Objective To explore the effect of Angiopoietin-2(ANG-2) and Endostatin (ENS) in patho genesis and development of endometriosis. Methods Twenty-five endometriosis cases and 24 controls were enrolled. ANG-2 and ENS in endometrium were detected by Immunohistochemistry. The concentrations of ANG-2 and ENS in peri-toneal fluid were measured with enzyme linked immunoabsorbant assay. Results In the EMs group, ANG-2 positive rate in eutopic endometrium was significantly higher than that of ectopic endometrium. The ANG-2 positive rate in ectopic endometrium was significantly higher than that of control group; in EMs group, the ENS positive rate in ec-topic endometrium was significantly higher than that in eutopic endometrium. The ENS positive rate in eutopic endo-metrium was significantly higher than that in control group. There was no significant difference of ANG-2 or ENS between stage Ⅰ～Ⅱand stage Ⅲ～Ⅳ. In peritoneal fluid, ANG-2 and ENS concentration and ratio of ANG-2/ ENS in endometriosis were significantly higher than that in controls. Conclusion The effect of angiopoietin-2 and endostatin in angiogenesis of endometriosis are important play a role in pathogenesis and development of the endo-metriosis.

Objective To explore the effect of Angiopoietin-2(ANG-2) and Endostatin (ENS) in patho genesis and development of endometriosis.Methods Twenty-five endometriosis cases and 24 controls were enrolled. ANG-2 and ENS in endometrium were detected by Immunohistochemistry. The concentrations of ANG-2 and ENS in peritoneal fluid were measured with enzyme linked immunoabsorbant assay.Results In the EMs group,ANG-2 positive rate in eutopic endometrium was significantly higher than that of ectopic endometrium. The ANG-2 positive rate in ectopic endometrium was significantly higher than that of control group;in EMs group,the ENS positive rate in ectopic endometrium was significantly higher than that in eutopic endometrium.The ENS positive rate in eutopic endometrium was significantly higher than that in control group. There was no significant difference of ANG-2 or ENS between stageⅠ～Ⅱand stage Ⅲ～Ⅳ. In peritoneal fluid,ANG-2 and ENS concentration and ratio of ANG-2/ENS in endometriosis were significantly higher than that in controls. Conclusion The effect of angiopoietin-2 and endostatin in angiogenesis of endometriosis are important play a role in pathogenesis and development of the endometriosis.

Twenty Newcastle disease virus(NDV)strains were isolated from diseased chicken and geese in field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi,and the antigenic analysis of the all NDV isolates had been done based on the reaction spectrum with a panel of monoclonal antibodies to the HN glycoprotein.The entire ORFs encoding HN protein of these NDV isolates were amplified by RT-PCR successfully,cloned and sequenced.The resultant sequences of HN genes of 13 isolates of chicken origin and 7 isolates of goose origin were gained and analyzed.The results of reaction spectrum showed that there were some distinct differences in the antigenic epitopes among the 20 NDV isolates.And the sequences revealed that the coding regions of the HN genes of these isolates all consisted of 1716 nt characteristic of virulent strains of NDV,coding for 571 amino acids.Neucleotides sequence homology were found to be from 94.8%to 100%among 18 NDV isolates of genotypeⅦ,and the neucleotides sequence homology between all the isolates and the other genotypeⅦstrains of recent years in China ranged from 92.1%to 99.6%.The deduced amino acid sequences and the receptor-binding regions of HN proteins between the NDV isolates of chicken origin and of goose origin were compared and analyzed.The results showed that some unique amino acid substitutions were found in the genome of the NDV isolates,and the close genetic similarity provided evidence for epidemiological linkage between the NDV isolates of chicken origin and of goose origin in the same period.

ORF1 and ORF2 gene of porcine circovirus type 2 were cloned by PCR with the specific primers designed according to genome of PCV2 (AY035820). Following extraction and digestion, PCR products were subsequently inserted into universal transfer vector plECMV (deleted partial gE and gI of pseudorabies virus) to generate recombinant transfer plasmid pIEORF1-ORF2. The genomic DNA of PRV TK-/gE- /LacZ+ strain and pIEORF1-ORF2 were co-transfected into IBRS-2 cells with lipofectin, and recombinant virus TK- /gE- /gI- /ORF1-ORF2+ was selected by PCR with ORF1 gene and ORF2 gene primers respectively. The recombinant virus was analyzed with Southern blotting and Western blotting. The results indicated that ORF1 and ORF2 gene of PCV2 had been inserted into the genome of TK- /gE- /LacZ+ strain and the expressed ORF1-ORF2 fusion protein could react with PCV2 positive sera. Result of virus titers detection showed the insertion of ORF1 and ORF2 gene did not influence propagation of recombinant virus.
Animals ; Cell Line ; Circovirus ; classification ; genetics ; Gene Transfer Techniques ; Genes, Viral ; Herpesvirus 1, Suid ; genetics ; Open Reading Frames ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Swine

OBJECTIVETo investigate the anti-inflammatory effect of Herba Siegesbeckiae extracts on mouse rheumatoid arthritis induced by arthrogen-CIA monoclonal antibody.

METHODSThe rheumatoid arthritis was induced by arthrogen-CIA arthritogenic monoclonal antibody in mice. The sandwich enzyme-linked immunosorbent assay was used to determine the concentration of IL-1βin mouse serum,and the content of IL-6,IL-17 and MMP-3 in supernatant of tissue homogenate of hind limb below the stifle of mice. One-way ANOVA was used for data analysis.

RESULTSThe toe swelling was attenuated in Siegesbeckiae group than that in model group [(0.218 ± 0.0307)cm(3) compared with (0.2545 ± 0.0179)cm(3), P<0.05]. The serum IL-1β level in Siegesbeckiae group was lower than that in model group [(63.74 ± 21.74)pg/ml compared with (104.96 ± 31.22)pg/ml, P<0.01]. The contents of IL-6, IL-17 and MMP-3 in tissue supernatants of Siegesbeckiae group were all lower than those of model group [(171.10 ± 48.35)pg/ml compared with (249.64 ± 75.08)pg/ml, P<0.05; (115.42 ± 56.52)pg/ml compared with (208.40 ± 88.54)pg/ml, P<0.05;(3660.31 ± 1680.99) pg/ml compared with (5420.79 ± 1201.43)pg/ml, P<0.05, respectively].

CONCLUSIONThe extract of Herba Siegesbeckiae has anti-inflammatory effect on mouse rheumatoid arthritis induced by mixed arthrogen monoclonal antibody.

Animals ; Anti-Inflammatory Agents ; therapeutic use ; Antibodies, Monoclonal ; Arthritis, Experimental ; drug therapy ; Arthritis, Rheumatoid ; chemically induced ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Interleukin-17 ; metabolism ; Interleukin-1beta ; blood ; Interleukin-6 ; metabolism ; Matrix Metalloproteinase 3 ; metabolism ; Mice ; Mice, Inbred BALB C

This study was purposed to explore the effects of Tanreqing injection on the biologic activities of human acute T lymphoblastic leukemia cell line Molt4 in vitro and its mechanism. Tanreqing injection was proportionally diluted and divided into 9 groups of different concentrations, including 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, 1:256 and 1:512. Molt4 cells were treated with those different concentrations of Tanreqing injection, and the cell growth status at various time points of different concentrations was observed under microscope. CCK-8 assay was employed to detect ability of cell proliferation, growth curve was drawn, the inhibition ratio and 50% inhibiting concentration (IC(50)) were calculated. Flow cytometry with PI and PI/Annexin V double stainings were used to detect the cell cycle and apoptosis of Molt4 cells after the treatment of Tanreqing injection respectively. Caspase-3 and Bcl-2 mRNA expression of Molt4 cells were determined by real-time quantitative PCR. The results showed that Tanreqing injection displayed an inhibitory effect on the proliferation of Molt4 cell line. In 1:2, 1:4, 1:8 and 1:16 concentration groups, great cytotoxicity was observed and numerous cells were dead. The inhibitory effect of Tanreqing injection was dose-dependent. IC(50) was 1:142 dilution concentration. In the 1:32 concentration group, S phase cell quantity remarkably decreased (p < 0.05) and apoptosis rate significantly increased (p < 0.05) at 72 hours after Tanreqing injection treatment. Simultaneously, caspase-3 mRNA expression increased and Bcl-2 mRNA expressions was downregulated (p < 0.05). It is concluded that the Tanreqing injection has an inhibitory effect on Molt4 cell proliferation and promotes its apoptosis. These biological effects of Tanreqing injection are partly related to cell reduction in S phase, downregulation of bcl-2 gene and upregulation of caspase 3.
Apoptosis ; drug effects ; Caspase 3 ; genetics ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics

The aim of this study was to investigate the curative effects of amifostine (AMF) combined with recombinant human beta-erythropoietin (rhbeta-EPO) on patients with pure erythroid aplasia (PEA). Two patients with PEA were treated with amifostine and rhbeta-EPO. The therapeutic regimen was adopted with AMF 0.4 g/day given by intravenous injection for 5 days first, then after a break of 2 days it went on for 3 weeks consecutively, that was considered as one treatment cycle. The rhbeta-EPO 6 000 U was used by subcutaneous injection for 3 times per week. The results showed that the red cell count, hemoglobin and reticulocyte count of two patients obviously increased after treatment. The erythroid ratio in bone marrow increased. Bone marrow biopsy showed that the erythroid proliferation improved. Intervals of red cell transfusions (RCT) in the two patients who live by red cell transfusion were prolonged after AMF treatments, and the amounts of each RCT was decreased obviously. The main side effect of amifostine was discomfort of digestive system, but was tolerated by all patients. In conclusion, amifostine plus rhbeta-EPO may be a new, effective and safety method especially for the elder PEA patients. The long-term curative effects and mechanism of amifostine still need further evaluation.
Aged, 80 and over ; Amifostine ; adverse effects ; therapeutic use ; Anemia, Aplastic ; drug therapy ; Drug Therapy, Combination ; Erythropoietin ; administration & dosage ; therapeutic use ; Humans ; Male ; Recombinant Proteins ; Treatment Outcome

OBJECTIVETo investigate the stress distribution in three ferrule designs in a maxillary central incisor restoration using PFM crown with post, and evaluate the biomechanical mechanism of the ferrule effect in the post crown by 3D finite element dynamic analysis.

METHODSThe 3D finite element model of a maxillary central incisor restored with post and PFM crown was established. By simulating three types of ferrule effect [crown wrapping dentine (A), core collar wrapping (B), and contrabevel (C)] under dynamic loading, the dentin stress was analyzed.

RESULTSDuring dynamic loading, the stress distribution tended to increase from the cervical part to root middle and radical part of the tooth, and greater high stress area was found around the apex of the post, where the peak stress was observed value. The stress of the labial dentin of the root inferior segment increased obviously. The high stress areas were invariable at every loading step during dynamic loading. The peak stress was sigma(vonA)

CONCLUSIONSWhen the ferrule is 2 mm, the ferrule effect maintained by crown wrapping cervical dentine (A) and core collar wrapping (B) is greater than that maintained by contrabevel (C),but there is no significant difference between A and B. The post with higher elastic modules may cause restore failure due to the high stress at the post apex and labial dentin.

Compressive Strength ; Crowns ; standards ; Dental Stress Analysis ; methods ; Finite Element Analysis ; Humans ; Incisor ; Post and Core Technique

OBJECTIVETo evaluate the effect of irbesartan on the proliferation, apoptosis, and VEGF mRNA expression of human umbilical vein cell line EA.hy926 in vitro.

METHODSThe human umbilical vein cell line EA.hyY926 were treated with various concentrations of irbesartan for 24 h. The cell proliferation after the treatment was detected by CCK8 assay, flow cytometry and FITC Annexin V/PI kit were used to detect changes in the cell apoptosis. RT-PCR was used to evaluate the expression of VEGF mRNA.

RESULTSThere were no changes in cell shape with various concentration of irbesartan. CCK-8 assay showed a greater rate of the cell proliferation in irbesartan group than that in control group with a dose-independent manner after 24 h treatment. After incubation with irbesartan, cell proliferation rate was significant (P < 0.05). FCM analysis showed no significantly changes in the cell apoptosis. Irbesartan increased the proliferation of EA.hy926 cells. At concentration of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) mol/L, VEGF mRNA expression enhanced either (P < 0.05).

CONCLUSIONIrbesartan could promote the proliferation and up-regulated VEGFmRNA expression in EA.hy926 cell line. This result suggested that in addition to antihypertensive effect, angiotensin receptor antagonist might be a novel therapeutic approach to chronic ischemic heart disease as heart failure.

Apoptosis ; drug effects ; Biphenyl Compounds ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Humans ; RNA, Messenger ; genetics ; Tetrazoles ; pharmacology ; Umbilical Veins ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effect of decreased expression of high mobility group Box-1 on the proliferation and apoptosis of HepG2 cells.

METHODSThree specific siRNAs of HMGB1 were designed and synthesized, and were transiently transfected into HepG2 cells by Lipofectamine 2000. The HMGB1 expression in HepG2 cells was detected by RT-PCR and Western blotting respectively. The proliferation activity in vitro was assessed by MTT assay. In situ apoptosis was evaluated by terminal deoxynucleotidyl transferase-deoxyuridine triphosphate nick end labeling (TUNEL) assay.

RESULTSAll of these specific HMGB1-siRNAs (1, 2, 3) efficiently and specifically inhibited the expression of the HMGB1 gene, and the levels of HMGB1 mRNA were 1.147+/-0.024, 1.014+/-0.042, 0.435+/-0.055, respectively, in HMGB1-siRNAs transfection group, which were significantly lower than that in Lipofectamine 2000 alone group (1.411+/-0.065, P < 0.01). Correspondingly, all of these specific HMGB1-siRNAs (1, 2, 3) could efficiently and specifically inhibit the expression of the HMGB1 protein, and the levels of HMGB1 protein were 0.369+/-0.035, 0.340+/-0.028, 0.097+/-0.020, respectively, in HMGB1-siRNAs transfection group, which were significantly lower than that in Lipofectamine 2000 alone group (0.553+/-0.051, P < 0.01). Of the 3 specific HMGB1-siRNAs, HMGB1-siRNA-3 (siRNAH3) had the highest inhibition rate (80%). The proliferation of HepG2 cells was markedly inhibited by siRNAH3 transfection. Compared to mock-transfection, siRNAH3 transfection dramatically suppressed the proliferation of HepG2 cells (P < 0.01). Moreover, siRNAH3 can induce apoptosis (P < 0.01).

CONCLUSIONsiRNA targeting HMGB1 mRNA can specifically reduce HMGB1 gene and protein expression. siRNAH3 can effectively suppress the proliferation and induce apoptosis of HepG2 cells.

Apoptosis ; Cell Proliferation ; HMGB1 Protein ; genetics ; Hep G2 Cells ; Humans ; RNA, Small Interfering