Objective:To observe influence of up‐regulated expression of myocardial heat shock protein (HSP) 70 in‐duced by heat stress on myocardial calcium‐activated potassium channel (KCa ) 3.1 expression in rabbits with atrial fibrillation (AF) caused by rapid atrial pacing (RAP) .Methods :A total of 24 New Zealand white rabbits were ran‐domly divided into sham operation group (n=8 ,only implant electrode without pacing ) ,pacing group (n=8 ,right atrium (RA) received RAP at 600 times/min for 6h) and heat stress pacing group (heat stress group ,n=8 ,received heat stress preconditioning ,then the same RAP as pacing group ) .Results:Compared with sham operation group and pacing group ,there were significant up‐regulation of HSP70 mRNA and protein expression in different sites of heart [HSP70 protein ,left atrium (LA):(39.00 ± 3.21) vs .(39.75 ± 2.82) vs .(69.75 ± 3.45) ,RA: (38.38 ± 2.92) vs .(39.50 ± 3.89) vs .(69.00 ± 2.93) ,left atrial appendage (LAA):(37.75 ± 3.28) vs .(39.00 ± 3.89) vs . (68.63 ± 3.23) ,right atrial appendage (RAA): (37.00 ± 3.85) vs .(38.38 ± 3.74) vs .(68.75 ± 2.82)] in heat stress group , P<0. 01 all ,but there were no significant difference between pacing group and sham operation group , P>0.05 ;compared with pacing group with down‐regulation of KCa3.1 mRNA and protein expressions ,there were significant up‐regulation of KCa3.1 mRNA and protein expressions in different sites of heart [KCa3.1 protein ,LA:(21.25 ± 1.67) vs .(24.00 ± 2.62) ,RA :(21.13 ± 1.96) vs .(23.75 ± 1.83) ,LAA :(21.00 ± 2.07) vs .(23.75 ± 1.67) ,RAA:(20.88 ± 2.03) vs .(23.50 ± 2.45)] in heat stress group ,P<0.05 all ,and there were no significant difference between heat stress group and sham operation group , P>0. 05. Conclusion:Heat stress may induce up‐regulated expression of myocardial HSP 70 of myocardium ,and HSP 70 may inhibit down‐regulation of KCa 3. 1 mR‐NA and protein expressions in rabbits with atrial fibrillation.

Objective:To explore influence of thrombus aspiration combined bivalirudin during emergency PCI on my- ocardial tissue perfusion and clinical prognosis in patients with acute ST elevation myocardial infarction (STEMI). Methods:A total of 102 patients with acute STEMI,who were confirmed with thrombus burden by CAG in our hos- pital from Jan 2012 to Jun 2014,were selected.According to random number table,they were randomly divided into thrombus aspiration + bivalirudin group (n=52,thrombus aspiration group)and heparin group (n=50,routine PCI group).TIMI blood flow grade 3 rate,TIMI myocardial perfusion grade (TMPG)after PCI,ST segment re- gression rate 2h after PCI,peak value and peak time of cTnI after PCI,LVEF,LVEDd,incidence rates of bleeding and major adverse cardiovascular events (MACE)on one week and one month after PCI were compared between two groups.Results:Compared with routine PCI group,there were significant rise in postoperative TMPG grade 3 rate (56.00% vs.88.46%),TIMI grade 3 rate (58.00% vs.88.46%)and ST segment regression rate (52.00% vs. 76.92%)in thrombus aspiration group,P<0.05 or <0.01. Compared with routine PCI group one month after PCI,there was significant rise in LVEF [(53.76±5.24)% vs.(57.95±5.51)%],and significant reductions in LVEDd [(53.70±3.39)mm vs.(50.63±1.24)mm],peak value [(16.00±4.28)μg/L vs.(13.81±4.00)μg/L]and peak time [(14.00±2.80)h vs.(13.00±2.23)h]of cTnI in thrombus aspiration group,P<0.05 or <0.01. Incidence rate of mild bleeding in thrombus aspiration group was significantly lower than that of routine PCI group (1.9% vs.16.0%),P<0.05,but there was no significant difference in incidence rate of MACE between two groups,P>0.05. Conclusion:Thrombus aspiration combined bivalirudin during emergency PCI is safe and fea- sible for acute STEMI patients,it can effectively reduce incidence rate of bleeding,remove coronary thrombus,im- prove myocardial tissue perfusion and doesn't increase incidence rate of MACE.

Objective To investigate the influence of hedysaryum polysaccharide (HPS)in the kidney function and expressions of Glut-1 mRNA and protein in kidney tissue of db/db mice with diabetic nephropathy (DN)and to elucidate its possible action mechanism.Methods 10 db/m mice were taken as normal control group(n=10);50 fueling animal model db/db mice with DN were randomly divided into model group,enalapril group and the low, middle and high doses of HPS groups(n=10).The mice in noral control group and model group were given physioloical saline by gavege;and the mice in the other groups were respectively given 10 mg·kg-1 ·d-1 enalapril, 100,200 and 400 mg·kg-1 ·d-1 HPS by gavage;lasted 8 weeks.Picric acid method was used to determine the serum creatinine(SCr)level of the mice,enzyme coupling rate method was used to determine the blood urea nitrogen (BUN)level,ELISA method was used to determine the urinary microalbumin(UMALB)level,RT-PCR method was performed to detect the expression of Glut-1 mRNA, and Western blotting and immunohistochemical methods were used to detect the expression of Glut-1 protein.Results Compared with model group,the levels of SCr, BUN, UMALB, the mRNA and protein of Glut-1 expressions were decreased, especially in 400 mg·kg-1 ·d-1 HPS and enalapril groups(P<0.01).The HE and Masson staining results showed that less inflammatory cells infiltration in glomerular of the mice were found, capillary lumens were unobstructed, and the collagen deposition was not obvious in 400 mg·kg-1 ·d-1 group.Conclusion HPS could improve the kidney function of the db/db mice and inhibit the Glut-1 mRNA and protein expressions obviously, which indicates that HPS could delay the development of DN by inhibiting the Glut-1 expression in the glomerular mesangial cell membrane.

Objective To analyse MMACHC mutations for 45 pedigrees with combined methylmalonic aciduria and homocyctinuria by Sanger sequencing, and to discuss the utility of prenatal genetic diagnosis for these pedigrees.Method Peripheral blood was collected from 45 probands and their parents from 2012-2015 in Genetic Counselling Clinic of the First Affiliated Hospital of Zhengzhou University, and the DNA were extracted from the blood.Then the coding sequence of MMACHC gene was amplified by PCR, and the PCR products were further sequenced to detect mutations for each pedigree.For 12 families, chorionic villus sampling was performed on the pregnant women to make prenatal genetic diagnosis.Result There were 14 distinct mutations detected in the 45 pedigrees, and the most frequent mutations are c.609G>A(W203X),c.658-660delAAG(K220del)and c.80A>G (Q27A).Two of those mutations have not been reported before:one is a splicing site mutation c.81+1G>A;while the other is a missense mutation c.665A>G,p.Y222C.Most mutations were found in exon 4.Among the 12 pedigrees who received prenatal diagnosis, 2 fetuses were normal, 7 fetuses were carriers of heterozygous mutation, and the other 3 fetuses were patients with compound heterozygous mutation or homozygous mutation.The couples whose fetuses were normal or carriers continued the gestation, while the couples whose fetuses were patients decided to terminate the pregnancy.After delivery, the outcome of the fetuses was the same as the prenatal diagnose results.Conclusion Two novel mutations of MMACHC were identified and prenatal genetic diagnosis helps to avoid the delivery of combined methylmalonic aciduria and homocyctinuria patients.

Objective To study the value of the technique of "Push-pull traction-relax homingrepeatedly confirmed" in the prevention of bile duct injury in LC. Methods From March 2001-August 2009, we applied this technique in 4800 cases of LC. The technique of "Push-pull traction" showed the structures of in the Calot's triangle. The technique of "relax homing" was to restore the cystic duct,hepatic duct and common bile duct to their original anatomical positions. The technique of "repeatedly confirmed" repeatedly identified the positions of the cystic duct, the common hepatic duct and the common bile duct. Results There was no bile duct injury. Conversion to open surgery happened in 118patients due to difficulties in identifying the Calot's triangle structures, bile duct stones, gallbladder cancer, and gallbladder-duodenal fistula. Conclusions The "Push-pull traction-relax homing-repeatedly confirmed" technique could effectively prevent bile duct injury in LC. The method is simple, easy to master and worthy of promotion.

Objective To investigate the value of prenatal diagnosis in identifying the etiology and predicting the prognosis of fetal pleural effusion (FPE).Methods Forty-two cases of FPE were recruited in this study from January 2012 to September 2016.Ultrasound scan and genetic tests were performed on all fetuses.Seven fetuses with severe FPE were given pleurocentesis.Pregnancy outcomes of all the fetuses were followed up.Results FPE was commonly accompanied with other abnormalities,such as ascites,hydrops,hydramnion,hygroma colli,abnormal posturing,joint contractures,arrhythmia and micromandible.Chromosomal abnormality was detected in 11 fetuses (26.2％),of which ten were further confirmed by karyotype analysis,including six with 45,X,three trisomy 21 and one trisomy 18,and one was detected with a 9.83 Mb uniparental disomy (UPD) located at 12q24.21q24.31 by gene chip.One fetus was diagnosed with--SEA/--SEA thalassemia.All of the 12 families decided to terminate the pregnancies after genetic counseling.Among the other 30 fetuses,seven with severe FPE and normal karyotype underwent pleurocentesis.Five of the seven cases were with favorable outcomes,one with progressive hydrops was aborted and one neonate with severe hydrops died after birth.Spontaneous regression of FPE with good outcome was found in two cases.Parents of the other 21 fetuses chose to terminate the pregnancies.Conclusions Prenatal diagnosis is important to identify the etiology and predict the outcome of FPE.Chromosomal abnormality is a relatively common cause of FPE,and 45,X and trisomy 21 are the most common abnormalities.Intrauterine intervention is beneficial for FPE without chromosomal or other definite genetic abnormalities.Genetic test may be of great value for pregnant counseling.

OBJECTIVETo induce Mycobacterium tuberculosis (MTB) resistance with ofloxacin (Ofx) of stepwise increasing concentration in vitro, investigate stability to fluoroquinolone (FQs) antibiotic of MTB, and analyze the molecular mechanism and mutation specialty of drug resistance preliminarily.

METHODSMTB Standard strain H37RV and 24 clinical isolates susceptible to Ofx were selected and experimentally serially subcultured in liquid culture medium containing increasing concentration of Ofx and induced the drug resistance to Ofx. Variety of Minimal Inhibitory Concentrations (MICs) to FQs drugs were detected by microwell-MIC-test method. Mutations of quinolone resistance determining region (QRDR) of gyrA gene were sequenced and identified. Relationship of different mutation sites and drug resistant degree were analyzed. A total of 6 MTB clinical isolates resistant to Ofx and induced drug resistant isolates in vitro were serially subcultured in liquid culture medium without drug. Variety of drug resistant stability, including MIC and mutation of gyrA gene were detected.

RESULTSMIC values of 21 Ofx susceptible isolates after induction were eight times higher than before, which were induced to drug resistant strains successfully and also resistant to Lfx and Mfx. Hot mutations of QRDR of gyrA gene were detected by sequencing, except one strain. Mutation of codon 94 occurred in 60% (12/20) of the strains with mutations and corresponding value of 50% Minimal Inhibitory Concentrations(MIC50) was ≥ 8 µg/ml. In all, 4 of 6 MTB clinical isolates resistant to Ofx harbored mutation of codon 90 (67%) , but the corresponding value of MIC50 was 2 µg/ml. After 21 serially subcultured in liquid culture medium without drug, MIC values of 6 clinical isolates resistant to Ofx were not changed obviously and mutations were also not changed. After 11 times serially subcultured in culture medium without drug, MIC values of induced drug resistant strains were also not changed obviously, but new mutations were detected in QRDR of 3 isolates.

CONCLUSIONMTB strains resistant to three kinds of FQs antibiotic were obtained by induction in vitro with Ofx. Codons 88, 94 mutations of QRDR of gyrA gene were related to the high level FQs drug resistance of MTB. Drug resistant stability of MTB to FQs was strong, and it is difficult for MTB to resume susceptibility.

Antitubercular Agents ; pharmacology ; DNA Gyrase ; genetics ; Drug Resistance, Bacterial ; genetics ; Microbial Sensitivity Tests ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Ofloxacin ; pharmacology

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.
Cell Proliferation ; drug effects ; Cells, Cultured ; Cloning, Molecular ; Drug Synergism ; Escherichia coli ; genetics ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stem Cell Factor ; biosynthesis ; genetics ; Umbilical Cord ; cytology

OBJECTIVETo describe serum homocysteine distribution and its associated factors in population of urban and rural areas in Beijing.

METHODSThe study population was represented by a randomly selected sample with 1 168 subjects, including both males and females aged 35 - 64. The levels of serum homocycteine were compared and the correlation with other risk factors were analyzed statistically.

RESULTS(1) Geometric mean of serum homocycteine was 15.4 micromol/L in males and 12.2 micromol/L in females (P < 0.001). (2) There was a significant difference in homocysteine levels between urban population and rural population. Men from rural area had 1.5 times higher homocyteine than from urban (18.0 micromol/L vs 12.0 micromol/L, P < 0.001), while the rural women had 1.3 times higher homocysteine level than urban women did. (3) The prevalence rate of hyperhomocysteinemia was 15.3% in population aged 35 - 64 in Beijing area. (4) Gender, residential location (urban or rural), smoking and education had independent effects on level of serum homocysteine by multivariate analysis.

CONCLUSIONPopulation in Beijing had higher serum level and prevalence rate of homocysteine than some western countries. Gender, geographic distribution, smoking and education had some influence on homocysteine level.

Adult ; Age Factors ; China ; epidemiology ; Female ; Homocysteine ; blood ; Humans ; Hyperhomocysteinemia ; epidemiology ; Male ; Middle Aged ; Risk Factors ; Rural Health ; statistics & numerical data ; Sex Factors ; Urban Health ; statistics & numerical data