Gene expression and immunohistochemical localization of basic fibroblast growth factor in renal cell carcinoma.

Renal cell carcinoma is known as a neoplastic condition of renal tubular cells and usually shows a hypervascular tumor in angiographic examination. We examined the presence of basic fibroblast growth factor (bFGF) in human renal cell carcinoma. To determine if alterations in bFGF gene expression are present in human renal cell carcinoma, paired samples of normal and neoplastic renal tissue from 6 patients were analyzed for bFGF mRNA content by Northern blot hybridization. In 4 out of 6 patients, tumor tissue expressed bFGF mRNA 2 to 4 times greater than corresponding normal tissue. Two patients showed minimal elevation of tumor bFGF mRNA. The localization of bFGF in the renal cell carcinoma tissue was also examined using immunohistochemical staining, and it was found that bFGF was positively stained at the nuclei of tumor cells and the cell surface. These results suggest that increased expression of bFGF may be associated with neoplastic growth in renal tubular epithelial cells and neovascularization.     

麝鼠香腺发育及腺细胞培养分离的初步研究

目前麝鼠香是麝香最好的天然替代物,麝鼠香来源于麝鼠生殖系统中的香腺,已发现其香腺中分泌细胞和支持细胞是麝鼠泌香的关键。本研究通过组织形态、HE染色、免疫组织化学染色及免疫荧光鉴定方法,初步描述了麝鼠香腺的发育过程。麝鼠香腺的形态结构显示其由小到大再至非泌香期萎缩;HE染色及组化结果显示,香腺在发育初期富含颗粒饱满的腺泡,雄激素分泌处于较低水平,分泌细胞数量较少,随着进一步发育,雄激素水平及分泌细胞数量逐渐升高,在两个月时达到最高,腺泡逐渐变大成熟并开始释放麝鼠香等分泌物;在泌香末期,腺泡逐渐被结缔组织取代。以上结果将有助于麝鼠香腺发育及分泌机制的研究,为提高麝香产量及实现体外泌香建立基础。另外,我们成功分离并鉴定了分泌细胞和支持细胞,以期为后续建立体外泌香体系奠定基础。     

麝鼠前列腺在繁殖期和非繁殖期转录组差异分析

麝鼠(Ondatra zibethicus L.)是季节性繁殖动物。成年雄性麝鼠在尿生殖孔上方肌肉与背皮之间有一对香腺,在繁殖期能分泌麝鼠香。与其他鼠类相比,麝鼠在繁殖期时前列腺—精囊腺组织极其发达。研究发现,经腹腔注射麝鼠香后,能够明显促进雄性小鼠的前列腺—精囊腺发育,表明两者之间可能存在一定的相关性。本研究利用RNA-seq技术对麝鼠繁殖期和非繁殖期的前列腺样品进行了转录组测序,对差异表达的基因进行GO分析和KEGG通路分析。结果显示,共筛选出1629个显著差异表达基因,涉及多种信号转导和能量代谢相关基因,其中OBP2、Bcl-2家族和肿瘤坏死因子受体超家族成员的差异表达提示麝鼠前列腺发育受多种机制调控,由香腺分泌的麝鼠香可能参与了前列腺的发育调节。     

Establishment and characterization of human pancreatic adenocarcinoma cell line SOJ producing carcinoembryonic antigen and carbohydrate antigen 19-9

A new tumor cell line derived from a human pancreatic exocrine adenocarcinoma was established in tissue culture and was transplantable in a nude mouse. In tissue culture, the neoplastic cells grew as epithelial-like, mucin-producing cells with a population doubling time of 50-70 hrs. Chromosomes ranged from 63 to 186 with a modal number of 77. Subcutaneous injection of 1 x 10(6) cultured neoplastic cells into nude mice resulted in tumor formation histologically closely resembling the original neoplasm. Ultrastructurally, the cell line showed characteristic ductal epithelium. Immunohistochemically, carcinoembryonic antigen (CEA). Carbohydrate Antigen 19-9 (CA19-9) and DU-PAN-2 antigen were demonstrated in the original tumor, the culture cells and the transplanted tumor. The cells secreted CEA (48.7 ng/1 x 10(5) cells/24 hrs) and CA19-9 (325 U/1 x 10(5) cells/24 hrs) in spent medium as well as sera of the nude mouse. This cell line has been passaged 30 times in vitro and maintained for more than one year. These characteristics will make the cell line SOJ a valuable tool in studying various aspects of biology of human pancreatic cancer.     

Uterine adenocarcinoma in a captive sika deer

A uterine adenocarcinoma with widespread visceral implantation and lymphatic metastases is described in a 4-yr-old captive nulliparous sika deer (Cervus nippon). The tumor was histologically characterized by proliferation of irregularly shaped acini and tubules lined by single- or multi-layered pleomorphic epithelial cells, which were sometimes multinucleated and contained PAS positive mucin. The neoplastic tissue was invasive and showed a marked desmoplastic reaction. A relationship between the tumor and a hormonal imbalance could not be ruled out. This is the first report of a tumor of the reproductive tract in a cervid.     

Relationship between the changes in the concentration of endoxan and the activity of kininforming enzymes within the Guérin tumor in rat.

The activity of rat kininforming system and endoxan level in certain organs and in the neoplastic tissue was studied. It was found that trypsin increases markedly the kininforming activity and endoxan level in the Guérin tumor, and slightly in the liver. A direct correlation between the level of endoxan when given with trypsin and the kinin activity were observed. The authors suppose that the selective accumulation of endoxan in the tumor depends on the trypsin -- induced activation of kininforming system within the neoplastic tissue.     

Fetal antigens as tumor growth enhancing factors in Yoshida's tumor rats

Malignant neoplastic cells have been shown to have some antigenic features identical to those of embryonic cells. Since several antigens are likely to be shared by both embryonic cells and neoplastic tissue, we tried to understand the meaning of the appearance of such antigens and the type of effect that the immunization with embryonic antigens would have on the survival of Yoshida's tumor rats. Wistar rats were immunized with fetal antigens by fetal cells (1.5 x 10(6)) suspended in 0.5 ml of Hanks solution plus an equal volume of Freund adjuvant, were injected in hind footpads, i.p. and i.m., respectively, for active immunization. Rabbit antigen sera were used for passive immunization. All animals presented ascites and tumor growth. Animals immunized by means of fetal cell antigens showed a mean survival rate after neoplastic transplant of 14 days. Animals that received rabbit immune serum showed a mean survival rate after neoplastic transplant of 17 days. The immunization by means of fetal antigens elicited a scanty effect on the survival of Yoshida's tumor transplanted rats. It can be concluded that antibodies, which are able to cross react with neoplastic cells, do not have cytotoxic effect and do not interfere with the survival of the neoplastic transplanted animals. Therefore, fetal antigens are likely able to carry out an immunosuppressive action. The fact that they appear on neoplastic cells could be seen as a metabolic modification effect or as a growth enhancing factor.     

Subcutaneous malignant mast cell tumor in a Japanese macaque (<Emphasis Type="Italic">Macaca fuscata</Emphasis>)

The histopathological, immunohistochemical, and ultrastructural morphologic characteristics of a tumor in the subcutaneous tissue of the chest of a 19-year-old female Japanese macaque were investigated. Consequently, the mass was diagnosed as a malignant mast cell tumor (MCT). Tumors were present in both mammary gland portions of the anterior thorax. Both tumors showed the same histopathological findings. The tumor tissue was defined by the presence of delicate connective tissue, and the tumor cells grew in a cord-like or cobblestone pattern. The tumor cell cytoplasm was very clear. The nuclei were relatively uniform and the cells showed a low nucleus-cytoplasm ratio. The cytoplasmic granules stained blue with Alcian blue and eosinophils had infiltrated into the tumor tissue. Furthermore, immunohistochemical analysis revealed that the tumor cell membrane was positive for the anti-c-kit antibody. In ultrastructural morphologic analyses, all tumor cells showed a rich cytoplasm and, occasionally, granules wrapped in a limiting membrane of high electron density. The tumor cells had metastasized to the axillary lymph nodes, the kidney, and the peritoneum. Based on these results, the mass was diagnosed as a malignant MCT originating from the subcutaneous tissue of the chest. Since cases of MCTs in macaques are very rare, this report presents important new knowledge of neoplastic lesions in this species.     

Human colon tissue in organ culture: preservation of normal and neoplastic characteristics

Normal and neoplastic human colon tissue obtained at surgery was used to establish conditions for organ culture. Optimal conditions included an atmosphere of 5% CO₂ and 95% O₂; tissue partially submerged with mucosa at the gas interface; and serum-free medium with 1.5 mM Ca²⁺ and a number of growth supplements. Histological, histochemical, and immunohistochemical features that distinguish normal and neoplastic tissue were preserved over a 2-d period. With normal tissue, this included the presence of elongated crypts with small, densely packed cells at the crypt base and mucin-containing goblet cells in the upper portion. Ki67 staining, for proliferating cells, was confined to the lower third of the crypt, while expression of extracellular calcium-sensing receptor was seen in the upper third and surface epithelium. E-cadherin and β-catenin were expressed throughout the epithelium and confined to the cell surface. In tumor tissue, the same disorganized, abnormal glandular structures seen at time zero were present after 2 d. The majority of cells in these structures were mucin-poor, but occasional goblet cells were seen and mucin staining was present. Ki67 staining was seen throughout the abnormal epithelium and calcium-sensing receptor expression was weak and variable. E-cadherin was seen at the cell surface (similar to normal tissue), but in some places, there was diffuse cytoplasmic staining. Finally, intense cytoplasmic and nuclear β-catenin staining was observed in cultured neoplastic tissue.     

Peroxisomes in human colon carcinomas. A cytochemical and biochemical study.

The presence of peroxisomes and their enzymic content were investigated and compared in healthy and neoplastic human colon epithelial cells using cytochemical studies at the ultrastructural level as well as biochemical analyses. Catalase-positive organelles were found to be more numerous in normal than in colonic neoplastic cells. Biochemical assays revealed that no D-aminoacid oxidase or L-alpha-hydroxyacid oxidase activity was detected in normal or tumor tissues. The specific activities of catalase, fatty-acyl CoA oxidase and enoyl-CoA hydratase/3 hydroxyacyl-CoA dehydrogenase (the so-called peroxisomal bifunctional enzyme of the beta-oxidation system) were found to be diminished in carcinoma cells compared with the control tissue. The fall in catalase activity correlated well with tumor stage according to Dukes, suggesting that this peroxisomal enzyme could be used as a potential prognostic marker.     

Human lymphomas transplanted in nude mice

This study was carried out to determine the take rate of malignant lymphomas transplanted subcutaneously in nude mice. Lymphoid tissue from patients with a variety of lymphoproliferative and lymphomatous disorders was employed, but only four (8.3% of total) non-Hodgkin's lymphomas of various types grew in the nude mice. These transplants were from patients with lymphoplasmacytic, lymphoblastic, immunoblastic and centroblastic lymphomas; two of them were serially transferred. High grade malignant lymphomas seem easier to transplant than low grade ones. In each case, the tumor growing in nude mice was similar to the patient's tumor. In one case, metastases were found in mesenteric lymph nodes and lung. Electron microscopic study showed that neoplastic cells were associated with peculiar dark cells the nature of which is discussed. Furthermore, in two transplanted lymphomas, neoplastic cells contained C type virus particles, indicating contamination by the murine xenotropic type C virus.     

INHIBITION OF EHRLICH TUMOR GROWTH IN MICE BY ELECTRIC INTERFERENCE THERAPY (IN VIVO STUDIES)

A study of solid tumor growth retardation by employing extremely low frequency (ELF) electric fields has been carried out. ELF electric fields were generated in tumor tissue in mice by the interference of two high frequency sinusoidal waves with the beat frequency centered at the tumor core. The results indicated a pronounced decrease in tumor growth rate in animals exposed to a 5-Hz interferential frequency for 1 hr daily. The 1 hr/day treatment produced a greater retardation effect than the 1 hr/week treatment. This indicates that treatment duration at the applied field frequency appears to play an important role in tumor growth delay. The dielectric properties of the tumor cells showed higher permittivity and conductivity values than homologous normal tissue. The permittivity of tumor cells treated daily with 5 Hz reaches nearly the same value as control tissue. Moreover, histological studies show that tumor tissues treated daily with the same frequency undergo partial regression and shrinkage of the aggregates of neoplastic cells leaving very little of them. We conclude that this new interferential technique is promising for tumor treatment in which a resonating electric field affects cell-to-cell communication.     

Identification of differentially expressed proteins in colorectal cancer by proteomics: down-regulation of secretagogin

To identify proteins with colorectal cancer-specific regulation, comparative 2-DE of individual-matched normal and neoplastic colorectal tissue specimens was performed. We found 15 protein spots with concordantly increased and 20 protein spots with concordantly decreased intensity in tumor tissue (expression regulation more than fivefold). Nine of these proteins were identified by MS/MS. Interestingly, one of the proteins, which exhibited a marked down-regulation in colorectal cancer tissues, was the recently identified endocrine cell-expressed protein secretagogin. The reduction of the secretagogin content in colorectal cancer tissues was confirmed by comparative immunoblotting (n = 17) and RT-PCR (n = 22) as well as by immunohistochemistry (n = 45) of individual-matched neoplastic and normal colorectal tissue specimens. Immunohistochemistry revealed absence of secretagogin-expressing cells in most of the colorectal cancer tissue specimens. However, some colorectal cancers were characterized by secretagogin-expressing cells. In normal mucosa, positively stained cells exhibited a neuroendocrine cell-characteristic morphology and mucosal location. In colorectal cancer tissues, secretagogin-expressing cells were characterized by a malignant morphology. Our findings might represent the basis for the clinical application of secretagogin as a biomarker for a distinct subgroup of colorectal cancers.     

Effects of cryosurgery in experimental carcinoma on lectin binding and keratin distribution

Histochemical alterations of lectin binding and keratin distribution in experimental carcinomas of the hamster cheek pouch were obtained following cryotreatment. Cryotreated carcinoma cells showed a characteristic reduction in lectin binding and keratin staining shortly following cryosurgery. Tumor tissue, on the 2nd and 3rd days after cryotreatment, displayed destruction and necrosis with almost a complete loss of lectin binding and keratin staining. The remaining neoplastic cells located in the deeper layer showed positive reaction for both lectin binding and keratin, which is indicative of tumor recurrence. Histochemical staining of lectin binding and keratin proteins were useful markers in cryotreated tumor cells to identify either destruction and necrosis or vital activity of neoplastic growth.     

T-cell lymphosarcoma in a female African green monkey (Cercopithecus aethiops)

A female African green monkey developed a generalized lymphosarcoma spontaneously that clinically and pathologically resembled malignant lymphoma in human beings infected with human T-lymphotropic virus type I. The monkey was serologically positive for simian T-lymphotropic virus. Immunocytochemical analysis of routinely fixed and paraffin embedded tumor tissue demonstrated neoplastic cells that stained positive with antihuman monoclonal UCHL1 which recognized a T-cell-restricted isotype of leukocyte common antigen.     

Fine needle aspiration cytology in recurrent amelanotic melanoma: a case report

BACKGROUND: Amelanotic melanoma can mimic a wide variety of epithelial and nonepithelial malignant tumors. Varied cytomorphology of melanoma has been described on exfoliative and fine needle aspiration cytology (FNAC). We report a case of recurrent amelanotic melanoma to highlight its varied cytomorphologic features, which may cause diagnostic problems on cytologic and on histologic examinations. CASE: A 63-year-old male presented with nodular swellings in the right anterior chest wall, right axilla and back. A nodule in the chest had been excised 6 months earlier. Clinically, the lesion was interpreted as recurrent soft tissue sarcoma. FNAC revealed malignant cells with highly varied morphology with plasmacytoid and pleomorphic malignant cells with occasional fibrocollagenous tissue strands showing adherent neoplastic cells. A cytologic diagnosis of pleomorphic malignant tumor was suggested, and the original histologic slides were reviewed; they showed a striking alveolar pattern that vaguely resembled an alveolar rhabdomyosarcoma. However, on immunohistochemistry, the tumor cells were S-100 and melan-A positive and desmin negative. A final diagnosis of amelanotic melanoma was made. CONCLUSION: Awareness of the highly varied cytomorphology of amelanotic melanoma minimizes the diagnostic difficulty on fine needle aspiration smears. Suitable immunohistochemical markers are of great value in difficult situations.     

Raman spectroscopy can differentiate malignant tumors from normal breast tissue and detect early neoplastic changes in a mouse model

Raman spectroscopy shows potential in differentiating tumors from normal tissue. We used Raman spectroscopy with near-infrared light excitation to study normal breast tissue and tumors from 11 mice injected with a cancer cell line. Spectra were collected from 17 tumors, 18 samples of adjacent breast tissue and lymph nodes, and 17 tissue samples from the contralateral breast and its adjacent lymph nodes. Discriminant function analysis was used for classification with principal component analysis scores as input data. Tissues were examined by light microscopy following formalin fixation and hematoxylin and eosin staining. Discriminant function analysis and histology agreed on the diagnosis of all contralateral normal, tumor, and mastitis samples, except one tumor which was found to be more similar to normal tissue. Normal tissue adjacent to each tumor was examined as a separate data group called tumor bed. Scattered morphologically suspicious atypical cells not definite for tumor were present in the tumor bed samples. Classification of tumor bed tissue showed that some tumor bed tissues are diagnostically different from normal, tumor, and mastitis tissue. This may reflect malignant molecular alterations prior to morphologic changes, as expected in preneoplastic processes. Raman spectroscopy not only distinguishes tumor from normal breast tissue, but also detects early neoplastic changes prior to definite morphologic alteration.     

The distribution of interleukin-19 in healthy and neoplastic tissue

The influence of interleukin (IL)-19, a recently discovered cytokine in the IL-10 family, on tissue is still unclear. Our aim was to determine the distribution of IL-19 expression and to delineate the cell types that express IL-19 in healthy and neoplastic tissue, because this information will significantly facilitate the exploration of its pathophysiological functions. We used tissue microarray technology and an immunohistochemical survey with an anti-IL-19 monoclonal antibody to examine the expression of IL-19 in 28 healthy and 15 neoplastic tissues. IL-19 protein was positively stained in 15 healthy tissue types and three major cell types: epithelial cells, endothelial cells, and macrophages. We also found that several types of tumor cells were positively stained for IL-19, especially in squamous cell carcinoma (SCC) of the skin, tongue, esophagus, and lung. SCC of the oral cavity expressed IL-19 mRNA and its receptors. In two cell lines derived from SCC of oral cavity tumor tissue, IL-19 specifically activated an intracellular signal and induced proliferation of the cells, which indicated that IL-19 may act in an autocrine manner in SCC tumors. This study provides important references for further investigation of the biological functions and clinical implications of IL-19 in humans.     

Bioavailability and Efficacy of a Gap Junction Enhancer (PQ7) in a Mouse Mammary Tumor Model

The loss of gap junctional intercellular communication is characteristic of neoplastic cells, suggesting that the restoration with a gap junction enhancer may be a new therapeutic treatment option with less detrimental effects than traditional antineoplastic drugs. A gap junction enhancer, 6-methoxy-8-[(2-furanylmethyl) amino]-4-methyl-5-(3-trifluoromethylphenyloxy) quinoline (PQ7), on the normal tissue was evaluated in healthy C57BL/6J mice in a systemic drug distribution study. Immunoblot analysis of the vital organs indicates a reduction in Cx43 expression in PQ7-treated animals with no observable change in morphology. Next the transgenic strain FVB/N-Tg(MMTV-PyVT) 634Mul/J (also known as PyVT) was used as a spontaneous mammary tumor mouse model to determine the biological and histological effects of PQ7 on tumorigenesis and metastasis at three stages of development: Pre tumor, Early tumor, and Late tumor formation. PQ7 was assessed to have a low toxicity through intraperitoneal administration, with the majority of the compound being detected in the heart, liver, and lungs six hours post injection. The treatment of tumor bearing animals with PQ7 had a 98% reduction in tumor growth, while also decreasing the total tumor burden compared to control mice during the Pre stage of development. PQ7 treatment increased Cx43 expression in the neoplastic tissue during Pre-tumor formation; however, this effect was not observed in Late stage tumor formation. This study shows that the gap junction enhancer, PQ7, has low toxicity to normal tissue in healthy C57BL/6J mice, while having clinical efficacy in the treatment of spontaneous mammary tumors of PyVT mice. Additionally, gap junctional intercellular communication and neoplastic cellular growth are shown to be inversely related, while treatment with PQ7 inhibits tumor growth through targeting gap junction expression.