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 investigate the changes and roles of myocardial adenosine triphosphate enzyme(ATPase) in the mechanism of cardiac dysfunction after blunt chest trauma（BCT）. Methods Thirtysix rabbits were divided into 6 groups with random number table, control group, 2 h group, 4 h group, 8 h group, 12 h group and 24 h group, 6 in each group. The models of BCT were established with BIMⅡ biological impact machine, catheterization technique was used through the right jugular artery into the left ventricle measure its pressure. The hemodynamics and the activities of ATPase in myocardial cell plasm, homogenate and mitochondria were measured at preinjury(control group), 2 h, 4 h, 8 h, 12 h and 24 h postinjury. Results Left ventricular endsystolic pressure(LVESP), the maximal ascending rate of left intraventricular pressure(+dp/dtmax), isovolemec pressure(IP) and the maximal physiological velocity(Vpm) decreased significantly at 2 h group after BCT(Plt;0.05）, and recovered to preinjury level in 4 h, 8 h and 12 h group during 4-12 h after BCT; isovolumic relaxation phase left ventricular pressure descending time constant (T). Left ventricular enddiastolic pressure(LVEDP) and the maximal descending rate of left intraventricular pressure(-dp/dtmax) were significantly higher （Plt;0.05, 0.01）. The activity of ATPase in homogenate, mitochondria and cell plasm decreased significantly at 2 h group and 4 h group after BCT（Plt;0.05, 001, respectively）, and 8 h group and 12 h group recovered after BCT. There was negative correlations between [CM(159mm]LVEDP and -dp/dtmax and the decrease of the activity of Na+-K+-ATPase in homogenate（r=-0.674, -0.691, Plt;0.05）, the Ca2+-ATPase in homogenate（r=-0.613,-0.642, Plt;0.05）, the Na+-K+-ATPase in mitochondria（r=-0.622,-0.616, Plt;0.05）,the Ca2+-ATPase in myocardial cell plasm（r=-0.672,-0.658, Plt;0.05）, the Na+-K+-ATPase in myocardial cell plasm（r=-0.627,-0.632,Plt;0.05）,and the Mg2+-ATPase in myocardial cell plasm（r=-0.677,-0.661, Plt;0.05）. Conclusion The left ventricular function is impaired obviously after BCT, especially in diastolic phase. The decrease of the activity of ATPase in myocardial cells may be one of the reasons of cardiac dysfunction after BCT.
Objective To investigate the effect of S-adenosylmethionine (SAM) on mitochondrial injury that was induced by ischemia-reperfusion in rat liver. Methods Fifty-four rats were randomly divided equally into 3 groups: control group, ischemia-reperfusion group (I/R group), and SAM-treated group (SAM group). Hepatic ischemia had been only lasted for 30 min by obstructing the blood stream of hepatic portal vena (the portal vena was only separated but not obstructed in control group). The rats of SAM group received SAM intraperitoneally 2 h prior to ischemia. Blood samples of each group were collected from the inferior cava vena at 0, 1 and 6 h after reperfusion and the serum levels of AST and ALT were detected. Mitochondrial super oxidedismutase (SOD), malondialdehyde (MDA), adenosine triphosphate (ATP) and energy charge (EC) in samples of liver tissue were detected, and the mitochondrial ultrastructure was observed with electronmicroscope. Results The serum levels of AST, ALT and mitochondrial MDA at 0, 1 and 6 h after reperfusion in the I/R group were significantly higher than those in the control group, whereas the levels of mitochondrial SOD, ATP and EC were significantly lower than those in the control group (P＜0.01). Except the value of 0 h, when it comes to SAM group, the levels of AST, ALT and mitochondrial MDA were significantly lower (P＜0.05) and the levels of mitochondrial SOD, ATP and EC were significantly higher (P＜0.05, P＜0.01) than those in the I/R group, respectively. The mitochondrial ultrastructure was injured obviously in I/R group when compared with that in control group. The number of mitochondria decreased and the mitochondria swelled, making the crista became obscure and the density of matrix became lower. The above changes in SAM group were less obvious when compared with those in I/R group. Conclusion SAM may protect mitochondrion against hepatic ischemia injury, since it may prevent mitochondrial lipid peroxidation, increase ATP, and eventually improve energy metabolism after ischemia-reperfusion.
Objective To study the distribution of P2 Y2 receptor in spine cord, dorsal root ganglia and sciatic nerve in rat, and to provide the basis for clarifying the mechanism of the effect of adenosine triphosphate(ATP) on the peripheral nerve regeneration. Methods Six specimens of the spine cord, dorsal root ganglia and sciatic nerve from SD rats were fixed rapidly in 4% paraformaldehyde which included DEPC, imbedded by paraffin and made into ultrathin section. According to the sequence of P2 Y2 receptor’s gene, DNA needle was adopted to detect the distribution of P2 Y2 receptor by hybridization technique in section under the light microscope after theyhad been stained in NBT liquid（50 mg/ml） and BCIP liquid （75 mg/ml）. In thecontrol group, the ultrathin section was only covered with hybridism buffer solution. The result of staining was observed. ResultsHybridization in section showed that P2 Y2 receptor was distributed mainly in the anterior horn cell of spine cordgray matter and Schwann cell of the dorsal root ganglia. No P2 Y2 receptor was observed in the sciatic nerve of both groups. Conclusion P2 Y2 receptor is located mainly in the spine cord and the dorsal root ganglia. Extracellular ATP can affect the cell of spine cord, dorsal root ganglia through P2 Y2 receptor.
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 investigate the regulatory effects of miR-26a-5p on the osteogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs) by regulating cAMP response element binding protein 1 (CREB1). Methods The adipose tissues of four 3-4 weeks old female C57BL/6 mice were collected and the cells were isolated and cultured by digestion separation method. After morphological observation and identification by flow cytometry, the 3rd-generation cells were subjected to osteogenic differentiation induction. At 0, 3, 7, and 14 days after osteogenic differentiation induction, the calcium deposition was observed by alizarin red staining, ALP activity was detected, miR- 26a-5p and CREB1 mRNA expressions were examined by real-time fluorescence quantitative PCR, and CREB1 protein and its phosphorylation (phospho-CREB1, p-CREB1) level were measured by Western blot. After the binding sites between miR-26a-5p and CREB1 was predicted by the starBase database, HEK-293T cells were used to conduct a dual-luciferase reporter gene experiment to verify the targeting relationship (represented as luciferase activity after 48 hours of culture). Finally, miR-26a-p inhibitor (experimental group) and the corresponding negative control (control group) were transfected into ADSCs. Alizarin red staining, ALP activity, real-time fluorescent quantitative PCR (miR-26a-5p) and Western blot [CREB1, p-CREB1, Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN)] were performed at 7 and 14 days after osteogenic induction culture. Results The cultured cells were identified as ADSCs. With the prolongation of osteogenic induction culture, the number of calcified nodules and ALP activity significantly increased (P<0.05). The relative expression of miR-26a-5p in the cells gradually decreased, while the relative expressions of CREB1 mRNA and protein, as well as the relative expression of p-CREB1 protein were increased. The differences were significant between 7, 14 days and 0 day (P<0.05). There was no significant difference in p-CREB1/CREB1 between different time points (P>0.05). The starBase database predicted that miR-26a-5p and CREB1 had targeted binding sequences, and the dual-luciferase reporter gene experiment revealed that overexpression of miR-26a-5p significantly suppressed CREB1 wild-type luciferase activity (P<0.05). After 7 and 14 days of osteogenic induction, compared with the control group, the number of calcified nodules, ALP activity, and relative expressions of CREB1, p-CREB1, OCN, and RUNX2 proteins in the experimental group significantly increased (P<0.05). There was no significant difference in p-CREB1/CREB1 between the two groups (P>0.05). Conclusion Knocking down miR-26a-5p promoted the osteogenic differentiation of ADSCs by up-regulating CREB1 and its phosphorylation.
Abstract： Objective To investigate the protective effects of adenosine (ADO) on lung ischemia/reperfusion injury following heart-lung transplantation in canine. Methods Canine heart-lung transplantation was performed．Canines were divided into two groups: transplant control groupand ADO group. The changes of arterial partial pressure of oxygen(PaO2) after reperfusion in two groups at 30，60，90，120 min were observed．The tissue contents of nitric oxide (NO) were measured at 10 min before ischemia, 10 min and 120 min after ischemia; 10 min and 60 min after reperfusion．The lung tissue samples were obtained 1h after reperfusion．The tissue myeloperoxidase(MPO) activity,content of malondialdehyde(MDA), content of superoxide dismutase(SOD), wet/dry ratio of lung(W/D) were measured.Microscopic examination of lungs was also conducted. Results (1)In ADO group，PaO2 were significantly higher than that in control group at 30，60，90 and 120 min after reperfusion (Plt;0.05)．(2) The tissue contents of NO at 120 min after ischemia, 10 min and 60 min after reperfusion were significantly lower than that at 10 min before ischemia(Plt;0.05). In ADO group，the tissue contents of NO at 120 min after ischemia, 10 min and 60 min after reperfusion were higher than that in control group respectively(Plt;0.05). (3)The tissue MPO activity, content of MDA, W/D in ADO group were significantly lower than those in corresponding control group. The content of SOD in ADO group were higher than that in control group(Plt;0. 05)．(4)The microscopic examination showed that there were severe leukocyte infiltration and edema formation in the alveolar space in control group, but the changes were less severe in ADO group. Conclusion Administration of ADO in canine heart-lung transplantation can protect the donor lung against ischemia／reperfusion injury．
ObjectiveTo explore the clinical value of age/pleural fluid adenosine deaminase (age/ADA) ratio and serum lactate dehydrogenase/pleural fluid adenosine deaminase ratio (Cancer Ratio, CR) in the diagnosis of malignant pleural effusions (MPE). MethodsThe study collected 44 patients with MPE and 48 patients with benign pleural effusion (BPE) to compare the differences in age, gender, carcinoembryonic antigen (CEA), age/ADA ratio and CR between the groups. The receiver operating characteristic (ROC) curve of CEA, age/ADA and CR was constructed and the area under the ROC curve (AUC), sensitivity and specificity was calculated to identify the diagnostic performance of the three indicators alone or in combination in MPE. ResultsCEA, age/ADA and CR were significant higher in the MPE group than those in the BPE group (all P<0.05), the AUCs of CEA, age/ADA and CR were 0.768, 0.837 and 0.866, respectively; the sensitivity was 61.36%, 88.64% and 81.82%, the specificity was 85.42%, 75.00%, 83.33%, respectively. The AUCs of CEA combined with age/ADA, CEA combined with CR, age/ADA combined with CR, CEA combined with age/ADA and CR were respectively 0.892, 0.911, 0.837 and 0.907; the sensitivity was 81.82%, 86.36%, 88.64% and 90.91%, the specificity was 79.17%, 79.17%, 75.00% and 77.08%, respectively. ConclusionsAge/ADA and CR demonstrated good diagnostic performance in MPE, moreover, the diagnostic performance can be further improved when combined with the traditional tumor marker CEA, and more research about its diagnostic value is needed in the future.