Objective To study the expression of vascular endothelial growth factor(VEGF)-C, VEGF-D and their receptor-3(VEGFR-3)in patients with colorectal cancer and their clinicopathological value.Methods Eighty specimens of the colorectal cancer and thirty normal adjacent bowels were stud- ied.The expression of VEGF-C,VEGF-D and VEGFR-3 proteins and mRNAs in specimens of colorectal cancers and normal colorectal tissues was studied by Strept-avidin-biotin complex method and RT-PCR. Clinicopathological data and survival of each patient were recorded and analyzed.Results①The staining of brown or filemot in cytoplast were observed as the positive expression of VEGF-C,VEGF-D and VEGFR-3 proteins.The positive rate(48.8%,56.3%,38.8%)and mean value(1.09?1.20,1.13?1.09,0.90?1.19)of VEGF-C,VEGF-D,VEGFR-3 expressions in specimens of colorectal cancer were significantly higher than those of the normal bowel tissues(P＜0.05).The expression of VEGF- C,VEGF-D and VEGFR-3 mRNAs by RT-PCR was correlated with that of VEGF-C,VEGF-D and VEGFR-3 proteins in colorectal carcinomas and normal bowel tissues.②Significant correlation between VEGF-C(P=0.0069),VEGF-D(P=0.0024)and VEGFR-3 expression was observed in colorectal cancers;moreover,no correlation between VEGF-C and VEGF-D.③The expression of VEGF-C, VEGF-D and VEGFR-3 in colorectal cancers was not correlated with age,gender,site and dimension of lesion,types of gross and histological,degree of differentiation and liver and pulmonary metastasis,but correlated significantly with Dukes' stage(P=0.0234,P=0.0003,P=0.0429)and lymph node me tastasis(P=0.0059,P＜0.01,P=0.0068).The increased death rate(P=0.0374,P=0.0127) and poor survival(P＜0.01,P＜0.01)were observed in the colorectal cancer patients with positive ex- pression of VEGF-C and VEGFR-3 when comparing with the patients of the negative expressions,but the expression of VEGF-D in colorectal cancers was not correlated with prognosis of the patients.Con- clusions Colorectal cancer cells may secrete lymphangiogenetic growth factors VEGF-C,VEGF-D and their receptor VEGFR-3,which induce the growth of lymphatic vessel endothelium and lymphangiogene- sis by VEGF-C,VEGF-D/VEGFR-3 signaling pathway,further accelerate lymphatic metastasis of colo- rectal cancers.VEGF-C,VEGF-D and VEGFR-3 might be acted as molecular phenotypes of lym- phangiogenesis in coiorectal cancers and important markers for evaluating lymphatic metastasis and prog- nosis in patients with coloreetaI carcinoma.

OBJECTIVETo evaluate the clinical significance of detection on lymphatic microvessel, lymphatic microvessel density (LMVD) and vascular endothelial growth factor-C (VEGF-C) in patients with colorectal carcinoma.

METHODSEighty tissue specimens of the colorectal carcinoma and the peritumoral tissue and thirty of adjacent normal bowel tissue were collected. The lymphatic microvessel and LMVD were determined by 5'-nucleotidase histochemical staining. The expression of VEGF-C protein and VEGF-C mRNA in specimens of colorectal carcinoma and normal colorectal tissues were studied by RT-PCR and immunohistochemical methods utilizing strept-avidin-biotin complex. Clinicopathological data and survival of each patient were obtained and analyzed.

RESULTS(1) The brown or filemot stained lymphatic microvessels were observed in specimens from the colorectal carcinoma, the peritumoral tissue and the normal bowel. Collapsed, nonfunctional lymphatic vessels were observed in the intratumoral tissue, and plenty of lymphatic vessels with large lumen referred as functional lymphatic vessels were observed in the peritumoral tissue. (2) The mean value of LMVD in the peritumoral tissue was significantly higher than that in the normal bowel tissue (9.76+/-2.85 vs. 5.49+/-1.43, t=8.220, P<0.01) and tumor tissue (9.76+/-2.85 vs. 2.13+/-0.96, t=15.118, P<0.001). (3) The positive rate (48.8% vs. 0, P<0.01) and mean value (1.09+/-1.20 vs. 0, P<0.01) of the VEGF-C protein expression in colorectal carcinoma specimens were significantly higher than that of the normal bowel tissue. The expression of VEGF-C protein was consistent with the expression of VEGF-C mRNA. The VEGF-C expression in intratumoral tissue demonstrated significant correlation with LMVD in the peritumoral tissue of colorectal carcinoma. (4) Both LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue correlated significantly with Dukes' stage (P<0.0001 and P=0.0234), lymph node metastasis (P<0.0001 and P=0.0059), and survival (P<0.0001 and P<0.0001), but not with age, sex, location and dimension of lesion, gross and histological type. Also, there was a positive significant correlation of LMVD in the peritumoral tissue with degree of differentiation (P=0.0168) and metastasis to the liver or the lung (P=0.0088).

CONCLUSIONSLymphatic microvessels in the peritumoral tissue are functional. The functional lymphatic microvessels, increased LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue may act as the morphological features and the molecular phenotype of lymphangiogenesis in colorectal carcinoma, and also as important predictive markers for evaluating lymphatic metastasis and prognosis in patients with colorectal carcinoma.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Microvessels ; Middle Aged ; Neoplasm Staging ; Prognosis ; Vascular Endothelial Growth Factor C ; metabolism ; Young Adult

OBJECTIVETo study the characteristics of thyroglossal tract cyst epidemiology and summarize how to improve the operation therapeutic effect.

METHODSThe clinic appearance, the pathological characteristics and the therapeutic effect of the 60 thyroglossal tract cyst cases were analyzed.

RESULTSThyroglossal tract cyst mostly occurred in children and teenagers. The incidence of female patients was more than that of male patients. Most of thyroglossal tract cyst located in the middle line of the neck and were explored tubular-objects and fibroropes. The histopathological feature was variform.

CONCLUSIONThe clinical appearance and pathology characteristic of thyroglossal tract cyst are variform. Thyroglossal tract cyst may be recurred easily if it is not operated well.

Adolescent ; Child ; Female ; Humans ; Male ; Thyroglossal Cyst

OBJECTIVETo isolate, identify and culture human spermatogonial stem cells (SSC) and then obtain purified and enriched human SSCs for research and application.

METHODSWe detected the expression of CD90 in the human testis using the immunofluorescence technique and isolated human testicular spermatogenic cells by two-step enzymatic digestion, followed by differential plating and magnetic-activated cell sorting (MACS) with CD90 as an SSC marker. Then we identified the isolated CD90-positive spermatogenic cells by RT-PCR and immunocytochemistry, and meanwhile cocultured them with Sertoli cells in SG medium in vitro.

RESULTSThe isolated CD90-positive cells showed a relatively homogeneous characteristic in size and morphology and expressed the genes specific for human SSCs, with high expressions (90.5%) of GFRA1, GPR125, and UCHL1. After coculture with Sertoli cells in the SG medium for 2 weeks, the isolated CD90-positive cells maintained a good activity.

CONCLUSIONCD90 can be regarded as a speci- fic marker for human SSCs and used to obtain highly enriched human SSCs by differential plating and MACS. Furthermore, the isolated human SSCs can be cultured in SG medium in vitro.

Adult Stem Cells ; cytology ; Biomarkers ; metabolism ; Cell Separation ; methods ; Cell Shape ; Cell Size ; Coculture Techniques ; Glial Cell Line-Derived Neurotrophic Factor Receptors ; metabolism ; Humans ; Immunohistochemistry ; Male ; Receptors, G-Protein-Coupled ; metabolism ; Sertoli Cells ; Spermatogonia ; cytology ; Testis ; metabolism ; Thy-1 Antigens ; isolation & purification ; metabolism ; Ubiquitin Thiolesterase ; metabolism

OBJECTIVETo establish immortalized cell line from the urothelium of the urinary bladder and identify the characteristics of the cell line.

METHODSHuman papillomavirus 16 (HPV-16) plasmid was used to transfect urothelium of infant urinary bladder in vitro with the help of Fugene-6, and this plasmid contained E6 and E7 genes of HPV-16. We also identified the existence of HPV-16 E6 and E7 genes and the biological characteristics of the cell line by PCR, immunohistochemistry, and the biology identification.

RESULTSBLTR-4 cell line, produced from the transfection of HPV-16K plasmid, was a cell line from urothelium with the expression of HPV-16 E6 and E7 genes. It had been cultured more than 70 passages, and the characteristics of growth was similar to the immortalized cell line as reported.

CONCLUSIONSBLTR-4 cell line is an immortalized cell line from urothelium of the urinary bladder, which contains HPV-16 E6 and E7 genes. BLTR-4 cell line is a good experimental model to investigate the relationship of the infection of high risk HPV and transitional cell carcinoma (TCC) in vitro.

Cell Line, Transformed ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; virology ; Plasmids ; genetics ; Repressor Proteins ; genetics ; Transcription, Genetic ; Transfection ; Tumor Virus Infections ; virology ; Urinary Bladder ; cytology ; Urinary Bladder Neoplasms ; virology

BACKGROUNDTo study the relationship of Epstein-Barr virus (EBV) and T cell lymphoma.

METHODSSixty cases of T cell lymphomas were examined for the presence of EBV using in situ hybridization for EBV encoded RNA (EBERs).

RESULTSEBERs were detected in tumor cells in 37(69.8%) of 53 cases with peripheral T cell lymphoma, but in none of seven cases of precursor T lymphoblastic lymphoma. The total detected EBERs were 37(61.6%) in 60 cases of T cell lymphomas. By Revised European-American Lymphoma(REAL) classification, EBERs were detected in 2/2 angioimmuno-blastic T cell lymphoma,17/18 angiocentric lymphoma, 4/6 anaplastic large cell lymphoma and 14/27 peripheral T cell lymphoma, unspecified (51.9%). The frequency of EBERs among the extranodal peripheral T cell lymphoma was higher than the nodal (P less than 0.01) there was no significant correlation with the sex, age and clinical stage.

CONCLUSIONSThis study indicated that high incidence of EBV was observed in peripheral T cell lymphoma, with predilection for angiocentric lymphoma and extranodal presentation.

Adult ; Aged ; DNA, Viral ; genetics ; Epstein-Barr Virus Infections ; pathology ; virology ; Female ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Lymphoma, T-Cell ; pathology ; virology ; Male ; Middle Aged ; Tumor Cells, Cultured

AIM:To investigate the synergistic effect of imperatorin on enhancing the anti -tumor effect of TNF-related apoptosis-inducing ligand(TRAIL)on breast cancer and the mechanisms.METHODS:T-47D and MCF-7 breast cancer cells were divided into control group ,imperatorin group,TRAIL group,imperatorin+TRAIL group and imperatorin+TRAIL+death receptor 5(DR5)siRNA group.The viability of T-47D and MCF-7 cells was measured by MTT assay. The apoptosis and mitochondrial membrane potential in T-47D cells were analyzed by flow cytometry.Western blot and flow cytometry analysis were performed to evaluate the expression of DR 5 on T-47D cell surface and the activation of caspase-8 and caspase-3.RESULTS:Imperatorin significantly enhanced the inhibition of cell viability induced by TRAIL of T -47D and MCF-7 cells,and significantly increased the apoptosis of T-47D cells induced by TRAIL.Imperatorin treatment ob-viously induced upregulation of DR5 expression and production of reactive oxygen species in the T-47D cells.In addition,imperatorin enhanced the TRAIL-induced damage of mitochondrial membrane potential and activation of caspase -8 and caspase-3.CONCLUSION:Imperatorin enhances the anti-tumor effect of TRAIL on breast cancer via upregulating the ex-pression of DR5.

OBJECTIVE: To investigate the expression of c-fos in rats' skin during wound healing. METHODS: Immunohistochemistry was conducted on paraffin section from incised wounding model of rat skin. RESULTS: Fos protein improved from the time of 10 min after wounding in the wound edge, then it reached peak at 3 h. 24 h after injury, the quantity of Fos expression had no difference with that of normal skin. CONCLUSION Fos is sensitive after wound, but should be used with other criteria in wounding interval estimation as it's unstediness.
Animals ; Genes, Immediate-Early ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-fos/biosynthesis* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Time Factors ; Wounds and Injuries/metabolism*

OBJECTIVETo investigate the association of changes in immune function with enterovirus 71 (EV71) cases with different severity of the disease.

METHODForty-six EV71-infected patients and 12 age-matched healthy children were enrolled in this study. The patients were divided into four groups according to critical degree of enterovirus 71 infection: hand-foot-and-mouth disease (HFMD); central nervous system disease (CNSD); autonomic nervous system dysregulation (ANSD) and pulmonary edema (PE). We analyzed CD14+ monocyte HLA-DR expression, lymphocyte immunophenotypes, the proportion of CD4+CD25+ Foxp3high regulatory T cells (Treg cells) and Th17 cells, cytokines (IL-1beta, TNF-alpha, IL-10, TGF-beta, IL-6, IL-17A), evaluated the mRNA levels of Foxp3 and ROR-gammat, and serum immunoglobulin and complements.

RESULT(1) Serum concentrations of IL-1beta and TNF-alpha elevated in mild cases, while declined in severe cases, and were lower in PE group (P<0.05). Serum concentrations of IL-10 and IL-10/TNF-alpha ratio gradually raised with the aggravation of the disease, and higher in PE group (P<0.05). (2) Circulating CD14+ monocyte HLA-DR expression, CD3+T cells, CD4+T cells, CD8+T cells, and NK cells gradually decreased, and lower in PE group (P<0.05). There was no significant difference in B cells, immunoglobulin and complement among the four groups. (3) The proportion of CD4+CD25+ Foxp3high Treg cells, mRNA level of Foxp, and serum concentrations of TGF-beta gradually decreased with the aggravation of the disease, while the proportion of Th17 cells, serum concentrations of IL-17A, mRNA level of ROR-gammat, and IL-6 gradually increased with the aggravation.

CONCLUSIONImmune function changed with different illness phases. The mild cases presented systemic inflammatory response syndrome status, while critically ill cases presented compensatory anti-inflammatory response syndrome or mixed antagonist response status. Immunoregulatory treatment of patients with EV71 infection should emphasize different methods at different stage and individualization.

Adolescent ; CD4-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Child ; Child, Preschool ; Enterovirus A, Human ; Enterovirus Infections ; immunology ; metabolism ; pathology ; Female ; HLA-DR Antigens ; immunology ; Humans ; Inflammation ; Interleukin-10 ; metabolism ; Lymphocyte Count ; Male ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo observe the exercise single photon emission computed tomography (SPECT) myocardial perfusion imaging of patients with myocardial bridge and assess the association between myocardial ischemia and extent of myocardial systolic compression.

METHODSSeventeen patients with myocardial bridge diagnosed by coronary angiogram were included and underwent exercise SPECT myocardial perfusion imaging.

RESULTSAbnormal SPECT perfusion imaging was evidenced in 12 out of 17 patients with myocardial bridge (2 out of 6 patients with systolic compression induced stenosis < 50%, 3 out of 4 patients with systolic compression induced stenosis between 50% - 75% and 7 out of 7 patients with the systolic compression induced stenosis between 75% - 100%).

CONCLUSIONExercise stress SPECT myocardial perfusion imaging could detect myocardial ischemia in patients with myocardial bridge and abnormal perfusion is positively related to the extent of systolic compression induced stenosis.

Adult ; Aged ; Coronary Angiography ; Exercise Test ; Female ; Heart ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Myocardial Bridging ; diagnostic imaging ; Myocardium ; Technetium Tc 99m Sestamibi ; Tomography, Emission-Computed, Single-Photon ; methods