Objective:To establish the method for the expression,purification and characterization of metallo-?-lactamases(IMP-1) and to explore the influence of experimental conditions on its separation and purification.Methods:pET9a/d-IMP-1 plasmid was transformed into E.coli competent BL21(DE3) cell and inoculated in LB medium.The supernatant were applied to SP Sepharose Fast Flow column,following by Sephadex G-75 column,and the purified enzymes were identified by SDS-PAGE and MALDI-TOF-MS and their activity was determined using Nitrocefin as substrate.Results:The purified enzyme showed high catalytic activity in the hydrolysis of most antibiotics.Its Michaelis constant K_m9.28?mol/L,and molecular weight Mr equalled 25111.9(determined by MALDI-TOF-MS).Conclusion:The method was established for the expression,purification and characterization of metallo-?-lactamases(IMP-1),which can be applicable to other metallo-?-lactamases.

Beginning with the authors' experience in analytic-chemistry bilingual teaching,this article probes into some problems of analytic-chemistry in undergraduate education.And the authors give some beneficial suggestions in the teaching material construction,teachers training and teaching quality control.

The present situation and problems in experiment teaching of analytical chemistry were explained,and innovation was explored and practiced in the faculty of laboratory medicine,including the opitimization of teaching contents,teaching methods and score system evaluation reformation.

The academic teaching quality and experimental teaching quality of sanitary chemistry will be improved,through enriching the teacher with specialized knowledge and cultivating the creative ability of thacher and updating the teaching methods.

Sample preparation and separation is a very crucial and complicated process in proteomics research.Based on the actual condition of university and students,this article explored the way to improve the teaching quality of sample preparation and separation in proteomics course for medical post-graduates aiming at cultivating and enhancing the students' scientific thought,making them have a better understanding of the rules,technologies,strategies in sample preparation and separation procedure.

A liquid chromatography-quadrupole-time-of-flight mass spectrometer(LC-Q/TOF-MS) based urinary metabolomic approach was employed to assess the toxicity-alleviation effect of Huangqi oral solution(HOs) on cisplatin-exposed rats and explore its possible mechanisms. Rat toxicity model was developed by multiple intraperitoneal injection of low-dose cisplatin, while HOs was orally administrated to rats simultaneously for 16 consecutive days to attenuate or reduce the cisplatin-induced toxicity. 24-hour urine samples on day 18 were collected and analyzed using LC-Q/TOF-MS to obtain the dataset of urinary metabolites. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to assess the quality of the dataset and screen the potential toxicity-alleviation biomarkers. The serum level of rat creatinine and urea nitrogen on day 20 was determined, and the results showed that successive administration of HOs significantly reduced the cisplatin-induced increase of creatinine and urea nitrogen. PCA cluster analysis clearly demonstrated that HOs could partly improve the CDDP-induced abnormality of metabolic profiling. 35 urinary metabolites were finally screened as the potential biomarkers associated with the toxicity-attenuation effect of HOs, according to the combination of the analysis results of OPLS-DA, t-test and fold change analysis. Further metabolic pathway analysis revealed that HOs could restore the metabolic disorders of amino acid, energy and nucleotide, thereby exerted its toxicity-alleviation effect.

Objective To construct microRNA (miRNA) -mediated gene regulation of ovarian cancer by establishing regulatory networks of miRNA and long non-coding RNA (lncRNA) database based on bioinformatic analysis.Methods Ovarian cancer-related regulatory miRNAs were obtained via miRwalk 2.0and HMDD v2.0.According to the frequency of miRwalk 2.0software, we screened and identified the corresponding target gene expression.Upon a matching analysis, ovarian cancer matched lncRNAs searched from the lncRNA database were performed to determine miRNAs interacted with lncRNA, and starBase v2.0was used to comprehensively analyze the obtained miRNA and lncRNA.Finally, miRNA-mediated gene regulatory networks of ovarian cancer were mapped by the Cytoscape software.Results MiRNA-mediated gene regulation of ovarian cancer was successfully established regulatory networks of miRNA and lncRNA databases.Hsa-let-7a, hsamir-21, hsa-miR-16, hsa-mir-17, hsa-mir-19b, hsa-mir-29aand hsa-mir-92awere high frequency miRNA of ovarian cancer.According to matching analysis, lncRNA gene symbol X inactive-specific transcript (XIST) played the central role in lncRNA-miRNA interaction in regulation of ovarian cancer.Conclusion The miR-NA-mediated gene regulating network of ovarian cancer was successfully established according to bioinformatic analysis.Databases like miRwalk, HMDD, starBase could be used as effective tools to analyze relationship between miRNA and disease.

Objective To investigate the regulatory effects of wedelobata on apoptosis and secretion of inflammatory factors in the alveolar epithelial cells infected by Streptococcus pn e um on i a e.Methods Alveolar epithelial cells A549 were divided into infection group(1×108/CFU/mL cultured cells of Streptococcus pneumoniae),control group(no treatment),infection+wedelolactone low-dose group,middle-dose group and high-dose group(pretreated with wedelolactone at 10,20 and 40 μmol/L and then cultured cells of Streptococcus pneumoniae at 1×108/CFU/mL).Alveolar epithelial cells A549 were divided into infection group(1×108/CFU/mL cultured cells of Streptococcus pneumoniae),control group(no treatment),infection+wedelolactone low-dose group,middle-dose group and high-dose group(pretreated with wedelolactone at 10,20 and 40 μmol/L and then cultured cells of Streptococcus pneumoniae at 1×108/CFU/mL).Results Compared with control group,the apoptosis rate,the relative expression levels of Bax,Caspase-3 protein,IL-6,IL-1β and TNF-α mRNA were higher in infection group,infection + wedelolide low dose group,medium dose group and high dose group,while the expression level of Bcl-2 protein was lower(P＜0.05).Compared with the infected group,the apoptosis rate,the relative expression levels of Bax,Caspase-3 protein,IL-6,IL-1β and TNF-α mRNA and the expression levels of Bcl-2 protein were lower in the infected + wedelolide low dose,medium dose and high dose groups.Moreover,the apop-tosis rate,the expression levels of Bax,Caspase-3 protein,IL-6,IL-1β and TNF-α mRNA were the highest in the infected + wedelactone high-dose group,and the expression levels of Bcl-2 protein were the lowest(P＜0.05).Conclusion The apoptosis rate of alveolar epithelial cells infected by Streptococcus pneumoniae decreased and the secretion of inflammatory factors decreased after the intervention of wedelia lactone.

OBJECTIVETo summarize the clinical characteristics of peripheral blood, immune phenotypes, fusion genes and cytogenetics of patients with t(8;21) acute myeloid leukemia(AML) through the retrospective analysis of 586 patients with t(8;21) AML from 15 blood disease research centers in Northern area of China.

METHODSThe factors affecting prognosis of patients with t(8;21) AML were investigated by using univariate and multivariate COX regression.

RESULTSThe immune type of t(8;21) AML patients was mainly with HLA-DR, CD117, CD34, MPO, CD38, CD13and CD33(>95%), part of them with CD19and CD56; the most common accompanied mutation of t(8;21) AML patients was C-KIT mutation (37.8%); in addition to t(8;21) ectopic, the most common chromosomal abnormality was sex chromosome deletions (38.9%). The univariate analysis revealed a significant survival superiority of OS and PFS in t(8;21) AML patients of WBC≤3.5×10/L without C-KIT mutation, the newly diagnosed ones achieved HSCT(P<0.05), only survival superiority on OS in t(8;21) AML patients with extramedullary infiltration and CD19 positive; the results of multivariate analysis showed a significant survival superiority on OS and PFS in t(8;21) AML patients with WBC≤3.5×10/L(P<0.05).

CONCLUSIONThe clinical features of t(8;21) AML patients in China are similar to those in other countries, WBC≤3.5×10/L is a good prognostic factor while the C-KIT mutation is a poor one in t(8;21) AML patients.