With the development of the prenatal medicine, the survival rate of the premature has im-proved obviously. However,doctors should focus on the ethics problem whether treat the extreme survival and critical diseased premature is necessary. During the process,the attitude of parents and the medical care personnel impact the premature survival rate. Therefore, the role excellent communication skills and the hospital ethics committee to improve the premature rate and avoid the medical conflict should not be ignored.

Purpose To indicate the expression of nattokinase (NK) in Pichia pastoris , an emulsion was prepared with the purified NK to prepare polyclonal antibody. In order to establish sandwich enzyme-linked immunosorbent assay (ELISA) to assay NK in organism, furthermore to lay the foundation for researching in vivo metabolism and function of NK. Methods The NK gene was cloned into a Pichia pastoris expression vector pHBM905A to construct the recombinant plasmid pPRONK.The recipient cell of Pichia pastoris GS115 was transformed with pPRONK which had been cut by restriction enzyme Sal I , under the induction of methanol. The expressed production is purified by salting out and ultrafiltration membrane. An emulsion was prepared with the purified NK and injected into rabbits to prepare polyclonal antibody. Results NK was expressed and identified by SDS-PAGE.The molecular mass of expressed production is about 27 kD.The fibrin plate assay indicated that the NK protein can cleavage fibrin effectively. ELISA analysis indicated that the polyclonal antibody titer is about 1:8 000. Western blot demonstrated that there was a special strap nearby 27 kD. Conclusion NK was successfully expressed in Pichia pastoris , the production can cleavage fibrin effectively and it had great immunogenicity.

Objective To evaluate the capability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry ( MALDI-TOF MS) for rapid identification of methicillin-resistant Staphylo-coccus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) strains.Methods Twenty-five MRSA and thirty MSSA isolates were collected from the Second Affiliated Hospital of Zhejiang University School of Medicine as the experiment group .Twelve MRSA and twenty-two MSSA clinical strains were isola-ted as the control .All strains were identified by MALDI-TOF MS and the results were further analyzed by ClinProTools 3.0 software.Results Four algorithms including support vector machine (SVM), genetic al-gorithm ( GA) , supervised neural network ( SNN) and quick classifier ( QC) showed similar results in dis-tinguishing MRSA from MSSA isolates .The sensitivity of GA was 100 .0%and the sensitivities of other algo-rithms were all greater than 95.0%.The specificities of GA, SVM and QC were all greater than 90.0%. The areas under the receiver operating characteristic ( ROC) curves of four characteristic peaks at mass-to-charge ratios (m/z) of 3279, 6485, 6555 and 3299 m/z were all greater than 0.9.The virtual gel view showed that the bands generated by MSSA isolates at 3279 , 6485 and 6555 m/z were obviously deeper in color than those generated by MRSA isolates .However , the bands of MSSA isolates at 3299 m/z were appar-ently lighter in color than those of MRSA isolates .83.3%of MRSA and 90.0%of MSSA isolates from the control group were correctly identified by the GA model .Conclusion MALDI-TOF MS could rapidly and accurately identify MRSA from MSSA isolates under the strictly controlled experimental conditions with the advantages of less time-consuming, high sensitivity and high specificity .The accurate identification of MRSA from MSSA isolates could be applied for the prevention and treatment of MRSA infection .

Objective To compare the safety and efficiency of laparoscopic cholecystectomy (LC) + laparoscopic common bile duct exploration (LCBDE) with endoscopic retrograde cholangiopancreatography (ERCP) /endoscopic sphincterotomy (EST) +LC for patients with concomitant gallstones and common bile duct stones.Methods The Pubmed,EMBASE and CNKI were searched for randomized controlled trials published from 1990 to March,2012.Revman 5.1 was used to perform the meta-analysis.The main outcomes were stone clearance rate,treatment morbidity,operative time and length of hospital stay.Results 9 studies with 1021 patients were included into the final analysis.Meta-analysis demonstrated that LC+LCBDE was similar to ERCP/EST+LC in the stone clearance rate (OR 1.55,95％CI 0.95～2.52,P=0.08),treatment morbidity (OR 1.12,95％CI 0.75～1.67,P=0.58),operative time (WMD-54.44,95％CI-107.7～-1.17,P=0.05) and length of hospital stay (WMD-0.22,95％CI-2.6～2.16,P=0.86).The P value of the stone clearance rate was approaching 0.05 with increased number of studies in the analysis.There was no significant difference in the operative time,probably because of small sample size.Conclusions LC-LCBDE was equivalent to ERCP/EST+ LC in stone clearance rate,treatment morbidity,operative time and length of hospital stay.There is a possibility that LC+LCBDE can be superior to ERCP/EST+LC in the stone clearance rate and the operative time with increase in sample size.

Objective To investigate the linezolid resistance mechanisms and molecular epidemiology of clinical isolates of methicillin-resistant coagulase-negative staphylococci (MRCoNS).Methods Seventeen MRCoNS,including 10 S.capitis,4 S.cohnii,2 S.haemolyticus,and 1 S.sciuri with various levels of linezolid resistance were isolated from intensive care units in our hospital from March to August 2011. Minimal inhibitory concentration (MIC) was determined by E-test method. Pulsed-field gel electrophoresis was performed to analyze the molecular epidemiology.PCRs and DNA sequencing were preformed to investigate the mechanisms of linezolid resistance in MRCoNS.Results Nine S.capitis with linezolid MIC of ＞256 μg/ml were indistinguishable,and another S.capitis with linezolid MIC of 4 μg/ml was closely related.Four S.cohnii with linezolid MIC of ＞256 μg/ml were belonged to the same clonal strain.MIC of linezolid for S.sciuri was 64 μg/ml,and were 4 μg/ml and 6 μg/ml for 2 S.haemolyticus,respectively.A commom G2576T mutation and a novel C2104T mutation were identified in 9 S.capitis with linezolid MIC of ＞256 μg/ml by DNA sequence analysis of domain V of the 23S rRNA gene.cfr gene was deteeted in all staphylococci except a S.sciuri whose 23S rRNA gene contained the G2576T mutation.Conclusion It is the first report of linezolid-resistant clinical isolates of staphylococci in China.Linezolid resistance in MRCoNS is related to the presence of DNA mutation in domain V of the 23S rRNA gene and cfr gene.It's a clonally dissemination of linezolid-resistant MRCoNS in intensive care units of our hospital.

While Choledocholithiasis is concomitant with gallstones in approximately 9.2％ ～ 14.3％ of the patients,and its treatment methods are mainly by surgery,including:(1)Open cholecystectomy and open choledocholithotomy 、T-tube drainage (OC-OCHTD) ; (2) Endoscopic retrograde cholangiopancreatography (ERCP) and Endoscopic sphincterotomy (EST),and followed by Laparoscopic cholecystectomy (LC) second stage (ERCP/EST + LC) ; (3) Laparoscopic cholecystectomy combined with laparoscopic common bile duct exploration (LC +CBDE).These three methods all have their own characteristics.The last two methods are minimally invasive surgery treatment.Practice has proved that LCBDE and ERCP + EST + LC treatment methods are feasible and show certain advantages,compared with OC-OCHTD reflecting the development of minimally invasive surgery.Practice has proved that LCBDE and ERCP + EST + LC treatment methods are feasible and show certain advantages,compared with OCHTD reflecting the development of minimally invasive surgery.But the indications of operation are need to be summarized constantly in clinical practice,while operation skills are also need to be improved.At present,many reports of the two methods of minimally invasive treatment have been published because of the controversy,such as surgical indications,hospitalization expense,operation time,therapy danger,complications and the length of hospital stay.Both LCBDE and ERCP + EST + LC show some advantages and disadvantages compared with each other.On proper indications,LCBDE embodies the superiority of minimally invasive surgery technology as a one-stage operation,which is preferred for patients of choledocholithiasis with concomitant gallstones.For patients,no matter what kind of operation method,less invasion,lower risk and less complications is the key for best treatment choise.

Objective To evaluate the effect of three different choices of indwelling urethral catheter clamping. Indwelling urethral catheter clamping choices included according to patients′complaints to clamp the catheter (Group A), released as schedule (Group B),and smooth passway of indwelling urinary catheter all the way (Group C). Methods We searched RCT about the effect of different choices of indwelling urethral catheter clamping from CNKI, VIP database,Wanfang database, PubMed on line. Literatures were searched from the database had been established till now. Two reviewers evaluate the literatures independently according to the RCT authentic assessment manual of Cochrane Collaboration. Other than different results, negotiation took as the measure to attach an agreement. No agreement reached, consult the opinion of the third person. RevMan5.2 was used to analyses the result of the study. Results Statistical difference between three clamping choice on the rate of successful in autonomous urination after catheter removed: both group A and group B preceded group C (Z=2.97, Z=4.34, P < 0.05), mean well group A preceded group B (Z=5.30, P < 0.05). Statistical difference did not found on the rate of urinary tract irritation after catheter removed between three choices. Conclusion Clamping indwelling urethral catheter according to patients′ complaints seems useful in bladder reconditioning before indwelling urethral catheter removal.

Humans are exposed to the ubiquitous radiofrequency (RF, 100 kHz-300 GHz) electromagnetic fields because of the mushroom development of wireless communications,raising concerns over the possible hazards of RF radiations.Epidemiological investigation has showed that chronic use of cellphones increases the risk of brain tumors.Since genetic damage is closely related to tumors, researchers have been trying to find out whether cellphones and other RF devices are genotoxic.However, the investigations have yielded both negative and positive results.This review summarized the recent in vitro and in vivo researches about genotoxicity of RF radiations and proposed a possible mechanism by which of RF radiations cause genetic damage.

Objective To investigate the water infectivity in key water regions,and analyze the transmission risk of schisto-somiasis in suspicious and high-risk environments in Hubei Province in 2016. Methods Schistosome-endemic areas of the Yangtze River,the Hanbei River and the Fu River were chosen as the surveillance and forecast sites. The water infectivity was detected by using the sentinel mice during the flood season. The infection status of residents in the villages around the surveil-lance sites and the activities of human beings and domestic animals were surveyed. The emergency response system was initiated when the water infectivity areas were detected. Results From May to June,among the ten surveillance sites of the first batch, two positive spots with infected mice were found,accounting for 20％. All the recovered mice(totally 200)were dissected:five mice were infected,with a total infection rate of 2.5％. Totally five schistosome adult worms were collected,with mean worm bur-den of 1 worm per infected mouse. The site with cercariae-infected water body started the emergency response and no epidemic occurred. From August to September,among the eight surveillance sites of the second batch,the recovered mice(totally 160) were dissected,and no infected sentinel mice were found. Conclusion The sentinel mice method plays an important role in an-alyzing the schistosomiasis transmission risk in Hubei Province.

Objective To evaluate the effect of four carbapenems combined with EDTA for metallobeta-lactamases (MBLs) detection.Methods Thirty MBLs-producing gram-negative bacteria ( 16 strains of Enterobacteriaceae and 14 strains of Non-fermentative),9 KPC-producing Enterobacteriaceae and 10 OXA-producing Acinetobacter baumannii strains were collected from the Second Affiliated Hospital of Zhejiang University.A double disk screening test with EDTA was performed for MBLs detection,comparing the changes of inhibition zone diameter with or without EDTA.Results When the inhibition zone diameter difference of ≥5 nn as standard,the sensitivity of panipenem (PAN) group was relatively high (66.7％,20/30),followed by meropenem group (MEM) (63.3％,19/30) and imipenem (IPM) group (60.0％,18/30),etrapenem (ETP) group was the wont (43.3％,13/30).When the inhibition zone diameter difference of ≥4 mm as standard,the sensitivity of meropenem and panipenem group was 80.0％ (24/30)respectively.When the inhibition zone diameter difference of ≥ 3 mm as standard,the sensitivity of imipenem and meropenem group was 90.0％ (27/30) respectively,while 83.3％ ( 25/30 ) for etrapenem and panipenem group.Specificity of four groups under these three interpretation was 100％ respectively.Results of screening test for 16 MBLs positive strains showed that the sensitivity of panipenem group was the highest (75.0％,12/16).While using ≥ 5 mm as the MBLs positive interpretation,the sensitivity of panipenem group was the highest (75.0％,12/16).While using ≥4 mm as the MBLs positive interpretation,the sensitivity of panipenem group (93.8％,15/16) was much higher than that of imipenem (75.0％,12/16),meropenem (68.8％,11/16) and ertapenem (68.8％,11/16).Conclusions Detection of MBLs with EDTA is easy to operate,and it shows a low false positive rate against gram-negative bacteria.Ertapenem is not recommended to screen for MBLs,the best method for screening for MBL production in gram-negative bacteria is the MEM-EDTA or PAN-EDTA with a breakpoint of ≥4 mm.As for Enterobacteriaceae,PAN-EDTA with a breakpoint of ≥4 mm is better than the other three carbapenems in screening for MBL production,but panipenem is not recommended for MBLs screening tests for nonfermentative gram-negative bacteria.