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.

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

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

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 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

BACKGROUNDMostly because of the limited number and proliferative ability of the transplanted hepatocytes, hepatocyte transplantation offers only temporary support to the hepatic function with rather poor functional replacement of the damaged liver parenchyma. This study aimed to observe the therapeutic effect of human thioredoxin (hTrx) gene-modified hepatocytes on experimental acute liver failure in rats.

METHODShTrx cDNA was obtained by reverse transcription-polymerase chain reaction (RT-PCR) from human osteosarcoma 143 (TK-) cells to construct the recombinant retrovirus vector pLEGFP/hTrx, which was packaged into PA317 cells to collect the recombinant retrovirus containing hTrx gene. After titration and characterization, the recombinant retrovirus was applied to primary cultured rat hepatocyte for infection to generate hTrx gene-modified rat hepatocytes, whose viability and antioxidative capacity were examined by immunohistochemistry and MTT assay, respectively. In a Sprague-Dawley (SD) rat model of acute liver failure, the modified hepatocytes were injected into the spleen, and the hepatic function and survival rate of the recipient rats were evaluated at different time points after the transplantation.

RESULTSNIH3T3 cells infected by the recombinant retrovirus were capable of expressing bioactive hTrx in the form of fusion proteins. Immunohistochemistry demonstrated normal function of the hTrx gene-modified hepatocytes, which possessed strong antioxidative capacity as shown by MTT assay. Transplantation of the modified hepatocytes in rats with acute liver failure resulted in significantly lowered serum alanine aminotransferase (ALT) and total bilirubin (TBIL) levels (P < 0.05). The hepatocytes exhibited long-term survival and efficient proliferation after transplantation. Fourteen days after the operation, the rat models receiving hTrx gene-modified hepatocytes had significantly higher survival rate than those without the transplantation.

CONCLUSIONhTrx gene-modified hepatocyte transplantation can effectively alleviate acute liver failure in rats.

Animals ; Hepatocytes ; metabolism ; transplantation ; Humans ; Liver Failure, Acute ; therapy ; Mice ; NIH 3T3 Cells ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Thioredoxins ; genetics

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