelial cells and evaluate the effect of transplantation of cultivated human limbal epithelial cells on ocular surface reconstruction in limbal stem cell deficiency rat model.Methods Human limbal cells were isolated and cultivated in vitro. Cytokertins 3, 12, and 19 (K3, K12 and K19) and p63 were detected by immunofluorescent staining or RT-PCR. BrdU labelling test was used to identify the slow cycling cells in the cultures. Limbal stem cell deficiency was established in rat cornea by alkali burn.Two weeks after injury, the rats received transplants of human limbal stem cells cultivated on amniotic membrane carrier. The therapeutic effect was evaluated by slit lamp observation, Hemotoxin and Eosin (HE) staining and immunofluorescent staining.Results On day 7 in primary culture, p63 and K19 were strongly expressed by most cells but only a few cells expressed K3. On days 14 and 21, p63 and K19 were still expressed by a majority of cells, but the expressive intensity of p63 decreased in a number of cells, while the proportion of K3 positive cells increased slightly and some cells coexpressed p63 and K3. RT-PCR showed that gene expression of both p63 and K12 were positive in cultivated limbal cells, but in mature superficial epithelial cells, only K12 was detected. BrdU labelling test showed that most cells were labelled with BrdU after 7 days‘ labelling and BrdU label retaining cells were observed after chasing for 21 days with BrdU free medium. For in vivo test, slit lamp observation, HE staining and immunofluorescent staining showed that the rats receiving transplant of human limbal stem cells cultivated on amniotic membrane grew reconstructed corneas with intact epithelium, improved transparency and slight or no neovascularization. A majority of epithelial cells of the reconstructed cornea were positive to antihuman nuclear antibody and cells expressing K3 were found mainly in superfacial epithelium.Conclusions Limbal stem cells can be cultivated in vitro: the cells are characterized by high proliferation and slow cycling and identified as p63/K19 positive and K3/K12 negative. During culture, some stem cells can proliferate and differentiate into mature cornea epithelial cells. Amniotic membrane is a suitable carrier for limbal stem cells. Transplantation of human limbal stem cells cultivated on amniotic membrane can functionally reconstruct rat cornea with limbal stem cell deficiency.     

USE OF SANDWICH ENZYME IMMUNOASSAY WITH GLASS BEADS TO DETECT LUNG CANCER CEA AND AFP

The double antibody sandwich enzyme im- munoassay (EIA) method with glass beads as solid phase is used to detect carcinoembryonic antigen (CEA) and a -fet.oprotein (AFP) in sera of patients with bronchogenic carcinoma. The enzyme labelled antibodies are made from puri- fied rabbit antibodies against human CEA and AFP linked with peroxidase using O-phenylene diamine as a substrate. The color react.ion was examined for quantitative analysis by electro- photometer. Elevated serum CEA levels ove:r 5 ng/ml were found in 2.9 0f the 42 patient (69'70) with lung cancer and 8 0f 25 cases (32'7c》 with benign lung diseases, but none of the latter had a level over 8 ng/ml. Only 3 0f the 46 normal controls (6.5%) had positive CEA higher than 5 ng,rml. Serial observations of 23 cases of broncho- genic carcinoma treated with Polyporus Umbel- latus 猪苓多糖 combined with Chemotherapeu- tic drugs were also examined. In 18 cases (78%), serum CEA concentration c:orresponds to the therapeutic effect after treatment, so it appears that the serum CEA level may be used for pro- gnostic assessmenL On the other hand only 2 of 30 cases with bronchogenic carcinoma showed positive serum AFP levels.     

Immunohistochemical study in dural arteriovenous fistula and possible role of ephrin-B2 for development of dural arteriovenous fistula

Background Although there were several clinical and experimental studies discussing the pathogenesis of dural arteriovenous fistula (DAVF), the pathological process leading to intracranial DAVF so far remains unknown. In this study, we investigated the expression of vascular growth factors in order to elucidate the possible role of these factors for the development of DAVF and to study the biological activity of this uncommon lesion.Methods We examined the histological features, proliferative and angiogenic capacities of the tissue specimens obtained from 6 patients who underwent surgery at our institution. Immunohistochemical staining for vascular endothelial growth factor (VEGF), its receptors FIk-1 and Fit-1, ephrin-B2, MIB-1 and proliferating cell nuclear antigen (PCNA) was performed using standard immunohistochemical techniques.Results A positive immunostaining was found for all antibodies studied except MIB-1, whereas nuclear endothelial expression of PCNA was observed in only 3/6 cases. VEGF stained positive in all of the available specimens (6/6). FIk-1 showed a positive immunoreaction in only 2/6 cases and Fit-1 in 4/6 cases. Ephrin-B2 was expressed in the majority (5/6) of the cases.Conclusions These results support the hypothesis that DAVFs might be acquired dynamic vascular malformations with low biological activity. Vascular growth factors like VEGF and ephrin-B2 might play a pivotal role in the formation of DAVF.     

Effect of amlodipine on apoptosis of human breast carcinoma MDA-MB-231 cells

Objective： To elucidate the effects of amlodipine on the proliferation and apoptosis of human breast carcinoma MDA-MB-231 cells. Methods： Light microscopy was used to determine the effects of amlodipine on cell morphology; Flow cytometry was used to quantitate cells undergoing apoptosis; the expression of a cell cycle-related protein, proliferating cell nuclear antigen （PCNA） and an antiapoptosis protein, Bcl-2 were assessed by immunocytochemistry. Results： Amlodipine concentration of 8.25umol/L （1/2 of ICs0） affected the morphology, decreased the expression of PCNA and Bcl-2 and induced apoptosis of human breast carcinoma MDA-MB-231 cells. Conclusion： The effect of amlodipine on the antiproliferation of human breast carcinoma MDA-MB-231 cells is related to inducement of apoptosis, and the decrease of the expression of Bcl-2 and PCNA may be the possible mechanism for proliferation inhibitory and inducement of apoptosis.     

Detection of herpes simplex virus type 1 in rheumatic valvular tissue

Background Rheumatic heart disease (RHD) is the most important sequela of rheumatic fever (RF):evidence that streptococcal infection is aetiological is prominent, but sometimes contradictory. Acute HSV-1 infection in mouse leads to carditis and valvulitis whereas recurrent infection results in inflammatory granulomatous lesions that resemble Aschoff bodies. Cells containing HSV-1 inclusions or virus infected giant cells appear similar to Anitschkow cells or Asehoff cells respectively. We hypothesized that HSV-1 infection also may be involved in RHD.Methods Formalin-fixed, paraMn-embedded valvular tissue samples from 32 patients with RHD were investigated for evidence of HSV-1 infection. HSV-1 antigen was detected by immunohistoehemistry, using HSV-1-specific monoclonal and polyclonal antibodies. HSV-1 glyeoprotein D gene sequences were amplified by nPCR,using β-globin gene amplification in the same samples as internal control. Valvular tissue from 5 cases of sudden death and 3 cases died of neisseria meningitis without a history of valvular disease was used for comparison. HSV-1-infected lung tissue was used as positive control.Results HSV-I antigens were detected in valvular tissues from 21 of 32 (65.6%) patients. Fifteen of these 21 (46.9% of cases), but no antigen-negative sample, were positive also for HSV DNA. Nueleotide sequence of PCR products was homologous to the targeted region of the HSV-1 glycoprotein D gene. HSV-1 antigen was present also in one case of sudden death but viral DNA was not found in any tissue sample from tile comparison group. Results from reagent and positive controls were as anticipated.Conclusions This is the first study to show the presence of HSV-1 antigen and genomic DNA in valvular tissues from patients with RHD and provides evidence for an association of HSV-1 infection with some cases of rheumatic valvular disease.     

食管基底细胞样鳞形细胞癌临床病理研究

目的：进一步阐明少见的食管基底细胞样鳞形细胞癌（BSC）的临床病理学特征。方法：收集21例食管BSC分别作苏木精-伊红（H-E）染色及组织化学、免疫组化染色（S-P法），其中7例作电镜观察，并进行随访。结果：BSC最主要的组织学特征是基底细胞样癌呈实体小叶状、筛网状排列，癌巢中央见粉刺状坏死，并恒伴有鳞形细胞癌或癌巢内伴有小灶性鳞状成分，表面黏膜可伴有异型增生或原位癌。免疫组化染色，肿瘤内的基底样细胞示：细胞角蛋白（CK）14／16（＋），上皮膜抗原（EMA）15／16（＋），波形蛋白（Vimentin）14／16（＋），S-100蛋白7／16（＋），癌胚抗原（CEA）和平滑肌肌动蛋白（SMA）阴性。电镜下示基底样癌细胞分化差，细胞器少，细胞间有少量桥粒，癌细胞梁索间可见特征性的多层折叠或指纹状的基膜。15例获随访结果，其中13例Ⅱ期以上者有10例于手术后2年内死于肿瘤复发或转移。结论：食管BSC是一种新的独立的临床病理类型，因分化低、恶性程度高，预后较差．     

20例睾丸和附睾腺瘤样瘤的临床病理、特染及免疫组化观察

目的：用特殊染色,免疫组化的方法探讨性生殖系统腺瘤样瘤的组织发生,特性及鉴定诊断。方法：收集睾丸和附睾腺瘤样瘤 病例20例,选用细胞角蛋白（CK）,波纹蛋白（VIM）,上皮膜抗原（EMA）,癌胚抗原（CEA）,S－100蛋白,血管VIII因子相关抗原（FVIII）,平滑肌肌红蛋白（SMA）,溶菌酶LYS,神经原纤维（NF）作为第一抗体,应用免疫组化方法（SP法）以及特殊染色,AB－PAS染色,网状纤维染色,Masson三色染色,观察其临床病理特征,结果：免疫组化结果显示20例睾丸和附睾腺瘤样瘤的KCK及VIM均呈强阳性,EMA8例阳性（40％）S－100蛋白2例阳性（10％）,SMA10例阳性,CEA,EMA,FVIII,MYO及NF均无表达,AB－PAS染色20例匀有阳性反应,网状纤维染色见瘤细胞周围网状纤维少,Masson三色染色部分间质呈红色,结论：男性生殖系统腺瘤样瘤间为皮起源,附睾及其附近为最常见部位,CK,VIM及AB－PAS染色是睾丸及附睾 瘤样瘤的阳性标记物,特染及免疫组化结果可作为诊断及鉴别诊断的重要参考依据。其生物学行为良性,预后良好。     

Basaloid squamous carcinoma of the esophagus:clinicopathological,immunohistochemical and electron microscopic studies of 16 cases

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涎腺肿瘤免疫组化研究

应用6种抗体(CEA、MAK-6、EMA、S-100、GFAP、HHF-35)对55例涎腺肿瘤和9例肿瘤旁涎腺组织进行免疫组化研究,结果表明:肿瘤旁涎腺组织的免疫光谱,以腺管上皮细胞为一端,肌上皮细胞为另一端,润管细胞和腺泡细胞位于光谱中间段,多形性腺瘤起源于同一类细胞,而不是混合性起源。简略介绍HHF-35的性质和应用价值,在识别肌上皮细胞方面,HHF-35比GFAP更为敏感。     

食管基底细胞样鳞状细胞癌临床病理分析

目的探讨食管基底细胞样鳞癌(BSC)的临床病理特点.方法回顾分析10年间食管癌手术病理档案,对符合基底细胞样鳞癌的病例行免疫组化标记.结果2*!960例食管癌中10例为食管基底细胞样鳞癌,占0.3%.其中8例曾误诊为低分化鳞癌、腺鳞癌、腺样囊性癌和小细胞癌.肿瘤主要由核大浆少的基底细胞样癌组织与多少不等的鳞癌或(和)原位鳞癌构成,两者间无移行.1例部分细胞免疫组化癌胚抗原阳性,2例局部癌组织突出素、嗜铬素阳性,1例灶性角蛋白17、角蛋白8阳性;无一例表达S100和肌动蛋白.结论食管基底细胞样鳞癌主要由基底样细胞组成,并具有鳞癌的变异型,具有高侵袭性又易在病理上误诊,因而需要对其临床和病理特征进行透彻研究和进一步提高认识.     

免疫组化在恶性间皮瘤鉴别诊断中的应用价值

目的 对恶性间皮瘤进行免疫组化的对比性研究,以进一步评价它们在恶性间皮瘤鉴别诊断中的价值。方法 采用免疫组织化学染色(LSAB)法对22例恶性间皮瘤和20例对照组腺癌组织进行EMA、CEA、CK、vimentin、间皮细胞(M-cell)等5种单克隆抗体的标记。结果 恶性间皮瘤的阳性率分别为 M-cell 91％,CEA 9％,vimentin 73％,CK82％,EMA64％;而腺癌的阳性率为分别为M-cell5％,CEA 85％,vimentin 15％,CK100％,EMA 100％。结论 在恶性间皮瘤和腺癌的鉴别诊断中,M-cell和CEA是较理想的标记物,尤其是M-cell,CEA和vimentin三种抗体联合应用更具有价值。而在恶性间皮瘤和其他梭形细胞肿瘤的鉴别诊断中,还应联合使用CK和EMA。     

Ⅳ型胶原与层粘蛋白在口腔鳞状细胞癌中的表达及其意义

目的 探讨口腔鳞状细胞癌(Oral Squamous Cell Carcinoma,OSCC)中Ⅳ型胶原(Col-Ⅳ)和层粘蛋白(LN)的表达与肿瘤临床参数的关系。方法 采用免疫组织化学SP法对45例口腔鳞状细胞癌标本,9例正常口腔粘膜标本中的Col-Ⅳ和LN的表达进行检测。结果 ①Col-Ⅳ与LN在口腔鳞状细胞癌中的表达具有高度一致性。②Col-Ⅳ和LN的异常表达与肿瘤分化程度、颈淋巴结转移呈负相关。③Col-Ⅳ和LN的表达与肿瘤的临床分期和肿瘤发生部位无关。结论 Col-Ⅳ与LN表达明显减少和缺失可做为口腔鳞状细胞癌浸润扩散能力强的生物学指标,有助于判断口腔鳞状细胞癌的恶性程度和生物学行为。     

分泌型脑膜瘤的临床和病理特点

目的探讨分泌型脑膜瘤的临床病理和免疫表型特点。方法对9例分泌型脑膜瘤分别进行HE、过碘酸-希夫（PAS）和PAS酶消化后（PAS—D）染色；用免疫组织化学SP法检测ER、PR、CEA、CK、EMA和MIB-1等标记；1例肿瘤进行电镜观察。结果9例分泌型脑膜瘤分别位于碟骨嵴、左顶部或额部；5例有明显的肿瘤周围水肿；9例中7例随访时间6～88月，未见复发；组织学特点：细胞质内或细胞间均可见大小不等的透明包涵体，血管周细胞增生；9例包涵体PAS染色及PAS—D染色均呈阳性；免疫表型检测显示9例肿瘤细胞PR阳性，8例ER阴性，7例包涵体和周围细胞CEA、CK、EMA阳性。1例MIB-1阳性率为5％，余8例MIB阳性率不超过2％。电镜观察见细胞间腔隙形成及含有粘液的细胞。结论分泌型脑膜瘤是一种少见的颅内脑膜瘤亚型，显示腺上皮分化，复发率低，预后较好。     

肾上皮样血管平滑肌脂肪瘤的临床病理和免疫组化特征

目的：探讨肾上皮样血管平滑肌脂肪瘤(AML)的临床病理特点及鉴别诊断。方法：对2例肾上皮样AML的临床资料、随访结果及组织病理进行分析，应用免疫组化染色观察了黑色素瘤相关抗原(HMB45)、(Pan)melanoma markerAb-1、角蛋白(CK)、平滑肌肌动蛋白(SMA)、上皮膜抗原(EMA)、波形蛋白(VIM)、肿瘤抑癌基因(p53)、增殖细胞核抗原(PCNA)、细胞增殖标记(Ki67)等在肾上皮样AML的表达，并与国外报道的28例进行比较。结果：组织学特点：瘤细胞呈上皮样或短梭形，细胞质为嗜酸性，核大，较深染、异形，可见病理性核分裂相。瘤细胞排列密集成片，伴大片坏死，瘤组织中散在多核巨细胞。免疫组化：HMB45和(Pan)Melanoma Marker Ab-1阳性，上皮标记EMA、CK阴性。结论：在未出现远处转移前，肾上皮样AML一般属潜在恶性病变。免疫组化对鉴别诊断有重要意义。     

腹膜原发性腺癌临床病理分析及鉴别诊断

目的 探讨腹膜原发性腺癌的临床病理特点和鉴别诊断.方法 回顾性分析8例腹膜原发性腺癌的临床病理资料,患者均为女性,平均年龄55.7岁,以腹胀、腹水为主要临床表现;其中浆液性乳头状腺癌7例,黏液腺痛1例.对肿瘤进行光镜观察,并做免疫绀化染色.结果 免疫组化染色显示肿瘤广谱CK、CK7、CK20、CA125、CEA、EMA、波形蛋白、钙网膜蛋白、P53 阳性比例分别为8/8、8/8、1/8、7/8、7/8、8/8、1/8、1/8、8/8.结论腹膜原发性腺癌被认为是起源于第二苗勒氏管系统的恶性肿瘤,起病隐匿、发病率低,与卵巢肿瘤相似有多种组织类型,因此容易误诊,免疫组化染色有助于鉴别诊断.