Objective To investigate prevalence of resistance genes for β-lactams in clinically isolated multidrug-resistant Acinetobacter baumannii (MDRAB)in a hospital.Methods 22 clinically isolated MDRAB strains were performed antimicrobial susceptibility testing,resistance genes forβ-lactams (TEM,SHV,CTX-M,PER,DHA, IMP ,VIM ,SIM,OXA-23,OXA-24,and OXA-58)in these strains were detected with polymerase chain reaction. Results The resistant rates of 22 isolates of MDRAB to ceftazidime,cefotaxime,cefepime,gentamycin,amikacin, ciprofloxacin,and compound sulfamethoxazole were all 100.00%;to imipenem,meropenem,and cefoperazone/sul-bactam were 50.00%,40.91 %,and 31 .82% respectively,intermediated rates were 31 .82%,36.36%,and 31 .82% respectively.The carriage rates of OXA-23,TEM,IMP ,and PER were 100.00% (n =22),72.73 %(n=16),54.55% (n = 12),and 18.18% (n =4)respectively.Detection results of SHV,CTX-M,DHA,VIM , SIM,OXA-24,and OXA-58 were all negative.Conclusion Carriage of IMP ,TEM,PER,and OXA-23 resistance genes are the major resistance mechanisms of MDRAB to β-lactamase antimicrobial agents in this hospital.

An HPLC-DAD-MS/MS method was developed for rapid analysis and identification of degradation products of buagafuran. Buagafuran and degradation products were separated on a Zorbax C8 column (5 microm, 4.6 mm x 150 mm) using acetonitrile-water (78 : 22) as mobile phase. The elutes were detected with diode array detector and tandem mass spectrometer via electrospray ionization source in positive ion mode. According to analysis of the retention time, UV spectra and MS, MS/MS data, combined with the possible degradation reaction of buagafuran, the structures of main degradation products were inferred. The results showed that six main degradation products were oxidation or peroxidation productions of buagafuran. Degradation product A was a double bond epoxidation product of buagafuran, degradation products B, C, D and E were the further oxidation products of degradation product A, degradation product F was a peroxidation product of buagafuran. The results indicated that the established method was effective in the rapid identification of the degradation products of buagafuran.

Objective To identify an suspected Vibrio cholerae isolated from Xiangyang Central Hospital and characterize the strain in terms of antibiotic resistance and relevant virulence genes.Methods Pathogen identification and susceptibility testing were completed with MicroScan WalkAway 40 Automated Microbiology System.Slide agglutination was used for serotyping. PCR and sequencing technology were employed for 16s RNA gene analysis.PCR technique was used to detect six major viru-lence genes.Results This suspectedVibrio cholerae isolate was confirmed as non-O1 and non-O139 Vibrio cholerae .Suscep-tibility testing results indicated that the strain was sensitive to ampicillin,chloramphenicol,trimethoprim-sulfamethoxazole, and tetracycline.16s RNA gene sequence analysis showed 100% homologous with the registered sequence in National Center for Biotechnology Information database.Virulence genes rtxC and toxR were identified.The other virulence genes such as tcpAET,ctxA,hlyA,and tcpACL were negative.Conclusions This suspected Vibrio cholerae isolate is confirmed as non-O1 and non-O139 Vibrio cholerae .The pathogenic factors may be related to the virulence genes rtxC and toxR.

The factor analysis method applied in imaging mass spectrometry data analysis was studied. The imaging mass spectrometric data were obtained by air flow-assisted ionization imaging mass spectrometry method. The sample contained some symbols which were drawn on slides using three different inks ( red, blue, black) . The imaging data analyzed by factor analysis method were divided into the background, black, blue and red factor. The results showed that the scores of m/z=443. 2, 478. 4, 322. 2(344. 2) in red, blue, black factor respectively were much larger than others. Therefore, they were markers of three inks. The results accorded with actual condition well and proved that the application of factor analysis in imaging mass spectrometric data analysis was feasible. The data analysis results of factor analysis and principal component analysis were compared. The results showed that the target sample markers could be extracted by factor analysis simply and quantitatively. It was of great potential in biomarker extraction, diseases diagnose and pharmacological analysis.

To screen the harmful substance 5-hydroxymethyl furfural content in commercially available traditional Chinese medicine injection which are commonly used, and to preliminarily evaluate the quality of these injections, 5-hydroxymethyl furfural was taken as an index. The contents of 5-hydroxymethyl furfural in 56 samples which consist of 23 kinds of traditional Chinese medicine injections and glucose injection were determined using LC-MS/MS, and 5-hydroxymethyl furfural was detected in 52 of these samples. The minimal content was 0.0038 microg x L(-1) and the maximum content was 1420 microg x mL(-1). The contents of 5-hydroxymethyl furfural were significantly different in traditional Chinese medicine injection which came from different kinds, manufacturers or batches. The results showed the quality difference of commercially available traditional Chinese medicine injection is significant taking 5-hydroxymethyl furfural content as assessment index. More attention should be paid to the safety of 5-hydroxymethyl furfural in traditional Chinese medicine injection, and unified limitation standard should be set to improve medication safety of traditional Chinese medicine injection.

Objective To research a simple and sensitive K-ras gene mutations detection method in order to be suitable for the routine mutation detection.Methods The corresponding detection locus oligonucleotide probe was designed.By the connection,amplification,labeling and ELISA reaction in probe,the mutation locus genotype was determined by the ELISA reaction detection value.With the six point mutations of G12S,G12R,G12C,G12D,G12A and G12V in 12 codons of K-ras gene as the detection objects,the plasma circulation DNA sample in 72 cases of lung cancer was detected,then the results were compared with those obtained by the direct sequencing.Results Three samples were identified as the G12S,G12R and G12A mutatins by the established method.But no K-ras mutations were detected in the samples by using the direct sequencing,indicating that the direct sequencing had lower sensitivity and was not suitable for the mutation detection of heterogeneous samples such as circulating DNA.Conclusion The simple and sensitive K-ras gene mutation detection method is established and can conduct the routine mutation detection for the heterogeneous samples.

OBJECTIVETo screen methylations of CpG islands in prostate cancer using restriction landmark genomic scanning (RLGS).

METHODSThe DNA was extracted from homogeneous cells captured by laser capture microdissection in 20 prostate cancer and 18 benign prostatic hyperplasia (BPH) tissues for scanning the CpG islands using RLGS. The methylation status of each CpG island was compared between the cancer and BPH samples to screen the genes involved in prostate cancer development. The screened genes were uploaded to DAVID database for GO analysis, and the genes with the most significant methylation were analyzed by pyrosequencing.

RESULTS AND CONCLUSIONAmong all the tested CpG islands, 10245 (37.2%) in prostate cancer and 8658 (30.3%) in BPH samples were found to be abnormally methylated, and >60% of the methylated CpG islands were in the promoter region. Compared with BPH samples, the prostate cancer samples showed differential methyation in 735 CpG islands, including 458 hepermethyated and 256 hypomethelated ones. Seven genes (DPYS, P16, APC, GSTP1, TMEM122, RARB, and ARHGAP20) in prostate cancer were identified to have distinct methylations. Bioinformatics analysis suggested that these genes were associated with several biomolecular and biological processes, and among them DPYS gene was involved in 13 GO anotated biologic functions, development of 50 diseases and 47 protein interactions. Pyrosequencing of 7 sites of the CPG island in DPYS gene showed a methylation frequency of 32.7%, suggesting the importance of DPYS gene in the carcinogenesis and progression of prostate cancer.

CpG Islands ; DNA Methylation ; DNA, Neoplasm ; genetics ; Genomics ; Humans ; Male ; Polymerase Chain Reaction ; Prostatic Hyperplasia ; genetics ; Prostatic Neoplasms ; diagnosis ; genetics

Three-dimensional(3D)cell spheroid models combined with mass spectrometry imaging(MSI)enables innovative investigation of in vivo-like biological processes under different physiological and patho-logical conditions.Herein,airflow-assisted desorption electrospray ionization-MSI(AFADESI-MSI)was coupled with 3D HepG2 spheroids to assess the metabolism and hepatotoxicity of amiodarone(AMI).High-coverage imaging of＞1100 endogenous metabolites in hepatocyte spheroids was achieved using AFADESI-MSI.Following AMI treatment at different times,15 metabolites of AMI involved in N-desethylation,hydroxylation,deiodination,and desaturation metabolic reactions were identified,and according to their spatiotemporal dynamics features,the metabolic pathways of AMI were proposed.Subsequently,the temporal and spatial changes in metabolic disturbance within spheroids caused by drug exposure were obtained via metabolomic analysis.The main dysregulated metabolic pathways included arachidonic acid and glycerophospholipid metabolism,providing considerable evidence for the mechanism of AMI hepatotoxicity.In addition,a biomarker group of eight fatty acids was selected that provided improved indication of cell viability and could characterize the hepatotoxicity of AMI.The combination of AFADESI-MSI and HepG2 spheroids can simultaneously obtain spatiotemporal infor-mation for drugs,drug metabolites,and endogenous metabolites after AMI treatment,providing an effective tool for in vitro drug hepatotoxicity evaluation.

OBJECTIVETo compare the safety, feasibility and efficacy of transperitoneal and retroperitoneal laparoscopic partial nephrectomy (LPN) in the treatment of renal tumors with R. E. N. A. L score more than 7.

METHODSThe clinical data were collected from 62 patients undergoing transperitoneal LPN (32 cases) and retroperitoneal LPN (30 cases) for a complex renal mass (R.E.N.A.L. score≥7) between January 2012 and March 2014. The surgical and early postoperative outcomes and complications were analyzed to evaluate the efficacy of the treatments. The mean operative time, estimated blood loss, warm ischemia time, surgical complications, blood transfusion rate, tolerating regular diet time, postoperative hospital stay and surgical margin were compared between the two groups.

RESULTSThe operations were completed successfully in all cases except for 1 case in transperitoneal group and 3 in retroperitoneal group that required conversion to open surgery. No significant differences were found in age, body mass index, ASA score, Charlson comorbidity index, tumor size or R.E.N.A.L. nephrometry score (P>0.05), nor in estimated blood loss, warm ischemia time, intraoperative complication, blood transfusion rate or surgical margin between the two groups (P>0.05, respectively). The transperitoneal LPN group had a shorter mean operative time than retroperitoneal LPN group (210.4∓59.2 vs 252∓58.3 min, P<0.05) but showed longer tolerating regular diet time (47∓10 h vs 23∓6 h, P<0.05) and postoperative hospital stay time (8.4∓1.9 days vs 6.5∓1.6 days, P<0.05).

CONCLUSIONBoth transperitoneal LPN and retroperitoneal LPN are safe, feasible and effective for surgical management of complex localized tumors, but the transperitoneal procedure offers larger operative space with better exposure; the retroperitoneal procedure better promotes postoperative recovery of the patients.

Humans ; Kidney Neoplasms ; diagnosis ; surgery ; Laparoscopy ; Length of Stay ; Nephrectomy ; Operative Time ; Retroperitoneal Space ; Retrospective Studies ; Treatment Outcome

Tumors are spatially heterogeneous tissues that comprise numerous cell types with intricate structures.By interacting with the microenvironment,tumor cells undergo dynamic changes in gene expression and metabolism,resulting in spatiotemporal variations in their capacity for proliferation and metastasis.In recent years,the rapid development of histological techniques has enabled efficient and high-throughput biomolecule analysis.By preserving location information while obtaining a large number of gene and molecular data,spatially resolved metabolomics(SRM)and spatially resolved transcriptomics(SRT)approaches can offer new ideas and reliable tools for the in-depth study of tumors.This review provides a comprehensive introduction and summary of the fundamental principles and research methods used for SRM and SRT techniques,as well as a review of their applications in cancer-related fields.