Objective To investigate the genotypes of extended spectrum β-lactamases (ESBLs) and their carrying modes in Escherichia coli (E.coli) isolates,and to analyze the mechanism of protein phosphorylation and ESBLs gene expression induced by β-lactam antibiotics or inhibited by histidine kinase inhibitors.Methods The predominant genotypes of ESBLs (KPC,TEM,SHV and CTX-M) and their carrying modes were identified by PCR and sequencing analysis.E-test and micro-tube dilution method were applied to measure minimal inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs).Immobilized metal ion affinity chromatography,bacterial protein phosphorylation detection kit and real-time fluorescent quantitation RT-PCR were performed to analyze the enhancing effects of 1/4 MIC penicillin or cefotaxime or the inhibitory effects of histidine kinase inhibitors (closantel,bromized or iodized methylimidazol) on protein phosphorylation and the expression of ESBLs at mRNA level in E.coli isolates.Results In 183 β-lactam antibiotics-resistant E.coli isolates,TEM and CTX-M genes (83.1％ and 77.1％) were highly expressed than other two ESBLs genes with a prevalent carrying mode of coexisting (65.0％) (P＜0.05).Penicillin or cefotaxime at 1/4 MIC induced the protein phosphorylation and promoted the expression of TEM,SHV and CTX-M at mRNA level (P＜0.05).Closantel (200 μmol),bromized methylimidazol (2 or 10 μmol) or iodized methylimidazol (20 or 50 μmol) could neither kill E.coli isolates nor inhibit their growth,but could inhibit the protein phosphorylation induced by above mentioned antibiotics and enhance the expression of ESBLs at mRNA level (P＜0.05).Moreover,the susceptibility of antibioticresistant E.coli strains to penicillin and cefotaxime were increased (P＜0.05).Conclusion TEM and CTX-M were the predominant genotypes of ESBLs carried by β-lactam antibiotics-resistant E.coli strains isolated from Zhejiang province,which were mostly found in a TEM plus CTX-M carrying mode.Sublethal dose of β-lactam antibiotics could up-regulate the expression of ESBLs genes in E.coli isolates via TCSS,but it could be inhibited by histidine kinase inhibitors.

Objective To investigate the expression levels of interferon -inducible genes (MNDA)in the peripheral blood mononuclear cells (PBMCs)of patients with systemic lupus erythematosus (SLE),and the relations between the gene expression levels and disease activity were explored.Methods Reverse transcription polymerase chain reaction(RT -PCR)method was used to detect the expression levels of MNDA in 30 patients with SLE and 25 controls.The associations between the expression levels of MNDA and urine protein,complement C3,C4,anti -double stranded DNA antibody and SLEDAI scores in patients with SLE were analyzed.Results (1)The expression level of MNDA mRNA in the SLE patients was significantly higher than that of the normal controls(t =6.99,P <0.01).(2)The expression level of MNDA in the urinary protein positive patients was significantly higher than that of the urinary protein negative patients (t =3.08,P <0.01),they were all significantly higher than the normal controls (t =9.32,4.87,all P <0.01).(3)The expression levels of MNDA mRNA were positively correlated with anti -double stranded DNA antibody and SLEDAI scores(r =0.534,0.508,all P <0.05),and not correlated with comple-ment C3 and C4(r =-0.472,-0.349,all P >0.05 ).Conclusion The expression levels of MNDA mRNA in patients with SLE are significantly higher than those of the normal controls.It is linked to kidney damage and disease activity,and helpful in evaluating SLE disease activity and severity.

AIM:To explore the association between single nucleotide polymorphism in exon 33 (E33SNP) of thyroglobulin gene and Graves ’ disease ( GD) relapse after antithyroid drug ( ATD) withdrawal .METHODS:The healthy controls (232 cases) and GD patients with discontinued treatment (243 cases) were selected.According to the time of re-lapse, the GD patients were divided into A, B and C subgroups.The A group contained 77 cases of relapse within 1 year, B group contained 86 cases of relapse 1~2 years after treatment and C group contained 80 cases without recurrence within 2 years.The genotypes of E33SNP were identified by RT-PCR.The genotype ratio of thyroglobulin between control group and observation group was comparatively analyzed , and the levels of thyroid-stimulating hormone ( TSH) , free triiodothyro-nine (FT3), free thyroxine (FT4) and thyrotropin receptor antibody (TRAb), ophthalmopathy and goiter size in A , B and C subgroups in different genotype GD patients were investigated .Moreover , cumulative efficiency for patients with different genotypes in the observation group after ATD treatment within 2 years were analyzed .RESULTS:The genotype of E33SNP between observation group and control group had no significant difference , but a significant difference between A , B and C subgroups was observed (P<0.05).The levels of TSH, FT3 and FT4, and goiter size of the patients with different geno-types had no significant difference , while the TRAb levels and ophthalmopathy presented a significant difference ( P <0.05).In addition, the cumulative efficiency within 2 years for GD patients with E33SNP T/T, E33SNP T/C and E33SNP C/C genotypes was 61.8%, 42.6% and 21.3%, respectively, all with significant differences (P<0.05).CONCLU-SION:The GD patients with E33SNP C/C genotype have significantly higher TRAb level and ophthalmopathy rate than those in the patients with E33SNP C/T and E33SNP C/C genotypes, and are more likely to relapse after ATD treatment . The GD patients with E33SNP T/T genotype show a lower recurrence rate .Therefore, combination treatment or other treat-ment modalities may be more reasonable for the GD patients with E 33SNP C/C genotype.

Objective To observe the apeptosis in liver injury following limbs ischemia-reperfusion(IR) in rats and the protective effects of taurine.Methods The model of limbs ischemia-reperfusion injury was established.30 Wistar rats were randomized into 3 groups: control group,IR group and tanrine + reperfusion group (TR group) (n = 10 for each group).The levels of malondialdehyde (MDA) and xanthineoxidas (XOD), calcium and myeloper-oxidase (MPO) in the liver tissue were measured.DNA fragmentation was observed and analyzed by agarose gel e-lectrophoresis.Apoptosis was detected by TUNEL methods.The morphologic changes were observed with HE stai-ning.Results Compared with control group,the values of MDA, XOD, MPO, calcium in liver tissue were increased significantly in IR group (P<0.01), but the values of those in TR group were lower than in IR group (P<0.01).The percentage of apeptosis cell was higher in IR group than in control group(P<0.01).Compared with IR group, the percentage of apoptosis cell was lower in TR group (P<0.01).IR group presented DNA ladder pattern, while TR group showed no specific DNA ladder pattern in agarose gel electrophoresis.Conclusion Apoptosis participates in the liver injury after limb ischemia-reperfusion.Taurine can mitigate the liver injury and apoptosis after limb is-chemia-reperfusion injury in rats.

Objective To observe BQ123 on lung tissue injury following ischemia-reperfusion of hind limbs in rats.Methods A total of 30 male SD rats were randomly divided into control,I-R,and BQ123 groups,with 10 rats in each group. After 4- hour of ischemia and 4-hour of reperfusion to the hind limbs, ET-1 was measured by radioimmunoassay, levels of myeloperoxidase (MPO) and malondialehyde (MDA) were examined by biochemical method,and the content of P-selectin was examined by ELISA. Immunohistochemical method was used to detect the expressions of Fas,Bcl-2 and Caspase-3 in lung tissues. Pulmonary apoptosis was examined by means of TdT-mediated dUTP nick end labeling ( TUNEL) . Results Compared with the control group,the levels of ET-1,MPO,MDA and P-selectin in lung tissues were all increased significantly in I-R group ( P<0. 01). The expressions of Fas,Bcl-2 and Caspase-3 were 0. 294±0. 003,0. 108±0. 005,and 0. 174±0. 003,significantly up-regulated in the I-R group. The apoptosis rate [(18. 83±2. 86)%] was significantly increased in the I-R group (P<0. 01). Compared with I-R group,the tissue contents of ET-1,MPO,MDA and P-selectin were significantly lowered in BQ123 group (P<0. 01). The expression levels of Fas (0. 115±0. 007) and Caspase-3 (0. 159±0. 006) were decreased,but the expression of Bcl-2 was increased (0. 128±0. 005). The apoptosis rate in BQ123 group was significantly lower [(10. 67±2. 16)%,P<0. 01].Conclusion BQ123 may have protective effects on lung tissue after limb ischemia-reperfusion in rats by means of improving neutrophil aggregation and reducing the expression of proteins related to cell apoptosis.

OBJECTIVETo construct a prokaryotic expression system of groEL gene of Leptospira interrogans serogroup Icterohaemorrhagia serovar Lai strain Lai, and to determine the immunoprotective effect of recombinant GroEL protein (rGroEL) in LVG hamsters.

METHODSThe groEL gene was amplified by high fidelity PCR and the amplification products were then sequenced. A prokaryotic expression system of groEL gene was constructed using routine genetic engineering technique. SDS-PAGE plus Bio-Rad Gel Image Analyzer was applied to examine the expression and dissolubility of rGroEL protein while Ni-NTA affinity chromatography was used to extract the expressed rGroEL. The immunoprotective rate in rGroEL-immunized LVG hamsters was determined after challenge with L.interrogans strain Lai. The cross agglutination titers of sera from immunized hamsters with different L.interrogans serogroups were detected using MAT.

RESULTSThe nucleotide and amino acid sequences of the cloned groEL gene were the same as those reported in GenBank. The constructed prokaryotic expression system of groEL gene expressed soluble rGroEL. The immunoprotective rates of 100 and 200 μg rGroEL in LVG hamsters were 50.0 % and 75.0%, respectively. The sera from the rGroEL-immunized LVG hamsters agglutinated all the L.interrogans serogroups tested with different levels.

CONCLUSIONThe GroEL protein is a genus-specific immunoprotective antigen of L.interrogans and can be used to develop an universal genetically engineering vaccine of Leptospira.

Agglutination Tests ; Animals ; Antigens, Bacterial ; immunology ; Chaperonin 60 ; genetics ; immunology ; Cricetinae ; Gene Expression ; Leptospira interrogans ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology

Objective:To understand the influencing factors of the elderly' s demand for the pension mode of medical-nursing combination in Urumqi. Methods: A questionnaire survey was conducted among 300 elderly people randomly selected in Urumqi. Results:Univariate analysis showed that there was significant difference between retirement monthly income, life self-care ability, type of chronic disease, the degree of understanding on the pension mode of medical-nursing combination, family support and the demand rate of the elderly for the pension mode of medical-nursing combination ( P< 0 . 05 ) . Two Logistic regression analysis showed that life selfcare ability, types of chronic diseases, the degree of understanding on the pension mode of medical-nursing combination, and family support factors were statistically significant. Conclusion:It should change the traditional concept of single pension mode, improve the medical-nursing combination service system to meet the needs of all levels of the elderly, establish a long-term care insurance system, and improve the training mechanism of the elderly.

OBJECTIVE To investigate the characteristics and laws of changes in electrically evoked auditory brainstem responses(EABR),electrically evoked stapedius reflex(ESRT) after cochlear implantation in children with inner ear malformation.METHODS 88 cases of Australian 24 multi-channel cochlear implants were divided into normal group and inner ear malformation group,The EABR and ESRT thresholds were measured at different periods within one year after operation.Behavior auditory responses,T-level and C-level were measured 1 year postoperatively to analyze their characteristics and changes.RESULTS The EABR and ESRT thresholds in the patients with inner ear malformation were significantly higher than those in the normal cochlear group(P＜0.05).The changing trend of EABR and ESRT were the same in the 2 groups,the overall trend is that low frequency value is low,high frequency value is higher.EABR,ESRT threshold increased gradually after 1 year,EABR was significantly correlated with T-level,and ESRT was significantly correlated with C-level.CONCLUSION The changes of EABR and ESRT thresholds in the cochlear implantation of the inner ear malformation group were similar to those of the normal cochlear group,the thresholds can be used to guide mapping for the inner ear malformation of the cochlear implant.

OBJECTIVETo use combined comparative genome hybridization (array-CGH) and conventional karyotype analysis to study the relationship between ultrasonographic abnormalities of fetuses and chromosomal aberrations.

METHODSOne hundred twenty two fetuses with ultrasonographic abnormalities in middle and late trimesters suspected with chromosomal abnormalities were collected between March 2012 and February 2013.

RESULTSThe pregnant women had an average age of 31 yr (22-38), among whom 35 were above the age of 35. The average gestational age was 27(+5) weeks (18-37 weeks), and the most common abnormal findings have involved heart, central nervous system and bones. Multiple malformations were found in 49 cases. The success rate of the combined methods was 100%. In 24 (19.7%) of the cases, a chromosomal abnormality was detected. Among all cases, 16 (13.1%) were detected by the combined method (12.3%). Seventeen cases (13.9%) of chromosomal abnormalities and 4 cases (3.3%) of polymorphic variation were detected by karyotype analysis, and 23 cases (8.9%) of abnormalities were detected by array-CGH. Meanwhile, 7 cases (5.7%) of abnormalities were detected by array-CGH, but the results of karyotype analysis were normal. One case (0.8%) with low level of chromosome chimerism detected by the karyotype analysis was missed by array-CGH.

CONCLUSIONThe results suggested that multiple congenital deformity of the fetus has a strong correlation with chromosomal abnormalities. For fetuses with ultrasonographic abnormalities, array-CGH can improve the detection sensitivity of the chromosomal disease.

Adult ; Chromosome Aberrations ; Chromosome Banding ; methods ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; Comparative Genomic Hybridization ; methods ; Female ; Fetal Diseases ; diagnosis ; diagnostic imaging ; genetics ; Gestational Age ; Humans ; Karyotyping ; Male ; Pregnancy ; Prenatal Diagnosis ; methods ; Ultrasonography, Prenatal ; methods ; Young Adult