Immunohistochemical Identification of Decorin and Biglycan in Human Dentin: A Correlative Field Emission Scanning Electron Microscopy/Transmission Electron Microscopy Study

Decorin and biglycan, two small leucine-rich proteoglycans, have been proposed to play important roles in matrix-mediated formation of mineralized tissues, and their three-dimensional arrangement in human dentin is still not completely understood. The aim of this study was to immunohistochemically analyze the distribution of decorin and biglycan in human predentin/dentin organic matrix under a high-resolution field emission in-lens scanning electron microscope (FEI-SEM) and a transmission electron microscope (TEM). Tooth dentin specimens were submitted to either a preembedding or a postembedding immunolabeling technique using primary antibodies antidecorin and antibiglycan and gold-conjugated secondary antibodies. Correlative FEI-SEM/TEM observations showed that the two antibodies yielded a similar labeling pattern over the processes of odontoblasts and the predentin. Decorin and biglycan were mainly associated with the collagen fibers within the predentin layer, revealing a moderate immunoreaction that was significantly higher compared to the one observed on dentin. Thus, a generally weak labeling for decorin was found in dentin, which, however, was significantly higher on odontoblast processes within dentinal tubules than in intertubular dentin. On the other hand, biglycan immunolocalization on dentin revealed few gold particles rather uniformly distributed, without showing significant differences between tubular and intertubular regions. In conclusion, this study reveals distinct distribution patterns of decorin and biglycan and their relation with collagen. Decorin’s and biglycan’s precise roles within prematrix and mineralized matrix in human teeth should be further clarified.     

Squamous Cell Carcinoma of Parotid Gland Associated with Concurrent Lymphoepithelial Cysts and Lymphoepithelial Lesion: Case Report and Proposed Histogenesis

Lymphoepithelial cyst and lymphoepithelial lesion have similar histologic features and an affinity for the parotid gland. Though considered as different entities, both conditions arise from heterotopic salivary epithelial rests or inclusions in intra- or peri-parotid lymph nodes. We present a case of squamous cell carcinoma of parotid gland associated with concurrent lymphoepithelial cyst and lymphoepithelial lesion in a patient who was not infected with human immunodeficiency virus. We propose that lymphoepithelial cyst and lymphoepithelial lesion have a similar histogenesis.     

人肾细胞癌原位移植裸鼠肺转移模型的建立

目的：建立肾癌原位肺转移动物模型,研究动物模型肾癌移植瘤和肺部转移瘤的生长情况,确定肾癌肺转移的初始时间。方法：人肾癌细胞株SN12-PM6种植于裸鼠右肾实质内,连续观察裸鼠肾脏肿瘤及肺部转移瘤的生长情况。所有采用标本10％甲醛溶液固定,常规石蜡包埋,切片HE染色后分析结果。结果：肾脏的成瘤率为100％（39／39）,第5周开始观察到肿瘤的肺转移灶,肺部转移率50％（4／8）,在随后3周其肺部转移率分别为75％（3／4）,100％（8／8）和100％（7／7）,肠道、肝脏、脾脏等脏器未发现明显的转移灶。结论：成功构建人肾细胞癌原位裸鼠肺转移动物模型,为肾癌特别是肾癌肺部转移提供了一个非常理想的研究模型。     

Incidence and Clinical Significance of Lymph Node Metastasis Detected by Cytokeratin Immunohistochemical Staining in Ductal Carcinoma In Situ

Background This study explored the long-term prognosis of patients with ductal carcinoma-in-situ (DCIS) and lymph node metastasis detected by cytokeratin immunohistochemical stains (CK-IHC).Methods Using the Columbia University breast cancer database, we identified all DCIS patients who had eight or more axillary nodes dissected and free of metastasis. Five-micrometer sections from all paraffin blocks containing lymph node tissue were stained with an anticytokeratin antibody cocktail (AE1/AE3 and KL1). The results of the CK-IHC and updated database were anonymized and merged. Survival of CK-IHC–positive and –negative patients was compared by using Kaplan-Meier curves and log-rank tests.Results CK-IHC was performed on 301 DCIS patients, who had an average of 16.7 axillary nodes dissected. Eighteen (6%) of 301 patients tested positive by CK-IHC. Seventy patients with bilateral breast cancer and 2 patients without any follow-up data were excluded, for a final study population of 229 patients. Among the 216 patients with negative CK-IHC, 18 patients died, compared with 1 of 13 patients with positive CK-IHC. The median follow-up for the study group was 127 months. Kaplan-Meier overall and breast cancer–specific survival estimates were similar for CK-IHC–positive and –negative patients (P = .81 and P = .73, respectively).Conclusions CK-IHC increases the incidence of positive nodes by 6% in DCIS patients. A positive node by CK-IHC does not seem to affect survival in these patients. These results raise concerns regarding the clinical significance of positive nodes by CK-IHC in DCIS patients.     

中药土贝母对人肾细胞癌影响的实验研究

土贝母制剂对体外培养的人肾颗粒细胞癌细胞系ＧＲＣ－１和裸鼠移植性人肾透明细胞癌ＲＬＣ－３１０的生长具有明显的抑制作用，并可诱导癌细胞凋亡。应用流式细胞仪（ＦＣＭ）分析药物作用前后癌细胞细胞周期的变化，结果提示土贝母可阻止ＧＲＣ－１及ＲＬＣ－３１０细胞由Ｇ０／Ｇ１期向Ｓ期进展，抑制ＤＮＡ合成，并使ＤＮＡ指数下降（Ｐ＜０．０１）。本研究为土贝母应用于临床治疗肾癌提供了重要的实验依据。     

Comparative Cytokeratin Expression in the Different Cell Types of Salivary Gland Mucoepidermoid Carcinoma

Mucoepidermoid carcinoma is the most common malignant salivary gland tumor, composed of several different cell types, with controversial histogenesis. The aim of this study was to assess the expression of cytokeratins in mucoepidermoid carcinoma, comparing to cytokeratin expression in normal salivary glands, in order to establish a possible correlation between tumor cells immunostaining and mucoepidermoid carcinoma histogenesis and differentiation. Eighty cases of salivary gland mucoepidermoid carcinoma were immunohistochemically examined with the use of antibodies against cytokeratins 6, 7, 8, 13, 14, 18, and 19. Cytokeratin expression varied according to the cellular type: squamous cells presented high expression of cytokeratins 6, 7, 8, 14, 18, and 19; intermediate and mucous cells of cytokeratin 7; clear and columnar cells of cytokeratins 6, 7, 8 and the latter also expressed cytokeratin 18. Cytokeratin 13 expression was low in all cell types. Cytokeratin immunoexpression in mucoepidermoid carcinoma was variable according to the cellular type; but regardless of the cellular type studied, cytokeratins 7 and 13 were, respectively, constantly high and low expressed. The immunoprofile of the normal salivary glands was variable according to the component but, in general, cytokeratin profile in mucoepidermoid carcinoma showed similarity to the immunoexpression on the excretory duct unit of normal salivary glands.     

Optical Microscopy and Electron Microscopy for the Morphological Evaluation of Tendons: A Mini Review

The morphological characteristics of tendons have been thoroughly evaluated via microscopy. Optical microscopy and electron microscopy are the most commonly used techniques for tendon tissue observation. According to the principles of both microscopy types, preparation and evaluation methods vary. Simple optical microscopy is commonly used in the observation of cells and extracellular matrix, and many stains, including hematoxylin–eosin, Van Gieson, Prussian blue, Alcian blue, and toluidine blue, are used for evaluating cells, collagen fiber arrangement, and noncollagenous proteins. Histological scoring systems have been used in many studies for semi‐quantification. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) are the most commonly used electron microscopy types, and special consideration is needed for the fixation and embedding protocols. Glutaraldehyde followed by osmium is most commonly used in the chemical fixation of tendon tissue, followed by epoxy resin embedment. Longitudinal sections captured in SEM images show the arrangement of collagen fibrils and the cells and lipid drops among them, while cross sections captured in TEM images show the diameter and distribution of collagen fibrils. SEM and TEM are used together for comprehensive evaluations. This mini review is focused on the preparation methodology and related evaluation indexes for the morphological evaluation of tendons.     

Acantholytic Squamous Cell Carcinoma in Upper Aerodigestive Tract: Histopathology, Immunohistochemical Profile and Epithelial Mesenchymal Transition Phenotype Change

Acantholytic squamous cell carcinoma is a rare variant of squamous cell carcinoma in the mucosa of upper aerodigestive tract. Histomorphologically, acantholytic squamous cell carcinoma may lose the typical features of conventional squamous cell carcinoma and mimic other epithelial or mesenchymal malignancies due to advanced acantholysis and dyskeratosis. Because of its rarity, information of prognosis, pathologic features and immunohistochemical profiles is limited. We have studied clinicopathologic features and immunohistochemical profiles of four acantholytic squamous cell carcinoma cases arising from upper aerodigestive tract. Clinical results indicate an aggressive biologic behavior. Morphologically, all tumors revealed significant acantholysis with separation of tumor cells and intratumoral spaces. The tumor cells were highly pleomorphic and growth patterns were variable. In immunohistochemical studies, all tumor cells revealed positive reactions for AE1/AE3 and p63 supporting a squamous epithelial origin. In contrast to conventional aerodigestive squamous cell carcinoma, acantholytic squamous cell carcinoma showed significant reductions of cytokeratin19, E-cadherin and concomitant up-regulation of vimentin expression. Both morphologic features and immunohistochemical profiles indicate that acantholytic squamous cell carcinoma has acquired an epithelial mesenchymal transition phenotype. However, in contrast to other solid malignant tumors, the epithelial mesenchymal transition phenotype change in acantholytic squamous cell carcinoma is not limited to the invasive front of the peripheral tumor but, rather, diffusely involves entire neoplastic lesion. In addition, because cytokeratin 19 staining is attenuated, this would be an insensitive marker for following up and/or in detecting disseminated tumor cells in cases of acantholytic squamous cell carcinoma in upper aerodigestive tract.     

Carbonic Anhydrase IX in Renal Cell Carcinoma: Implications for Prognosis,Diagnosis, and Therapy

Context

The clinical management of patients with renal cell carcinoma (RCC) remains difficult, and the development of new diagnostic, prognostic, and therapeutic tools is still required.

Objective

To review the current knowledge on the RCC-associated antigen carbonic anhydrase IX (CAIX) and provide evidence for how this antigen may aid in the clinical management of RCC.

Evidence acquisition

Clinical papers describing diagnostic, prognostic, and/or therapeutic applications of CAIX in RCC were selected from the Pubmed database. The search was manually augmented by reviewing the reference lists of articles.

Evidence synthesis

Expression of CAIX is regulated by the Von Hippel Lindau (VHL) protein (pVHL). Because of the invariable VHL mutational loss in clear-cell RCC (ccRCC) patients, CAIX expression is ubiquitous in ccRCC. Determination of CAIX expression in nephrectomy specimens of RCC patients improves prognostic accuracy; high CAIX expression appears to correlate with a favourable prognosis and a greater likelihood of response to systemic treatment for metastatic disease. Therefore, CAIX expression might be used to stratify metastatic ccRCC (mRCC) patients for systemic treatment. When incorporated into the RCC nomogram, CAIX expression seems to improve diagnostic accuracy for primary RCC as well as mRCC patients, but further evidence is required. Clinical studies with the CAIX-specific monoclonal antibody (mAb) cG250 have provided unequivocal evidence that ccRCC lesions can be imaged with radiolabeled cG250. Results are awaited of a large, randomised trial that aims to establish the value of cG250 imaging for primary RCC. The outcome of another large, placebo-controlled study is awaited to establish the usefulness of CAIX-targeted therapy in the adjuvant setting. Therapeutic trials with high-dose radiolabeled cG250 and CAIX-loaded dendritic cells in mRCC patients are still in phase 1 or 2.

Conclusions

CAIX improves diagnostic accuracy and is an attractive target for imaging of and therapy for ccRCC.     

Evidence of interepithelial seminoma spread into the rete testis by immunostaining of paraffin sections with antibodies against cytokeratin and vimentin

Summary Seminomas are tumors of high proliferative activity and show a marked tendency towards local invasion with the capacity for interepithelial spread within the seminiferous tubules as well as into the rete ductules. Immunohistologic investigations were carried out on paraffin sections of 47 typical seminomas. Immunostaining with antibodies against cytokeratin and vimentin allows the convenient detection of even small rete residuals in cases of subtotal rete destruction as well as the identification of discrete interepithelial seminoma spread within the rete ductules, thus facilitating seminoma staging.     

人肾癌细胞系RLC-310的建立和生物学特性

新建成的RLC-310细胞系来自人原发性肾透明细胞癌组织,在体外经过2年的连续培养,已传代130次。细胞可重叠生长,群体倍增时间32小时,集落形成率44％,具有超二倍体核型,染色体众数64。细胞没有支原体污染,有特异性的LDH同功酶谱,培养上清中可测得高浓度的白细胞介素6(IL-6)活性。细胞接种Balb/c裸鼠可形成移植瘤,并具有原发肾癌的组织学结构。     

Application of the Interferon Minipellet to Human Renal Cell Carcinoma in Nude Mice

Background The interferon (IFN) minipellet is a sustained-release formulation of human lymphoblastoid IFN, with atelocollagen used as the carrier material. We evaluated the antitumor effect of the IFN minipellet on an established human renal cell carcinoma cell line (KU-2) transplanted in nude mice.
Methods The treatment was started when tumor nodules had grown to 6 to 8mm in diameter. The IFN minipellet, or an aqueous solution of IFN, was given by subcutaneous injection, or peritumor injection, on days 1 and 10. Antitumor effects were evaluated according to tumor weights calculated as (long diameter) × (short diameter)²/2 in 7 groups consisting of 6 mice each.
Results IFN levels remained detectable in both tumor tissue and serum up to 10 days after peritumor injection of the IFN minipellet. Administered by the peritumor route, the IFN minipellet inhibited growth of the tumor significantly as compared with tumor growth in the untreated mice. The IFN minipellet showed greater inhibition of tumor growth by peritumor injection compared to subcutaneous injection. The aqueous solution of IFN was not effective either by subcutaneous or by peritumoral injection.
Conclusion Results indicate that the IFN minipellet is useful in the treatment of renal cell carcinoma.     

Simple Enucleation for the Treatment of PT1a Renal Cell Carcinoma: Our 20-Year Experience

Objectives

To evaluate the safety and efficacy of simple enucleation as a conservative treatment for pT1a RCC, and to report on the incidence of major complications, local recurrence, and progression-free and disease-specific survival rates.

Methods

We retrospectively reviewed the data of 232 patients who had nephron-sparing surgery (NSS) by simple enucleation between 1986 and 2004 for sporadic, unilateral, pathologically confirmed pT1a RCC. The patients’ status was evaluated last in September 2005. The mean (median, range) follow-up was 76 (61, 12–225) months.

Results

The mean (SD, median, range) tumor greatest dimension was 2.8 (0.78, 2.85, 0.6–4) cm. The histopathologic review according to the International Union Against Cancer and American Joint Commission for Cancer (1997) classification revealed 198 clear cell (85.3%), 18 papillary (7.8%), 15 chromophobe (6.5%) and one (0.4%) collecting duct RCCs. There were no major complications, such as prolonged acute tubular necrosis/chronic renal insufficiency and bleeding requiring open reoperation. One patient developed postoperative late retroperitoneal fluid collection consistent with urinoma, which required aspiration, drainage position and JJ stenting for 3 weeks. The 5- and 10-year cancer-specific survival were 96.7% and 94.7%, respectively. The 5- and 10-year progression-free survival were 96% and 94%, respectively. Overall, 13 (6.4%) patients had disease progression, three of whom had local recurrences alone (1.5%) elsewhere in the kidney; none had local recurrence at the level of the enucleation bed.

Conclusions

Simple tumor enucleation is a safe and acceptable nephron-sparing treatment that provides excellent long-term local control and cancer-specific survival rates.