ObjectiveTo find out the antimicrobial resistance of clinical sequential isolates of Serratia marcescens,investigate the primary antimicrobial resistant mechanism of Serratia marcescens to β-lactams antibiotics.MethodsReview the antimicrobial resistance data of 247 Serratia marcescens isolates collected sequentially from different clinical wards during 2007 to 2010 in the First Hospital of Ningbo,which their antimicrobial susceptihility testing was got by using Vitek2-Compact system and matching products of gram-negative susceptibility card (GNS).The antimicrobial resistant genes of 20 carbapenems resistant isolates were detected by PCR.Results The Serratia marcescens resistant rates to ceftriaxone,aztreonarn and ciprofloxacin in our hospital were 70.4％ ( 174/247 ),64.8％ ( 160/247 ),57.4％ ( 142/247),respectively,the resistant rates were lower to amikacin,gentamicin,imipenem and meropenem,which were 3.5％ ( 8/229 ),5.4％ ( 13/241 ),5.9％ ( 14/237 ),8.1％ ( 20/247 ),respectively.PCR experiment showed that the expression levels of the AmpC gene in 4 strains were higher than that of the negative reference strains.The expression levels were 98.3,102.3,121.5,87.3 times compared to the negative reference strains,respectively.Twelve strains (strain no.2,3,5,6,9,10,14,15,16,17,18 and 19)produce bothblaCTX-MandblaKPC-2 enzymes.Highly detecedhlaCTX-Mof Serratia marcescens in our hospital included CTX-M1,CTX-M2,CTX-M9.Isolates no.7 and 18 were carrying blaSHV gene,Isolates no.8 and 13 were carrying blaSME,Isolates no.11 and 20 were carrying blaTEM.There were 5 strains (no.3,4,5,7 and 16) lose the outer membrane protein (OMP) genes ompC and ompF.Two strains( no.1 and 12 ) lose OMP gene ompF only,and one strain ( no.20 ) was lose OMP gene ompC only.ConclusionsThe cause of β-lactam antibiotics resistance of Serratia marcescens was complicated,and the most important mechanism is producing β-1actams and loss of OMP.Understanding the evolution and drug resistant mechanisms will help for best use of antibiotics and reducing the selection of antibiotics to resistant isolates.

Autophagy is a crucial biological process of eukaryotes, which is involved in cell growth, survival and energy metabolism, while the premise of the autophagy function is activated autophagic flux. It has been confirmed that impaired autophagic flux promotes pathogenesis of many chronic inflammatory diseases, especially cancer, neurodegenerative disease and tissue fibrosis, therefore the analysis of autophagic flux state is important for revealing autophagy function and the mechanism of autophagy related diseases. Given that autophagy is a dynamic process with multiple steps, it is very hard to observe the real state of autophagic flux. Summarized here is the novel concept and current approach to detect autophagic flux. This knowledge is crucial for the researching of the biological function of autophagy, and may provide some strategies for developing autophagy-related drug.

Two-factor designs are very commonly used in scientific research. If the two factors have interactions, research designs like the factorial design and the orthogonal design can be adopted; however, these designs usually require many experiments. If the two factors have no interaction or the interaction is not statistically significant on result in theory and in specialty, and the measuring error of experimental data under a certain condition (usually one of the experimental conditions that are formed by the complete combination of the levels of the two factors) is allowed in specialty, researchers can use random block design without repeated experiments, balanced incomplete random block design without repeated experiments, single factor design with a repeatedly measured factor, two-factor design without repeated experiments and two-factor nested design. This article introduces the last two design types by examples.

To evaluate the safety and efficacy of transrectal high-intensity focused ultrasound (HIFU) in the treatment of benign prostatic hyperplasia (BPH), serial studies were conducted in 150 BPH patients before and 30 min, 1, 2, 6 and 12 month(s) after Sonablate-500 HIFU treatment. A silicon-coated indwelling 16F latex catheter was placed during the determination of the therapy zone. Preoperative and postoperative evaluations were made by using the international prostate symptom score (IPSS), quality of life (QOL), uroflowmetric findings and transrectal ultrasound, and incidence of complications. The cystourethrography was done in 23 patients within 1 year postoperatively. The results showed that after HIFU treatment, IPSS and QOL scores were significantly decreased at 1, 2, 6 and 12 month(s) (P<0.01). Maximum urine flow rate (6.0 to 17.2 mL/s, P<0.01), PVR (75.0 to 30.3, P<0.01) and prostatic volume (65.0 to 38.1 mL, P<0.05) were significantly improved 12 months after the operation. Recurrent urinary retention (n=2) and urethrorectal fistula (n=1) occurred at the 15(th) postoperative day. The duration of the HIFU prostate ablation was 25-90 min. The mean time for an indwelling catheter was 3-19 days. These data demonstrate that treatment of BPH with Sonablate-500 HIFU is safe and effective.
Prostatic Hyperplasia/diagnosis ; Prostatic Hyperplasia/*therapy ; Ultrasound, High-Intensity Focused, Transrectal/adverse effects ; Ultrasound, High-Intensity Focused, Transrectal/instrumentation ; Ultrasound, High-Intensity Focused, Transrectal/*methods

Maternal-fetal ABO blood type incompatibility is an autoimmune disease.This disease can lead to miscarriage,stillbirth,children edema,early-onset neonatal jaundice,hemolytic anemia,kemicterus and neonatal death,etc.The author overviewed the etiology and pathogenesis of Matemal-fetal ABO blood type incompatibility,as well as the TCM therapeutic methods and effects in treating this disease at pre-pregnancy period and pregnancy.

Objective To provide reference for expanding the employment positions of graduates in medical infor-mation management and information system of Chongqing Medical University by analyzing their employment situa-tion and trends.Methods The jobs of 171 graduates from Chongqing Medical University Medical Information School in 2008-2013 were registered according to the agreements of graduates, Chongqing Medical University Medical In-formation School, and employing units.Results The employment rate of graduates in medical information manage-ment and information system was relatively high .Their main employing units were medical and health organizations and associated enterprises in the City of Chongqing and Sichuan Province.The employment rate was low in coastal regions.The number of employed graduates in medical and health organizations reached its peak in 2013, and re-mained unchanged in medical and health associated enterprises.Conclusion Although the employment situation of graduates in medical information management and information system is good., the number of employing units and regions is relatively less.The literacy and ability of graduates in medical information management and information system should be improved, their employment channels should be expanded, and their professional education should be further perfected with gradual saturation of demand for graduates in employing units.

How to choose an appropriate design type to arrange research factors and their levels is an important issue in scientific research. Choosing an appropriate design type is directly related to the accuracy, scientificness and credibility of a research result. When facing a practical issue, how can researchers choose the most appropriate experimental design type to arrange an experiment based on the research objective and the practical situation? This article mainly introduces the related contents of the design of one factor with two levels and the design of one factor with k (k≥3) levels by analyzing some examples.

Autophagy is a crucial biological process of eukaryotes, which is involved in cell growth, survival and energy metabolism, while the premise of the autophagy function is activated autophagic flux. It has been confirmed that impaired autophagic flux promotes pathogenesis of many chronic inflammatory diseases, especially cancer, neurodegenerative disease and tissue fibrosis, therefore the analysis of autophagic flux state is important for revealing autophagy function and the mechanism of autophagy related diseases. Given that autophagy is a dynamic process with multiple steps, it is very hard to observe the real state of autophagic flux. Summarized here is the novel concept and current approach to detect autophagic flux. This knowledge is crucial for the researching of the biological function of autophagy, and may provide some strategies for developing autophagy-related drug.
Autophagy ; Fibrosis ; Humans ; Inflammation ; pathology ; Neoplasms ; pathology ; Neurodegenerative Diseases ; pathology

ObjectiveTo determine whether aadA2 gene can be translated from the ATG triplet,which there was no plausible ribosome binding site preceding it,and synthetized a functional protein in class 1 integron.MethodsSite-specific mutagenesis was used to construct aadA2 gene cassette with different start codons,together with their upstreamed promoters of variable regions were cloned into plasmid pACYC184 respective.The constructed plasmids were then transfored into Escherichia coli JM109,Western blot was used to detect the translation products of aadA2 gene with different start codons.Broth microdilution method was used to detect the minimal inhibitory concentrations to streptomycin in Escherichia coli JM109 containing aadA2 gene with different start codons.ResultsaadA2 gene can initiate translation from both ATG and GTG triplets in aminoacyl -3-adenylyltransferase protein synthesis,though there was no plausible ribosome binding site preceding the ATG triplet.Besides GTG and ATG triplets,there was other start codon downstream of the GTG triplet in aadA2 gene.The translated products that initiated from the start codons that described above were all functional AAD(3) proteins that can be detected by anti- aminoacyl -3-adenylyltransferase polyclonal antisera in Western blot and conferred different resistance levels to streptomycin in E.coli.ConclusionWhen inserted as the first gene cassette in class 1 integron,aadA2 gene can initiate translation from ATG triplet and synthetized a functional protein,though there was no plausible ribosome binding site preceding it.This structural characterization of class 1 integron can initiate translation of the open reading frame harbored in gene cassette that integrated into class 1 integron,though there was no plausible RBS preceding the start codon.This make class 1 integron be more convenience to express the genes that capture from environment.

ObjectiveTo prepare antiserum specific to aminoacy1-3″-adenylyltransferase [ AAD (3″) ],and to explore the application value of the prepared antiserum in detecting the expression levels of aadA2 gene that downstream of 8 different promoters (PcS,PcH2,PcH1,PcW,PcS-P2,PcH2-P2,PcH1P2 and PcW-P2 ) of variable regions in class 1 integron.MethodsaadA2 gene was amplified by polymerase chain reaction(PCR) and cloned into the expression plasmid pET19b.After inducing,the recombined aminoacy1-3″-adenylyltransferase[ AAD(3″)] with His-tag was expressed,purified and immunized rabbits to get anti- AAD(3″) specific serum.The prepared antiserum was used to detect the translation levels of aadA2 gene that downstream of different promoters of variable regions in class 1 integron by Western blotting (WB).Broth microdilution method was used to detect the minimal inhibitory concentrations (MIC) to streptomycin in Escherichia coli JM109 with aadA2 gene downstream of different promoters of variable regions.ResultsRecombined AAD (3″) expression plasmid pET19b-aadA2 was constructed successfully and was verified by sequence analysis.After transformed into E.coli BL21 ( DE3 ),a resoluble recombined AAD(3″) high expression strain was obtained.After fermentation,recombined AAD(3″) was purified and immunized rabbits.The anti- AAD(3″) specific serum was obtained with titer ＞ 1∶100 000.WB was used to detect the expression levels of AAD (3″),the translation product of aadA2 gene,that downstream of 8 different promoters of variable regions.The relative expression level of AAD (3″) that downstream of PcW was assumed to be 1,then the relative expression levels of AAD(3″),which all were detected 3 times independently,that downstream of PcS,PcH2,PcH1,PcS-P2,PcH2-P2,PcH1-P2 and PcW-P2 were 12.9±2.3,9.1±1.0,2.0±0.4,16.0±1.3,14.1 ±1.3,10.5±0.7 and 8.9 ±1.7 respective.Very different expression levels of AAD (3″) that downstream of different promoters of variable regions were obtained( F =32.421,P ＜ 0.01 ).The mean values of MIC,which all were detected 3 times independently,to streptomycin in E.coli JM109 with aadA2 gene downstream of PcS,PcH2,PcH1,PcW,PcS-P2,PcH2P2,PcH1-P2 and PcW-P2 were 256,256,64,128,32,128,4 and 64 mg/L respective.These results indicated the different expression levels of aadA2 gene that downstream of different promoters of variable regions can confer their host bacteria different resistance levels to streptomycin.Conclusions Resoluble recombined AAD(3″) is purified successfully and high titer anti- AAD(3″) specific antiserum is obtained from the immunized rabbits.This laid foundation for further investigation on the correlationship between the expressions of intI1 gene and the gene cassettes within variable regions.The expression levels of antibiotic gene cassettes that downstream of different promoters of variable regions are very different,so are the very different antibiotic resistance levels of the host bacteria.Therefore more attentions should be paid to the researches on the classification of promoters of variable regions when molecular epidemiology studies on the class 1 integrons in clinical isolates were conducted.