OBJECTIVETo investigate the hypermethylation status of blu gene promoter in nasopharyngeal NK/T cell lymphoma and its role in the tumorigenesis and molecular diagnosis of this lymphoma.

METHODSTwenty cases of paraffin-embedded nasopharyngeal NK/T cell lymphomas tissues were studied by using methylation specific PCR (MSP).

RESULTSHypermethylation of blu gene promoter is detected in 6 of 20 (30%) nasopharyngeal NK/T cell lymphoma. The 6 positive cases were 4 in 15 nasopharyngeal NK cell lymphomas, 1 of 2 NK like T cell lymphoma and 1 in 3 peripheral T cell lymphomas (unspecified type).

CONCLUSIONHypermethylation of blu gene promoter in nasopharyngeal NK/T cell lymphoma indicated the inactivation of blu gene and its possible role in the tumorigenesis of this lymphoma. blu gene methylation could be detected in paraffin-embedded tissue and used as a new molecular marker for the diagnosis of this lymphoma.

DNA Methylation ; Genes, Tumor Suppressor ; Humans ; Lymphoma ; genetics ; pathology ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Polymerase Chain Reaction ; methods ; Tumor Suppressor Proteins

Biopsy, Fine-Needle ; Carcinoma ; diagnosis ; pathology ; surgery ; Carcinoma, Medullary ; diagnosis ; pathology ; surgery ; Carcinoma, Papillary ; diagnosis ; pathology ; surgery ; Carcinoma, Papillary, Follicular ; diagnosis ; pathology ; surgery ; Diagnosis, Differential ; Goiter, Nodular ; diagnosis ; pathology ; therapy ; Hashimoto Disease ; diagnosis ; pathology ; therapy ; Humans ; Lymphoma ; diagnosis ; pathology ; surgery ; Thyroid Nodule ; pathology ; surgery ; Thyroidectomy

OBJECTIVETo investigate the biological effects of overexpression of the human DNA polymerase (pol-beta) on cellular response to DNA damage.

METHODSThe cell strain HLFbeta from the stable overexpression of the human pol-beta was contaminated with methyl methanesulfonate (MMS) for investigating the effects of the pol-beta on the cellular responses to DNA damage on the aspects such as the DNA damage, the cell cycle and the induced mutation rate.

RESULTSThe cell HLFbeta from the stable overexpression of the human pol-beta was obtained through the screening. The cellular response to DNA damage of HLFbeta induced by the MMS in the intermediate and high dosage group (ranging from 0.5 to 0.8 mmol/L) was significantly lower than that in the control group. The analysis for the cell cycle distribution showed that both the two types of cells contaminated by MMS had retardation at G(2) phase. In the HLFbeta group, the cells had the obvious G(2) phase retardation and 49.0% of the cells were retarded at G(1) phase as well when the MMS was increased to 0.5 mmol/L while in the control, only 20.1% of the cells were retarded at the G(1) phase when the same dosage of MMS was administered. Moreover, the MMS-induced mutagenesis in HLFbeta was increased from 4.5 x 10(-6) to 8.2 x 10(-6), significantly higher than that in the control group (P < 0.05).

CONCLUSIONHigh Pol-beta level decreases cellular DNA damage induced by MMS. Nevertheless, the overexpression of Pol-beta can also increase error-prone DNA synthesis during DNA repair process.

Cell Cycle ; drug effects ; genetics ; Cell Line ; DNA Damage ; genetics ; physiology ; DNA Mutational Analysis ; DNA Polymerase beta ; biosynthesis ; genetics ; DNA Repair ; Dose-Response Relationship, Drug ; Humans ; Methyl Methanesulfonate ; toxicity ; Mutagens ; toxicity ; Mutation

Objective To investigate the expression and clinical value of immunohistochemistry in endometrial stromal tumors.Methods Immunohistochemical technique(Envision method)was applied to de- tect the expression of CD_(10),SM-MHC,h-caldesmon,AE1/3,CD_(99),Ki-67,CD_(34),c-kit,ER and PR in 15 cases of endomertrial stromal sarcoma and 3 metastases.The clinical pathological data,including the histological characteristics,histochemical and immunohistochemical staining features,complication,differential diagnosis and prognosis of endometrial stromal tumours were analyzed.Results Among the 18 cases of endometrial stromal tumor,17 cases had shown positive for CD_(10),including 13 cases diffuse positive and 4 muitifocal,7 cases with smooth muscle differentiation,3 cases with epithelial differentiation,7 cases with sex-cord differ- entiation.13 cases of ER and 16 cases of PR were positive expression in endometrial stromal sarcoma.Ki-67 in range 36 %～78 %.Conclusion Endometrial stromal tumour can display multi-differentiation.They show various pathomorphological features,Smooth muscle and sex-coed differentiation,the most common types. CD_(10) can be expressed consistently in endometrial stromal tumors.CD_(10) with h-caldesmon and SM-MHC can be used to make differential diagnosis between the endometrial stromal tumors and cellular leiomy0ma.ER and PR should be routinely estimated and be a prognostic predictor for endometrial stromal sarcoma.

Background The incidence of retinal vascular diseases increase annually,such as diabetic retinopathy,retinopathy of prematurity and age-related macular degeneration.The key of treatment for these diseases is how to evaluate retinal vascular change effectively and objectively.Retro-orbital injection of fluorescein isothiocyanatedextran (FITC-dextran) is a simple and effective method for observing C57BL/6J mouse retinal vessels.But,whether it is suitable for other mice and rats is seldom reported.Objective This experiment was to assess the feasibility of the observation of retinal vessels by retro-orbital injection of FITC-dextran in different genus of mouse and offer the reference for relevant study.Methods Twelve animals of C57BL/6J mice,Kunming mice,SD rats and Wistar rats were selected,respectively and divided into the experimental group and control group at average.The right eyes of the animals of the experimental group received the retro-orbital injection of 9 ml/kg FITC-dextran,and the right eyes of animals of the control group received PBS solution at the same volume and way.All the animals were sacrificed 10 seconds after injection and both eyes of each animal were obtained for retinal stretched preparation.The retrobulbar tissue and whole-mount retina were viewed under a fluorescence microscope.The use of the animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Retinal blood vessels labeled by FITC-dextran could be observed in both eyes of C57BL/6J mice and Kunming mice to present with a green fluorescence in experimental group under a fluorescence microscope,but no any fluorescence-labeled retinal blood vessel was exhibited in the control mice.The retinal blood vessel could not be observed in all eyes of SD rats and Wistar rats after the injection of FITC-dextran both in the experimental group and the control group under a fluorescence microscope.The surrounding tissues of the right eyes of mice and rats dyed with green fluorescence of FITC-dextran in the experimental group,however,green fluorescence could not be seen in the surrounding tissues of the left eyes of mice and rats.Conclusions Retro-orbital injection of FITC-dextran is a suitable method of observing the retinal vessels of mouse but not rat.

OBJECTIVETo develop a new vaccine expressing membrane-bound heat shock protein 70 (mbHSP70) and further study its antitumor therapeutic effect.

METHODThe pre- established vector expressing mbHSP70 was transfected into CT26 cells of colorectal cancer. After the CT26 cells were incubated with 900 microg/ml G418, the sub-clones resistant to G418 were harvested and the HSP70 positive clones were selected by limiting dilution. The clones were amplified and inactivated, thereby the vaccine expressing mbHSP70 was prepared. Lymphocyte proliferation stimulated by the vaccines, NK and CTL activity was observed. The antitumor efficacy of vaccine was observed in BALB/c mice model with colorectal cancer.

RESULTSThe laser confocal microscopy and flow cytometry showed that there existed much HSP70 on the vaccine membrane surface. The HSP70 gene-modified vaccine displayed augmented lymphocyte proliferation and higher NK and CTL activity in vitro,and marked tumor suppression and prolonged survival time of the vaccinated micein vivo, when compared with its counterpart. Furthermore, mbHSP70-expression vaccine elicited lymphocyte proliferation most intensively, generated the highest NK and CTL activity as well as the strongest antitumor effect, and prolonged survival time of the vaccinated mice.

CONCLUSIONA new vaccine expressing mbHSP70 has more potent antitumor immunity and better therapeutic efficacy than HSP70 gene-modified vaccine did.

Animals ; Cancer Vaccines ; therapeutic use ; Cell Membrane ; immunology ; metabolism ; HSP70 Heat-Shock Proteins ; immunology ; Immunotherapy ; Mice ; Mice, Inbred BALB C ; Neoplasms, Experimental ; therapy ; Transfection

OBJECTIVETo study the influence of transfection with human transforming growth factor-beta1 (hTGF-beta1) gene on the osteogenic potential and differentiation of the cultured human gingival fibroblast (GF).

METHODSEnzyme kinetics method was used to measure the effects of the transfection on the alkaline phosphatase (ALP) activity. Immunohistochemistry stain and image analysis were applied to evaluate the alteration of the content of osteopontin (OPN), bone sialoprotein (BSP), osteonectin (ON), osteocalcin (OC) in the GF with transfection. Mineralization nodules formation in vitro was also used.

RESULTSThe ALP activity of the GF after transfection was higher than the GF without transfection significantly (P<0.05), and was close to that of the PDLCs (P>0.05). The content of OC in GF was not improved after transfection, was similar with that of PDLCs (P>0.05). Under immunohistochemistry stain, the contents of OPN, ON, BSP expressed in GF with transfection were higher than those of GF without transfection (P<0.05), but similar to those of PDLCs (P>0.05). In the mineralized cultured medium, the nodules were observed in the GF with transfection and PDLCs after 21 days and 24 days alternatively. After von Kossa stain, purple mineralization nodules were observed.

CONCLUSIONThe GF transfected with pcDNA3-hTGF-beta1 could express some osteogenic characters, though these characters are restricted.

Alkaline Phosphatase ; Cell Differentiation ; Fibroblasts ; Gingiva ; Humans ; Integrin-Binding Sialoprotein ; Osteocalcin ; Osteonectin ; Osteopontin ; Transfection ; Transforming Growth Factor beta1

OBJECTIVETo investigate the frequencies of HLA-Alu repeat polymorphisms (AluMICB, AluTF, AluHJ, AluHG and AluHF) in Chinese Lisu and Nu ethnic populations.

METHODSThe frequencies of HLA-Alu repeat polymorphisms in above populations were determined with polymerase chain reaction (PCR). The associations between HLA-Alu repeat polymorphisms and HLA-A, HLA-B and HLA-C alleles were also analyzed. Phylogenetic trees were constructed with genetic distance calculated from the frequencies of HLA-Alu repeat polymorphisms.

RESULTSFrequencies of AluTF*2 and AluHF*2 were different between the two populations (P< 0.05), while those of other three insertions were similar. The strength of association between HLA-Alus and HLA alleles were different (P< 0.05) in the two populations. Although AluMICB*2 were associated with HLA-B*56:01 in both populations, the association was stronger in Lisu population (74.0%) but moderate in Nu population (30.7%). HLA-Alus were associated with particular HLA subtypes, e.g., AluHG*2 with certain HLA-A*02 subtypes. By phylogenetic analysis, Lisu and Nu were clustered together with southern Chinese and Thai populations.

CONCLUSIONThe distribution of HLA-Alus and the strength of associations between HLA-Alus and HLA class I alleles have varied between the two populations. Study of this association may facilitate identification of origins, evolution, progenitor haplotypes and recombination within the HLA class I region.

Adolescent ; Adult ; Aged ; Alleles ; Alu Elements ; Asian Continental Ancestry Group ; genetics ; Child ; Female ; Genes, MHC Class I ; Humans ; Male ; Middle Aged ; Phylogeny ; Polymorphism, Genetic ; Young Adult

OBJECTIVETo investigate the frequencies of chemokine (C-C motif) receptor 5 gene (CCR5)Δ32 deletional mutation of in Han and Dai populations from Yunnan province. Immortalized cell lines were derived from a family carrying the CCR5Δ32 mutation.

METHODSBlood samples of 346 Han and 355 Dai individuals were collected for genotyping. The coding regions of CCR5 gene were amplified with PCR followed by agarose gel electrophoresis. Suspected mutations were verified with DNA sequencing. Immortalized cell lines were constructed by using Epstain Barr virus and cyclosporine A. The difference between the cell lines and original blood samples was verified with PCR.

RESULTSOne ethnic Han individual was confirmed to be heterozygous for a deletional mutation by sequencing, which has led to discovery of a family with CCR5Δ32. Nine immortalized cell lines were established from this family, and no difference between the cell lines and original blood samples was detected by PCR.

CONCLUSIONTogether with previous reports, this study has indicated a significant difference in CCR5Δ32 among different ethnic groups in China. Established immortalized cell lines can also provide material for future research.

Base Sequence ; China ; Ethnic Groups ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Receptors, CCR5 ; genetics ; Sequence Deletion

The sialyl Lewis X (SLex) antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin (E-selectin).The combination of SLex antigen and E-selectin represents an important way for malignant tumor metastasis.In the present study,the effect of the SLeX-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels.The SLex expression in HepG2 cells treated with different concentrations of SLeX-binding DNA aptamer was detected by flow cytometry.Besides,the adhesion,migration,and invasion of HepG2 cells were measured by cell adhesion assay,and the Transwell migration and invasion assay.The results showed that the FUT7 expression was up-regulated at both mRNA and protein levels in HepG2 cells.SLeX-binding DNA aptamer could significantly decrease the expression of SLex in HepG2 cells.The cell adhesion assay revealed that the SLeX-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLex in the HepG2 cells.Additionally,SLeX-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1.The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5,10,20 nmol/L SLeX-binding DNA aptamer than those in the negative control group (P＜0.01).Our study demonstrated that the SLeX-binding DNA aptamer could significantly inhibit the in vitro adhesion,migration,and invasion of HepG2 cells,suggesting that the SLeX-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.