Objective: To investigate the protective effect of human lipoxin A4 (LXA4) on N2a cell damage induced by β-amyloid protein 25-35 (Aβ25-35) and the underlying mechanism. Methods: Aβ25-35 was used to treat N2a cells to establish Alzheimer's disease (AD) cell injury model. Meanwhile, LXA4 was added to the experimental group at different concentrations (50, 100 and 200 nmol·L-1 ). MTT assay was used to detect the activity of N2a cells. The apoptosis was detected by Hoechst 33258-PI staining, the expression of P62 and TRAF6 mRNA was detected by RT-PCR, and the expression of P62 and TRAF6 protein was detected by Western blot. Results: Compared with that of the model group, the cell survival rate of LXA4 protective group (50,100 and 200 nmol·L-1 ) increased (P <0. 01) and the apoptosis of N2a cells induced by Aβ25-35 was reduced by LXA4 (100 and 200 nmol·L-1 ) . Compared with that of the model group, the expression of P62-mRNA and protein-P62 of N2a cells treated with Aβ25-35 increased (P <0. 05 or P <0. 01) and the expression of TRAF6-mRNA and protein-TRAF6 of N2a cells treated with Aβ25-35 were reduced (P <0. 05 or P <0. 01). Conclusion: LXA4 has protective effect on N2a cell damage induced by Aβ25-35 , and its mechanism may be related to the up-regulation of P62 gene and down-regulation of TRAF6 gene.

Biomarkers ; blood ; Child ; Chorionic Gonadotropin ; blood ; Follicle Stimulating Hormone ; blood ; Growth Disorders ; etiology ; Humans ; Klinefelter Syndrome ; complications ; diagnosis ; genetics ; Magnetic Resonance Imaging ; Male ; Mediastinal Neoplasms ; complications ; diagnosis ; surgery ; Puberty, Precocious ; diagnosis ; etiology ; Teratoma ; complications ; diagnosis ; surgery ; Testis ; pathology

OBJECTIVETo investigate serum level and gene polymorphisms of matrix metalloproteinase 9 (MMP-9), and platelet glycoprotein VI (GPVI) in patients with acute coronary syndrome (ACS).

METHODSIn a prospective study of 179 patients with documented ACS and 164 controls, we measured baseline serum MMP-9 levels using ELISA and determined the MMP-9/C-1562T and MMP-9/G5564A genotypes using PCR-restriction fragment length polymorphism. Fib serum level was measured by Clauss assay. We also analyzed the Fib/Bbeta-148C/T and GPVI/T13254C polymorphisms.

RESULTSSerum levels of MMP-9 and Fib in ACS patients were significantly higher than in controls (P < 0.001), and serum level of Fib in the acute myocardial infarction group was higher than in patients with unstable angina (P < 0.05). No significant difference between ACS patients and controls was found in frequencies of MMP-9/C-1562T, MMP-9/G5564A, Fib/Bbeta-148C/T, and GPVI/T13254C genotypes and alleles (P > 0.05). The T allele of the Fib/Bbeta-148T polymorphism was associated with increased plasma Fib level (P < 0.05). There was a strong positive correlation between serum level of MMP-9 and Fib (r = 0.289, P < 0.01).

CONCLUSIONSerum levels of MMP-9 and Fib were independent risk factors of ACS. There was an obvious relationship between the Bbeta-148C/T mutation and high Fib level. No significant difference between controls and ACS patients was found in the frequencies of MMP-9 C-1562T and G5564A, Fib Bbeta-148C/T and GPVI T13254C genotypes and alleles (P > 0.05).

Acute Coronary Syndrome ; genetics ; Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Matrix Metalloproteinase 9 ; blood ; genetics ; Middle Aged ; Platelet Membrane Glycoproteins ; genetics ; Polymorphism, Single Nucleotide

Congresses as Topic ; Humans ; Rhinitis ; Rhinitis, Allergic, Perennial

OBJECTIVETo investigate the effect of different preparation method on the quality of Shen-mai injection.

METHODThe Shen-mai injection samples were prepared using three different methods. Fingerprints of Shen-mai extracts red ginseng, and its intermediates were obtained using an HPLC analytical procedure. The contents of ginsenoside Rg1, Rc and Rb1, and the gross saponins of Shen-mai extract were quantitatively mensured with HPLC procedures.

RESULTThere was significant difference in fingerprints and chemical contents of the injections prepared by the three different methods.

CONCLUSIONThe quality of Shen-mai injection was greatly influenced by the preparation method. HPLC fingerprinting method can be applied for the determination of the Shen-mai preparations.

Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Ginsenosides ; analysis ; Injections ; Ophiopogon ; chemistry ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Saponins ; analysis ; Technology, Pharmaceutical ; methods

OBJECTIVETo express and purify the fusion proteins of glutathione S-transferase (GST)-N-terminal of histone deacetylase4 (HDAC4-N') (1-1952 bp) and GST- C-terminal of HDAC4 (HDAC4-C') (1708-3255 bp) in E.coli.

METHODSThe DNA fragments (HDAC4-N' and HDAC4-C') amplified by PCR were ligated into GST fusion vector (pGEX-6P-1) to construct the recombinant plasmids. After identification with restriction digestion and DNA sequencing, the recombinant plasmids were transformed into E.coli BL21 and induced by IPTG for their expression. After identification by SDS-PAGE and Western blotting, the target proteins were purified by glutathione sepharose 4B.

RESULTSThe results of restriction digestion and DNA sequencing confirmed successful construction of the recombinant plasmids. The relative molecular masses of the fusion proteins were approximately 110500 and 93080 as shown by SDS-PAGE. Western blotting demonstrated that the fusion proteins could be recognized by the specific anti-HDAC4 antibody.

CONCLUSIONWe have successfully constructed the recombinant expression vectors of pGEX-6P-1/HDAC4-N' and pGEX-6P-1/HDAC4-C' and induced the expression of the fusion proteins, which may facilitate functional studies of HDAC4 with other proteins.

Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Histone Deacetylases ; biosynthesis ; genetics ; Humans ; Peptide Fragments ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Repressor Proteins ; biosynthesis ; genetics

Objective To investigate the effect of autophagy on the apoptosis of H 9C2 cardiomyocytes.Methods H9C2 cardiomyocytes were incubated with different concentrations of high glucose and high lipids ( HGHL ) for different time.The surface area of cardiomyocytes was measured after HE staining .The cell viability and apoptotic rate were measured by flow cytometry.Western blot was used to detect the expression levels of LC3Ⅱ, p62, Beclin-1,LAMP2 and cleaved caspase-3.Results After treatment with HGHL, the cells appeared hypertrophy in a concentration-and time-dependent manner , the cell hypertrophy was most obvious under the condition of HGHL(500 μmol/L,36 h)(P<0.001).Cell apoptosis increased in a concentration-and time-dependent manner, the apoptotic rate was nearly 50%under the condition of HGHL ( 500 μmol/L,36 h) ( P<0.001 ) .After treatment with HGHL for 24 h, compared with the control group , the expression of LC3Ⅱ was very significantly increased ( P<0.01) , the expression of Beclin-1 and LAMP 2 were significantly decreased ( P<0.05) , but the expression of p 62 was significantly increased ( P<0.01 ) .Compared with the control group and the intervention group , the expression of cleaved caspase-3 was significantly increased ( P<0.01 ) after 1 h of chloroquine pretreatment .Conclusions HGHL induced H9C2 cardiomyocytes hypertrophy promote H 9C2 cardiomyocytes apoptosis in a concentration-and time-dependent manner .HGHL may inhibit autophagy formation and degradation of H 9C2 cardiomyocytes at the same time , leading to abnormal autophagy accumulation of cardiomyocytes , and promote apoptosis , suggesting that inhibition of autophagy may be an important cause for promote apoptosis .

To establish a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency.

OBJECTIVETo investigate the effects of rabbit limbs ischemia/reperfusion on myocardial necrosis and apoptosis in vivo.

METHODSThirty-six healthy new zealand rabbits were randomly divided into 3 groups: (1) Sham group; (2) I/R(Ischemia/reperfusion) group; (3) RPostC (remote postconditioning) group. The activity of blood serum creatine kinase (CK) and lactate dehydrogenase (LDH) were measured at baseline, the end of ischemia after 60 min and 120 min of reperfusion respectively. The extent of myocardial ischemia and the scope of myocardial infarction were assessed by evans blue and Triphenyl tetrazolium chloride (TTC). The myocardial cell's apoptosis at the area of myocardial ischemia was estimated by Tunel. Protein expression of caspase-3, Bcl-2 and Bax in myocardial ischemic area were analyzed with the method of immunohistochemistry.

RESULTSCompared with I/R group, the myocardial infarct size and the CK activity were significantly reduced in RPostC group. The Tunel positive index of RPostC group in ischemic myocardium was significantly lower than that in I/R group (21.79% +/- 1.07% vs 35.81% +/- 1.10%, P < 0.05). Caspase-3 positive cells index was calculated with randomly selected five regions in each slide and then the positive cells in per hundred cells were calculated. The RPostC group of caspase-3 positive cells was significantly lower than that in I/ R group(25.03% +/- 1.16% as 39% +/- 2.43%, P < 0.05). Compared with the sham group, the Bax protein expression index and the Bcl-2 protein expression index of I/R group and RPostC group were increased. The Bax/Bcl-2 ratio of RPostC group decreased, while it was increased in I/R. Compared with the I/R group, the Bax protein expression and Bax/Bcl-2 ratio of RPostC group significantly reduced, but the expression index of Bcl-2 ratio was significantly increased, the differences were statistically significant.

CONCLUSIONLimbs ischemia/postconditioning could significantly reduce necrosis and apoptosis of ischemia/reperfusion myocardium. The mechanism of reducing the myocardial cell apoptosis may have relation to inhibiting the activation of pro-apoptotic gene caspase-3 and increased expression of Bcl-2.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Creatine Kinase ; blood ; Ischemic Postconditioning ; L-Lactate Dehydrogenase ; blood ; Lower Extremity ; Male ; Muscle, Skeletal ; blood supply ; Myocardial Reperfusion Injury ; pathology ; Necrosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rabbits ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo study the prediction value of age and congestive heart failure (CHF) for occurrence of multiple organ dysfunction syndrome elderly(MODSE) in old patients with hypertension.

METHODSMedical history of 19,996 cases (aged over 60 year) admitted to PLA General Hospital because of hypertension or developing hypertension during hospital stay from Jan 1993 to Dec 2008 were analyzed retrospectively. According to age the patients were divided into four groups: 60-69 year group; 70-79 year group; 80-89 year group; > or = 90 year older group. The incidence of CHF and the morbidity of MODSE induced by CHF at different ages and different boundary ages were investigated.

RESULTS1. The incidence of MODSE in CHF cases was higher than that in the non-CHF cases (7.43% versus 3.05%, Chi(2) 195.15, P < 0.01), showing CHF were the important factor in happening of MODSE. 2. The incidence of CHF and the morbidity of MODSE were 10.60% versus 18.88% versus 30.11% versus 60.57%, P <0.05, P < 0.05 and 1.6 versus 7.0 versus 17.08 versus 25.47% , in 60-69 year group; 70-79 year group; 80-89 year group; > or =90 year older group, P < 0.05. Occurrence of CHF and that of MODSE were positively correlated with age (r = 0.696 - 0.987, P < 0.01). High risk population of MODSE induced by CHF were old patients with hypertension above 69 year old.

CONCLUSIONThe age is valuable for early prediction of MODSE induced by CHF in old patients with hypertension. The distinctly boundary age for the incidence of MODSE induced by CHF in old patients with hypertension is 69.

Age Factors ; Aged ; Aged, 80 and over ; Female ; Heart Failure ; epidemiology ; Humans ; Hypertension ; complications ; Incidence ; Male ; Middle Aged ; Multiple Organ Failure ; epidemiology ; Retrospective Studies