Objective To investigate the role of hypoxia inducible factor(HIF)-prolyl hydroxylase 1 (HPHl)and factor inhibiting HIF-1(FIH-1)in placentas in the pathogenesis and development of severe pre-eclampsia.Methods RT-PCR and western blot analyses were used to detect the HPH1 and FIH-1expression levels in placentas of 34 patients with severe pre-eclampsia and 24 cases of term pregnancy (normal pregnancy group)and their correlations with symptoms were analyzed.Results (1)The HPHI mRNA and protein expression levels in placentas of severe pre-eclampsia group were 0.40±0.04 and 59.5±3.4 separately,significantly lower than those of normal pregnancy group,0.84±0.12 and 71.6±1.7(P<0.01).The FIH-1 mRNA and protein expression levels in placentas of severe pre-eclampsia group wereQ 31 ±0.05 and 45.6±2.4 separately,significantly lower than those of normal pregnancy group,0.43±0.04 and 54.9±2.1(P<0.01).(2)The mRNA and protein expression levels of HPH1 and FIH-1 in severe pre-eclampsia group were all negatively correlated with mean arterial pressure(MAP)[the Spearman correlation coefficient was-0.854(P<0.01)],urinary protein per 24 hours[the Spearman correlation coefficient was-0.936(P<0.01)1 and the occurrence of fundus oculi artery spasm[the Spearman correlation coefficient was-0.854(P<0.01)].(3)rrhe expression of HPHl mRNA in placentas of all the 58 cases WBB 0.58±0.27.higher than the expression of FIH-1 mRNA,which was 0.39±0.10.There was a positive correlation between them.The pearson correlation coefficient was 0.686(P<0.01).The expression of HPH1 protein in placentas of all the 58 cases was 64.5±6.7,higher than the expression of FIH-1,which was 49.4±5.2.There was a positive correlation between them.The Pearson correlation coefficient was 0.947(P<0.01).Conclusion The expression imbalance of HPH1 and FIH-1in palcenta may play an important role in the pathogenesis and development of severe pre-eclampsia through inhibiting HIF-1a.

Objective To evaluate different expressions of high mobility group box 1(HMGB1)and receptor for advanced glycation end products(RAGE)in placentas and their relationship with preeclampsia.Methods Fifteen early-onset pre-eclaraptic women(early-onset pre-eclampsia group),22 late-onset pre-eclamptic women(late-onset pre-eclampsia group)and 12 normotensive women(control group)in the third trimester were recruited at the Shengjing Hospital of China Medical University from March 2006 to March 2007.The localization and levels of HMGB1 and RAGE in placentas of the three groups were detected by the strept avidin biotin-peroxidose method.Results (1)Immunoreactivities to HMGB1:positive immnnostaining for HMGB1 was observed in trophoblast,macrophages,decidual cells,vascular muscle cells,endothelial cells and placental mesenchymal cells in the placentas from the pre-eclamptic women,while a low level of immunoreactivities was observed in the placentas from healthy pregnancies;the staining was observed within both the nuclei and the cytoplasm,mainly in the cytoplasm.The cytotrophoblast,especially the nuclei was extensively positive for HMGB1 in early-onset pre-eclampsia. (2)Immunoreactivities to RAGE:positive immunostaining for HMGB1 was observed in syncytiotrophoblast,macrophages and endothelial cells in the placentas from the preeclamptic women,while a low level of immunoreactivities was observed in the placentas from healthy pregnancies:the staining was in the cytoplasm and(or)cell membrane.The trophoblast was extensively positive for RAGE in early-onset pre-eclampsia.(3)Positive rate of HMGB1 expression:the expression of HMGB1 in early-onset group(73%,11/15)and late-onset group(64%,14/22)was significantly higher than that in normal group(17%,2/12;P<0.05),but no significant difference was found in early-onset group and late-onset group(P>0.05).(4)Positive rate of RAGE expression:the expression of RAGE in early-onset group(80%,12/15)and late-onset group (82%,18/22)was significantly higher than that in normal group(25%,3/12;P<0.05),but no significant difference was found in early-onset group and late-onset group(P>0.05).Conclusions The increased expression of HMGB1 and RACE in the placenta may play an important role in the pathogenesis of pre-eclampsis.The different locations may be associated with the occurrence of different onset types of pre-eclampsia.

The fruit of Pandanus tectorius (PTF) has a long history of use as a folk medicine to treat hyperlipidemia in Hainan province, South China. Our previous studies have shown that the n-butanol extract of PTF is rich in caffeoylquinic acids and has an adequate therapeutic effect on dyslipidemic animals induced by high-fat diet. In this work, seven caffeoylquinic acids isolated from PTF were screened for the lipid-lowering activity in HepG2 hepatoma cells. Oil-Red O staining, microscopy and intracellular triglyceride (TG) and total cholesterol (TC) quantification showed that 3-O-caffeoylquinic acid (3-CQA), 3, 5-di-O-caffeoylquinic acid (3,5-CQA), and 3,4,5-tri-O-caffeoylquinic acid (3,4,5-CQA) significantly inhibited lipid accumulation induced by oleic acid and decreased intracellular levels of TC and TG in a dose-dependent manner. These three caffeoylquinic acids showed no significant cytotoxicity at concentrations of 1 -50 μmol x L(-1) as determined by MTT assay. Realtime quantitative PCR revealed that 3-CQA and 3, 5-CQA significantly increased the expression of lipid oxidation-related genes PPARα, CPT-1 and ACOX1 while 3-CQA, 3, 5-CQA and 3,4,5-CQA decreased the expression of lipogenic genes SREBP-1c, SREBP-2, HMGR, ACC, FAS. Overall, 3-CQA, 3, 5-CQA and 3, 4, 5-CQA may be the principal hypolipidemic components in PTF which can decrease intracellular lipid accumulation through up-regulating the expression of lipid oxidative genes and down-regulating the expression of lipogenic genes.
Carcinoma, Hepatocellular ; metabolism ; China ; Cholesterol ; metabolism ; Gene Expression Regulation ; Hep G2 Cells ; Humans ; Lipid Metabolism ; Liver Neoplasms ; metabolism ; Oleic Acid ; Pandanaceae ; chemistry ; Quinic Acid ; analogs & derivatives ; chemistry ; Sterol Regulatory Element Binding Protein 1 ; Triglycerides ; metabolism

Objective:To prepare monoclonal antibodies specifically against an immunogenic fragment in ectodomain of prostate-specific membrane antigen ( PSMA ).Methods: An polypeptide immunogenic fragment in the ectodomain of PSMA was predicted by biological information technology,and then it was expressed prokaryotically.BALB/c mice were immunized with the prokar-ytically expressed recombinant polypeptide antigen,to prepare the monoclonal antibodies specifically against an immunogenic fragment in ectodomain of PSMA by hybridoma technology,purification of monoclonal antibody by affinity chromatography,characterization of the monoclonal antibodies by Western blot.The radioimmunoimaging in prostate cancer model was performed by using the labeled McAb.Results:Throught the software analysis,we got the antigen fragment in the ectodomain of PSMA containing 310aa sequences higher specificity, artificially synthesized gene sequence of the region, and constructed a prokaryotic expression vector pET-32a-r-ectodomain-PSMA,by prokaryotic expression we obtained the 50 kD target antigen,after hybridization,the three positive hybridoma cell lines (5E6,4A5 and 4D7) were selected by ELISA using target antigen,the isotypes of 5E6 and 4A5 were IgG2a,the isotypes of 4D7 were IgG1,the titer of three monoclonal antibodies was above 1∶256 000.Western blot results showed that the prepared monoclonal anti-bodies could binding specifically to the antigen in the ectodomain of PSMA.Radioimmunoimaging in prostate cancer animal model results further confirm that the prepared monoclonal antibodies could combinate with the antigen in the ectodomain of PSMA in the animal body, and make the tumor imaging.Conclusion: The prepared monoclonal antibodies can specifically recognizes the PSMA antigen,which laid the foundation for the immunodiagnosis and immunotherapy of prostate cancer.

Objective To study effects of different degree of hypothyroidism in severe preeclampsia (S-PE) pregnant women on renal function and the correlation between them.Methods 46 S-PE patients with subclinical hypothyroidism (SCH) registered for treatment in the Shengjing Hospital of China Medical University from May 2011 to March 2013 were selected into SCH group,and 23 S-PE with overt hypothyroidism (OH) were selected into OH group,and 109 S-PE with normal thyroid stimulating hormone (TSH) levels were selected into simple group.Thyroid hormone and kidney function tests were analyzed in pregnant women with S-PE.We made an analysis of the relative risk of the detection rate of abnormal renal function and also the relationship between the levels of thyroid hormone and serum uric acid,serum urea and creatinine in patients with S-PE.Results (1) In SCH group serum TSH was (6.1±3.2) mU/L,free triiodothyronine (FT3) was (4.0±0.6) pmol/L,free thyroxine (FT4) was (11.8± 1.5) pmol/L; in OH group serum TSH was (5.2± 1.3) mU/L,FT3 was (3.7±0.6) pmol/L,FT4 was (9.3±0.5) pmol/L; in simple S-PE group serum TSH was (1.9±0.8) mU/L,FT3 was (4.0±0.8) pmol/L and FT4 was (11.9±1.9) pmol/L.TSH in SCH group was significantly higher than that in simple S-PE group (P＞0.01),the difference of in SCH and OH group were not statistically significant (P＞0.05).The difference of FT3 in three groups were not statistically significant (P＜0.05) ;FT4 in OH group was significantly lower than thoes in SCH and simple groups (P＜0.05).(2)Serum uric acid,creatinine and urea levels in OH group was (436± 114),(75± 15) μmol/L and (6±3)mmol/L,in simple S-PE group they were (378± 114),(65 ±22) μmol/L and (5±3) mmol/L.In comparison,the differences was statistically significant(P＜0.05).The differences were not statistically significant in SCH and OH groups (P＞0.05).(3)The abnormal detection rate of uric acid was significantly higher in SCH than that in OH group [46％ (21/46) versus 22％ (5/23),OR=3.0,P＜0.05].The comparison of remaining index has no statistical significance(P＞0.05).(4)In SCH group there was a significant inverse correlation of serum FT3 with serum urea levels,serum creatinine and serum uric acid (r=-0.32,-0.58,-0.35,P＜0.05).There was not a correlation of serum TSH,FT4 with indicators of renal function (P＞0.05).In OH group there was a negative correlation between FT3 and serum creatinine concentrations (r=-0.40,P＜0.05).In OH group there was not a correlation of FT3 with serum uric acid and urea (P＞0.05).There was a positive correlation between TSH and serum creatinine in simple S-PE group (r=0.20,P=0.04).There was not a correlation between TSH and serum urea(r=0.04,P=0.65),and serum uric acid (r=0.12,P=0.20).Conclusions There was effect of different hypothyrosis state in pre-eclampsia patients on renal function.Serum uric acid,urea and creatinine concentrations in S-PE pregnant women with OH were significantly higher than those in simple S-PE group with normal TSH.There was a negative correlation between FT3 and serum creatinine in S-PE.Hence the thyroid function should be regularly monitored in S-PE patients to find damage of renal function and management hypothyrosis.

OBJECTIVE: To access application value of multi-slice spiral computed tomography (MSCT) and coronary artery calcium scoring (CACS) in investigation the coronary artery disease (CAD), and to explore the effective way of virtual autopsy to evaluate the sudden death due to CAD. METHODS: Nine cases of sudden cardiac death were collected to analyze MSCT before the autopsy. The quantitative analysis of the degree of coronary artery calcium was made by Agatston's method. The CACS of all the subjects were calculated based on the diagnostic criteria for CAD, in which calcium scoring was more than 400. The results of CACS were compared with that of the autopsy. RESULTS: Only 2 cases got the high calcium scoring which were more than 400 in the 9 cases died of CAD confirmed by the autopsy. The prediction rate of CACS for CAD was only 22.2%. Pulmonary edema of different severity was found in both autopsy and MSCT. There was a higher morbidity rate in the left anterior descending of coronary artery than the other branches. CONCLUSION Obvious calcification of coronary artery can be detected by MSCT and calculating CACS. To detect subtle calcification needs other technologies such as postmortem angiography.
Autopsy ; Coronary Angiography ; Coronary Artery Disease/diagnostic imaging* ; Death, Sudden/pathology* ; Humans ; Multidetector Computed Tomography/methods* ; Predictive Value of Tests ; Vascular Calcification/diagnostic imaging*

OBJECTIVETo assess the association between G894T (Glu298Asp) mutation in exon 7 of the endothelial nitric oxide synthase gene and premature coronary heart disease (P-CHD).

METHODSHospital-based case-control study was conducted. Newly-diagnosed CHD patients were recruited as study subjects. 132 CHD patients diagnosed at/before age 55 for males and 65 for females were assigned to P-CHD case group with other 172 CHD patients as the control group. Polymerase chain reaction with Ban II restriction enzyme digestion was performed to detect the G894T mutation.

RESULTSG894T mutant genotypes in P-CHD group (TT, GT and GG frequencies were 6.06%, 20.45% and 73.48%, respectively) were significant higher than those in control group (TT, GT and GG frequencies were 1.74%, 11.63% and 86.63%, respectively) (P = 0.01). Mutant T allele frequency in P-CHD group was also significantly higher than that in control group (16.29% versus 7.56%, P = 0.001, OR = 2.38, 95% CI: 1.38 - 4.16). Stepwise multiple logistic regression analysis at 0.05 significant level with sex, smoking, alcohol drinking, and overweight covariates indicated that G894T mutation also having significant effect on P-CHD (P = 0.01, OR = 2.25, 95% CI: 1.19 - 4.26).

CONCLUSIONThis study suggested that G894T mutation in endothelial nitric oxide synthase gene might serve as a major risk factor to the pathogenesis of P-CHD in this study population.

Age Factors ; Case-Control Studies ; Coronary Disease ; enzymology ; genetics ; Exons ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Middle Aged ; Nitric Oxide Synthase Type III ; genetics ; Point Mutation ; Risk Factors

Objective:To determine the optimal condition for mouse embryonic stem cells (mESC) culture with stirred bioreactor,and to develop a method for mass production of embryoid bodies (EB). Methods:The different initial cell concentrations of mESC and the initial stirring speed of bioreactor were investigated to determine the optimal condition for EB formation. Induced by ascorbic acid,the differentiation of EBs formed in stirred bioreactor into cardiomyocytes was compared with EBs formed in Petri dish. Immunofluorescence staining and RT-PCR were used to identify the cardiomyocytes derived from mESC. Results:The formation of a large number of uniform relatively EBs was achieved in stirred bioreactor when mESC were seeded initially with 1?105~3?105 cells/ml and stirring speed was set to 15~30r/min. Most of cells in the EBs formed in bioreactor were viable. EBs produced in bioreactor differentiated into cardiomyocytes more efficiently compared with EBs from Petri dish. The cardiac specific genes were expressed in ESC-derived cardiomyocytes. Conclusions:Stirred bioreactor culture could enhance the efficiency of EB formation and differentiation into cardiomyocytes,which may be a more ideal culture system for EB formation.

Objective :To explore plasma expression levels of human maternal expression gene 3 (MEG3) and urotheli-al carcinoma antigen 1 (UCA1) in patients with acute myocardial infarction (AMI) and their clinical significance . Methods : A total of 90 AMI patients treated in our hospital were enrolled as AMI group ,and 50 healthy subjects un-dergoing physical examination in our hospital simultaneously were treated as healthy control group .Plasma expres-sions of MEG3 and UCA1 ,serum levels of creatine kinase isoenzyme MB (CK-MB) and cardiac troponin I (cTnI) were observed and compared between healthy control group and AMI group on 1h ,3h ,6h and 12h after onset . Spearman correlation analysis was used to analyze their correlation .Results : Compared with healthy control group , there was significant rise in plasma MEG3 expression [ (0-002 ± 0-001) vs .0-017 ± 0-003)] ,and significant reduc-tion in UCA1 expression [ (0-027 ± 0-005) vs .(0-017 ± 0-002)] in AMI group on 1h after onset , P＝0-001 both ;after 3h ,there were significant rise in serum levels of CK-MB [ (20-01 ± 3-05) IU／L vs.( (32-10 ± 4-40) IU／L] and cTnI [ (1-01 ± 0-87) ng／L vs.(2-10 ± 0-91) ng／L] in AMI group , P＝0-001 all.Spearman correlation analy-sis indicated that in plasma MEG3 expression was significant positively correlated with serum CK-MB and cTnI levels ( r＝0-351 ,0-368 , P＜0-05 both) ,and plasma UCA1 expression was significant inversely correlated with serum levels of CK-MB and cTnI (r＝ －0-416 ,－0-425 , P＜0-01 both) AMI patients .Conclusion : Plasma MEG3 level significant rises ,and UCA1 level significantly reduces in AMI patients during early onset period .Both are signifi-cantly correlated with CK-MB and cTnI levels ,which may be used as new markers diagnosing early AMI .

Objective:To study therapeutic effect of levosimendan combined milrinone on severe refractory heart failure (SRHF) and its influence on serum levels of brain natriuretic peptide (BNP) and uric acid (UA).Methods:A total of 156 SRHF patients were enrolled,randomly and equally divided into levosimendan group and milrinone group,both groups re-ceived corresponding medication based on routine treatment for 7d.Heart rate,blood pressure,serum BNP and UA levels,left ventricular end-systolic dimension (LVESd),left ventricular end-diastolic dimension (LVEDd) and left ventricular e-jection fraction (LVEF) before and after treatment,total effective rate and incidence of adverse reactions were observed and compared between two groups.Results:Compared with milrinone group after treatment,there were significant reduc-tions in heart rate [ (73.79 ± 7.61) beats/min vs.(70.39 ± 7.45) beats/min],systolic blood pressure [ (128.84 ± 13.11) mmHg vs.(121.86 ± 12.53) mmHg],scores of lung wet rales [ (2.05 ± 0.33) scores vs.(1.53 ± 0.21) scores],difficulty breathing [ (2.11 ± 0.36) scores vs.(1.60 ± 0.25) scores] and lower extremity edema [ (2.03 ± 0.34) scores vs.(1.50 ± 0.18) scores],serum levels of BNP [ (459.62 ± 46.27) μg/L vs.(248.73 ± 25.91) μg/L] and UA [ (355.97 ± 36.47) μmol/L vs.(282.75 ± 28.61) μmol/L],LVESd [ (41.62 ± 4.52) mm vs.(36.87 ± 3.71) mm] and LVEDd [ (51.89 ± 5.37) mm vs.(47.85 ± 4.83) mm],and significant rise in 24h urine volume [ (3204.59 ± 321.52) ml vs.(3695.78 ± 370.62) ml] and LVEF [ (42.36 ± 4.31)% vs.(47.85 ± 4.86)%] in levosimendan group,P＜0.01 all.Total effective rate of levosimendan group was significantly higher than that of milrinone group (89.74% vs.71.79%),and incidence rate of adverse reactions was significantly lower than that of milrinone group (5.13% vs.28.21%),P＜0.01 both.Conclusion:Levosimendan therapy can significantly reduce serum BNP and UA levels,improve cardiac function in SRHF patients.It possesses significant therapeutic effect,and it＇s safe and reliable,which is better than milrinone.