目的 探讨喉鳞状细胞癌(LSCC)中人乳头瘤病毒(HPV)及细胞周期相关蛋白p16的表达及意义.方法 收集北京同仁医院2005-01-01-2011-12-31 LSCC病例资料332例,回顾性分析其临床病理学特点,采用聚合酶链反向点杂交(PCR-RDB)方法检测HPV16 DNA,构建组织芯片并采用免疫组织化学法染色观察细胞周期相关蛋白p16的表达,比较HPV阳性组和HPV阴性组临床病理学特点的差异,分析高危型HPV16与p16的关系;采用RNAscope方法检测HPV16 E6/E7 mRNA的表达,观察在LSCC中是否具有转录活性的HPV,从而明确LSCC是否与HPV有关联.结果 在332例LSCC中有45例(45/332,13.6％)检测到高危型HPV,其中40例(40/332,12.0％)检测到HPV16,HPV 16阳性与淋巴结内肿瘤囊性转移有关联,χ2=9.774,P=0.004;T1患者HPV16阳性率高于T4患者,P=0.011;组织学分级也对患者是否表达HPV16 DNA有一定的影响,P=0.049.与LSCC患者年龄、性别、是否吸烟/饮酒、TNM分期及肿瘤大小等均无统计学意义的关联,均P>0.05.在332例LSCC中36例(36/332,10.84％)p16阳性表达,其中女性阳性率高于男性,差异有统计学意义的关联,χ2=4.841,P=0.04;但与年龄、是否吸烟/饮酒、TNM分期、肿瘤部位、组织学类型、组织学分级及淋巴结转移无统计学意义的关联,均P>0.05.HPV16表达与p16表达有统计学意义的关联,χ2=3.944,P=0.047.但是p16与HPV16的表达并不完全一致,其敏感性仅为20％(8/40).332例LSCC均检测了HPV 16 E6/E7 mRNA的表达,染色结果均为阴性.结论 部分LSCC中伴有HPV16的感染,但HPV16并不是一个主要的致癌因素,且LSCC中p16表达并不是HPV16感染的替代性指标.

Objective To compare the consistency of immunohistochemical staining between the two commercial secondary antibodies. Methods Eighteen common immunohistochemical primary antibodies were selected and positive and negative controls were set up according to the recommendations from the AD Hoc Committee of International Experts. Under the same experimental conditions, the DAKO automatic immunohistochemical staining platform was used to test two different secondary antibodies for immunohistochemical staining. The standard group for the secondary antibody was provided by the DAKO polymer system (DAKO EnVision FLEX, High pH), and the experimental group for the secondary antibody was provided by the Power-StainTM kit (Power-StainTM 1.0 Poly HRP DAB Kit for Mouse+Rabbit). Subsequently, the images were captured. A single-blind, positioning, qualitative and semi-quantitative scoring criterion was used for describing the positive stains by the experienced pathologist. Absorbance corrected values, measured area values and positive integral absorbance were detected by the digital pathology quantitative measurement in the same areas from the two groups. Then, the mean absorbance was calculated. Results The stains of all the samples from the two groups showed accurate location and consistent qualitative evaluation. No significant differences were found between the two groups in all the semi-quantitative scoring, including stain intensity, positive stain percentages and mean absorbance. Conclusion The two commercial secondary antibodies have strong consistency in the immunohistochemical staining.

Hepatoid adenocarcinoma is a peculiar but aggressive type of extrahepatic adenocarcinoma that morphologically resembles hepatocellular carcinoma. To date, only a handful of cases have been described in the colon. Here, such a case is reported and pertinent literature is reviewed. Characteristic features of the tumor include large polygonal cells with marked nuclear atypia, prominent nucleoli and eosinophilic granular or clear cytoplasm arranged in trabeculae and solid nests or sheets. The tumor was immunohistochemically positive for AFP, CDX-2 and hepatocyte paraffin 1.

OBJECTIVE:To investigate the role of human papillomavirus (HPV) in laryngeal squamous cell carcinoma (LSCC) and analyse the relationship between HPV and the expression of cell cycle-related proteins. DESIGN:The study consisted of LSCC between 2005 and 2011 in Tongren Hospital. Clinical data such as age, sex, smoking/alcohol consumption, and TNM stage were collected. HPV DNA and cell cycle-related proteins were assessed in terms of clinical features. SETTING:Single-centre study. PARTICIPANTS:A total of 332 LSCC patients were included in the study. MAIN OUTCOME MEASURES:The presence of genotype-specific HPV DNA was evaluated using PCR-RDB in formalin-fixed paraffin-embedded tissues. All samples were also evaluated for p16INK4A , p21WAF1/CIP1 , P53, Cyclin D1, and Ki67 immunohistochemical staining by tissue microarray. RESULTS:HPV DNA was detected in 45 of 332 (13.55%) patients with LSCC, with HPV-16 being the predominant genotype. The presence of HPV-16 DNA was significantly associated with basaloid squamous cell carcinoma and cystic lymph node metastasis (P < 0.05). Of the 332 patients, 36 (10.84%) were scored as p16INK4A positivity and they were more likely to be female (P < 0.05). Cyclin D1-positivity and p21WAF1/CIP1 -positivity were observed in 60.24% (200/332) and 40.66% (135/332), respectively. In 114 cases (34.33%), LSCCs had moderate- to -strong p53 accumulation, which was correlated with TNM stage (P < 0.05). HPV-16 DNA was correlated with p16INK4A and manifested a higher Ki-67 labelling index and p21WAF1/CIP1 expression than HPV-16-negative tumours (P < 0.05). No relationship was observed between Cyclin D1or P53 expression and HPV-16 infection (P > 0.05). CONCLUSION:HPV DNA was detected in 13.55% patients with LSCC, with HPV-16 being the predominant genotype and it was correlated with p16INK4A and manifested a higher Ki-67 labelling index and p21WAF1/CIP1 expression than HPV-16-negative tumours.

AIMS To investigate the frequency and transcriptional activity of HPV and its correlation to p16 and p21 expression in basaloid squamous cell carcinoma (BSCC) of the larynx. METHODS We evaluated tissues from 29 patients with BSCC of the larynx for the expressions of p16 and p21 proteins by immunohistochemistry (IHC) and for HPV E6 and E7 mRNA by RNA in situ hybridization (ISH). The presence of genotype-specific HPV DNA was evaluated using PCR-RDB in formalin-fixed paraffin-embedded tissues. P16 and p21 expression and HPV DNA status were correlated with clinicopathological features. RESULTS HPV DNA was detected in 8 of 29 (27.59%) patients, with HPV-16 being the predominant genotype. P16 and p21-positivity were observed in 7/29 (24.14%) and 8/29 (27.59%) patients, respectively. HPV was not correlated with p16 expression (P > 0.05). However, p21 expression was significantly higher in HPV-positive tumors than in HPV-negative tumors (P < 0.05). No cases exhibited transcriptionally active HPV in our series. CONCLUSION Our findings suggest that a small fraction of BSCC of the larynx is HPV DNA-positive in this Chinese population, p21 expression was significantly higher in HPV-positive tumors, and no cases were HPV transcriptionally active in this small cohort. Further research of HPV and its role in BSCC of the larynx are warranted.

Objective To investigate the clinicopathological features of the carcinoma arising in phyllode tumors of the breast.Methods Morphological observation and immunohistochemistry using Envision method were performed on 5 cases of carcinoma arising in phyllodes tumors of the breast from January 2000 to may 2017.Results All 5 patients were female,aged 26 ～ 58 years (median age 53),and all the masses increased rapidly in short time.2 cases arose from benign phyllode tumors including DCIS,LCIS and ILC;3 DCIS arose from borderline phyllode tumors.Immunohistochemistry showed all malignant epithelial components were diffusely positive for ER and PR,and DCIS was positive for p120 and Ecadherin,LCIS was positive for p120 and negative for E-cadherin;mesenchymal component all expressed CD34 and 2 cases were also Bcl-2 positive.All patients were alive without recurrence and metastasis followed by 4 ～ 72 months.Conclusion Carcinoma arising from phyllode tumors of the breast is rare and characterized by in situ and invasive carcinoma in a background of phyllode tumor of the breast.The immunohistochemical features of malignant epithelial component are similar to ordinary breast cancer,but with better prognosis.Because of its rarity,there is no standardized treatment prescription for this disease.

Objective: To study the significance of HPV and cell cycle related proteins in basaloid squamous cell carcinoma (BSCC) of the larynx. Methods: Twenty-nine cases of laryngeal BSCC from Beijing Tongren Hospital from January 2005 to December 2011 were reviewed. HPV typing by polymerase chain reaction-reverse dot blot (PCR-RDB) and p53, Ki-67, p16, p21 and cyclin D1 expression by immunohistochemistry were performed. The relationship between these indicators, various pathologic parameters (TNM, tumor size, tumor site and lymph node metastasis) and HPV status was analyzed. Results: There were 27 male and 2 female patients. The median age was 62 years. Lymph node metastasis and supraglottic tumor location were slightly higher than that of "usual" SCC, but not statistically significant (P>0.05). HPV DNA was detected in 27.6% (8/29) of the laryngeal BSCC, and all were HPV16. The expression of HPV was not related to age, alcohol consumption, tumor stage and tumor size. p53 was expressed in 31.0%(9/29) of laryngeal BSCC, and these cases were more likely supraglottic and had lymph node metastases (P<0.05). p16 staining was seen in 24.1% (7/29) of laryngeal BSCC, and these cases showed slightly higher rate of lymph node metastasis compared to p16 negative cases. The expression rates of p21 and cyclinD1 were 27.6% (8/29) and 69.0%(20/29), respectively, which were not related to age, tumor size, stage, lymph node metastasis, smoking and drinking. There were only 3 p16+ /HPV+ cases, which showed higher p21 and Ki-67 index compared to the HPV negative group (P<0.05). Conclusion: Some laryngeal BSCC expresses HPV DNA, possibly indicating an association with HPV; but p16 expression is not a reliable indicator for HPV infection.

目的 分析散发型淋巴管肌瘤病(S-LAM)的临床及病理特点,提高对该病的认识.方法 对11例经病理证实的S-LAM进行回顾性分析,采用SMA、vimentin、HMB45、Ki-67、CD34、D2-40、ER和PR等抗体进行免疫组化染色,并复习文献.结果 11例均为女性,表现为乳糜性胸或腹水,有胸闷、呼吸困难、气促等症状;病程中均有多个部位累及,包括肺、腹膜后、淋巴结.胸部CT示两肺散在或弥漫分布囊腔影,直径0.2 ～2cm,囊壁薄而清晰;腹膜后受累者CT显示囊性低密度影.11例均有月经不调,其中3例合并子宫肌瘤,1例合并卵巢畸胎瘤、肾血管平滑肌脂肪瘤及肝血管瘤,1例合并小肠胃肠间质瘤,2例合并肝囊肿.免疫组化显示11例HMB45均(+),5例ER灶性(+),8例PR局灶(+).随访5～ 60个月,患者均存活,且症状缓解.结论 LAM的临床表现无特异性,CT表现有一定的特异性,最终确诊依赖于组织病理,熟悉这些特点有助于早期诊断,但预后差.

Purpose To assess the clinical and pathological features of small gastrointestinal stromal tumours (sGIST).Methods To reevaluated the clinical,histological and immunohistochemical parameters of 21 sGISTs.The standard immunohistochemical panel antibodies were studied on the tumor sections.All data were compared with clinical sGIST.Results There were a total of 7 females and 14 males of sGISTs.The median age was 63 years old.The tumors were predominantly located in the stomach showing a spindle cell morphology and the tumor sizes ranged from 0.5 cm to 1.5 cm.9 sGISTs combined with malignant tumors,which were gastric cancer have been incidentally detected during surgery.As the lesions were small in size,with infrequent bleeding,necrosis,mucosal invasion,ulceration and less mitotic index,sGISTs reoccurred less compared with clinical sGIST.p53,Ki-67 labeling index and microvascular density (MVD) in sGIST were significantly lower than clinical sGIST (P ＜ 0.05).Conclusion sGIST may occure with digestive tract cancer synchronously.p53,Ki-67 labeling index and MVD were lower than clinical GIST,which means better prognosis.

目的 探讨肾移植后淋巴组织增生性疾病(PTLD)的临床病理学特征,提高对该疾病的诊治水平.方法 报道2例肾移植后PTLD,采用CD20、CD3、CD45RO、CD79a、bcl-6、CD10、Mum-1和Ki-67等抗体进行免疫组化染色,并进行EBV原位杂交检测,同时结合文献复习进行讨论.结果 2例患者肾移植术后均采用三联免疫抑制疗法,PTLD诊断时间分别为3年和16年.2例病理组织学类型均为单型性PTLD,其中1例为外周T细胞性淋巴瘤,1例为弥漫性大B细胞性淋巴瘤,均无特异性临床表现.确诊后均将免疫抑制剂减量,其中1例辅以利妥昔单抗治疗.1例患者确诊后短期内死亡,另1例患者生存.结论 PTLD是发生在器官移植后具有独特形态和临床特征的淋巴组织增生性疾病,预后较差,采用免疫抑制剂减量和抗利妥昔单抗治疗有效.

AIMS:To study the clinicopathologic features of Stewart-Treves syndrome (STS) in postmastectomy patients including the epidemiology, presentation, morphology, differentiation, pathogenesis and therapeutic options.METHODS AND RESULTS:Ten cases of STS in postmastectomy patients were retrospectively identified in our archives, and immunohistochemistry for CD34, CD31, D2-40, HHV-8, CK, EMA and Ki-67 was performed. All ten patients presented with lymphedema after mastectomy as the first sign. Physical examination revealed multiple raised, pinkish-red papulo-vesicular lesions or ulceration as the early evidence of tumor in the field where radiation therapy was introduced. Microscopic examination revealed infiltrative proliferation of vessels and the heteromorphic tumor cells expressed CD34, CD31 and D2-40. Despite the various treatment modalities, 5 patients died in an average of 19 months, 4 patients survived to the last follow-up (9-31 months), and 1 patient got lost.CONCLUSIONS:STS is a fatal complication of postmastectomy lymphedema. Patients with STS have very poor prognosis. The key to improve patient's survival is the early diagnosis through a high alert of this disease by primary care physicians and comprehensive physical examination of patients with pertinent history and suspicious clinical presentations followed by prompt biopsy for definitive diagnosis.

目的 探讨黏液样脂肪肉瘤的影像学及病理学特征.方法 采用免疫组化法对15例黏液样脂肪肉瘤进行染色,观察其影像学特征,并复习相关文献.结果 15例患者术前CT显示肿物为界限清楚的低密度影,MRI显示SE T1WI上为低信号,而T2WI上为明显高信号,类似于水的密度或信号,影像学检查缺乏特异性诊断,不易分辨肿物性质.组织学形态:由不同成熟阶段的脂肪母细胞、黏液样基质及毛细血管网组成,部分区域具有“肺水肿样”特征.免疫表型:脂肪母细胞S-100阳性,免疫组化标记CD34可见“鸡爪样”的小血管.结论 黏液样脂肪肉瘤影像学特征往往表现为低密度水肿样影像,诊断无特异性,病理检查可明确诊断.

Purpose To investigate the significance of TKTL1 expression in invasive ductal carcinoma(IDC) of breast.Methods The expression of TKTL1 was detected in 54 cases of IDC and 10 cases of ductal carcinoma in situ(DCIS) by immunohistochemical staining.The relationship between the TKTL1 expression and pathologic features(ER,PR,histological grade,tumor size,vessel invasion,nerve invasion and lymph node metastasis) were also analyzed.Results(1) The expression of TKTL1 had no significant difference between IDC and DCIS(P0.05).(2) The expression of TKTL1 was positively correlated with lymph node metastasis(P=0.047),but had no relationship with ER,PR,tumor size,vessel invasion,nerve invasion and histological grade(P0.05).Conclusions Overexpression of TKTL1 may correlated with invasion and lymph node metastasis in IDC.

Objective To investigate the expression of Bcl-2,P53,CyclinD1 in B-cell lymphoma and their relevance with the development of B-cell lymphoma.Methods The preparation of high-throughput sample was made by method of tissue microarray.The expression of Bcl-2,P53,CyclinD1 in B-cell lymphoma was examined by using immunohistochemical staining.Results The expression of Bcl-2 and P53 was significantly higher in DLBCL,FL,MALT,MCL and SLL than that in normal tissues.CyclinD1 was only detected in DLBCL and MCL.Conclusions The coordination expression of Bcl-2 and P53 and the abnormal expression of CyclinD1 were two important mechanism of the occurrence of DLBCL.

Purpose To explore the clinicalpathological features of the tumor and tumor-like lesions of the internal auditary canal (IAC) and cerebellopontine angle (CPA).Methods The four cases of tumor of the IAC and CPA were studied by immunohistochemical staining and light microscopy,combined with MRI and literature review.Results In four cases,one was lipochoristoma (lipomatous tumors) of the acoustic nerve,two were low-grade papillary adenocarcinoma of endolymphatic sac origin,and another one was juxtaoral organ of Chievitz presenting clinically as a tumor mass.Conclusions Lipomatous tumors in internal auditory canal and cerebellopontine angle are very rare; low-grade papillary adenocarcinoma of endolymphatic sac origin (Albeit locally aggressive) is rarely reported and has low malignant potential. Juxtaoral organ of Chievitz presents clinically as a tumor and may be mistaken as perineural invasion of carcinoma if the surgical pathologist is unaware of its existence.

[Objective] To investigate the clinicopathologic, immunohistochemical and molecular biological features, and to emphasize in studing and summarizing special kinds of primary cutaneous lymphoma.[Methods]There were 40 cases which were previously diagnosed as primary cutaneous lymphoma from 1983 to 2001 in Beijing Railway Hospital (including clinical data and tissue blocks). All cases were investigated by using HE and immunohistochemical methods, and TCRβ/γ and IgHFr2A / Fr3A gene assay were employed in 25 cases. The follow-up information was obtained for all cases. [Results]Median age was (41.53±20.14) years(range 2~73 years). The lesions showed in generalize, especially in trunk and upper limb. Nodule was much more than others. Some various subtypes (for example, PCCD30 positive/negative large-cell lymphoma, primary cutaneous diffuse large B cell lymphoma, primary cutaneous blastoma and subcutaneous panniculitic T cell lymphoma) had different clinical and histological features. Immunohistochemical method and gene rearrangement were helpful to diagnosis. [Conclusion]①Primary cutaneous lymphoma was not rare, and most of them were primary cutaneous T-cell lymphoma. ②There were some special subtypes in primary cutaneous lymphoma that were different from nodal or extranodal lymphoma (except skin). The revised classification (integration of EORTC and WHO2001) should be used.③CD30 was a primary sign of prognosis of primary cutaneous large-cell lymphoma. CD30-positive was good, whereas poor.④Two or more antibodies had better use in diagnosis. ⑤Gene rearrangement assay was helpful in distinguishing from early or borderline pathological changes of primary cutaneous lymphoma.

Objectiveand To analyze the clinical,pathologic and radiological features of endolymphatic sac tumor(low-grade papillary adenocarcinoma)and to investigate its characteristics.Methods To analyze the data of endolymphatic sac tumor(low-grade papillary adenocar cinoma),and the clinical manifestations of recurrent right ear's hearing drop and right side facial nerve paralysis.MRI and CT display the right Temporal Bone occupying and permeatal destruction of bone.Results Clinical signs at presentation range from a mass in the external ear canal to sensorineural deafness and cranial nerve palsy.Imaging studies reveal a destructive lesion of the petrous bone which was heterogeous on CT/MRI scanning.Light microscopy reveals two chief patterns:a follicular pattern and a papillary/solid pattern.Conclusion Endolymphatic sac tumor is a rare tumor with an endolymphatic sac origin,which may extend into the cerebellopontine angle.It must be differentiated from middle ear adenoma,jugular paraganglioma,choroid plexus papilloma,meningioma and metastatic carcinoma of the thyroid.

Aim: To investigate the effect of type I insulin like growth factor (IGF-1) on the wound healing of rats, so as to provide theoretical and experimental evidence for clinical application of IGF-1 to accelerate the process of wound healing. Methods: The experiment was conducted in the Experimental Animal Laboratory of Beijing Shijitan Hospital from May to June 2002. (1) Thirty-two clean grade Sprague-Dawley rats in either gender with body mass of 180 to 200 g were selected and randomly divided into IGF-1 group, IGF-1 and recombinant human growth hormone (rhGH) group, rhGh group and normal saline (negative control) group with 8 rats in each group. (2) The round wound models with diameter of 1.6 to 1.8 cm were made in the back skin of rats and given interventions after hemostasis by compression. (3) IGF-1 group: Every wound was given liquid of 0.1 mL with IGF-1 concentration of 1 mg/L; IGF-1 and rhGH group: The dosage was 0.1 mL with IGF-1 concentration of 1 mg/L and rhGH concentration of 0.2 U/mL; rhGH group: The dosage was 0.1 mL with rhGH concentration of 0.2 U/ mL; Negative control group: 0.1 mL physiological saline was applied in every wound. (4) The specimens in the wound site were harvested after 1, 4, 7, 11, 14 and 17 days. The same dosage medicament was applied in the residuary wounds. The healing tissue was sampled at 14 and 17 days after the wound site was healed after 11 and 13 days. The samples were fixed with 10% formalin for 10 hours, differentiated and dehydrated in alcohol. (5) Immunohistochemistry (IHC) and Image analysis was employed to detect the relationship between the related factors of wound healing: Proliferating cell nuclear antigen (PCNA) and Type I collagen in the process of wound healing after the application of ectogenous IGF-1 and rhGH. Results: Thirty-two rats were involved in the result analysis. (1) Changes of expression of type I collagen after IGF-1 and rhGH were applied in wound: The expression of type I collagen was found in the cytolymph in IGF-1 group, IGF-1 and rhGH group after 1 day of wound, the expression kept the trend of increase in 11, 14 and 17 days after wound. (2) Changes of expression of PCNA: The expression of PCNA in the cell nucleus in the IGF-1 group, IGF-1 and rhGH group reached the peak value at day 4. In the rhGH group and negative control group, the expression of PCNA gradually increased and arrived the peak value at day 7, then reduced step by step to the normal range after wound healed at days 14 and 17. (3) Expression of IGF-1: In the IGF-1 group, IGF-1 and rhGH group, the expression of IGF-1 in the cytolymph and cell nucleus always kept the peak value. In the rhGH group and negative control group, the expression of IGF-1 was found after 1 day and arrived the peak value at day 7, reduced from day 11 and to the normal range after wound healed in 14 days and 17 days. (4) The t-test was used in the comparison among different groups. Conclusion: Application of ectogenous IGF-1 in wound will help to the splitting, proliferation and excretion of fibroblast; it is the direct promoter for proliferation of fibroblast and will accelerate the process of wound healing. On the other hand, the effect of rhGH used in wound is not powerful.

AIM: To investigate the effects of angiotensin-Ⅱ (Ang-Ⅱ) on the proliferation and collagen synthesis in rat cardiac fibroblasts and the expression of Ang-Ⅱ receptor on fibroblasts. METHODS: [3H]-TdR and [3H]-prolin at different concentrations were incubated with the confluent fibroblasts of newborn rats stimulated by Ang-Ⅱ. Receptor of Ang-Ⅱ on fibroblasts and intracellular free calcium ions were examined by autoradiograms and fluorescence technique. RESULTS: In the presence of Ang-Ⅱstimulation, the increase in incorporation of [3H]-TdR and [3H]-prolin was much higher than that of control in a dose-dependent manner. Autoradiograms showed that Ang-Ⅱ was uniformly distributed over the membrane of the fibroblasts. The silver grains on fibroblasts were obviously decreased after adding unlabeled Ang-Ⅱ by the autoradiograms. The concentration of free calcium ions was increased in fibroblast. CONCLUSION: These findings suggest that Ang-Ⅱ promotes fibroblast proliferation and the syntheses of collagen protein, in which Ang-Ⅱ binds to the membrane receptor of fibroblasts to activate the signal system in cells, such as intracellular free calcium ions. [