目的 探討聚腺苷二磷酸核糖聚合酶-1（poly ADPribose polymerase-1,PARP-1） mRNA在重癥急性胰腺炎(severe acute pancreatitis，SAP)大鼠腎臟中的表達及意義。方法 48只Wistar大鼠按隨機數字表法分為SAP組和假手術組(SO)組，分別于造模術后1、3、6及12 h測定血清肌酐，觀察胰腺和腎臟組織病理變化，并以RT-PCR法檢測PARP-1 mRNA在腎臟中的表達水平。結果 SAP組大鼠術后血清肌酐逐漸升高，于3、6及12 h明顯高于SO組(Plt;0.05)。SAP組大鼠術后胰腺出現腺體破壞、腺泡壞死、出血、炎性細胞浸潤等病理損害，且呈進行性加重； SO組各時相胰腺組織基本正常。SAP組大鼠術后出現腎小管上皮細胞變性、壞死、腎小球瘀血、缺血等改變，并隨時間延長逐漸加重，其損傷程度在3、6及12 h明顯較SO組嚴重(Plt;0.05)。SO組大鼠腎臟組織僅表達少量PARP-1 mRNA，而SAP組大鼠隨病程延長腎臟組織中PARP-1 mRNA表達逐漸增加，自3 h時起明顯高于SO組（Plt;0.01）。結論 在SAP發病過程中，PARP-1 mRNA的表達在腎臟組織中逐漸增加，PARP-1可能參與了SAP相關腎損傷過程。
Objective To evaluate the inhibiting effect of adenosine on rat retinal ganglion cells (RGC) death induced by P2X7 and N-methyl-D-aspartate (NMDA) receptor. Methods (1) Long-Evan neonatal rats were back labeled with aminostilbamidine to identify RGC. The viability of RGC affected by P2X7 excitomotor BzATP (50 mu;mol/L), glutamate receptor excitomotor NMDA (100 mu;mol/L) and adenosine (300 mu;mol/L) was detected. (2) RGC from the retinae of unlabeled neonatal rats were cultured in vitro. After labeled with Fura-2 methyl acetate, an intracellular calcium indicator, the effect of BzATP, NMDA and adenosine on intracellular Ca2+ level was detected byCa2+ imaging system. Results Both BzATP (50 mu;mol/L) and NMDA(100 mu;mol/L) could kill about 30% of the RGC. Cell death was prevented by adenosine (300 mu;mol/L) with the cell viability increased from (68.9plusmn;2.3)% and (69.9plusmn;3.2)% to (91.2plusmn;3.5)% (P＜0.001) and (102.1plusmn;3.9)% (P＜0.001), respectively. BzATP (50 mu;mol/L) led to a large, sustained increase of intracellular Ca2+ concentration to (1183plusmn;109) nmol/L. After the adenosine intervened, Ca2+ concentration increased slightly to (314plusmn;64) nmol/L (P＜0.001). Conclusion Adenosine may prevent RGC death and increase of intracellular Ca2+ concentration from P2X7and NMDA receptor stimulation. (Chin J Ocul Fundus Dis, 2007, 23: 133-136)
Objective To investigate the role of mitochondrial adenosine triphosphatesensitive potassium channel(mitoKATP) in immature myocardial ischemic preconditioning, and to provide evidence for immature myocardial protection. Methods Langendorff isolated heart infused model was used in the experiment. Twentyfour rabbits (aged from 14 to 21 days) were randomly divided into 4 groups：ischemiareperfusion group（I/R group）, myocardial ischemic preconditioning group(E1 group), 5hydroxydecanoate(5-HD) group (E2 group) and Diazoxide (Diaz) group(E3 group). Hemodynamics recovery rate, myocardial water content(MWC), the leakage rates of serum creatine kinase and lactate dehydrogenase, adenosine triphosphate content, superoxide dismutase activity, malondialdehyde content, myocardial cell Ca2+ content and myocardial mitochondrial Ca2+ content, myocardial mitochondrial Ca2+-ATPase activity, the adenosine triphosphate(ATP) synthesizing ability of myocardial mitochondria were tested, and myocardial ultrastructure was observed via electron microscopy. Results The hemodynamics recovery rate, myocardial water content(P＜0.05), adenosine triphosphate content, superoxide dismutase activity, myocardial mitochondrial Ca2+-adenosine triphosphyatase(ATPase) activity and the ATP synthesizing ability of myocardial mitochondria of the rabbits in E1 and E3 group were significantly better than that in I/R group and E2 group(P＜0.05). Malondialdehyde content, the leakage rates of serum creatine kinase and lactate dehydrogenase, myocardial cell Ca2+ content and myocardial mitochondrial Ca2+ content of the rabbits in E1 group and E3 group were significantly lower than that in I/R group and E2 group (P＜0.05). The myocardial ultrastructure injury in E1 and E3 group were significantly reduced compared with that in I/R and E2 group. Conclusion Myocardial ischemic preconditioning has significant protective effects on immature myocardium. Its mechanism may be related to the activation of mitoKATP.
ObjectiveTo summarize functions and mechanisms of poly ADP-ribose polymerase (PARP) inhibitors and its application in germline BRCA mutated breast cancer.MethodThe literatures about the PARP inhibitors and their applications in the treatment of germline BRCA mutated breast cancer at home and abroad in recent years were collected to make a review.ResultsAs a DNA repair enzyme, the PARP played an important role in the DNA repair pathway. Based on this mechanism, the PARP inhibitors had been developed and widely used in the clinic. On the other hand, the previous studies had shown that the PARP inhibitors marked the synthetic lethal effect in the cancers with homologous recombination deficiency mechanism. By inhibiting the PARP activity in the tumor cells with BRCA mutation, all the DNA damage repair pathways were blocked, which could induce the cell apoptosis or increase the sensitivity of tumor cells to chemoradiotherapy, resulting in the cell death.ConclusionIn patients with germline BRCA mutated breast cancer, PARP inhibitors can selectively kill breast cancer cells and show a high potential for individualized treatment.
Objective To study the effects of adenosine 2A receptor activation on activation, proliferation, and toxicity of T lymphocytes stimulated by phytohemagglutinin (PHA) in vitro. Methods A model of activated T cells was established by stimulating the cells with PHA. Those T cells were treated with different concentrations of adenosine 2A receptors agonist (0.01 μmol/L, 0.1 μmol/L, 1 μmol/L, and 10 μmol/L CGS21680). The expressions of CD69, CD25 and proliferation of T cells were measured by fluorescent antibody stain and flow cytometry. ELISA method was used to detect IL-2 and INF-γ levels. Results All concentrations of CGS21680 significantly inhibited the expressions of CD25 and CD69 on PHA-stimulated T cells surface and proliferation of T cells （Plt;0.05， Plt;0.01）. IL-2 and INF-γ secreted by T cells were significantly suppressed, too （Plt;0.01）. Conclusion Activation of adenosine 2A receptor can effectively inhibit the activation, proliferation, and toxicity of T cells in vitro.
目的:評價國產腺苷臨床應用安全性及腺苷負荷心肌灌注顯像對冠心病的診斷價值。方法:對51例臨床疑診冠心病患者,分別靜脈注射腺苷,注射3min末,靜脈注射核素顯像劑99TcmMIBI 925 MBq,1～1.5h行心肌灌注斷層顯像,分析患者在腺苷注入后的血流動力學改變、副作用發生情況。經半年以上隨訪排除或確診冠心病,評價腺苷負荷心肌灌注顯像對冠心病的診斷價值。結果:腺苷輸注后,86.3%(44/51) 患者出現各種副作用,停藥后均很快緩解。腺苷負荷試驗心肌灌注顯像診斷冠心病的敏感性90.9%,特異性71.4%,準確性88.2%,陽性預測值95.2%,陰性預測值55.6%。結論:國產腺苷可安全地應用于負荷心肌灌注顯像。腺苷負荷心肌灌注顯像診斷冠心病敏感性較高,具有重要的臨床應用價值。
Objective To study the effects of morroniside (MOR) on the proliferation and osteogenic differentiation of mouse MC3T3-E1 cells. MethodsThe 4th generation MC3T3-E1 cells were randomly divided into 6 groups: control group (group A), MOR low dose group (10 μmol/L, group B), MOR medium-low dose group (20 μmol/L, group C), MOR medium dose group (40 μmol/L, group D), MOR medium-high dose group (80 μmol/L, group E), and MOR high dose group (100 μmol/L, group F). The proliferation activity of each group was detected by cell counting kit 8 (CCK-8) assay; the bone differentiation and mineralized nodule formation of each group were detected by alizarin red staining; real-time fluorescence quantitative PCR (RT-qPCR) was performed to detect cyclin-dependent kinase inhibitor 1A (P21), recombinant Cyclin D1 (CCND1), proliferating cell nuclear antigen (PCNA), alkaline phosphatase (ALP), collagen type Ⅰ (COL-1), bone morphogenetic protein 2 (BMP-2), and adenosine A2A receptor (A2AR) mRNA expressions; Western blot was used to detecte the expressions of osteopontin (OPN), Runt-related transcription factor 2 (RUNX2), and adenosine A2AR protein. ResultsThe CCK-8 assay showed that the absorbance (A) values of groups B to F were significantly higher than that of group A at 24 hours of culture, with group C significantly higher than the rest of the groups (P<0.05). The MOR concentration (20 μmol/L) of group C was selected for the subsequent CCK-8 assay; the results showed that the A values of group C were significantly higher than those of group A at 24, 48, and 72 hours of culture (P<0.05). Alizarin red staining showed that orange-red mineralized nodules were visible in all groups and the number of mineralized nodules was significantly higher in groups B and C than in group A (P<0.05). RT-qPCR showed that the relative expressions of P21, CCND1, and PCNA mRNAs were significantly higher in group C than in group A (P<0.05). The expressions of ALP, BMP-2, COL-1, and adenosine A2AR mRNAs in groups B to E were significantly higher than those in group A, with the expressions of ALP, BMP-2, COL-1 mRNAs in group C significantly higher than the rest of the groups (P<0.05). Compared with group A, the expressions of OPN and RUNX2 proteins in groups B and C were significantly increased, while those in group D and E were significantly inhibited (P<0.05). There was no significant difference between groups B and C and between groups D and E (P>0.05). The relative expression of adenosine A2AR protein in groups B to E was significantly higher than that in group A, with group C significantly higher than the rest of the groups (P<0.05). Conclusion MOR can promote the proliferation and osteogenic differentiation of MC3T3-E1 cells; the mechanism of MOR may be achieved by interacting with adenosine A2AR.
ObjectiveTo systematically review the clinical efficacy and effects on pregnancy outcomes of S-adenosy-L-methionine combined with ursodesoxycholic acid in the treatment of intrahepatic cholestasis of pregnancy. MethodsDatabases such as PubMed, The Cochrane Library, CNKI, VIP, WanFang Data were searched for the studies about the clinical efficacy and effects on pregnancy outcomes of S-adenosy-L-methionine combined with ursodesoxycholic acid in the treatment of intrahepatic cholestasis of pregnancy up to December 31st, 2013. Two reviewers independently screened literature, extracted data and evaluated methodological quality. Then meta-analysis was conducted using RevMan 5.0.24 software. ResultsA total of 11 RCTs involving 776 patients were included. The results of meta-analysis showed that, combined medication reduced blood biochemical indexes inlcuding ALT (MD=3.63, 95%CI 0.63 to 6.64, P=0.02), TB (MD=3.70, 95%CI 1.45 to 5.96, P=0.001), and AST (MD=7.61, 95%CI 2.47 to 12.75, P=0.004). Combined therapy significantly decreased the rates of amniotic fluid contamination (OR=0.29, 95%CI 0.19 to 0.45, P=0.000 01), cesarean section (OR=0.53, 95%CI 0.36 to 0.79, P=0.002), postpartum hemorrhage (OR=0.32, 95%CI 0.12 to 0.90, P=0.03), preterm birth (OR=0.36, 95%CI 0.24 to 0.55, P < 0.000 01), fetal distress (OR=0.33, 95%CI 0.19 to 0.58, P=0.000 1) and neonates asphyxia (OR=0.30, 95%CI 0.19 to 0.47, P < 0.000 01). Combined therapy was also beneficial to improving pruritus symptoms (MD=0.20, 95%CI 0.08 to 0.31, P=0.000 08) and benefiting fetus growth (MD=0.45, 95%CI 0.23 to 0.66, P < 0.000 1). ConclusionThe combination of S-adenosy-L-methionine and ursodesoxycholic acid is superior to ursodesoxycholic acid alone in improving clinical symptoms and pregnant outcomes of patients with intrahepatic cholestasis of pregnancy.