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OBJECTIVE: To investigate the effect of Staphylococcal peptidoglycan (PGN-sa) on raw264.7 cells differentiating into osteoclasts.
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Objective To investigate the molecular mechanism of osteoclast differentiation induced by staphylococcal lipoteichoic acid (LTA-sa). Methods Raw264.7 cells were treated with LTA-sa in a concentration of 200 ng/mL for 0, 5, 10, 20, 40, 60 min and 0, 1, 2, 3 days respectively, and the proteins in signaling pathways associated with osteoclast differentiation were measured with western blot. In addition, Raw264.7 cells were treated with different concentrations of LTA-sa (100, 200 and 400 ng/mL) and PBS for 0, 1, 2, 3 days, the expression of TNF-α, IL-1α and IL-6 was detected with Enzyme linked immunosorbent assay (ELISA). Results (1)Western blot showed that, under stimulation of LTA-sa, IκB-α decreased at 5 min and 10 min, while the phosphorylation of nuclear factor κB increased at 10 min . In addition , NFATc1 increased in 2 and 3 days gradually. The above results were statistically analyzed, and the difference was significant in statistics (P < 0.001). (2)ELISA showed that the expression of IL-6 increased in 2 and 3 days along with the increasing concentration and prolonging stimulation time of LTA-sa. Data were statistically analyzed, the difference was significant in statistics (P < 0.001). Conclusion LTA-sa promotes osteoclast differentiation through the NF-κB signaling pathway and the secretion of IL-6.
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ObjectiveTo evaluate the effects of diagnosis of cervical insufficiency and different management on pregnancy outcomes.MethodsFrom June 2004 to May 2010, a retrospective analysis was carried out on 554 patients with cervical insufficiency in nine hospitals in Guangdong Province, China. The patients were divided into two groups, the cervical cerclage surgical treatment group (surgical group,n=357) and the expectant treatment group (n=197). These patients were then re-diagnosed according to the unified diagnostic criteria by the American College of Obstetricians and Gynecologists (2014), and divided into the definite diagnosis group (n=425) and the indefinite diagnosis group (n=129). The two independent samplest-test andChi-square test were used to compare pregnancy outcomes between the definite and indefinite diagnosis groups, and the different treatment groups.ResultsThe full-term delivery rate in the surgical group was significantly higher than that in the expectant treatment group [40.3% (144/357) vs 23.4% (46/197),χ2=16.254, P=0.000], and the late abortion rate was lower in the surgical group than in the expectant treatment group [22.4%(80/357) vs 40.1% (79/197),χ2=19.419,P=0.000]. In women with a definite diagnosis of cervical insufficiency, full-term delivery rate [44.7% (117/262) vs 20.9% (34/163),χ2=24.844,P=0.000], and newborn body weight were significantly higher in the surgical group [(2 664.3±762.2) vs (2 416.9±845.0) g,t=1.160,P=0.014] than in the expectant treatment group and the late abortion rate was significantly lower [21.4% (56/262) vs 41.1% (67/163),χ2=19.021,P=0.000]. Cervical cerclage in the indefinite diagnosis group did not resulted in raising the full-term delivery rate [28.4% (27/95) vs 35.3% (12/34),χ2=0.561], preterm delivery rate [46.3%(44/95) vs 29.4% (10/34),χ2=2.940], late abortion rate [25.3% (24/95) vs 35.3% (12/34),χ2=1.252] and newborn body weight [(2 526.5±761.8) vs (2 683.4±725.8) g,t=0.004] compared with expectant treatment group (allP>0.05). Pregnancy outcomes in the surgical treatment group in relation to twin pregnancies were not significantly different in the≥28 weeks delivery rate [81.4% (37/46) vs 69.2% (18/26),χ2=1.156], late abortion rate [19.6% (9/46) vs 30.8% (8/26),χ2=1.156] and newborn birth weight [(2 003.2±621.0) vs (1 807.5±609.4) g, t=0.057] compared with those in the expectant treatment group (allP>0.05).ConclusionsIn accordance with the diagnostic criteria for cervical insufficiency and indications for cervical cerclage in surgical cases, cervical cerclage can effectively improve pregnancy outcome. But cervical cerclage is not recommended in twin pregnancies with cervical insufficiency.
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Objective To explore the method of primary culture and biological characteristics of aortic vascular smooth mus-cle cells (VSMC)in mice,providing experimental material for cellular and molecular scientific research of vascular disease. Methods Thoracic and abdominal aortas in mice were isolated and VSMC were obtained by using improved method of tissue piece inoculation.Digested with trypsin and passaged,VSMC were purified with differential adherence method.The conditions of cellular morphology and growth were observed under inverted phase contrast microscope,and VSMC were identified with hematoxylin-eosin (HE)staining and immunofluorescence.Results VSMC were isolated successfully and grown vigorously with good bioactivity,the cells had a radial or typicalpeak-valleylike growth,and showed fusiform,abundant cytoplasm with large and round or oval nucle-us by HE staining,the expressions of specific cytoplasmicα-smooth muscle actin were positive by immunofluorescence stain.Conclu-sion It can isolate and cultivate VSMC with high purity and good activity under in vitro conditions with simple,economical,reliable method.
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The expression of Aurora B in normal endometria and endometrial carcinomas and its relation with clinicopathologic parameters of endometrial carcinomas were investigated. Streptavidin-biotin peroxidase (SP) immunohistochemical technique was used to detect the expression of Aurora B in 10 cases of normal proliferative phase endometria, 10 cases of normal secretory phase endometria and 72 cases of endometrial carcinomas respectively. According to the 1988 International Federation of Gynecology and Obstetrics (FIGO) grade, there were 37 patients in grade 1, 23 in grade 2 and 12 in grade 3 respectively. According to the FIGO stage, there were 59 patients in stage I-II and 13 patients in stage III-IV. Aurora B was expressed in both normal proliferative phase endometria, secretory phase endometria and endometrial carcinomas, but its positive labeling index (PLI) in proliferative phase endometria was significantly higher than that in secretory phase endometria (P1/2 myometrial invasion (all P<0.01). Aurora B exerts its functions in the replication of normal endometrial glandular cells; Expression of Aurora B is significantly correlated with biologic behavior of endometrial carcinoma, indicating that Aurora B may be a promising prognostic factor in endometrial carcinoma.
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Carcinoma/metabolism , Cell Proliferation , Endometrial Neoplasms/metabolism , Endometrium/metabolism , Gene Expression Regulation , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Prognosis , Protein Serine-Threonine Kinases/biosynthesisABSTRACT
To investigate the relationship between MDR1 and MDR3 gene and drug resistance to cisplatin of ovarian cancer cells. Two siRNAs (MDR1, MDR3) which specifically targeted MDR1 and MDR3 genes were transfered into A2780/DDP cells. Then double staining with Annexin- V-FITC/PI was used to detect cell apoptosis by the flow cytometry (FCM). A2780/DDP cell viability was determined by MTT. MDR1 and MDR3 mRNA were assessed by RT-PCR. Caspase-3 protein was detected by Western blotting. Transfection of MDR1 and MDR3 siRNA into A2780/DDP cells failed to reverse the drug-resistance of A2780/DDP cells to cisplatin (P0.05). No significant differ- ence in the apoptosis efficiency was observed between the MDR1 and MDR3 siRNA, pSuppressor- Neo vector transfection cells and untreated cells (P0.05). In the presence of cisplatin of different concentrations, the viability of A2780/DDP cells was not significantly decreased after the transfection. No changes in MDR1 and MDR3 mRNA were found in MDR1 and MDR3 siRNA-transfected A2780/DDP cells. As compared with pSuppressorNeo and untreated groups, no significant difference existed in the expression of MDR1 and MDR3 mRNA (P0.05). The expression of caspase-3 protein in MDR1 and MDR3 siRNA transfected A2780/DDP cells was not significantly increased. It is con- cluded that multidrug resistance induced by cisplatin in ovarian carcinoma cell lines is not due to overexpression of MDR1 and MDR3 gene. The drug resistance of ovarian carcinoma cells to cisplatin is not mediated by P-glycoprotein.
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The purpose of this paper is to investigate the reversal effect of small interfering RNA (siRNA) targeting MDR1 and MDR3 genes on the resistance of MCF-7/ADR cells to adriamycin. siRNA plasmid vector targeting MDR1 and MDR3 genes was transfected into MCF-7/ADR cells, and then was stained with Annexin-V FITC (fluorescein isothiocyanate conjugated) to detect the early stage cell apoptosis by flow cytometry (FCM). 50% inhibition concentration (IC50) of adriamycin for MCF-7/ADR cells was determined by MTT method. MDR1 and MDR3 mRNA was assessed by RT-PCR. Treatment of MCF-7/ADR cells with the two kinds of siRNAs resulted in a reversal of adriamycin resistance of MDR to different extents. 1) The apoptosis efficiency of MDR1 and MDR3 siRNA vector after transfection was (18.21+/-1.65) % and (9.07+/-2.16) % respectively (P<0.05), and there was significant differences in the apoptosis efficiency between pSuppressor Neo vector and the MDR1siRNA or MDR3 siRNA vector (P<0.01); 2) The reversal effect of MDR1 siRNA is higher than that of MDR3 siRNA (P<0.05); 3) The expression of MDRI and MDR3 mRNA can be restrained by pSuppressor Neo MDR1 and MDR3 siRNA respectively, and the reduction in the mRNA level was in a time-dependent manner (P<0.01). MDR1 and MDR3 gene silencing can enhance intracellular adriamycin accumulation in MCF-7/ADR cells, improve sensitivity of MCF-7/ADR cells to adriamycin, and induce cell apoptosis. The reversal effect of adriamycin resistance by siRNA of MDR1 was more effective than that of MDR3.
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The purpose of this paper is to investigate the reversal effect of small interfering RNA (siRNA) targeting MDR1 and MDR3 genes on the resistance of MCF-7/ADR cells to adriamycin.siRNA plasmid vector targeting MDR1 and MDR3 genes was transfected into MCF-7/ADR cells, and then was stained with Annexin-V FITC (fluorescein isothiocyanate conjugated) to detect the early stage cell apoptosis by flow cytometry (FCM). 50 ％ inhibition concentration (IC50) of adriamycin for MCF-7/ADR cells was determined by MTT method. MDR1 and MDR3 mRNA was assessed by RT-PCR. Treatment of MCF-7/ADR cells with the two kinds of siRNAs resulted in a reversal of adriamycin resistance of MDR to different extents. 1) The apoptosis efficiency of MDR1 and MDR3siRNA vector after transfection was (18.21±1.65) ％ and (9.07±2.16) ％ respectively (P＜0.05), and there was significant differences in the apoptosis efficiency between pSuppressor Neo vector and the MDR 1 siRNA or MDR3 siRNA vector (P＜0.01); 2 ) The reversal effect of MDR 1 siRNA is higher than that of MDR3 siRNA (P＜0.05); 3 ) The expression of MDR1 and MDR3 mRNA can be restrained by pSuppressor Neo MDR1 and MDR3 siRNA respectively, and the reduction in the mRNA level was in a time-dependent manner (P＜0.01). MDR1 and MDR3 gene silencing can enhance intracellular adriamycin accumulation in MCF-7/ADR cells, improve sensitivity of MCF-7/ADR cells to adriamycin, and induce cell apoptosis. The reversal effect of adriamycin resistance by siRNA of MDRlwas more effective than that of MDR3.
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Objective To study the effects of ropivacaine on the duration of labor and mode of delivery in the primigravidas using patient-controlled epidural analgesia (PCEA). Methods Retrospective analysis was performed. The 190 healthy, full-term, and single-fetus parturient primigravidas who received PCEA with 0.1% ropivacaine+fentanyl (1 ?g/ml ) were in the epidural analgesia group. Another 222 primigravidas who didnot receive PCEA were in the control group. The duration of labor and modes of delivery, and the neonatal Apgar scores in both two groups were recorded and evaluated. Results Those in the epidural analgesia group experienced a significantly longer first stage [(426?161) minutes], longer second stage [(54?27) minutes] and longer full duration of delivery [(489?166) minutes] than those in the control one [(409?170) minutes, (364?167) minutes and (37?22) minutes]. The rate of using pitocin in the epidural analgesia group (30.2 %) was significantly higher than that in the control group (4.1%). The cesarean section rate in epidural analgesia group (20.0 %) was lower than that in the control one (28.4%); while the rate of instrumental delivery in the epidural analgesia group (20.0%) was significantly higher than that in the control one (6.3%). In summary, there were significant differences between two groups in the duration of labor, the rate of using pitocin, the rate of instrumental delivery and the rate of cesarean section. But there were no differences found for those newborn who had Apgar scores less than 7 at the point of both one and five minutes (7.9% and 4.5%, 2.6% and 0.5% respectively). Conclusion Epidural ropivacaine labor analgesia lengthens the duration of labor and increases the rate of instrumental delivery, but it has no significant negative effects on the neonates.