Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Face ; microbiology ; Female ; Humans ; Middle Aged ; Mycobacterium avium Complex ; isolation & purification ; Mycobacterium avium-intracellulare Infection ; metabolism ; microbiology ; pathology ; Nasal Cavity ; microbiology ; Nose Diseases ; metabolism ; microbiology ; pathology ; Vimentin ; metabolism

Objective To report 6 cases of hydroa vacciniforme-like cutaneous lymphoma,and to inves- tigate the relationship between this disorder and Epstein-Barr virus(EBV)infection.Methods Pathological and immunohistochemical examinations were performed in the biopsy specimens obtained from all 6 patients. Skin lesions were subjected to EBV encoded RNA(EBER)detection by in situ hybridization.Serological assay and quantification of EBV DNA were performed.Results All the 6 patients had recurrent papules, papulovesicles,necrosis and variola-like scar with chronic intermittent fever;four of the patients also presented with edema of the face,hands and feet.Pathologically,there were multilocular vesicles in the epidermis,and large numbers of infiltrating lymphocytes through the dermis.The cells were atypical with mitotic figures. Immunohistochemical staining of the lesions of 4 patients showed large quantities of cells expressing CD56, scattered cells expressing CD3 and CD45RO,and cells expressing grazyme B and T cell intracellular antigen-1 (TIA-1);a diagnosis of hydroa vacciniforme-like cutaneous NK/T lymphoma was made in these 4 cases. In the lesions of another 2 patients,the cells expressing CD3 and CD45RO,but not CD56,were observed; the diagnosis of hydroa vacciniforme-like cutaneous T-cell lymphoma was made in them.EBER was detected in the tumor cells of all the 6 patients.The IgG titers of anti-Epstein-Barr viral capsid antigen increased in all patients(1:5120 in 2 cases,1:2560 in 2 cases,1:1280 in 2 cases).The copies of EBV DNA were increased in the peripheral blood of both the two detected cases.A chronic active EBV infection was confirmed in all patients.Conclusions Hydroa vacciniforme-like cutaneous lymphoma is clinically characterized by edema of face,hands and feet,vesicular eruptions and variola like scars;histologically,it is characterized by infiltrates of atypical cells consistent with lymphoma,and necrosis in the center of vessels.NK/T is the primary immunophenotype of this disease.There is a close association between chronic active EBV infection and hydroa vacciniforme-like cutaneous lymphoma.

Objective To investigate the analgesic effect of oxysophoridine(OSR)and the influence of verapamil(Ver)on the antinociception of OSR when two drugs were co-administrated in mice.Methods The number of writhing within 15 min after ip different doses of OSR was observed in painful mouse mo- dels caused by acetic acid.The hot plate method was used to assess nociceptive sensitivity of CaCl_2 and Ver before ip OSR.Nitric oxide(NO)in serum was measured by spectrophotometry.Results The number of writhing was decreased and the latency of licking the hind paws was prolonged in a dose-dependent manner after ip OSR.The antinociception of OSR could be antagonized by CaCl_2 and enhanced by Ver.No inter- ference was detected in serum volume of NO.Conclusion These results suggest that OSR can antagonize the acute pain caused by acetic acid and hot plate in a dose-dependent manner in mice.Calcium channel blocker could enhance the effect of OSR.

Objective To compare the expression of serum miR-100 in patients with esophageal cancer and healthy person ,and explore the value of miR-100 in diagnosis for esophageal cancer .Methods Real-time fluorescent quantitative polymerase chain reac-tion was used to detecting miR-100 in 40 esophageal cancer patients(study group) and 50 healthy person(control group) .Results The expression of miR-100 in the study group and control group were 6 .399 ± 3 .541 ,2 .625 ± 1 .515 respective ,the expression in the study group was significant higher than that of the control group(t= 9 .07 ,P< 0 .05) .The under area of receiver operating char-acteristic curve of miR-100 in diagnosis for esophageal cancer was 0 .832(95% confidence interval was 0 .731 - 0 .934) ,when the Cut off value was 5 .285 ,the sensitivity and specificity of miR-100 in diagnosis for esophageal cancer were 65% and 95% . Conclusion Serum miR-100 in esophageal cancer patients is higher than that in healthy person ,which might be a new molecular markers in diagnosis for esophageal caner .

The optimization on liquid fermentation and purification of the fibrinolytic enzyme from Stenotrophomonas maltophilia(DR-929) was investigated.The results showed the best fermentation are amidulin 2.0%,soya flour 1.0%,yeast extract 0.5%,NaCl 1.0%,CaCl2 0.02%,MgSO4 0.05%,inoculum of 36 hours,fermental time 4d,initial pH 8.0 or 9.0,temperature 25℃,volume of media 30ml,volume of inoculum 5% or 6%.The purification process includes the following steps: removing cells by the centrifugation,25%~70% saturation ammonium sulfate precipitation,HIC with Phenyl FF(high sub),IEC with Q-Sepharose FF,gel filtration chromatography with Superdex 75.SDS-PAGE electrophoresis was used to examine the purification effect,and the results indicated that homogeneous strap in SDS-PAGE and has a molecular weight around 28.3kDa.The purification factor and activity recovery of the fibrinolytic enzyme are 271.5 and 24.5%,respectively.

OBJECTIVETo construct four expression vectors carrying enhanced green fluorescent protein (EGFP) gene under the control of different HBV promoters, and to detect and analyze their expressions in hepatoma cell lines.

METHODSFour HBV promoters were amplified using PCR, and they were inserted into the T-vector and identified using restriction enzymes and sequencing, then cloned into the expression vector pEGFP-1. The four recombinant plasmids were transfected into human hepatoma cell line HepG2 by lipofectamine2000, and the positive cell clones were detected using fluorescence microscopy.

RESULTSAll target fragments were separately obtained and successfully cloned into the expression vector. The expressions of EGFP under the control of the four promoters were detected. The expressions of EGFP controlled by different promoters had some differences.

CONCLUSIONSReporter gene EGFP under the control of four HBV promoters can be specifically expressed in hepatoma cell line HepG2, and different promoters give different results; this may provide another option in gene therapy of liver diseases.

Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Eukaryotic Cells ; metabolism ; Genetic Therapy ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HeLa Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; pathology ; Promoter Regions, Genetic ; genetics ; Transfection

Alcohol Oxidoreductases ; genetics ; isolation & purification ; Cloning, Molecular ; Hep G2 Cells ; Humans ; Two-Hybrid System Techniques

Objective To design a new method to verify the position of multileaf collimator(MLC)leaf using a two-dimensional ion chamber array(2D-array).Methods 2D-array of PTW T10018 Seven29~(TM) was used to calibrate the accuracy of MLC leaf position of Elekta Precise accelerator.The edge function of the leaf position of MLC was measured and used as the reference value.The precision of MLC leaf was then evaluated through comparing the measured and reference values.Results The accuracy of MLC leaf position was found within?0.1 mm.Conclusion This method of verifying the accuracy of multileaf collimator leaf position is easy,simple and reliable

Objective　 To investigate the clinicopathologic features of cytotoxi c natural killer (NK)/T-cell lymphomas of the lymph nodes. Methods　 Clinicopatho logic observations and follow up on 5 cases of cytotoxic NK/T-cell lymphomas of the lymph nodes, immunohistochemical staining for CD45RO,CD8,CD56,CD30,CD20,TIA -1 and in situ hybridization for EBER1/2 were performed. Results　 (1) The clinico pathologic features of cytotoxic NK-T-cell lymphomas of the lymph nodes were s um marized; (2) The tumor cells were positive for CD45RO in 4 of the 5 cases, in wh ich 3 cases were also positive for CD56. One case was of null cell type. The tum or cells were positive for TIA-1 and EBER1/2 in all 5 cases. Conclusions　 The clin icopathological features and special immunophenotypes of cytotoxic NK/T-cell ly m phomas of the lymph nodes were present in all 5 cases. Clinically, this entity t ends to be an aggressive process with poor prognosis.

OBJECTIVETo investigate the effect of deletion of the La protein binding site of HBV RNA, caused by its mutation, on the HBV S-mRNA stability of S gene, to study the role of the site in hepatitis B virus life cycle, and to try to find a new anti-HBV target in the future.

METHODSA HBV vector with mutation related to the La protein binding site was constructed using molecular cloning and PCR based site directed mutagenesis, and the vector was named pHBV-mLa. The HBV RNA secondary structure of the site was calculated using a computer. Normal HBV vectors and mutant vectors were respectively transfected into HepG2 cells by Lipofectamine 2000. HBV S-mRNA levels in the two groups were analyzed using semi-quantitative RT-PCR, and HBsAg secretion into the culture media was tested using ELISA.

RESULTSA HBV vector with mutation related to the La protein binding site was successfully constructed, and it was identified and confirmed using restriction analysis and sequencing. The HBV RNA secondary structure of the mutant vector was completely different to the stem-loop structure of the normal HBV vector. Semi-quantitative RT-PCR and ELISA analyses showed that the level of HBV S-mRNA in the mutant vector group was significantly lower than that in the normal HBV vector group (t'=12.703, P<0.05), and the expression efficacy of HBsAg was reduced in the mutant vector group (t= 44.648, P<0.01).

CONCLUSIONSThe change of La protein binding site in the HBV RNA caused by the mutation in HBV DNA disorganizes the stem-loop structure in the HBV RNA site. With the structural change, the La protein cannot bind the site and stabilize the HBV RNA (HBV S-mRNA), as the cleavage site in the upstream of the stem-loop structure is exposed to endoribonuclease. This results in HBV S-mRNA decay and affects the expression of the S gene. This study shows that only the sequence of this site in the HBV DNA is reserved, then the stem-loop structure in the La protein binding site will remain intact, and the disorganization of the stem-loop structure affects the stability of the transcripted HBV RNA. The La protein binding site in HBV RNA and the special secondary structure of the site are crucial to the life cycle of the hepatitis B virus.

Binding Sites ; Cell Line, Tumor ; Gene Deletion ; Genetic Vectors ; Hepatitis B virus ; genetics ; Humans ; Mutation ; Nucleic Acid Conformation ; RNA Stability ; RNA, Messenger ; genetics ; RNA, Viral ; genetics ; Viral Envelope Proteins ; genetics