Adult ; Carcinoma, Renal Cell ; secondary ; Female ; Humans ; Kidney Neoplasms ; pathology ; Magnetic Resonance Imaging ; Vaginal Neoplasms ; secondary

OBJECTIVETo investigate the process and HPLC fingerprint of Charred Radix Scutellariae, and lay a foundation of Charred Radix Scutellariae quanlity control mode.

METHODSelect HPLC-UV fingerprint. Chromatogram condition: Hypersil C18 column (5.0 mm x 200 mm, 5 microm), mixtures of methanol, 0.4% phosphoric acid and acetonitrile as mobile phase in a gradient mode. Flow rate: 1.0 mL x min; Detection wavelength was set at 277 nm.

RESULTThere were no evident differences among Charred Radix Scutellariae that were normatively manufactured and processed.

CONCLUSIONThis test proves the process is feasible, this method recurs well and can be used to provide scieitific basis for the normative process and quanlity control mode of Charred Radix Scutellariae.

Charcoal ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Flavanones ; analysis ; Flavonoids ; analysis ; Hot Temperature ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Scutellaria baicalensis ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo find sensitive and specific micro-metastic markers for prostate cancer.

METHODSUsing nested reverse transcription-PCR, we examined the expression of PSA, hK2 and PSMA mRNA in peripheral blood mononuclear cells of 51 patients with prostate cancer, 33 patients with benign prostate hyperplasia (BPH) and 32 normal young people.

RESULTSThe expression rates of PSA, hK2 and PSMA mRNA were 52.9%, 43.1% and 64.7%, respectively in prostate cancer group, and 6.2%, 7.7% and 4.6%, respectively in control group (BPH patients and normal young people) with statistical significance (P < 0.01). Although the expression rate of PSA and hK2 mRNA increased with cancer progression, there was no statistical significance among patients in different stages. The expression rate of PSMA mRNA was higher than that of PSA and hK2 mRNA in each clinical stage.

CONCLUSIONPSMA mRNA expression detected by nested RT-PCR is of greater value for the diagnosis, therapy choice and prognostic evaluation of prostate cancer patients.

Aged ; Antigens, Surface ; blood ; Biomarkers, Tumor ; blood ; Glutamate Carboxypeptidase II ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; blood ; pathology ; Prostatic Neoplasms ; blood ; pathology ; Tissue Kallikreins ; blood

OBJECTIVETo observe the role of interleukin (IL) 21 alone, IL12 alone, and IL21 plus IL12 for inducing the antitumor activity of peripheral blood mononuclear cells (PBMCs) in patients with endometrial cancer.

METHODSPBMCs were isolated from peripheral blood in patients with endometrial cancer in vitro, and kept the culture with low-level IL2. IL2-stimulated PBMCs were cocultured under different conditions (with anti-IL21 antibody, IL21 alone, IL12 alone, or IL21 plus IL12) for 72 h. The cytotoxicity of PBMCs was then examined by lactate dehydrogenase(LDH) released assay. CD4(+) CD25(+) FOXP3(+) regulatory (Treg) cell and CD4(+) IL17A(+) T-helper (Th17) cell proportion were determined with flow cytometry. Cell proliferation and apoptosis were measured by cell counting kit-8(CCK-8)assay and flow cytometry, respectively.

RESULTSIn comparison to control group, both IL21 and IL12 significantly enhanced the cytotoxicity of PBMCs. The IL21 plus IL12 group had superior effect to IL21 alone and IL12 alone. IL21 and IL12 significantly decreased the percentages of Treg cells and the rate of PBMCs apoptosis. IL21 or IL12 had no significant effect on the differentiation of Th17 cells and the proliferation of PBMCs.

CONCLUSIONSIL21 and IL12 can enhance the cytotoxicity of PBMCs in patients with endometrial cancer, which can be further strengthened with treatment of IL21 plus IL12. Such effects may be achieved by inhibiting the differentiation of Treg cells and the apoptosis of PBMCs, but not by the differentiation of Th17 cell.

Adult ; Aged ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endometrial Neoplasms ; immunology ; pathology ; Female ; Humans ; Interleukin-12 ; pharmacology ; Interleukins ; pharmacology ; Leukocytes, Mononuclear ; drug effects ; immunology ; pathology ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo introduce statistical methods of time trend analysis on cancer rates.

METHODSCancer incidence data collected by the Shanghai Cancer Registry during 1991 to 1999 was used in the analysis to calculate the crude and age-adjusted rates, percent changes (PCs) and annual percent changes (APCs). APCs were estimated by a linear regression of the logarithm on the incidence rates during the nine years. It also introduced a method for partitioning a linear trend in age-adjusted rates into site-specific contributions to the overall floating trend. 95% confidence intervals for the APCs and contributions were described in the paper.

RESULTSA decreasing rates were observed for cancers of stomach and esophagus among both men and women in urban Shanghai from 1991 to 1999. The increasing rates among men would include cancers of colon, rectum, gall bladder, pancreas, prostate, urinary bladder, kidney and leukemia. The rates of cancers among women increased for colon, rectum, lung, breast, gall bladder, endometrium, ovary, urinary bladder and kidney. The changes of above cancers over time were statistically significant (P < 0.05 or P < 0.01), but rates for other cancer sites changed little. The APCs (weighted method) and contributions for the cancers of stomach, esophagus, colon, rectum and prostate were -2.99% and -65.72%, -2.90% and -17.07%, 12.30% and 21.46%, 2.94% and 18.62%, and 3.11% and 15.09% among men, and -6.05% and -39.55%, -1.08% and -35.19%, 2.81% and 28.64%, and 3.69% and 15.70% for the cancers of stomach, esophagus, breast and colon in women, respectively.

CONCLUSIONAPC, and related statistics could be used to describe and analyze the time trend of cancer rates rather than PC or/and graphical method alone.

Algorithms ; China ; epidemiology ; Female ; Humans ; Incidence ; Linear Models ; Male ; Neoplasms ; epidemiology ; Time Factors

OBJECTIVETo investigate the distribution of hepatitis B virus genotype in Hubei province (China) and its clinical significance.

METHODSSerum samples from 190 HBV DNA positive patients with chronic HBV infection,including 52 asymptomatic HBV carriers (ASC), 56 chronic hepatitis (CH), 32 fulminant hepatic failure (FHF), 22 liver cirrhosis (LC), and 28 hepatocellular carcinoma (HCC) patients were collected and tested for HBV genotypes by type-specific primers.

RESULTSA simple and precise genotyping system based on PCR using type-specific primers was developed for the determination of genotypes of hepatitis B virus (HBV). Of the 190 patients, 140 (73.7%) were genotype B and 42 (22.1%) were genotype C. Genotype B was more prevalent in the FHF and HCC patients than in the ASC patients; the ALT value was significantly higher in genotype B than in genotype C patients. The rate of anti-HBe was significantly higher in genotype B than in genotype C except in the patients of the ASC group.

CONCLUSIONThe system we used seems to be a useful tool for the molecular diagnosis of HBV infection and for large-scale surveys. Genotype B, genotype C and BC combination exist in Hubei province, and genotype B is the major genotype in this area especially in FHF and HCC patients.

Adult ; Carcinoma, Hepatocellular ; virology ; Carrier State ; virology ; China ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; complications ; virology ; Humans ; Liver Cirrhosis ; virology ; Liver Failure, Acute ; virology ; Liver Neoplasms ; virology ; Male ; Middle Aged

OBJECTIVETo investigate the effect of apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3G (APOBEC3G) mediated antiviral activity against hepatitis B virus (HBV) and duck hepatitis B virus (DHBV).

METHODSTotal RNA was extracted from peripheral blood mononuclear cells (PBMCs), RT-PCR product was cloned into the EcoR I/Hind III restriction sites of the CMV-driven expression vector fused with a hemagglutinin fusion epitope tag at its carboxyl terminal. Replication competent 1.3 fold over-length HBV was constructed with full-length HBV of ayw subtype. The mammalian hepatoma cell HepG2 was cotransfected with the replication competent 1.3 fold over-length HBV and various amounts of CMV-driven expression vector encoding APOBEC3G-HA. Levels of HBsAg and HBeAg in the media of the transfected cells were determined by ELISA, HBV DNA. RNA from intracellular core particles was examined using Northern and Southern blot analyses. Chicken hepatoma cell LMH was cotransfected with head-to-tail dimer of an EcoR I monomer of DHBV and various amounts of CMV-driven expression vector encoding APOBEC3G-HA. DHBV DNA from intracellular core particles was examined using Southern blot analysis.

RESULTSCMV-driven expression vector encoding APOBEC3G-HA and replication competent 1.3 fold over-length HBV were constructed. There was a dose dependent decrease in the levels of intracellular core-associated viral (HBV and DHBV) DNA and extracellular production of HBsAg and HBeAg. Levels of intracellular core-associated viral RNA were also decreased, but the expression of HBcAg remained almost unchanged.

CONCLUSIONAPOBEC3G suppresses HBV and DHBV replication and also suppresses HBsAg and HBeAg expression.

APOBEC-3G Deaminase ; Cytidine Deaminase ; genetics ; Hep G2 Cells ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B Virus, Duck ; physiology ; Hepatitis B e Antigens ; metabolism ; Hepatitis B virus ; physiology ; Humans ; RNA, Messenger ; genetics ; Virus Replication

OBJECTIVETo investigate the function of interferon alpha (IFNalpha) in a Chinese marmot model of hepatitis B, we expressed the Chinese marmot (Marmota himalayana) IFNalpha family gene (IFNA) in eukaryotic cells and prokaryotic cells.

METHODSEukaryotic and prokaryotic expression plasmids harboring Chinese marmot interferon alpha gene with different genotypes were generated using molecular cloning technology. We detected the biological activity of all expression products by viral protection assay, and analyzed their differences and species restriction of the biological activity.

RESULTSThe Chinese marmot functional genotype IFNalpha was expressed in the baby hamster kidney (BHK) cell line, and these products protected WH12/6 cells challenged by encephalomyocarditis virus (EMCV). The Chinese marmot IFN-alpha5 also expressed in E. Coli induced by IPTG, and purified fusion protein had antiviral biological activity. The biologic activity displayed differences among different subtype IFNalpha, and it had strict species restriction.

CONCLUSIONThe IFNalpha family gene of the Chinese marmot can be expressed in both eukaryotic and prokaryotic cells, and the expression products show antiviral activity in a protection assay. This study provides, for the first time, evidence that IFNalpha from the Chinese marmot has an antiviral function in vitro and can be used to improve the efficacy of current therapies for HBV infection in our Chinese marmot model.

Animals ; Eukaryotic Cells ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; Hepatitis B ; metabolism ; Interferon-alpha ; biosynthesis ; genetics ; physiology ; Marmota ; metabolism ; Prokaryotic Cells ; metabolism ; Signal Transduction ; Transcription Factors

OBJECTIVETo investigate the feasibility of endothelial cell-targeted therapy to cure post-burn hypertrophic scar.

METHODSA hypertrophic scar animal model was made. Intralesional injecting of VEGF monoclonal antibody was performed for three weeks. The changes of scar in volume and morphology were observed.

RESULTS1. The volume of scar decreased. 2. The number of the capillary, the amount of collagen I and collagen III decreased. 3. Transmission electron microscope examinations demonstrated many dead or apoptotic fibroblasts and endothelial cells. Fibrocytes were seen relatively common.

CONCLUSIONVEGF induces the growth and development of hypertrophic scar in that it induces excessive and uncontrollable angiogenesis, which favors excessive collagen synthesis. Endothelial cell-targeted therapy may be a promising method to cure post-burn hypertrophic scar.

Animals ; Apoptosis ; Burns ; complications ; Cicatrix, Hypertrophic ; chemically induced ; therapy ; Collagen Type I ; Collagen Type III ; Disease Models, Animal ; Endothelial Cells ; Feasibility Studies ; Neovascularization, Pathologic ; etiology ; Vascular Endothelial Growth Factor A

OBJECTIVESTo study the effects of tumor suppressor in lung cancer-1 (TSLC1) on human hepatocyte carcinoma cell line HepG2.

METHODSA full length of TSLC1 cDNA was amplified from RNA of normal human liver cells by RT-PCR, and it was cloned into a pCI-neo expression vector and transfected into human hepatocellular carcinoma cell line HepG2. The HepG2 cells transfected with this plasmid (experimental group) and those treated with pCI-neo vector (control group) and without any treatment (blank group) were compared. Cell morphology was studied microscopically and cell growth was analyzed with MTT assay. FACSort flow cytometry analysis was performed to assess the cell cycle distribution and apoptosis.

RESULTSA stable cell line expressing TSLC1 protein was successfully established. Morphologically, cells of the experimental group were tightly aggregated when compared with those of the control and blank groups. The growth of TSLC1-transfected cells was significantly suppressed in vitro compared with those of the control and blank groups. The amount of G0-G1 cells was 63.66%+/-3.83% (P less than 0.01) in the experimental group, while those of the control and blank groups were 47.45%+/-0.91% and 54.47%+/-0.96% respectively. The amount of S phase cells in the experimental group, 22.90%+/-6.04%, was significantly lower (P less than 0.05) than that of the control group (36.58%+/-0.61%) and the blank group (33.61%+/-2.99%), which suggested a G0-G1 cell cycle arresting. The number of cells in early and late phase apoptosis (17.09%+/-0.20% and 16.11%+/-0.40% respectively) were significantly higher than those of the control and blank groups (P less than 0.01).

CONCLUSIONSTSLC1 strongly inhibits the growth of HepG2 cells in vitro and induces apoptosis of the cells, suggesting that TSLC1 may have a tumor suppressor function in HCC.

Apoptosis ; genetics ; Cell Adhesion Molecule-1 ; Cell Adhesion Molecules ; Cell Proliferation ; Hep G2 Cells ; Humans ; Immunoglobulins ; genetics ; Membrane Proteins ; genetics ; Transfection ; Tumor Suppressor Proteins ; genetics