Objective To examine the expression of ATP-binding cassette transporter A1(ABCA1)in eukaryotie cells and the effect of arsenic resistance after the transfection of eukaryotic expression vector containing ABCA1 gene.Methods HeLa cells were transfected with the recombinant plasmid by lipofectaonmine 2000 (recombinant plasmid group),empty plasmid and untransfected HeLa cell as the control group.The level of the mRNA was examined by real-time PCR,and the expression of ABCA1 protein wag examined by Western blot,the change of cell survival rate was examined by methyl thiazolyl tetrazolium(MTT)after exposure in a series of arsenic [0(contro1),4,8,16,32,64,128 μmol/L]for 48 hours.Results Expression level of ABCA1 mRNA in recombinant plasmid,empty plasmid and untransfeeted groups was(2.09±0.08)×10-4,(0.09±0.02)×10-4,(0.08±0.02)×10-4,there was a significant difference between the groups(F=1499.23,P<0.01).The level of ABCA1 mRNA in recombinant plasmid group was higher than empty plasmid and untransfected group(all P<0.01).Western blot showed that specific protein straps existed at 254×103 in all the three groups,with a similar size to the ABCA1 protein.The amount of the recombinant plasmid group was higher than the other two groups.MTT shows that arsenic concentration at 4,8,16,32,64,128 μmol/L,the survival rates of recombinant plasmid group was(94.8±0.9)%,(86.5 ± 2.6)%, (77.8 ± 2.0)%, (56.0 ± 2.0)%, (23.8 ± 1.7)%, (18.6 ± 0.6)%, higher than that of empty plasmid group[ (85.3 ± 1.1)%, (78.7 ± 0.6)%, (67.8 ± 2.4)%, (43.2 ± 1.5)%, (14.5 ± 1.3)%, (8.0 ± 0.4)%], the difference of survival rate had a statistical signifieance(t = 18.985,6.689,5.922,9.504,9.481,32.634, all P < 0.01). Conclusions ABCA1 protein is over expressed in HeLa cells after transfect ABCA1 gene. ABCA1 protein increases resistance of arsenic in HeLa cells.

The glutamate dehydrogenase (EC.1.4.1.4) gene which amplified from the genome of Brevibacterium flavum GDK-9 by polymerase chain reaction was linked with pUCm-T for sequence alignment. Analysis of gdh sequences revealed that the whole sequence is 1927 bp, only one ORF existed, which used ATG as the initiation codon and coded a peptide of 448 amino acids with a calculated molecular weight of 48 kD. The comparability between the cloned gdh sequence to the reported sequence is high to 99.55%. Only the 1190th base mutation (C→A) lead to the change of amino acid sequence (Thr→Asn), the others are not. The recombinant plasmid pXG was then transformed into E. coli XL-Blue and Brevibacterium flavum GDK-9 which was induced by IPTG. SDS-PAGE analysis revealed that there was a clear induced protein band with molecular mass of 48.7 kD on expected position. Standard glutamate fermentations indicated that although the level of GDH increases the intracellular glutamate pool, the level of GDH has no influence on glutamate secretion.

Objective To study the influence of HBV-DNA with different load levels of HBsAg-positive among fathers on the rate of neonatal cord blood HBV-DNA.Methods Using HBsAg and HBV-DNA as screening indicators for pregnant women and their husbands from an obstetric clinic.161 pregnant women whose HBsAg and HBV-DNA were negative,but HBsAg was positive among their husbands and their newborns,were selected.Blood samples from those pregnant women,their husbands and their newborns were collected to detect the related indicators.Using ELISA to detect hepatitis B virus markers(HBVM),and FQ-PCR to detect the levels of HBV-DNA load.According to neonatal cord blood HBV-DNA detection guideline,newborns with cord blood HBV-DNA positive were selected as cases,others as controls.Results(1)Result of the study showed that there was a dose-response relationship between paternal serum HBV-DNA load levels and neonatal cord blood HBV-DNA positive rates in newborns(trend χ~2=64.117,P=0.000).The rate of vertical transmission of HBV from HBsAg-positive father to infant in the paternal serum HBV-DNA＞1.0×107 copies/ml group was significantly higher than HBV-DNA＜1.0×107 copies/ml group(χ~2=71.539,P=0.000).(2)There was a positive rank correlation between semen positive HBeAg and vertical transmission of HBV from HBsAg-positive father to infant(χ~2=6.892,P=0.009).Conclusion There was a dose-response relationship between paternal serum HBV-DNA load levels and neonatal cord blood HBV-DNA positive in newborns.Paternal serum HBV-DNA≥1.0×107 copies/ml and with HBeAg positive status were risk factors of vertical transmission of HBV from HBsAg-positive father to infant.

ABO Blood-Group System ; Erythroblastosis, Fetal ; epidemiology ; Humans ; Infant, Newborn

OBJECTIVETo study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.

METHODMTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.

RESULTBufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.

CONCLUSIONBufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Osteosarcoma ; pathology ; Proteomics

OBJECTIVETo explore whether there exists coincidence of the most appearing time of clinical features of liver cancer at different longitude and latitude, according to the law of field equation and the theory of warpage of space time by Einstein.

METHODSThree regions with different longitude and latitude were selected randomly and sampled. There were 36 items altogether, including 12 clinical items, which were used to imitate the yearly cycle cosine curve. The acorphases and the ratioes of amplitudes and means were compared to justifying whether they were in the same range.

RESULTSAll the acorphases of 36 items appeared between -90.1degrees to -207.5 degrees (from april to july), existing in one third of the same range, in which 13 items occurred rhythmly (P<0.05). The image acorphases of liver cancer at the early and middle stage and gamma-glutamyl transpeptidase acorphase appeared between -98.5 degrees to -148.2 degrees (from april to may), in which 5 items occurred rhythmly (P<0.05).

CONCLUSIONIt is the same mode of the yearly biologcal cycle for liver cancer malignant growth within the most appearing time (from april to july). It will increase the detecting rate of liver cancer at the early and middle stage during this time (especially from april to may).

Carcinoma, Hepatocellular ; pathology ; Cell Cycle ; physiology ; Chronobiology Phenomena ; Hepatocytes ; physiology ; Humans ; Liver Neoplasms ; pathology ; Mathematical Computing ; Periodicity

BACKGROUNDBrachial-ankle pulse wave velocity (baPWV) is a reliable method for measuring arterial elasticity, but the absence of reference value for baPWV has limited its wide use. We conducted an epidemical study in north China to investigate the reference value of baPWV for Chinese people and its influential factors.

METHODSA total of 974 identified healthy subjects were recruited in this study. The values of baPWV were evaluated noninvasively with an automatic device.

RESULTSFor healthy population, the mean value of baPWV was higher for male (P < 0.001). Multiple regression analysis demonstrated that both age and systolic blood pressure were positively associated with baPWV for male and female (P < 0.001). BaPWV value was higher in male than in female in younger group (< 50 years) but not in older group (P

CONCLUSIONSAging is the most important reason of arterial stiffness, but the effect of age on baPWV augmentation is greater for healthy female than their male counterpart. The reference values of baPWV by sex and age are very useful for clinical and preventive medicine.

Adult ; Age Factors ; Aged ; Ankle Brachial Index ; Asian Continental Ancestry Group ; Female ; Humans ; Male ; Middle Aged ; Reference Values ; Regression Analysis ; Sex Factors

Objectives: To investigate whether remote ischemic conditioning (RIC) applied to patients with ST-segment elevation myocardial infarction (STEMI) before percutaneous coronary intervention (PCI) could affect circulating miR-208b level or not. Methods:Patients diagnosed with STEMI undergoing PCI from January 2016 to July 2017 were enrolled from the Department of Cardiology, Affiliated Zhongshan Hospital of Dalian University.The patients were randomly allocated to two groups: (1) control group (n=25), PCI alone; (2) RIC group (n=50), PCI combined with RIC (three cycles of 5 min inflation and 5 min deflation of the right lower limb with blood pressure cuff performed before reperfusion). Serum miR-208b was measured before and immediately, at 24 h, and 48 h after PCI with real-time quantitative polymerase chain reaction. Results: The expression of miR-208b was significantly higher immediately post PCI than that before operation in the control group (84.1±9.0 vs 77.8±9.4; P=0.032), while it was significantly lower immediately post PCI than that before operationin RIC group (71.0±9.3 vs 77.4±8.8; P=0.028).miR-208b level was similar before PCI between the control and RIC groups (P=0.874), which was significantly reduced immediately post PCI in RIC group as compared with the control group (P=0.021).The peak value of creatine kinase isoenzyme (CK-MB) in the limb RIC group was significantly lower than that in the control group ([135.2±18.6] U/L vs [167.7±17.2] U/L; P=0.038).The area under the CK-MB curve of the RIC group was significantly smaller than that of the control group ([3 060.7±17.1] U/L vs [3 635.9±15.1] U/L); P=0.047]. The left ventricular ejection fraction (LVEF) in RIC group was significantly higher than that in the control group ([57.8±7.8]% vs [51.9±7.9]%; P=0.003) post PCI. The expression level of serum miR-208b was positively correlated with CK-MB AUC in RIC group (r=0.498, P<0.001). Conclusions: RIC of the lower limb prior to PCI could reduce miR-208b level and improve cardiac functionin STEMI patients.

Numerous studies have been done to explore the association between mannose-binding lectin two (MBL2) gene polymorphisms and the risk of tuberculosis (TB). However, the results are inconsistent. We performed a meta-analysis to investigate whether polymorphisms in the MBL2 gene were associated with TB risk. Databases including PubMed, Medline, Chinese Biomedicine Database, China National Knowledge Infrastructure, Wanfang Database, and Weipu Database were searched to find relevant articles published up to 2 October, 2012. Odds ratio (OR) with 95% confidence interval (CI) was used to evaluate the strength of association. All statistical tests were performed by using Revman 5.1 software and STATA 11.0 software. Six case-control studies including 1106 cases and 1190 controls were accepted in the meta-analysis. The results indicated that individuals carrying the MBL2 codon 54 B allele may have an increased risk of TB as compared with AA homozygotes (BB+AB vs. AA: OR=1.52, 95% CI: 1.22-1.88), whereas MBL2 +4 P/Q was possibly not associated with TB susceptibility in Chinese population.
China ; epidemiology ; Codon ; genetics ; Genetic Markers ; genetics ; Genetic Predisposition to Disease ; epidemiology ; genetics ; Humans ; Mannose-Binding Lectin ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Prevalence ; Risk Assessment ; Tuberculosis ; epidemiology ; genetics

Objective To investigate the general situations of gravida,pregnancy complications,childbirth and neonatal outcomes of twin pregnancies conceived by assisted reproductive techniques (ART) and those conceived spontaneously.Methods A retrospective analysis was carried out on the basic information,perinatal complications,delivery information and neonatal outcomes of twin pregnancies received by ART (ART group,n=518) and those conceived spontaneously (SC group,n=293).Results Gravida age was older in ART group than in SC group (P＜0.05),while the gravidity and parity in ART group was more than that in SC group (P＜0.05).There were more embryos and high proportion of diamniotic cyst in ART group than in SC group (P＜0.05).The gestational incidence of diabetes,abnormal placenta (placental adherence,placenta accrete,placenta previa) and postpartum hemorrhage were higher in ART group than in SC group (P＜0.05),while no statistically significant differences existed between the two groups in the incidence of gestational complications (such as gestational hypertension disease,cholestasis and amniotic fluid volume abnormality) and in neonatal outcomes including neonatal weight,incidence of asphyxia and congenital malformation and neonatal death (P＞0.05).Conclusion Twin pregnancy conceived by ART may lead to higher incidences of gestational diabetes mellitus and abnormal placenta and more postpartum hemorrhage,but no significant difference existed in the neonatal outcomes between twin pregnancies conceived by ART and those conceived spontaneously.