OBJECTIVETo study the roles of actin and transforming growth factor (TGF)-beta(1) in the injury repair and the development of emphysema.

METHODSWistar rats were randomly divided into two groups: the smoking and infection group (group SI) and the control group (group C). The rats of group SI received smoking irritation accompanying with repeated intranasal infection. Subgroups of the experimental animals were killed in the 2nd, 4th, 8th and 16th weeks respectively. The morphological changes of lungs were compared and PaO(2), PaCO(2) as well as the right ventricular systolic pressure (RVSP) were analysed. The lung sections were stained with immunohistochemistry for actin and TGF-beta(1).

RESULTSIn comparison with animals of group C, thickening of the bronchiolar walls, narrowing of bronchiolar lumens, and area of emphysema were much severe in animals of group SI (P < 0.05). The muscularization of intra-alveolar arteries in group SI in the 16th week was apparent in comparing with that in group C (P < 0.05). PaO(2) values in group SI were significantly decreased, and RVSP values in group SI were significantly increased in the 8th and 16th week (P < 0.05). Actin expression was increased in animals of group SI in the 4th and 8th week (0.24 +/- 0.06 and 0.25 +/- 0.05) in comparing with that of group C (0.09 +/- 0.03) (P < 0.05). Animals of group SI showed a significant increase of TGF-beta(1) in lung tissue in different periods as mentioned in above (33.33 +/- 12.11, 45.71 +/- 15.12, 71.43 +/- 16.76 and 86.25 +/- 20.66 respectively).

CONCLUSIONSThe increased expression of actin and TGF-beta(1) protein in small airways induced by smoking irritation and Klebsiella Pneumoniae may interfere with the repair response, and contributes to the development of emphysema.

Actins ; metabolism ; Animals ; Bronchi ; metabolism ; pathology ; Epithelial Cells ; metabolism ; Female ; Klebsiella Infections ; microbiology ; Klebsiella pneumoniae ; Lung ; pathology ; Male ; Pulmonary Disease, Chronic Obstructive ; metabolism ; microbiology ; Pulmonary Emphysema ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Smoking ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1

OBJECTIVETo investigate the correlation between matrix metalloproteinase 9 (MMP-9) expression and tumor invasion and metastasis as well, and to explore the potential application of controlled expression of target gene in tumor gene therapy.

METHODSOne self-contained tetracycline-regulated retroviral vector containing anti-sense cDNA of MMP-9 was constructed and transfected into a metastatic human melanoma cell line WM451 which expressed a high level of MMP-9. Techniques such as growth rate measurment, MTT assay, (3)H-thymidine incorporation, colony forming ability in soft agar, invasion assay in Boyden chamber, as well as zymography and Western blot were applied to analyze the expression of MMPs and behaviors of tumor cells in vitro before and after gene transfection. Tumorigenecity and spontaneous metastasis were tested in nude mice.

RESULTSIn the presence of exogenous tetracycline, the transfected antisense MMP-9 did not affect the endogenous level of MMP-9 in WM451 cells, and showed no significant changes in cell behaviors in comparison with that of the vector-transfected control cells. Nevertheless, withdrawal of tetracycline from the medium caused a significant down-regulation of expression and activity of MMP-9. The capacity of cell growth in vitro, colony forming ability in soft agar, invasion through Matrigel all were inhibited remarkably when compared with the controls. Spontaneous metastasis in nude mice was significantly inhibited.

CONCLUSIONSTransfection of anti-sense MMP-9 can down-regulate the invasion and metastasis of melanoma cells both in vitro and in vivo, further clarifying the important role of MMP-9 in tumor progression.

Animals ; Cell Division ; Cell Line, Tumor ; DNA, Antisense ; DNA, Complementary ; genetics ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Genetic Vectors ; Humans ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Melanoma ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Retroviridae ; genetics ; Tetracycline ; pharmacology ; Transfection

OBJECTIVETo investigate the ways of inactivation and expression of p16 gene mRNA and its protein as well, and their clinicopathological significance in non-small cell lung carcinomas (NSCLC).

METHODSComparative- multiplex PCR, in situ hybridization, and immunohistochemistry were used to detect the promotor methylation status, mRNA, and protein expression in 64 cases of NSCLC, respectively.

RESULTSPromoter methylation of p16 gene was detected in 36 (56.3%) of 64 NSCLC cases. This positive result of methylation showed a negative correlation statistically with p16 protein expression by immunohistochemistry (P < 0.05). By in situ hybridization, 13 cases (20.3%) showed positive results for mRNA and all of these positive cases (13/13) had also a positive result by immunohistochemistry. Thirty-seven cases (57.8%) showed a negative immunohistochemical result. The metastatic rate of lymph nodes of those NSCLC patients with either promoter methylation or negative protein expression was higher (P = 0.038), and 3-year survival rate was lower statistically (P = 0.002).

CONCLUSIONDysfunction of p16 gene in NSCLC is mainly caused by promoter methylation, and patients with p16 gene dysfunction may have a poor prognosis.

Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; secondary ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; DNA Methylation ; DNA, Neoplasm ; genetics ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Genes, p16 ; Humans ; Lung Neoplasms ; genetics ; pathology ; Lymphatic Metastasis ; Prognosis ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Survival Rate

OBJECTIVETo study the relationship between angiogenesis and lymphangiogenesis with the expression of vascular endothelial growth factor C (VEGF-C) and VEGFR-3 in human non-small cell lung cancer (NSCLC).

METHODSSamples of 76 NSCLC cases with the neighboring noncancerous tissue were studied using anti- VEGF-C, VEGFR-3 and CD34 antibodies. Assessment of lymphatic vessel density and microvessel density (MVD) were performed.

RESULTSVEGF-C expression in NSCLC was associating with the differentiation of tumor cells (P = 0.009). Expression of VEGF-C and VEGFR-3 was significantly associated with lymph node metastasis (P = 0.008 and P = 0.013 respectively) and lymphatic invasion (P = 0.027 and P = 0.020 respectively). A significant positive correlation was found between VEGF-C in cancer cells and VEGFR-3 in lymphatic endothelial cells (P = 0.009). The number of lymphatic vessels (P = 0.006) and microvascular (P = 0.046) in VEGF-C positive tumors was significantly larger than in VEGF-C-negative tumors. Lymphatic vessel density was closely related to lymph node metastasis (P = 0.010), lymphatic invasion (P = 0.019) and clinical stages (P = 0.015). MVD was closely related to blood metastasis (P < 0.001) and clinical stages (P < 0.001). Patients with positive VEGF-C expression had a worse prognosis than those with a negative VEGF-C expression (P < 0.001).

CONCLUSIONSVEGF-C/VEGF-D in NSCLCs, are related to lymphangiogenesis and angiogenesis, as well as to the occurrence and the development of lung cancers. VEGF-C promotes intratumoral lymphangiogenesis via VEGFR-3, resulting facilitated invasion of cancer cells into the lymphatic vessels. VEGF-C expression can be a useful predictor of poor prognosis in NSCLC.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microcirculation ; pathology ; Middle Aged ; Neoplasm Staging ; Prognosis ; Survival Rate ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor Receptor-3 ; metabolism

OBJECTIVETo analyze the loss of heterozygosity (LOH) at 5 loci on chromosome 3p in soft tissue leiomyosarcoma (LMS).

METHODSLOH was detected in 22 cases of LMS using PCR-silver staining targeting 5 microsatellite sites on 3p14.2-pter. Relation between LOH and LMS clinical pathological features was also analyzed.

RESULTSTen of 22 LMS samples showed LOH at more than one locus (45.4%). Among the 5 loci, LOH occurred more frequently at D3s1295 (36.8%) and D3s1289 (10.5%), but absent at D3s1293. No significant difference was found on LOH incidence between different grade, size and location of LMS.

CONCLUSIONSLOH on chromosome 3p14.2-23 region is relatively frequent in LMS. Region around D3s1295 and D3s1289 may harbor tumor suppressor gene relating to LMS.

Abdominal Neoplasms ; genetics ; pathology ; Chromosomes, Human, Pair 3 ; Extremities ; Genes, Tumor Suppressor ; Humans ; Leiomyosarcoma ; genetics ; pathology ; Loss of Heterozygosity ; Microsatellite Repeats ; Retroperitoneal Neoplasms ; genetics ; pathology ; Soft Tissue Neoplasms ; genetics ; pathology

OBJECTIVETo investigate the clinicopathological manifestations of early renal amyloidosis (AL) and its diagnostic criteria.

METHODSFifteen cases with early renal amyloidosis admitted from 1994 to 2001 were collected from the hospital, and their clinical and pathological features were reviewed. Of them, the initial diagnoses were not made by depending findings from the light microscopy (LM) and immunofluorescense (IF), but confirmed by electron microscopy (EM) afterwards. Immuno-electron microscopy (IEM) were applied for amyloidosis typing.

RESULTSMost patients of early renal AL were in the middle to old age. Nephrotic syndrome was the most prominent symptoms and signs accompanying with rare microscopic hematuria and hypertension. Most of them had a normal renal function. Pathological examinations of renal biopsies using LM and IF showed mild mesangial proliferation and mild thickening of glomerular basement membrane (GBM). Immunoglobulins and complements were negative or only scanty in certain cases, but in all cases there was a light chain protein deposition homogeously. There were 4 cases of minimal change glomerulopathy, 5 cases of mild mesangial proliferative glomerulonephritis, 5 cases of stage I membranous nephropathy, and 1 case of cast nephropathy diagnosed with LM. The amyloid fibrils (diameter 8 - 10 nm) were randomly distributed in the mesangium, along GBM and at the arteriolar wall under EM. Additionally, Congo red staining was positive. IEM demonstrated that amyloid fibrils labeled with colloid gold was combined with a kind of light chain protein which was confirmed as the light chain type of AL.

CONCLUSIONSThe diagnosis of early renal AL was occasionally neglected by depending only findings of LM and LF. However, special amyloid fibrils can be detected using EM. EM observation is an indispensable technique for the diagnosis of early renal AL and the typing of AL may further be determined by using IEM.

Adult ; Aged ; Amyloidosis ; metabolism ; pathology ; Basement Membrane ; metabolism ; Female ; Humans ; Immunoglobulin Light Chains ; metabolism ; Kidney Diseases ; metabolism ; pathology ; Kidney Glomerulus ; metabolism ; pathology ; Male ; Microscopy, Immunoelectron ; Middle Aged

OBJECTIVETo investigate the relationship between expression of c-erbB-2, CD44v6 and nm23 gene proteins and the clinicopathological features of colorectal carcinomas.

METHODSImmunohistochemical technique was adopted to detect the protein expression of c-erbB-2, CD44v6 and nm23 genes in 92 cases of colorectal carcinomas. Of the 92 cases, 28 cases were followed up for more than 10 years, in which the prognosis was analyzed.

RESULTSExpression of c-erbB-2 and CD44v6 was correlating with UICC staging (P < 0.05), and c-erbB-2 was associated with prominent peritumoral lymphocytic infiltration (P < 0.05). In the 28 cases with followed-up data, univariant analysis revealed that the survival rate was correlated with histological grading, UICC staging, tumor growth pattern, peritumoral lymphocytic infiltration, and expression of c-erbB-2, CD44v6 and nm23. However, Cox stepwise proportional hazards analysis showed that only UICC staging, c-erbB-2 and nm23 status retained significant independently in prospecting prognosis.

CONCLUSIONSBesides UICC III and IV stage, overexpression of c-erbB-2 and decreased expression of nm23 are considered acceptable as the parameters in predicting of poor prognosis in colorectal carcinoma.

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; Biomarkers, Tumor ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Follow-Up Studies ; Genes, erbB-1 ; Glycoproteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Immunohistochemistry ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Invasiveness ; Neoplasm Staging ; Nucleoside-Diphosphate Kinase ; metabolism ; Prognosis ; Proportional Hazards Models ; Receptor, ErbB-2 ; metabolism ; Rectal Neoplasms ; metabolism ; pathology ; Survival Rate

OBJECTIVETo study the histogenesis and pathological characteristics of gastrointestinal stromal tumors (GIST) and GIST type stromal tumor (ST) beyond the gastrointestinal tract.

METHODSA retrospective study was carried out on leiomyoma, leiomyosarcoma and neurilemoma (46 cases in gastrointestinal tract and l3 cases in urinary tract and perineal area). 4 antibodies (CD117, CD34, SMA, S-100) were used for immunohistochemical staining.

RESULTSAmong 45 cases of GIST, the positive rate of CD117 and CD34 was 93.3% and 88.9% respectively. Among 12 cases of GIST type ST beyond the gastrointestinal tract, the positive rate of CD117 and CD34 was 83.3% and 75.0% respectively. In 2 cases (1 in gastrointestinal tract) of leiomyomas, both CD117 and CD 34 were negative in tumor cells, while SMA was extensively positive.

CONCLUSIONSCD117 and CD34 positivity are the most valuable factors in diagnosing ST. Both GIST and GIST type ST beyond the gastrointestinal tract are considered originating from a proto-interstitial stem cell with disoriented differentiation.

Actins ; analysis ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; analysis ; Biomarkers, Tumor ; analysis ; Female ; Gastrointestinal Stromal Tumors ; chemistry ; pathology ; Humans ; Immunohistochemistry ; Leiomyoma ; chemistry ; pathology ; Leiomyosarcoma ; chemistry ; pathology ; Male ; Middle Aged ; Neurilemmoma ; chemistry ; pathology ; Pelvic Neoplasms ; chemistry ; pathology ; Perineum ; Proto-Oncogene Proteins c-kit ; analysis ; Retrospective Studies ; S100 Proteins ; analysis ; Urologic Neoplasms ; chemistry ; pathology

OBJECTIVETo explore the origin and differentiation of gastrointestinal stromal tumors (GISTs).

METHODSImmunohistochemistry staining and electron microscopy were adopted.

RESULTSIn 212 cases of primary GISTs, the positive rates of CD117, CD34, alpha-SMA, MSA, desmin, S-100, PGP9.5 were 96.7%, 77.3%, 19.3%, 15.6%, 1.9%, 16.3%, and 12.3% respectively. Among them, GISTs showed a diffuse and strong positivity for CD117. Electron microscopy of tumor cells demonstrated numerous mitochondria, prominent perinuclear Golgi complex, smooth and rough endoplasmical reticulum and intermediate filaments. Irregular caveolae, dense plaque, incontinuous basal lamina were observed occasionally. Cytoplasmic processes were often observed accompanying with local adhesion present between the processes or between the processes and the cell membrane.

CONCLUSIONSData from both immunophenotype and electron microscopy suggest that GIST might originate from the mesenchymal cells, differentiating to be ICC afterwards, and possessing myoid characteristics in various extent.

Cell Differentiation ; Gastrointestinal Stromal Tumors ; chemistry ; ultrastructure ; Golgi Apparatus ; ultrastructure ; Humans ; Immunohistochemistry ; Microscopy, Electron ; Proto-Oncogene Proteins c-kit ; analysis ; S100 Proteins ; analysis ; Stromal Cells ; chemistry ; ultrastructure ; Ubiquitin Thiolesterase ; analysis

OBJECTIVETo explore the histogenesis and neural differentiation of gastrointestinal stromal tumor (GIST).

METHODSThe ultrastructural morphology and neural differentiated antigen expression were studied in 20 cases of GIST using electron microscopy and immunohistochemistry.

RESULTSAll of the 20 cases mentioned were positive for c-kit expression. The ultrastructural features of neural differentiation were observed in 7 cases, while no neural or myogenic differentiation seen in 12 cases. Myogenic differentiation to smooth muscle was observed in one case. The ultrastructural features of neural differentiation included scattered or cluster distribution of dense core granules in cytoplasm and cytoplasmic processes; formation of synaptic construction of cell processes; and neurogenic-like processes. In some cases pinocytotic vesicles under the cell membrane and skenoid fibers were seen. Neural differentiation with dense core granules was seen in one case in benign, one case in borderline and five cases in malignant group. The positive reactivity of neural differentiated antigen NSE, CD99, S-100p and CD56 in cases of the neural differentiation group appeared in seven, seven, five and four cases respectively, which were significantly higher than that of the undifferentiated group.

CONCLUSIONSIt's rather difficult to differentiate GIST accompanying with neural differentiation from gastrointestinal autonomic nerve tumor if depending only on its histology and immunophenotype appearance, since many features were overlapping in both tumors. Examination of the neural ultrastructures and neural differentiated antigen in GIST might be helpful to clarify the neural differentiation and potential behavior of GIST.

12E7 Antigen ; Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Autonomic Nervous System Diseases ; metabolism ; pathology ; Biomarkers, Tumor ; metabolism ; CD56 Antigen ; metabolism ; Calcium-Binding Proteins ; metabolism ; Cell Adhesion Molecules ; metabolism ; Cell Differentiation ; Child ; Diagnosis, Differential ; Female ; Gastrointestinal Stromal Tumors ; metabolism ; ultrastructure ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism ; Stromal Cells ; ultrastructure ; Synapses ; ultrastructure