Bromodeoxyuridine, an analogue of thymidine, can be detected by means of monoclonal antibodies and utilized as a marker of the S-phase of the cell cycle. In vitro immunohistochemical application of the BrdU/anti-BrdU-MAb method permits a quantitative assessment of the proliferative activity of a tissue as well as the direct location of the actively replicating cells in histological sections. In this paper, a method for the detection of the labeling index of S-phase cells in normal and neoplastic tissues with in vitro BrdU labeling and standard immunohistochemical techniques using anti-BrdU-MAb and avidin-biotin peroxidase complex is described. We have employed this method in 47 human solid tumor samples, including squamous cell carcinomas of head and neck and cervix uteri, adenocarcinomas and malignant lymphomas, and also evaluated the possible application of the BrdU labeling index to estimate the cycling S-phase cells in neoplastic cell populations. In our data, the in vitro labeling index varied greatly in an individual case (3.56-29.2%) and from an area to an area within the same case. Squamous cell carcinomas of the head and neck showed higher LI than those of the cervix uteri. A case of metastatic carcinoma to the lung from ductal carcinoma of the breast had the highest LI (29.2%), in contrast to the low LI (3.6%) in the primary ductal carcinoma of breast.
Adenocarcinoma/immunology/pathology ; Antibodies, Monoclonal/*immunology ; Breast Neoplasms/immunology/pathology ; Bromodeoxyuridine/*immunology ; Carcinoma, Squamous Cell/immunology/pathology ; Cell Nucleus/immunology/*physiology ; Head and Neck Neoplasms/immunology/pathology ; Humans ; Immunohistochemistry ; *Interphase ; Lymphoma/immunology/pathology ; Neoplasms/immunology/*pathology

OBJECTIVE: To study the clinicopathologic features, immunophenotype, diagnosis and differential diagnosis of Sinonasal teratocarcinosarcoma (SNTCS). METHOD: The clinical findings, morphologic features and immunohistochemical markers in one case of SNTCS were studied, and the relevant literatures were reviewed. RESULT: The Tumor tissue is composed of three layers, with mature and immature squamous epithelium nests, neural epithelial cells and olfactory neuroblastoma-like cells derived of ectoderm; Sarcomatoid components and bone tissue derived of mesoderm; The glandular and tubular structures part of which is adenocarcinoma and respiratory epithelium derived of endoderm; The fetal clear cell squamous epithelium is typical. In addition, diffuse large cytoplasm-with high light and cytoplasm with dark light has no obviously boundery. Immunohistochemical staining showed immune markers of different germ layers corresponding, squamous epithelium, glandular epithelium and respiratory epithelium were positive for CK and EMA, neural epithelial cells and olfactory neuroblastoma-like cells were positive for S-100, NSE and Syn, sarcomatoid area was positive for Vim, light dye area was positive for Vim, CD99 and CK, dark area was positive for NSE and GFAP. CONCLUSION SNTCS is a rare malignant tumor with the features of teratoma and carcinosarcoma, its histopathological and immunohistochemical features were typical, should be more drawn and sliced to avoid misdiagnosis and missed diagnosis.
Adenocarcinoma ; Carcinosarcoma ; diagnosis ; immunology ; pathology ; Diagnosis, Differential ; Esthesioneuroblastoma, Olfactory ; diagnosis ; immunology ; pathology ; Humans ; Nasal Cavity ; Nose Neoplasms ; diagnosis ; immunology ; pathology ; Teratoma ; diagnosis ; immunology ; pathology

OBJECTIVETo construct a human phage antibody library and screen the single chain variable fragment (ScFv) antibudies to peroxiredoxin I (Prx I) of lung adenocarcinoma.

METHODSThe total RNA was isolated from the lymph nodes of lung cancer patients to amplify V(H) and V(L) genes by RT-PCR. V(H) and V(L) were linked with a DNA linker by SOE-PCR to construct the single chain variable fragment gene. The ScFvs were coloned into the phage vector pCANTAB5E. The insert ratio of the ScFv antibody library was identified by PCR, and the products were digested by SfiI/NotI and analyzed with 1% agarose gel electrophoresis. Three rounds of panning against lung adenocarcinoma cell line A549 and Prx I were performed, and the positive clones were identified for soluble expression. The soluble antibodies were identified by SDS-PAGE and Western blotting, and ELISA and immunocytochemistry were used to characterize the activity of the antibodies.

RESULTSA recombination phage antibody library was constructed. The insert ratio of ScFv gene was 77% (23/30), and enzyme digestion identified the target product. The sixth phage harvest resulted in a yield 180 folds of that of the first one. Positive reactions to A549 cells were detected in 6 of 10 random clones, with a positivity rate of 60%. The soluble human ScFvs against Prx I of lung adenocarcinoma were expressed in E. coli HB2151 and confirmed by SDS-PAGE and Western blotting. ELISA and immunocytochemistry demonstrated a relative specific affinity of the soluble antibodies to A549 cells.

CONCLUSIONScFv antibodies against lung adenocarcinoma have been acquired by phage display antibody library technique, and the soluble antibodies have a relative avidity specific to human lung adenocarcinoma A549 cells overexpressing PrxI.

Adenocarcinoma ; immunology ; pathology ; Antibodies, Neoplasm ; biosynthesis ; genetics ; immunology ; Antibody Specificity ; Cell Line, Tumor ; Humans ; Immunoglobulin Variable Region ; immunology ; Lung Neoplasms ; immunology ; pathology ; Peptide Library ; Peroxiredoxins ; immunology ; Single-Chain Antibodies ; biosynthesis ; genetics ; immunology

OBJECTIVETo study the expression of a glycoprotein of plant origin in normal, benign and malignant breast tissues.

METHODSExpression of a plant glycoprotein was examined in 5 samples of normal breast tissues, 20 fibro-adenoma and 136 breast cancer by SABC immunohistochemical staining and the results were analyzed by SPSS statistics software.

RESULTSNo positive staining was detected in normal breast tissues (0/5). Weak staining was observed in 4 of 20 (20.0%) breast fibro-adenoma. Positive staining was demonstrated in 116 out of 136 (85.3%) breast cancer specimens. The differences were statistically significant. The expression of plant-associated human cancer antigen was related to pathological grade (P < 0.05), tissue invasiveness (P < 0.01) and recurrence (P < 0.05), but not to patients' age, tumor size and c-erbB-2 expression.

CONCLUSIONThe plant glycoprotein studied may be a human cancer-associated antigen which might be a potential marker of breast cancer.

Adenocarcinoma, Mucinous ; immunology ; pathology ; Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Breast ; immunology ; Breast Neoplasms ; immunology ; pathology ; Carcinoma, Ductal, Breast ; immunology ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; immunology ; pathology ; Female ; Fibroadenoma ; immunology ; pathology ; Humans ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; metabolism ; Plants ; immunology ; Receptor, ErbB-2 ; metabolism

OBJECTIVETo study the relationship between Chinese medical syndrome types of bronchioloalveolar carcinoma (BAC) and Th1/Th2.

METHODSTotally 60 BAC patients were syndrome typed as qi and yin deficiency syndrome (QYDS) and qi stagnation and phlegm-blood stasis syndrome (QSPSS), 30 cases in each group. Meanwhile, 30 subjects with benign pulmonary nodules were recruited as the control group. The contents of interferon-gamma (INF-gamma), interleukin 4 (IL-4), IL-2, and IL-5 were detected using thoracoscopic technique.

RESULTSAs for Th1 (INF-gamma and IL-2), it was ranked from high to low as the control group > the QSPSS group > the QYDS group (P < 0.05). As for Th2 (IL-4 and IL-5), it was ranked from high to low as the QYDS group > the QSPSS group >the control group (P < 0.05). As for Th1/Th2 (INF-gamma/lL-4, IL-2/IL-5), it was ranked from high to low as the control group > the QSPSS group >the QYDS group (P < 0.05).

CONCLUSIONSCompared with the tissue of benign nodules, Th1 function in tumor tissue of BAC patients was weaker and Th2 function stronger. Chinese medical syndrome types of BAC had correlation with Th1/Th2. Patients of excess syndrome had stronger immunity with Th1/Th2 shifting left,while those of deficiency syndrome were predispose to humoral immunity with Thl/Th2 shifting right.

Adenocarcinoma, Bronchiolo-Alveolar ; diagnosis ; immunology ; pathology ; Adult ; Aged ; Female ; Humans ; Lung Neoplasms ; diagnosis ; immunology ; pathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Th1 Cells ; immunology ; Th1-Th2 Balance ; Th2 Cells ; immunology

OBJECTIVETo investigate the relationship between the immunity and invasiveness in differentiated thyroid cancer (DTC).

METHODSTotally 74 DTC who were treated in Peking Union Medical College Hospital from September 2012 to December 2012 were enrolled in this study. These 74 patients were divided into membrane invasion group (n=36) and without membrane invasion group (n=38); also, they were divided into distant metastasis group (n=18) and without distant metastasis group (n=56). Natural killer (NK) cells and T-cell subsets were chosen as indicators for cellular immunity to investigate the correlation between cellular immunity and invasiveness in DTC.

RESULTSUnivariate analysis showed that the membrane invasion (Χ(2)=12.175, P=0.000) and distant metastasis (Χ(2)=8.139, P=0.006) correlated with cell immunity, whereas distant metastasis correlated with lymphocytic thyroiditis (Χ(2)=7.094, P=0.008). Further investigation shows that distant metastasis was associated with the percentage of CD8+T cell subgroup (Χ(2)=5.429, P=0.020), and membrane invasion was significantly associated with NK cells (Χ(2)=2.445, P=0.018) and CD4/CD8 disorder subgroup (Χ(2)=8.079, P=0.002). Multivariate analysis showed that cell immunity disorder was a risk factor for membrane invasion [OR=5.701,95%CI(2.075~15.666), P=0.001] and distant metastasis [OR=5.063,95%CI (1.571~16.320), P=0.008]. Further analysis showed that CD8+T cell was a risk factor for metastasis [OR=2.236,95%CI( 1.084~4.613), P=0.029], and CD4/CD8 disorders were the risk factors for membrane invasion [OR=2.802,95%CI(1.257~6.244), P=0.012].

CONCLUSIONCell immunity in thyroid cancer has close relationship with membrane invasion and distant metastasis, especially when the percentage of CD8+T cells decreases and when the NK cells and CD4/CD8 are abnormal, which may lead to membrane invasion and distant metastasis.

Adenocarcinoma ; immunology ; pathology ; Adolescent ; Adult ; Child ; Female ; Humans ; Immunity, Cellular ; Killer Cells, Natural ; immunology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; T-Lymphocyte Subsets ; immunology ; Thyroid Neoplasms ; immunology ; pathology ; Young Adult

OBJECTIVETo investigate the distribution patterns and proliferative activity of lymphatic vessels in colorectal carcinomas (CRC) and their relationship with tumor metastasis and disease prognosis.

METHODSThe microlymphatic density (MLD) and microvascular density in tumoral and non-tumoral areas of 96 cases of CRC were evaluated by immunohistochemistry, using monoclonal antibodies for podoplanin and CD34 respectively. The Ki-67 expression of the lymphatic and blood vessels was detected by double-labeling immunohistochemistry. The relationship between MLD and clinicopathologic features and prognosis was analyzed.

RESULTSThe lymph vessels at central and superficia1 portions of CRC often had a reticular architecture with numerous tiny and ill-defined lumina, while those at the tumor borders had large and open lumina. The MLD at tumor borders (51.2 +/- 25.5) was significantly higher than that in normal colorectal mucosa (29.4 +/- 9.0) and other portions of CRC (P < 0.01). The Ki-67 labeling index of the lymphatic lining cells at tumor borders (0.23 +/- 0.17) was significantly higher than that in other portions of CRC (P < 0.05). The MLD significantly correlated with lymphatic involvement by tumor cells, regional lymph node metastasis and distant metastasis (P < 0.01). The 5-year survival rate was also significantly lower in patients with high MLD (P < 0.05).

CONCLUSIONSNeolymphatic vessels are commonly seen in CRC, especially at tumor borders. High MLD at tumor borders is associated with metastasis. The detection of MLD at tumor borders may thus be useful in predicting lymph node metastasis and prognosis in patients with CPC.

Adenocarcinoma ; immunology ; pathology ; physiopathology ; Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; immunology ; pathology ; physiopathology ; Endothelium, Vascular ; immunology ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Middle Aged ; Prognosis ; Survival Rate

BACKGROUND: Previous studies have shown that the neutrophil-to-lymphocyte ratio (NLR) has a significant impact on the prognosis of many malignant tumors such as gastric cancer, colorectal cancer and pancreatic cancer, but the study on the prognosis of patients with resectable lung adenocarcinoma is less. The aim of this study is to investigate the correlation between the NLR and the clinicopathologic features of adenocarcinoma of lung patients who underwent radical pneumonectomy. Furthermore, this study aimed to clarify the predictive and prognostic significance of NLR in patients who underwent pneumonectomy for lung adenocarcinoma. METHODS: This study reviewed the medical records of 163 patients with lung adenocarcinoma who underwent pneumonectomy. The receiver operating characteristic (ROC) curve and Youden index were used to determine the cut-off value of the NLR. Survival curves were described by Kaplan-Meier method and compared by Log-rank test. The univariate and multivariate analyses were performed with the Cox proportional hazard model to identify the prognostic factors. RESULTS: When the NLR value was 2.96, the Youden index was maximal, with a sensitivity of 77.5% and a specificity of 75.9%. The 5-year survival rate in the low NLR group was higher than that in the high NLR group (P<0.05). The univariate and multivariate analyses showed that TNM staging and NLR were independent factors in predicting survival rate. CONCLUSIONS The NLR value was a simple and useful tool to predict the prognosis of lung adenocarcinoma after radical pneumonectomy.
Adenocarcinoma ; diagnosis ; immunology ; pathology ; surgery ; Adenocarcinoma of Lung ; Aged ; Cell Count ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Lung Neoplasms ; diagnosis ; immunology ; pathology ; surgery ; Lymphocytes ; cytology ; Male ; Middle Aged ; Neoplasm Staging ; Neutrophils ; cytology ; Pneumonectomy ; Prognosis ; ROC Curve ; Retrospective Studies

CD99 is characteristically expressed in Ewing's sarcoma/primitive neuroectodermal tumor. Recently its immunoreactivity has also been reported in other tumors. However, the significance of CD99 isoforms expressed in these tumors has not been elucidated. In this study, we evaluated the expression of CD99 isoforms and its relationship with histopathologic parameters in gastric adenocarcinomas. Paraffin sections of 46 gastric adenocarcinomas were stained with an anti-CD99 monoclonal antibody, YG32. Twelve (26.1%) cases of 46 gastric adenocarcinomas showed immunoreactivity to YG32. The CD99 expression was also seen both in non-neoplastic foveolar epithelial cells and infiltrating lymphocytes. In addition, Western blot and RT-PCR analyses revealed that the type I is the predominant isoform of CD99 in non-neoplastic and neoplastic gastric tissues. The CD99 expression was usually seen in the intestinal type adenocarcinoma, while rarely in the diffuse type. The CD99 immunoreactivity decreased in MMP-2-overexpressing adenocarcinomas (p=0.028). Our results suggest that the type I is the major isoform of CD99 expressed in non-neoplastic gastric mucosa and gastric adenocarcinomas and its downregulation in gastric adenocarcinoma may be associated with cellular dedifferentiation and/or MMP-2 overexpression.
Adenocarcinoma/*immunology/pathology ; Adult ; Aged ; Antigens, CD/*analysis/genetics ; Cell Adhesion Molecules/*analysis/genetics ; Female ; Gastric Mucosa/cytology/immunology ; Humans ; Male ; Matrix Metalloproteinases/metabolism ; Middle Aged ; Protein Isoforms/analysis/genetics ; RNA, Messenger/genetics/metabolism ; Stomach Neoplasms/*immunology/pathology

To investigate the immunologic characteristics and cytotoxicity of the RetroNectin-activated cytokine-induced killer cells (CIK) against drug-resistant lung cancer cell lines DDP-A549 (DDP: Cisplatin). Peripheral blood mononuclear cells (PBMC) were collected from healthy donors and divided into two groups: group I and group II. Seeded samples of group I into culture flask precoated with RetroNectin and CD3MAb to induce the CIK cells while seeded the group II into culture flask precoated with CD3MAb. In both groups, IFN-gamma was put into the flask on the same day and then IL-2 on the second day. The proliferation of CIK cells was tested by cytometirc analysis. The cytotoxicity activity of CIK cells was determined by MTT assays. The phenotype changes of CIK cells were identified by flow cytometric analysis. Scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to view the cytotoxicity against DDP-A549 of CIK cells and the changes of DDP-A549. The total CIK cells significantly increased by 524.77 fold in cell proliferation number due to the activation to CIK cells of RetroNectin. The expression rate of CD3+CD56+ cells was (31.40 +/- 1.91)%. The cytotoxicity of CIK cells showed statistically significance between DDP-A549 and the sensitive strains of parental generation A549 (P < 0.01). There was no significant difference of CIK cells' cytotoxicity between two groups when the effector: target ratio was fixed (P > 0.05). RetroNectin can significantly improve the proliferation activity of CIK cells. There was no evident influence to the cytotoxicity of CIK cells. CIK cells may be used as the immuotherapy to lung adenocarcinoma owing to its significant inhibition to the proliferation of DDP-A549.
Adenocarcinoma ; immunology ; pathology ; Cisplatin ; pharmacology ; Cytokine-Induced Killer Cells ; cytology ; immunology ; Cytotoxicity, Immunologic ; Drug Resistance, Neoplasm ; Fibronectins ; metabolism ; pharmacology ; Humans ; Lung Neoplasms ; immunology ; pathology ; Recombinant Proteins ; pharmacology ; Tumor Cells, Cultured