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 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.

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.

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

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

Hyperpolarization activated cyclic nucleotide-gated cation (HCN) channels are involved in neuronal rhythm regulation, synaptic activity, membrane resistance and dendritic integration due to their voltage-gated physiological properties. Recent studies have shown that HCN channels play an important role in central nervous system diseases, such as temporal lobe epilepsy, neuropathic pain, learning and memory disorder, substance abuse addiction and other related diseases. In this paper, we summarize the research progress of HCN channels in central nervous system diseases in recent years.

OBJECTIVE: Based on the establishment of a rat model of trigger point, this study was to intervene with warm acupuncture, and to evaluate the effect on pathological morphology and pain-induced inflammation of the rat model by microscopic pathology and microdialysis. METHODS: Sixty-four SD rats were randomly divided into group A (blank control), group B (model control) and group C (model and intervention control). Groups A and B were divided into 3 groups (A0, A1, A2 and B0, B1, B2), the group C was divided into 2 groups (C1 and C2). The MTrPs model was established in both groups B and C, warm acupuncture intervention were given to the C1 group for 7 days and the C2 group for 15 days. Rats were sacrificed in batches. MTrPs were locally sampled and stained with hematoxylin-eosin after the preparation. The pathological changes were observed under light microscopy. The iocal interleukin-1β and prostaglandin E2 were detected by microdialysis technique. RESULTS: Microscopically, the muscle fibers of the model were arranged disorderly, broken, twisted, local fibrosis, contracture thickening and so on; macrophage and other inflammatory cell invasion in local area and a large area of adhesion occurred on the contracture nodule, the pathological state of local muscle fibers was significantly improved after warm needle intervention, local microvascular formation and maturation, local muscle fiber repair. After successful modeling, the amount of interleukin-1β and prostaglandin E2 in group B0 was significantly higher than that in group A0 before warm needle intervention (<0.01). After warming intervention for 7 days, there was no significant difference in the amount of interleukin-1β and prostaglandin E2 between group C1 and group B1 (>0.05). Group C1 and B1 were significantly higher than group A1 (<0.01); warm needle intervention for 15 days, the amount of interleukin-1β and prostaglandin E2 in group C2 were lower than those in group B2 (<0.05), but those in group C2 and B2 were significantly higher than group A2 (<0.01), and the amount of interleukin-1β and prostaglandin E2 in group C2 was lower than group C1 (<0.05). CONCLUSIONS The modeling method of exercise combined hitting used in this study was proved to be effective by histopathology; warm acupuncture can improve the pathological and inflammatory state of local muscle fiber in myofascial pain trigger of rat, promote local microvascular formation and maturation, and help the trigger point local muscle fiber repair.
Acupuncture Therapy ; Animals ; Inflammation Mediators ; Myofascial Pain Syndromes ; Rats ; Rats, Sprague-Dawley ; Trigger Points

BACKGROUNDLumiracoxib is a highly selective cyclooxygenase-2 (COX-2) inhibitor with antiinflammatory, analgesic and antipyretic activities comparable with class specific drugs, but with much improved gastrointestinal safety. No studies have examined lumiracoxib for antitumorigenic activity on human nonsmall cell lung cancer cell lines in vitro or its possible molecular mechanisms.

METHODSThe antiproliferative effect of lumiracoxib alone or combined with docetaxol on A549 and NCI-H460 lines was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Drug-drug interactions were analyzed using the coefficient of drug interaction (CDI) to characterize the interactions as synergism, additivity or antagonism. Morphological changes were observed by acridine orange fluorescent staining. Extent of apoptosis was determined by flow cytometry.

RESULTSLumiracoxib (15 - 240 micromol/L) has an inhibitory effect on the proliferation of A549 and NCI-H460 cell lines in concentration- and time-dependent manners with the IC50 values of 2597 micromol/L and 833 micromol/L, respectively. The synergistic effect was prominent when lumiracoxib (15 - 240 micromol/L) was combined with docetaxol (0.2 - 2 micromol/L) (CDI < 1). Fluorescent staining showed that lumiracoxib could induce apoptosis in A549 and NCI-H460 cells. Lumiracoxib treatment also caused an increase of the sub-G1 fraction in each cell line and resulted in an increase of G0/G1-phase cells and a decrease of S-phase cells.

CONCLUSIONSLumiracoxib had antiproliferative effect on the human nonsmall cell lung cancer cell lines A549 and NCI-H460 and had a significant synergy with docetaxol, which may be related to apoptotic induction and cell cycle arrest.

Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Diclofenac ; analogs & derivatives ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Taxoids ; pharmacology

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