OBJECTIVETo investigate the effect of component I from agkistrodon acutus venomon (AAVC-I) the migration of human umbilical vein endothelial cells (HUVECs), and to elucidate the possible anti-angiogenic mechanism of AAVC-I.

METHODSThe effect of AAVC-I on the migration of HUVECs which was cultivated in vitro and treated with AAVC-1 at four concentrations: 0, 20, 40, 80 microg/ml, was observed by methods of scratch wound-healing and Transwell assay. The expression level of mRNA and protein of P-selectin and intercellular cell adhension molecule-I (ICAM-1) were examined by RT-PCR and Western blot assay.

RESULTSCompared with the blank group, the migration ability of HUVECs in each AAVE-I treated group was reduced in a dose-dependent manner, and the expression level of the mRNA and protein of P-selectin and ICAM-1 were decreased.

CONCLUSIONAAVC-I inhibits the migration of endothelial cell, which is acted by down-regulation of the expression content of mRNA and protein of P-selectin and ICAM-1.

Cell Movement ; drug effects ; Cells, Cultured ; Crotalid Venoms ; pharmacology ; Down-Regulation ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; P-Selectin ; metabolism ; RNA, Messenger

OBJECTIVETo investigate the management and prognosis of thyroid well-differentiated carcinoma invading the upper aerodigestive tract.

METHODSA retrospective analysis of the management was performed done 62 patients with thyroid well-differentiated carcinoma invading the upper aerodigestive tract. The main method of surgery was shaving excision, and the other means including partial thyrochondrectomy, total laryngectomy, sleeve tracheal resection, sternocleidomastoid myoperiosteal flap and myodermal flap reconstruction, or simply palliative excision. Some patients received postoperative radioactive isotope therapy and radiotherapy. All patients were followed-up for 2 to 15 years with an average of 6.5 years.

RESULTSThe best curative effect was proved in the patients with local invasion, with the lumen uninvolved and their locoregional control rate was 100.0% (17/17). And the second choice was in patients with more extensive involvement of the upper aerodigestive tract structures. For them, extensive surgical management was done attempting to remove all gross disease followed by reconstruction, their locoregional control rate was 87.5% (7/8). And the third place was designated to patients with local invasion for which shaving excision was performed even though minor residual disease was left, their locoregional control rate was 55.6% (5/9). The poorest result went to simple palliative excision. For 17 patients with minor residual tumor, the locoregional control rate of those who were given postoperative radioactive isotope therapy was significantly higher than those without.

CONCLUSIONAccording to the limits and degree of invasion in the upper aerodigestive tract by thyroid well-differentiated carcinoma, different ways of surgery is indicated. For patients with residual disease, radioactive isotope therapy should be used to improve the result and life quality. Advanced lesions should be given postoperative radio therapy.

Adolescent ; Adult ; Aged ; Combined Modality Therapy ; Digestive System ; pathology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Respiratory System ; pathology ; Retrospective Studies ; Survival Rate ; Thyroid Neoplasms ; mortality ; pathology ; therapy

Objective The aim of this study was to investigate the mechanism and efficacy of tetramethylpyrazine-eluting stents (TES) on inhibiting neointima formation in porcine coronary artefies.Methods TES was prepared by tetramethylpyrazine spray-coated in bare metal stents(BMS).Pigs were implanted with TES or BMS(n=7 each),respectively.Quantitative coronary angiography(QCA)and intravascular ultrasound (IVUS) were performed before,immediately after stenting and at 28 days after stenting.Coronary arteries segments (5 cm) before and post stenting area (5 cm) as well as at stenting location were harvested at 28 days post stenting for histopathological examinations(inflammation,vascular smooth muscle cells proliferation and apoptosis).Results Follow up QCA at 28 days showed that percentage diameter stenosis were significandy lower in the,TES group than that in the BMS group[(10.0±2.1)%VS (60.2±23.5)%.P=0.01].Tlle lumen area determined by IVUS was similar between the two groups and there was no in-stent thrombosis in TES or BMS treated animals.Internal elastic lamina area was significantly largerwhile the neointimal ares [(1.51±0.45)mm2 vs(4.60±1.39)mm2,P=0.04] Was significantly smaller in the TES group than that in the BMS group.Histopathologieal assessments showed fewer inflammatory cells in the stented-coronary artery walls than those at the border zones of stenting in both groups.The number of proliferating cells were significantly decreased while apoptotic cells were significantly increased in the TES group compared with the BMS group (all P<0.05).Conclusion TES could effectively reduce in.stent restenosis in this porcine model by attenuating Vascular smooth muscle proliferation and enhancing vascular smooth muscle apoptosis post stenting.

Objective To investigate whether single nucleotide polymorphisms of catechol-O-methyl transferase (COMT) of Val158Met are associated with the risk of breast cancer.Methods Gene polymorphisms of COMT were detected using di-allele-specific-amplification with artificially modified primers combined with SYBR Green I real-time polymerase chain reaction in a case-control study,which included 96 breast cancer patients (treatment group) and 116 healthy women(control group).Results The frequency of allele G in COMT gene Val158Met was 65.10％ and 71.98％ in treatment group and control group,and the frequency of allele A were 34.90％ and 28.02％ respectively.There were no differences between the two groups in allele frequencies of COMT Val158Met among Guangxi Baise population (all P ＞ 0.05).COMT Val158Met G/G genotype frequency distribution of the treatment group (38.54％) was lower than that of control group (54.31％),A/G genotype frequency distribution of the treatment group was higher(53.13％)than that of control group (35.34％).The distribution frequency differences of the two groups homozygous wild-type and heterozygous were statistically significant (all P ＜ 0.05).A/A genotype frequency distribution was relatively similar in the treatment group and control group,and they were 8.33％ and 10.35％ respectively.The breast cancer risks of Guangxi Baise women with A/G heterozygous genotype increased by 2.118 times compared with that of G/G homozygous genotype.Conclusion Gene polymorphism of COMT Val158Met may be associated with the risk of breast cancer.

Many of the epiphytic Orchids are used as traditional Chinese medicine. The chemical components and pharmacology have been studied in recent 15 years. This article reviewed the studies which will be beneficial to reveal the relatives among these medicinal plants in the Orchid Family and be helpful to develop new drugs.
Adjuvants, Immunologic ; isolation & purification ; pharmacology ; Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Flavones ; isolation & purification ; pharmacology ; Humans ; Molecular Structure ; Orchidaceae ; chemistry ; classification ; Plants, Medicinal ; chemistry ; classification ; Platelet Aggregation Inhibitors ; isolation & purification ; pharmacology ; Polysaccharides ; isolation & purification ; pharmacology ; Stilbenes ; isolation & purification ; pharmacology

This study was purpose to investigate apoptosis pathway of leukemia K562 cells induced by anticoagulant fraction from Agkistrodon acutus venom (AVVC-1). The mitochondrial transmembrane potential (ΔΨm) of leukemia K562 cells was detected by flow cytometry with JC-1 single staining. The expression of cytochrome C in the mitochondrial of leukemia K562 cells was analyzed by Western blot after AVVC-1 treatment. The distribution of cytochrome C in leukemia K562 cells was measured by immuno-fluorescence test. The results showed that the potential of mitochondrial membrane decreased after treatment with different concentrations of AVVC-1 (12.5, 25, 50, 100 µg/ml) for 6 h (P < 0.01). The expression level of cytochrome C protein in mitochondria obviously declined after treatment with 30 µg/ml AVVC-1 for 48 h, and the fluorescent intensity of cytochrome C in cytosol was enhanced at the same time. It is concluded that AVVC-1-induced K562 cell apoptosis is related with mitochondrial damage, and cytochrome C may be a useful agent for investigating human leukemia therapy by using AVVC-1.
Agkistrodon ; Animals ; Apoptosis ; drug effects ; Cytochromes c ; metabolism ; Humans ; K562 Cells ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; metabolism ; Snake Venoms ; pharmacology

OBJECTIVETo study the effects of sirolimus (SRL) on the growth of transplanted human hepatocellular carcinoma (HCC) in nude mice.

METHODSHepG2 cells were Implanted into the liver of nude mice. The implanted mice were then treated with SRL and tacrolimus (FK506). The expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) was detected by immunohistology, microvessel density (MVD) was counted by immunostaining with anti-CD34 antibody for endothelial cells. Tumor apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay.

RESULTSThe tumor weight was (352+/-38) mg, (683+/-53) mg and (675+/-45) mg in SRL, FK506 and control group respectively. The tumor weight was significantly decreased in SRL group (P < 0.01), and there was no difference between FK506 group and control group. The expression of VEGF and PCNA protein was remarkably down-regulated in SRL group compared to control group (P < 0.05), and it was not significantly different between FK506 group and control group (P > 0.05). Compared to the control group, MVD was significanly decreased in SRL group, and the apoptosis index of tumor cell was significantly higher in SRL group (P < 0.01).

CONCLUSIONSRL inhibits transplanted HCC tumor growth by reducing tumor angiogenesis, inhibiting tumor proliferation and inducing tumor apoptosis.

Animals ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Immunohistochemistry ; Liver ; blood supply ; pathology ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Proliferating Cell Nuclear Antigen ; metabolism ; Sirolimus ; administration & dosage ; pharmacology ; Tacrolimus ; administration & dosage ; pharmacology ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays

Betulinic acid is a naturally occurring pentacyclic triterpenoid, which has antiretroviral, antimalarial, and anti-inflammatory properties. The purpose of this study is to investigate the HBV DNA replication inhibition in the mouse model with betulinic acid. Hydrodynamic injection method via the tail vein with the Paywl. 3 plasmid was used to establish the animal mode (n = 15), and the mice were randomly divided into the PBS control group (n = 5), Betulinic acid treatment group (n = 5) and lamivudine control group (n = 5). The day after successful modeling , the mice would have taken Betulinic acid (100 mg x kg(-1)), lamivudine (50 mg x kg(-1)), PBS drugs orally, once daily for 7 days, blood samples were acquired from the orbital venous blood at 3, 5, 7 days after the administering, HBsAg and HBeAg in serum concentration were measured by ELISA and the mice were sacrificed after 7 days, HBV DNA southern detections were used with part of mice livers. The results showed that betulinic acid significantly inhibited the expression of HbsAg in the mice model at the fifth day compared with the control group, and there was no significant differences between the effects of lamivudine and the PBS control group; both the betulinic acid and lamivudine groups had no significant inhibition for the HBeAg expression; the HBV DNA expressions of the liver tissue from the betulinic acid and lamivudine groups were inhibited compared with the control group. Taken together, these results reveal betulinic acid can inhibit the HBsAg expression and replication of the liver HBV DNA in the mouse model.
Acute Disease ; Animals ; Antiviral Agents ; pharmacology ; DNA Replication ; drug effects ; DNA, Viral ; biosynthesis ; Hepatitis B ; blood ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; immunology ; physiology ; Male ; Mice ; Plasmids ; genetics ; Triterpenes ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo establish fluorescence resonance energy transfer (FRET) assay method of detecting proteolytic activity of non-structural protein 3-4A (NS3-4A) serine protease of hepatitis C virus (HCV) for high throughput screening inhibitors against HCV in vitro.

METHODSHCV recombinant plasmid pMAL~c2/NS3-4A was transformed into the E.coli strain K12TB1. Maltose-binding-protein (MBP) NS3-4A fusion protein expression was induced by adding isopropyl-β-D-thiogalacto-pyranoside (IPTG) and purified by affinity chromatography. The proteolytic activity of MBP-NS3-4A protease was analyzed by FRET with the special protease substrate. The reaction system in this model was optimized, and the reliability of the model was evaluated.

RESULTSHigh throughput screening model for HCV NS3-4A protease inhibitors was established, and the best concentrations of enzyme and substrate were optimized. In the model, the Km value of protease was 4.74 μmol/L, Z factor was up to 0.80, and coefficient of variation (CV) was 1.91%. BILN 2061, one of the known HCV protease inhibitors, was measured with the Ki of 0.30 nmol/L.

CONCLUSIONThe assay model using FRET method for HCV NS3 4A serine protease is stable and reliable, and the model is suitable for high throughput screening for HCV NS3 4A protease inhibitors.

Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; Fluorescence Resonance Energy Transfer ; Hepacivirus ; enzymology ; High-Throughput Screening Assays ; methods ; Protease Inhibitors ; pharmacology ; Viral Nonstructural Proteins ; antagonists & inhibitors ; genetics

OBJECTIVETo study the effects of Rapamycin (RPM) on angiogenesis and tumor progression in human hepatocellular carcinoma (HCC) implantation mice.

METHODSTumor tissues of HCC were implanted into the liver of nude mice. Then, nude mice were treated with RPM and cyclosporine A (CsA). Real-time PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF). Immunohistochemical stain and image analysis were used to detect the protein expression of VEGF and proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) was counted by endothelial cells immunostained by anti-CD34 antibody. The concentration of VEGF in the peripheral blood was detected by ELISA.

RESULTS(1) The tumor weights were (372 +/- 35) mg, (769 +/- 39) mg and (751 +/- 42) mg in RPM, CsA and control group respectively. The tumor weight was significantly decreased in RPM group and no difference in CsA group compared with control group. (2) The expression of VEGF mRNA, VEGF and PCNA protein in tumor tissues and concentration of VEGF in the peripheral blood were remarkably down-regulated in RPM group compared with control group (P < 0.05) and were not remarkably different in CsA group from in control (P > 0.05).(3) Comparing with the control, the tumor MVD was remarkably decreased in RPM group (P < 0.05), and no difference in CsA group (P > 0.05).

CONCLUSIONRPM can inhibit angiogenesis and tumor progression of HCC by down-regulated the gene and protein expression of VEGF and inhibited the growth of tumor.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Nude ; Neovascularization, Pathologic ; drug therapy ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Messenger ; genetics ; Sirolimus ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Xenograft Model Antitumor Assays