OBJECTIVETo study the in vitro expression of 6 novel missense mutations (R270G, P275A, F121L, A156P, E183G, I324N) and a previously described R408Q mutation of phenylalanine hydroxylase (PAH) gene and explore the genotype-phenotype correlation through comparison of protein levels and residual enzyme activities.

METHODSSeven expression vectors containing PAH cDNA were constructed with a site-directed mutagenesis kit. The plasmids were extracted and sequenced to confirm the target mutations. pcDNA3.0 containing PAH cDNA was transfected into COS-7 cells and total proteins were extracted 48 h after transfection. The quantities of proteins and residual enzyme activities of the 7 mutants were assessed with the wild-type PAH gene as reference.

RESULTSRelative quantities of PAH proteins for R270G, P275A, F121L, A156P, E183G, I324N and R408Q were 10.5%, 56.6%, 54.3%, 8.7%, 8.5%, 67.3% and 85.4%, respectively. The residual enzyme activities were 7.7%, 27.6%, 19.0%, 10.4%, 9.1%, 50.6% and 40.2%, respectively.

CONCLUSIONPAH residual enzyme activities of 7 PAH mutants were all significantly reduced.

Amino Acid Sequence ; Animals ; COS Cells ; Cercopithecus aethiops ; Genetic Association Studies ; methods ; Humans ; Molecular Sequence Data ; Mutation, Missense ; Phenylalanine Hydroxylase ; genetics ; Sequence Alignment

OBJECTIVETo investigate the effect of periodontal infection on circulating C-reactive protein (CRP) in type 2 diabetes patients.

METHODS32 diabetes patients with advanced periodontitis participated in this study. They were compared to a group of 32 diabetes patients without periodontal disease, who were mathed with regard to age (+/- 3 years), gender and body mass index (+/- 1 kg/m2). The concentration of CRP on circulation was measured by ELISA.

RESULTSSignificant difference was found in the level of CRP and the percentage of subjects with elevated CRP levels > or = 3 mg/L on circulation between the two groups(P < 0.05).

CONCLUSIONPeriodontal infection results in higher circulating CRP in type 2 diabetes patients. This elevated inflammatory factor may exacerbate insulin resistance and increase the risk for great vessels complications of diabetes mellitus.

Adult ; C-Reactive Protein ; Diabetes Mellitus, Type 2 ; Female ; Humans ; Male ; Middle Aged ; Periodontal Attachment Loss ; Periodontal Pocket ; Periodontitis

OBJECTIVETo establish a method of iduronate-2-sulfatase (IDS) activity assay and mutation analysis of IDS gene for the prenatal diagnosis of mucopolysaccharidosis type II (MPSII).

METHODSPrenatal diagnosis of two cases was performed using cultured fetal amniotic fluid cells. Enzyme activity of IDS in cultured fetal amniotic fluid cells extracted from the two pregnant women at high risk of MPS II was measured. Meanwhile, genomic DNA was extracted for fetal gender testing and mutation analysis of the IDS gene.

RESULTSEnzyme activity assay showed that IDS activity in amniotic fluid cells was significantly decreased. IDS gene sequencing showed that the male fetus was hemizygous mutant, and the female fetus was carrier of heterozygous mutation. Therefore the male fetus was an MPS II patient and the female fetus was a mutation carrier.

CONCLUSIONDetermination of IDS activity in fetal amniotic fluid cells together with IDS gene mutation analysis is a rapid, sensitive and accurate method of prenatal diagnosis of MPS II. Using this method, prenatal diagnosis for pregnant women at high risk of MPSII can be achieved.

Base Sequence ; Child ; Child, Preschool ; Exons ; Female ; Glycoproteins ; genetics ; metabolism ; Humans ; Male ; Mucopolysaccharidosis II ; diagnosis ; genetics ; metabolism ; Mutation ; genetics ; Pregnancy ; Prenatal Diagnosis

AIMTo approach the effects of multi-site synchronous ventricular pacing on myocardial mechanics and cardiac work.

METHODSFive modes of multi-site synchronous ventricular pacing were randomly performed in 12 dogs with anesthetized, opened chest and artificial-ventilation. Some parameters were measured simultaneously including: the peak of left ventricular pressure rise and fall (+/- dp/ dt(max)), the time constant of left ventricular relaxation(tau), the muscle tensile force in left/right ventricular wall (V-tensile force, V-TF), SV, LVSW and RVSW.

RESULTSThe myocardial systolic mechanical parameters: +dp/dt(max) and LV-TF of cHisB-LVPL and RVA-LVPL pacing by biventricular pacing modes were increased than that of cHisB-RVA pacing in right ventricular bifocal pacing mode. +dp/dt(max) in above two groups of biventricular pacing was increased than that in cHisB-RVA pacing. Tau value of cHisB-LVPL and RVA-LVPL pacing modes were shorted than that of cHisB-RVA pacing. The above parameters of cHisB-RVA-LVPL and cHisB-RVA-LVA biventricular trifocal pacing were superior to that of cHisB-LVPL and RVA-LVPL biventricular pacing. The +dp/dt(max), LV-TF and RV-TF of cHisB-RVA-LVPL pacing were increased as compared with that of cHisB-RVA-LVA pacing (P > 0.05). The -dp/dt(max) in cHisB-RVA-LVPL pacing were increased by 6.0% and tau value was shorted by 3.7% compared with those in cHisB-RVA-LVA pacing (P > 0.05). SV, LVSW and RVSW of cHisB-LVPL and RVA-LVPL biventricular pacing were increased than those of cHisB-RVA bifocal pacing. The above parameters of cHisB-RVA-LVPL pacing were increased than that of cHisB-RVA-LVA and cHisB-LVPL pacing.

CONCLUSIONIt was explained that the cHisB-RVA-LVPL biventricular trifocal sites synchronous pacing mode would increase the velocity of ejection and filling during myocardial contraction and relaxation and enhance cardiac work by maintaining normal VSS.

Animals ; Cardiac Pacing, Artificial ; methods ; Dogs ; Female ; Heart ; physiology ; Heart Ventricles ; Male ; Myocardium

Objective Amnestic mild cognitive impairment (aMCI) and Alzheimer's disease (AD)used to be evaluated by an-alyzing the characteristics of the brain gray matter via diffuse tensor imaging (DTI).This study was to characterize the damage to the mi-crostructural integrity of the brain gray matter in aMCI and AD patients using the mean diffusivity ( MD) of DTI and to investigateits transformation rules. Methods This retrospective study included 30 cases ofaMCI, 30 cases of AD, and another 30 normal controls. We measured the MD values of the whole brain gray matter , established the MD network of the water molecules in the gray matter , char-acterized the small-world network, and performed correlation analysis by neuropsychological assessment . Results Compared with the normal controls, the MDnetworks of the aMCI and AD groupsexhibited abnormal small world characteristics , namely, a higher clustering coefficient and a longer path , which reflected the damage to themicrostructure of the gray matter , and theaverage connectivity in the aMCI patients was lowerthan that in thenormal controls but higher than that in the AD group ( P<0.05) .The MD values of the gray matter were negatively correlated with the Montreal Cognitive Assessment Scale scores in the 11 hub regions of the brain , including the bilateral hippo-campus and the limbic system ( P<0.05) . Conclusion The diffusion feature of the water molecules in the brain gray matter of the aMCIpatient may help to detect early ADand reflect the damage to the brain microstructure in patients with neurodegenerative diseases .

OBJECTIVETo compare the acute hemodynamic effects and safety of intravenous injection of recombinant human brain natriuretic peptide (rhBNP) versus intravenous nitroglycerin (NIT) in acute myocardial infarction (AMI) patients with heart failure.

METHODSOn top of standard therapy, 42 consecutive patients who suffered from anterior wall AMI with heart failure [pulmonary capillary wedge pressure (PCWP) > 16 mm Hg] within 12 to 24 hours from the onset of chest pain were randomized into rhBNP group (n = 21, 1.5 microg/kg bolus intravenous injection followed by 0.0075 microg.kg(-1).mn(-1) for the first 3 hours and 0.015-0.03 microg.kg(-1).mn(-1) infusion for following 21 hours) and NIT group (n = 21, 10 to 100 microg/mn intravenous infusion for 24 hours). The hemodynamic parameters were monitored by Swan-Ganz catheter at baseline, during drug infusion and 6 hours post infusion withdraw; total urine output was also obtained. The major adverse cardiac events (MACE) were observed up to 1 week after drug infusions.

RESULTSCentral venous pressure and systolic blood pressure remained unchanged after rhBNP or NIT infusion. Compared to baseline level, PCWP was significantly reduced by 48.9% (P < 0.01) at 30 minutes after rhBNP infusion and this effect remained up to 6 hours post infusion withdraw; PCWP reduced by 28.7% (P < 0.05) at 2 hours after NIT infusion and this effect remained to 6 hours before infusion withdraw. Cardiac index (CI) was increased by 27.1% (P < 0.05) at 1 hour after rhBNP infusion and remained till 6 hours post infusion withdraw; CI was significantly increased at 3 hour after NIT infusion and this effect disappeared after infusion withdraw. The PCWP and CI values were significantly higher in rhBNP group than that of NIT group at 30 minutes and 2 hours (P < 0.05). Heart rate was significantly reduced at 30 minutes (95.3 +/- 7.4 vs. 118.0 +/- 8.2 bpm, P < 0.05) and at 2 hour (92.8 +/- 6.8 vs. 109.2 +/- 7.6 bpm, P < 0.05) in rhBNP and NIT group, respectively and heart rate remained reduced during the whole infusion period in both groups. The total urine output for 30 hours in rhBNP group (1870 +/- 535 ml) tended to be higher than that in NIT group (1538 +/- 620 ml, P > 0.05). There was no symptomatic hypotension or other adverse events during drug infusion in both groups and MACE up to 1 week post drug infusion was also similar between the two groups.

CONCLUSIONIntravenous injection of rhBNP results in more rapid and long-lasting hemodynamic improvements than that of NIT in AMI patients with heart failure and it is also feasible and safe for clinic use in AMI patients with heart failure.

Aged ; Female ; Heart Failure ; complications ; drug therapy ; physiopathology ; Hemodynamics ; drug effects ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; complications ; drug therapy ; physiopathology ; Natriuretic Peptide, Brain ; therapeutic use ; Recombinant Proteins ; therapeutic use

BACKGROUNDThe incidence of no reflow phenomenon limits the clinical outcomes of percutaneous coronary intervention (PCI). This randomized controlled study was designed to evaluate the immediate protective effects of intensive statin pretreatment on myocardial perfusion and myocardial ischemic injury during PCI.

METHODSAltogether 228 patients with acute coronary syndrome (ACS) were randomly assigned to standard statin group (SS group, n = 115) and intensive statin group (IS group, n = 113). Patients in the SS group received 20 mg simvastatin and patients in the IS group received 80 mg simvastatin for 7 days before PCI. Thrombolysis in myocardial infarction (TIMI) flow grade (TFG), corrected TIMI frame count (CTFC) and TIMI myocardial perfusion grade (TMPG) of the intervened vessel were recorded before and after stent deployment. Creatine kinase (CK) isoenzyme MB, troponin I and plasma level of high sensitive-C reactive protein (hs-CRP), P-selectin and intercellular adhesion molecule (ICAM) were measured before and 24 hours after the procedure.

RESULTSThe TFG after stent deployment was significantly improved with less TIMI 0-1 and more TIMI 3 blood flow in the IS group than in the SS group (all P < 0.05). Patients with no reflow phenomenon were less in the IS group (P < 0.001). The CTFC was lower in the IS group than in the SS group (P < 0.001). TMPG was also improved in the IS group than in the SS group (P = 0.001). Although PCI caused a significant increase in CK-MB 24 hours after the procedure, the elevated CK-MB value was lower in the IS group than in the SS group (18.74 +/- 8.41 vs 21.78 +/- 10.64, P = 0.018). Similar changes were also found in troponin I (0.99 +/- 1.07 in the IS group vs 1.47 +/- 1.54 in the SS group, P = 0.006). CK-MB elevation occurred in 27.8% (32/115) of the patients in the SS group vs 15.9% (18/113) in the IS group (P = 0.030). Myocardial necrosis was detected in 4.4% (5/115) of the patients in the SS group, whereas 0.9% (1/113) in the IS group (P = 0.341). But no myocardial infarction was found. Similarly, the patients with increased level of troponin I were much more in the SS group (36.5%, 42/115) than in the IS group (19.5%, 22/113) (P = 0.04). Among them, myocardial necrosis was detected in 13.0% (15/115) of the patients in the SS group, while 4.4% (5/113) in the IS group (P = 0.021). Myocardial infarction was found in 4.4% (5/115) of the patients in the SS group and 0.9% (1/113) in the IS group (P = 0.213).

CONCLUSIONSIntensive statin pretreatment for 7 days before PCI can further improve myocardial blood perfusion, protect the myocardium from ischemic injury. These effects are associated with the lowered levels of hs-CRP, P-selectin and ICAM.

Acute Coronary Syndrome ; drug therapy ; pathology ; therapy ; Aged ; Angioplasty, Balloon, Coronary ; methods ; Anticholesteremic Agents ; therapeutic use ; Female ; Heart ; drug effects ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Simvastatin ; therapeutic use ; Treatment Outcome

OBJECTIVETo evaluate the influence of intracoronary administration of anisodamine on myocardial blush grade (MBG) and left ventricular regional and global systolic function and synchrony in the acute myocardial infarction (AMI) patients with no-reflow phenomenon post percutaneous coronary intervention (PCI).

METHODSForty-seven AMI patients who underwent PCI within 12 hours of onset and MBG was 0 - 1 were randomized to receive standard therapy [group B, n = 23, 18 males, mean age (62.72 +/- 11.48) years] or standard therapy plus intracoronary administration of anisodamine [200 microg/ml, group A, n = 24, 18 males, mean age (64.23 +/- 12.27) years]. The left ventriculography (LVG) was performed immediately and 6 months after PCI to measure the ventricular volume, LVEDP and wall motion score (WMS). Equilibrium radionuclide angiography (ERNA) was performed 1 week and 6 months after PCI to determine the parameters of left ventricular regional, global systolic function and systolic synchrony. Incidence of major adverse cardiac events (MACE) during the follow-up was analyzed.

RESULTSAnisodamine [(2530 +/- 340) microg/person)] was well tolerated by patients. The MBG remained unchanged in group B and significantly increased from grade 0.74 +/- 0.32 to grade 2.33 +/- 0.28 10 min after anisodamine injection in group B. Six months post PCI, LVESVI [(40.53 +/- 8.12) ml/m(2) vs. (50.32 +/- 8.26) ml/m(2)], LVEDVI [(80.13 +/- 9.74) ml/m(2) vs. (87.17 +/- 10.25) ml/m(2)], WMS [(8.24 +/- 1.31) vs. (10.23 +/- 1.82)] and LVEDP [(13.36 +/- 4.21) vs. (16.38 +/- 3.21) mm Hg, 1 mm Hg = 0.133 kPa] were significantly lower in group A compared with that in group B (all P < 0.05) while LVEF [(44.02 +/- 5.86)% vs. (38.52 +/- 5.18)%], PER [(1.86 +/- 0.09) EDV/s vs. (1.61 +/- 0.09) EDV/s] and PFR [(2.19 +/- 0.32) EDV/s vs. (1.78 +/- 0.17) EDV/s] measured by ERNA were significantly increased in group A compared with that in group B (all P < 0.05). (2) LrEF(2)-LrEF(8) in group A were higher by 13.96%, 25.02%, 30.36%, 22.86%, 27.67%, 22.07% and 18.71% respectively compared with that in group B. (3) Phase analysis showed that the left ventricular systolic synchrony parameters PS [(46.04 +/- 8.93) degrees vs. (53.19 +/- 162) degrees ], FWHM [(23.02 +/- 6.27) degrees vs. (25.02 +/- 5.31) degrees ] and PSD [(7.92 +/- 4.12) degrees vs. (11.76 +/- 4.11) degrees ] were also significantly lower in group A than that in group B (all P < 0.05). (4) During the 6 months of follow-up, the incidence of MACE in group A was significantly lower than that in group B (P < 0.05).

CONCLUSIONIntracoronary administration of anisodamine is safe and could partly attenuate the no-reflow phenomenon, improve the left ventricular systolic function and synchrony and reduce the incidence of MACE in patients with no-reflow phenomenon post AMI-PCI.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; physiopathology ; therapy ; Myocardial Reperfusion ; Solanaceous Alkaloids ; administration & dosage ; therapeutic use ; Ventricular Function

BACKGROUNDPrevious studies show that sleep-related breathing disorder (SRBD) is common in patients with heart failure (HF) and is associated with increased mortality. This study aimed to determine whether there was significant difference of subjective daytime sleepiness between HF patients with and without SRBD.

METHODSWe enrolled, prospectively, 195 consecutive HF patients with left ventricular ejection fractions (LVEF) < or = 45% and all subjects underwent polysomnography to measure the sleep structure between 2005 and 2008. Patients were then assigned to those with SRBD including obstructive and central sleep apnea (apnea-hypopnea index (AHI) > or = 5/hour of sleep) and those without SRBD (AHI < 5/hour) according to the sleep study. The subjective sleepiness was assessed with Epworth sleepiness scale (ESS).

RESULTSAmong 195 HF patients, the prevalence of obstructive sleep apnea (OSA) was 53% and of central sleep apnea (CSA) was 27%. There was no significant difference of ESS scores between patients without SRBD (NSA) and with SRBD (NSA vs OSA: 6.7 +/- 0.6 vs 7.6 +/- 0.4, P = 0.105 and NSA vs CSA: 6.7 +/- 0.6 vs 7.4 +/- 0.5, P = 0.235, respectively), indicating that SRBD patients had no more subjective daytime sleepiness. Compared with NSA, patients with SRBD had increased arousal index (ArI) (NSA vs OSA: 14.1 +/- 1.4 vs 26.3 +/- 1.5, P < 0.001 and NSA vs CSA: 14.1 +/- 1.4 vs 31.3 +/- 3.5, P < 0.001, respectively), more awake number after sleep onset (NSA vs OSA: 19.2 +/- 1.5 vs 26.2 +/- 1.4, P = 0.01 and NSA vs CSA: 19.2 +/- 1.5 vs 36.9 +/- 4.4, P < 0.001, respectively), and reduced proportion of slow-wave sleep (SWS) (NSA vs OSA: 13.8 +/- 1.7 vs 9.3 +/- 0.7, P = 0.024 and NSA vs CSA: 13.8 +/- 1.7 vs 8.9 +/- 0.9, P = 0.024, respectively).

CONCLUSIONSOSA and CSA remain common in patients with HF on optimal contemporary therapy. Patients with both HF and SRBD have no significant subjective daytime sleepiness compared with patients without SRBD, despite of significantly increased awake number, arousal and decreased proportion of deep sleep stages. It is not a credible way and means to exclude SRBD in patients with HF according to the absence of subjective daytime sleepiness.

Adult ; Aged ; Aged, 80 and over ; Female ; Heart Failure ; physiopathology ; Humans ; Male ; Middle Aged ; Polysomnography ; Sleep Apnea Syndromes ; epidemiology ; etiology

BACKGROUNDThe risk of radial artery occlusion (RAO) needs particular attention in transradial intervention (TRI). Therefore, reducing vascular occlusion has an important clinical significance. The aim of this study was to determine the appropriate puncture site during TRI through comparing the occurrence of RAO between the different puncture sites to reduce the occurrence of RAO after TRI.

METHODSWe prospectively assessed the occurrence of RAO in 606 consecutive patients undergoing TRI. Artery occlusion was evaluated with Doppler ultrasound in 2 days and 1 year after the intervention. Risk factors for RAO were evaluated using a multivariate model analysis.

RESULTSOf the 606 patients, the RAO occurred in 56 patients. Compared with TRI at 2-5 cm away from the radius styloid process, the odds ratio (OR) for occlusion risk at 0 cm and 1 cm were 9.65 (P = 0.033) and 8.90 (P = 0.040), respectively. The RAO occurred in the ratio of the arterial diameter to the sheath diameter ≤1 (OR = 2.45, P = 0.004).

CONCLUSIONDistal puncture sites (0-1 cm away from the radius styloid process) can lead to a higher rate of RAO.

TRIAL REGISTRATIONClinicalTrials.gov, NCT01979627; https://clinicaltrials.gov/ct2/show/NCT01979627?term = NCT01979627 and rank = 1.

Aged ; Arterial Occlusive Diseases ; etiology ; Cardiac Catheterization ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Punctures ; Radial Artery