To explore the destructive effect of residual uracil DNA glycosylase on dUTP incorporation to the cDNA PCR products,PAGE and DNA-EIA hybridization technique were applied to detect the efficiency of anti-contamination and the rate of DNA hybridization.In this study,it was found that:⑴Good effect could be achieved with the reaction of 2u of uracil DNA glycosylase(UDG)for 20min at 37℃.⑵UDG could be inactivated at 94℃ for 10min.The hybridization rate of control group of PCR products was 94 18% at room temperature for 65 hours.The hybridization rates of test groups at-20℃ for 65 hours were 87 69%,77 24%,76 83%,respectively.And the inactivation times of anti-contamination groups at 94℃ were“0”min,“10”min and “20”min,respectively;at room temperature for 65 hours,these hybridization rates were 74 77%,72 50%,70 29%,respectively.There were significant differences between control and rest groups(P0 05).Our study suggested that the hybridization rate of test groups kept at-20℃ decreased 19 41%,after 65 hours,the rate decreased 27 45%,but in control group only decreased 5 82% for 65h at room temperature.So that,we conclude that the residual UDP can cause above results.

Objective To investigate the safety,efficacy and effects of emergent percutaneous coronary intervention (PCI) in patients from Second Affiliated Hospital,Medical College,Zhejiang University with cardiogenic shock (CS) complicating acute myocardial infarction (AMI).Method Twenty-seven patients with CS complicating with AMI were treated by PCI with intraaortic balloon counterpulsation (IABP) support.The change of hemodynamics before and after IABP and PCI,the characteristics of PCI,the mortality during hospitalization, the major adverse cardiac events (MACE) and left ventricular ejection fraction at 30-day follow-up were observed.Results The hemodynamics were significantly improved after IABP.No patients died during PCI.Two patients died after PCI and the total mortality was 7.4% in hospital.During the period of 30-day follow-up, one patient died of heart failure.The left ventricular ejection fraction greatly improved at 30 days after PCI. Conclusions The data suggested that the use of IABP during PCI in patients with CS complicating AMI was safe, decreased mortality and improved prognosis.

Objective To explore the mechanism of hepatic autophagy inhibition induced by ischemia-reperfusion injury in the aging liver.Methods The healthy male Lewis rats aged 3 months (3M) and 24 months (24M) were selected,and then were randomly divided into 3M IRI group,3M sham operation group,24M IRI group,24M sham operation group.In the experimental group,noninvasive vascular clamp was used to clamp the left and middle hepatic lobes (about 70o％ hepatic ischemia).The liver was subjected to ischemia at 37 0.5℃ for 30min and reperfusion for 6h.The hepatic duodenal ligament was dissected only by sham operation.The serum levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured at 6 h after operation.Liver tissues of each group were examined by liver pathology and the number of autophagosome of LC3B in liver tissue of each group was observed under confocal microscopy.The changes of autophagyrelated protein were analyzed by Western blotting.Results The levels of serum ALT and AST in 24M IRI group were significantly higher than those in 3M IRI group,the difference was statistically significant (P＜0.05).Pathological analysis showed that 3M IRI group showed spotty necrosis,the 24M IRI group showed massive necrosis and the infiltration of the inflammatory cells;Confocal microscopy showed that the number of autophagosome in the liver tissue of the 24M sham group was slightly lower than that of the 3M sham operation group and the number of autophagosome in the 24M IRI group was significantly lower than that in the 3M IRI group (P＜0.05).The levels of autophagyrelated proteins (Beclin1 and ATG4B protein) in 24M IRI group were significantly down-regulated compare to 3M IRI group (P＜0.05).Conclusion The ischemia-reperfusion injury of liver in aged rats inhibits autophagy,and its mechanism may be related to the decrease of autophagy-related protein level in hepatic ischemia-reperfusion injury.

Remote organ ischemic preconditioning is to conduct a transient and sublethal ischemic adaptation in non-vital organs before occurring cerebral ischemia/reperfusion injury in remote vital organs. Remote organ iscbemic preconditioning has been studied for as long as 15 years in the field of myocardial iscbemia. However, only recently it has become a therapeutic strategy for the treatment of cerebrovascular diseases, This article briefly reviews the methods and mechanisms involved in the protective effects of cerebral ischemia of remote organ ischemic preconditioning.

0.05).The percentage of CD40 positive cells in CBMC-derived DC was lower than that in PBMC-derived DC[(34.80?7.77)% vs(54.37?9.57)%,P

AIM: To study effect of endogenous carbon monoxide on intracellular calcium concentration and explore the mechanism in brain protection of endogenous CO in focal cerebral ischemia in rats. METHODS: SD rats were divided into three groups randomly, which including hemin, ZnPP group and saline group as control. Respectively saline, hemin, ZnPP were injected intra-peritoneally twelve hours before middle cerebral artery was occluded. Twenty four hours after MCAO model was set up, the concentration of carbon monoxide in blood and intracellular calcium in neural cells was examined. RESULTS: Contrast to saline group, the concentration of CO in blood rose up while intracellular calcium in occluded side decreased in hemin group; the concentration of CO in blood went down while intracellular calcium in occluded side rose up in ZnPP group, there was significant difference among them (P0.05). CONCLUSIONS: It may be one of mechanisms on brain protection in ischemic cerebral tissue that carbon monoxide affected intracellular calcium concentration of neural cells by regulating Ca~(2+)-K~+ channel on cell membrane as a messenger gaseous molecular and neurotransmitter. [

OBJECTIVETo explore the effect of Bushen Gujin Recipe (BGR) on serum and synovial expression of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in knee osteoarthritis (KOA) model rabbits.

METHODSTotally 36 8-month-old healthy male New Zealand white rabbits were randomly divided into 4 groups, i.e., the normal control group, the model group, the Western medicine group (Meloxicam, at the daily dose of 6 mg/kg), and the TCM group (BGR, at the daily dose of 53 g/kg), 9 in each group. Modeling was performed in all rabbits except those in the normal control group by using Hulth A method. All medication was performed for 8 consecutive weeks. Contents of IL-1 and TNF-α were detected using ELISA from serum, partial synovial tissue of the front knee joint, cartilage and subchondral bone of the medial femoral condyle.

RESULTSThe joint space became narrowed in the Western medicine group, ranging between the model group and the TCM group. The articular surface was rough with obvious osteophytes. The joint space was slightly narrower in the TCM group; the articular surface was slightly rough with mild osteophytes. Compared with the normal control group, contents of IL-1 and TNF-α in serum and synovial increased in the model group (P < 0.01). Compared with the model group, contents of IL-1 and TNF-α in serum and the synovial fluid decreased in the two treatment groups (P < 0.01). There was no statistical difference in contents of IL-1 and TNF-α between the Western medicine group and the TCM group.

CONCLUSIONBGR promoted the synthesis of cartilage matrix and carti- lage repair through inhibiting the secretion of IL-1 and TNF-α, and prolonging cartilage degeneration.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-1 ; metabolism ; Knee Joint ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Rabbits ; Synovial Fluid ; Synovial Membrane ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Adult ; Angiomatosis ; immunology ; pathology ; surgery ; Antigens, CD ; metabolism ; Diagnosis, Differential ; Female ; Hemangioma ; immunology ; pathology ; Humans ; Hyperplasia ; Spleen ; immunology ; pathology ; Splenectomy ; Splenic Diseases ; immunology ; pathology ; surgery ; Splenic Neoplasms ; immunology ; pathology

Adenomatous Polyposis Coli Protein ; genetics ; Chromatography, High Pressure Liquid ; methods ; Codon ; genetics ; DNA Mutational Analysis ; Fibromatosis, Aggressive ; genetics ; pathology ; Humans ; Mutation ; Polymerase Chain Reaction ; beta Catenin ; genetics