Objective To explore the efficacy of calmodulin antagonist berbamine(BBM) in down-regulating survivin mRNA. Methods Human breast cancer cell line MCF7 and its adriamycin-resistant counterpart MCF7/ADR were used in the study. The cells were cultured with different concentration of BBM for 72 hours. The mRNA expression level of survivin gene in both MCF and MCF7/ADR cells was detected by semi-quantitative RT-PCR. Results After treating MCF7 and MCF7/ADR cells by 20?mol/L BBM, the mRNA expression level of survivin gene decreased from 0.43?0.02 to 0.21?0.04 in MCF7 cells, and from 0.57?0.05 to 0.45?0.04 in MCF/ADR cells(P

Objective To detect serum anti-Treponema pallidum specific antibody of 26 707 cases by Abbott I2000SR auto-matic chemiluminescent microparticle immunoassay analyzer,and treponema pallidum particle agglutination assay (TPPA) was regarded as a standard reference method which was used to detect anti-Treponema pallidum specific antibody.To analyze the false positive rate of Abbott I2000SR according to the TPPA.Methods Collected 26 707 serums from inpatients and outpatients of the hospital during September 1,2013 to March 5,2014.The subjects were asked to fasting conditions taking venous blood 3 ml,3 000 r/min centrifugal 10 min utes after the separation of serum,detected the Anti-TP by CMIA (Ab-bott I2000SR)and the TPPA testing,analyzed test results by statistical methods.Results There were 52 cases detected by I2000SR whose S/CO values of 26 707 cases of serum Treponema pallidum specific antibodies were 1 to 2,of which 9 cases were verified positive by TPPA,and the positive rate was 17.31%.There were 26 cases detected by I2000SR whose S/CO values of Treponema pallidum specific antibodies were 2 to 3,of which 9 cases were verified positive by TPPA,and the posi-tive rate was 34.62%.There were 26 cases detected by I2000SR whose S/CO values of Treponema pallidum specific anti-bodies were 3 to 5,of which 9 cases were verified positive by TPPA,and the positive rate was 34.62%.There were 25 cases detected by I2000SR whose S/CO values of Treponema pallidum specific antibodies were 5 to 7,of which 11 cases were veri-fied positive by TPPA,and the positive rate was 44%.There were 25 cases detected by I2000SR whose S/CO values of Treponema pallidum specific antibodies were 7 to 10,of which 17 cases were verified positive by TPPA,and the positive rate was 68%.There were 28 cases detected by I2000SR whose S/CO values of Treponema pallidum specific antibodies were 10to 13,of which 24 cases were verified positive by TPPA,and the positive rate was 85.71%.There were 23 cases detected by I2000SR whose S/CO values of Treponema pallidum specific antibodies were 13 to 17,of which 20 cases were verified posi-tive by TPPA,and the positive rate was 86.96%.There were 24 cases detected by I2000SR whose S/CO values of Trepone-ma pallidum specific antibodies were 17 to 21,of which 22 cases were verified positive by TPPA,and the positive rate was 91.67%.There were 29 cases detected by I2000SR whose S/CO values of Treponema pallidum specific antibodies were 21 to 26,of which 28 cases were verified positive by TPPA,and the positive rate was 96.55%.There were 104 cases detected by I2000SR whose S/CO values of Treponema pallidum specific antibodies were above 26,of which 104 cases were verified posi-tive by TPPA,and the positive rate was 100%.The total number of positive cases were 364,of which 254 were positive ca-ses,the positive rate was 69.78%.False positive rate was 0.42% and positive predictive value was 69.78%.Conclusion Abbott I2000SR automated chemiluminescent microparticle immunoassay analyzer has the feature of automated detection, closed reagents,simple operation,speed,and more accurate results and so on.Although high sensitivity but its results have false positive,so cannot diagnose based on the results of Abbott I2000SR,and need use of the TPPA to test and corroborate.

Objective To analyze the phenotype and genatics in a Chinese family with early-onset familial Alzheimer's disease(EOFAD). Methods Peripheral blood were collected in available members in the family and genomic DNA was extracted. PCR-sequencing of exon 16 and exon 17 of the amyloid precursor protein(APP) gene, presenilin 1 (PSEN1), and presenilin 2 (PSEN2) was performed. Results At age 40, two EOFAD patients (siblings) in the family developed an insidious onset of difficulties in memory. One ( Ⅱ3 in the pedigree) showed blinking. The other ( Ⅱ 5 ) showed irritability and bradykinesia.Progressive diffuse coritcal atrophy in bilateral temporal cortex was observed. Moderate diffuse cerebral dysfunction was observed in Ⅱ3 by the electroencephalogram study and neuropsychological assessments.Sequencing revealed that both patients were heterozygous for a mutation c. 2343 G ＞ A in exon 17 of APP,causing the amino acid substitution Val715Met. Four members ( Ⅱ1, Ⅱ 3, Ⅱ 5 and Ⅲ1 ) were homozygous for ApoE ε4 allele. Ⅱ9 was ε2/ε4. Conclusions This study identified a mutation, Val715Met in the APP gene in Chinese patients with EOFAD. We suggest screening for APP gene mutations in Chinese patients with EOFAD.

Objective To investigate the effects of cefoprazone/sulbactam(CPZ/Sul)and dexamethasone(DXM) on serum IL-8,IL-10 in SD rats with severe pneumonia due to Klebsiella pneumoniae.Methods A rat model of severe pneumonia was induced by intratra- cheal instillation of Klebsiella pneumoniae with a dose of 3.6×108CFU(0.3ml).60 male SD rats were random divided into control group,model group,cefoprazone/sulbactam(CF-L/Sul)group and cefoprazone/sulbactam plus dexamethasone(DXM+CPZ/Sul)group.15 rats in each group were then divided into three subgroups and 5rats in each subgroup were examined at the 6th,8th and 10th day,respectively.Arterial gas analysis.total WBC and PMN cell counts in BALF were examined.ELISA method was used to detect the level of IL-8 and IL-10 in serum.RESULTSIn model group,PaO2 were progressively decreased while total WBC and PMN cell counts in BALF were markedly increased.In CPZ/Sul group,all parameters were alleviated,but there were no significant difference of serum IL-8 and IL-10 between model group and CPZ/Sul group.In DXM+CPZ/SUl group,all parameters were significantly alleviated including serum IL-8 and IL-10.Conclu sions Severe pneumonia of rat due to Klebsiella pneumoniae was characterized by significant elevation of serum IL-8 and IL-10,which suggested the imbalance of inflammatory/anti-inflammatory mediators during the course of severe pneumonia.DXM codd markedly decrease the production of inflammatory mediators and slightly decrease the production of anti-inflammatory mediators,which could restore the balance and contribute to the relief of severe pneumonia.

Objective:To study the role of 14-3-3εand Cdc25B in germinal vesicle (GV)-stage arrest of mouse oocytes,and to pay foundation for further study on the molecular mechanism of PKA/Cdc25B/14-3-3εpathway in GV-stage arrest of mouse oocytes.Methods:The eukaryotic expression vectors of pcDNA3.1-ZEO-HA-14-3-3ε, pcDNA3.1-MYC-Cdc25B-WT, pcDNA3.1-MYC-Cdc25B-S321A, and pcDNA3.1-MYC-Cdc25B-S321D were transcribed into mRNA invitro.The mouse GV-stage oocytes were collected after superovulation and divided into no injection group,TE buffer microinjection group,14-3-3εmRNA injection group,14-3-3εmRNAs + Cdc25B-WT mRNA injection group,and 14-3-3εmRNA + Cdc25B-S321A mRNA injection group,14-3-3εmRNA+Cdc25B-S321D mRNA injection group.The protein expression levels of HA-14-3-3εand MYC-Cdc25B and the phosphorylation status of Cdc2-pTyr15 were observed by Western blotting method.The morphological changes and germinal vesicle breakdown (GVDB)rates of mouse oocytes were observed under phase-contrast microscope. Results:None of the oocytes in no injection group, TE buffer microinjection group, 14-3-3εmRNA injection group,14-3-3εmRNA + Cdc25B-WT mRNAs injection group and 14-3-3εmRNA + Cdc25B-S321D mRNA were able to undergo GVBD until at least 20 h after injection (P>0.05 );the GVBD rates of oocytes in 14-3-3εmRNA+Cdc25B-S321A mRNA group at 1 h (5.00%±0.68%),2 h (62.00%±3.56%)and 3 h (100.00%± 0.00%)after injection were significantly higher than those in no injection group and TE buffer injection group (P<0.01);the oocytes in 14-3-3εmRNA+ Cdc25B-Ser321A mRNA group at 20 h (79.00%±2.80%)after injection progressed to MII (P<0.01).Conclusion:14-3-3εcan regulate the transition from GV to GVBD of mouse oocytes by means of phosphorylation and dephosphorylation of S321-Cdc25B.

Objective:To explore the effect of sub-inhibitory concentration of imipenem on the bio-activities of methicillin resistant Staphylococcus aureus(MRSA) and illuminate the inhibitory effects of carbapenem antibioty on the activity of MRSA and their mechanisms,and to provide the basis for using the carbapenem antibiotics in the treatment of MRSA infection.Methods:Five ST239 type of MRSA clinical isolates were selected and were co-cultured with 1/10 and 1/2 minimal inhibitory concentration (MIC) of imipenem for 1.5,6.0 and 12.0 h,which were divided into control group(no imipenem),1/10MIC group(1/10MIC of imipenem),and 1/2MIC group(1/2MIC of imipenem).Fluorescent quantitative real-time PCR method was used to determine the relative mRNA expression levels of virulence-related genes fibronectin A(fnbA),staphylococcal protein A(spa),α-hemolysin(Hla),leukocidin D(lek-D),leukocidin E(lek-E),and enterotoxin A in various groups;Spectrophotometry was used to detect the proliferation activity of MRSA strains in various groups in vitro.Results:After co-culture for 6.0 and 12.0 h,the proliferation activities of 5 trains of MRSA in 1/2MIC group in vitro were significantly decreased compared with control group (P<0.01).The relative mRNA expression levels of 6 virulence-related genes of 5 strains of MRSA in 1/10MIC and 1/2MIC groups were significantly decreased compared with control group(P<0.01).After co-culture for 12.0 h,the mRNA expressions of all the tested virulence-related genes were not found.Conclusion:The sub-inhibitory concentration of imipenem shows obviously inhibitory effect on the mRNA expressions of multiple virulence-related genes of ST 239 type of MRSA strains,and higher concentration of imipenem can suppress the proliferation of MRSA strains in vitro.Imipenem shows the potential value in the treatment of the severe MRSA infected patients.

Objective To explore the efficacy of calmodulin antagonist berbamine(BBM)on multidrug resistance(MDR)reversal and its mechanism. Methods Human breast cancer cell line MCF7 and its adriamycin-resistant counterpart MCF7/ADR were used in the study.The cells were cultured with ADR and different concentration of BBM. MTT assay was used to analyze the effect of BBM on cell growth inhibition.According to the MTT assay,the 50% inhibitory concentration(IC 50 ),the multiples of drug resistance and increased sensitivity of ADR were calculated.The concentration of intracellular ADR and expression level of P-glycoprotein(P-gp)were detected by flowcytometry(FCM).The mRNA expression level of mdr1 gene was detected by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR)with ?-actin as internal reference. Results The IC 50 of ADR in MCF7 and MCF7/ADR cells were(0.98?0.06)?mol/L and(101.20?5.72)?mol/L,respectively.The resistant multiple of MCF/ADR cells to ADR was 103 folds higher than that of MCF7 cells.BBM increased the chemo-sensitivity of ADR in MCF7/ADR cells with dose-dependent relationship,i.e.when 5*!?mol/L ,10*!?mol/L and 20*!?mol/L BBM was added into the culture the chemo-sensitivity of ADR was increased to 2.76,5.88,and 28.26 folds(P

Objective To elucidate the difference between methicillin-resistant Staphylococcus aureus (MRSA)and methicillin-sensitive S.aureus (MSSA)in terms of genotypes and distribution of virulence genes with the clinical strains isolated from Hohhot,and explore the relationship between the changing resistance of S.aureus and the virulence transition.Methods Pulsed field gel electrophoresis (PFGE)and multi locus sequence typing (MLST)methods were employed to do molecular typing for 30 MRSA strains and 30 MSSA strains isolated from inpatients in Hohhot,Inner Mongolia.PCR method was used to profile the distribution of virulence genes among these strains.Results PFGE typing results showed that 60 S.aureus strains were classified into 19 major types.MSSA strains belonged to 16 types,mainly types I and H.MRSA strains mainly belonged to types of K and M.Among the 20 strains with different PFGE types,MRSA strains were mainly identified as ST-239 type.but the prevalence of sec ,seg ,sei,sem,sen,seo,fnbB ,ebpS and cap 5 was higher in MSSA strains than in MRSA strains (P<0.05).Conclusions The clinical strains of S .aureus isolated from Hohhot showed diverse genotyping features.ST-239 was the major PFGE type of MRSA strains.The prevalence of virulence genes was higher in MSSA strains than in MRSA strains. Characteristic cluster is found for specific virulence genes.The results also suggest that acquisition of specific antibiotic resistance may be associated with change of specific virulence feature in S.aureus.

Brucellosis is a systemic allergic zoonosis caused by Brucella infection.Its incidence is growing.New and recurrent cases have been reported annually.The infection of Brucella is mainly due to its different virulence factors and mechanism of immune escape.In this paper,the virulent factors,infection mechanism of Brucella and damage to human beings are reviewed,so as to provide a new strategy to the diagnosis and prevention of brucellosis.