Objective: To understand the medical staff' cognition of medicalethics.Method: Descriptive study and various analysis were performed. Results: Four aspects including the understanding of hospital Ethic Review Committee, the status of medical ethics education, the understanding of medical ethics in clinic and research, the requirement of medical ethics training were surveyed among the medical staff in five hospitals. Conclusion: The influence of medical ethics in clinical practice and medical research were greatly improved. The hospital Ethic Review Committee should be strengthened and a lot of works was needed for medical ethics education.

Objective To compare the clinical application effect of bi -level positive airway pressure ( BiPAP) and nasal continuous positive airway pressure ( CPAP) in the treatment of preterm infants with neonatal respiratory distress syndrome ( NRDS) .Methods 64 neonates with NRDS were divided into the BiPAP group ( n=35) and NCPAP group (n=29) according to the randomized controlled study method.The blood gas index before and after 2,12 hours of treatment,the total time for ventilation,length of stay,and the incidence rate of abdominal disten-sion,gastroesophageal reflux,bronchopulmonary dysplasia,intraventricular hemorrhage,ventilator-associated pneumo-nia in the two groups were compared.Results The levels of pH and PaO2 in the BiPAP group were higher than those in the NCPAP group in noninvasive respiratory support for 2,12h(t=2.391,2.556,2403,2.355,P=0.020,0.013, 0.019,0.022).The incidence rates of hypoxemia,apnea,re-intubation mechanical ventilation rate after 12h in the BiPAP group were lower than those in the NCPAP group(t=5.049,4.988,4.215,P=0.025,0.026,0.040).The length of stay and mechanical ventilation time in the BiPAP group were shorter than those in the NCPAP group( t=-2.096,-2.669,P=0.041,0.010).Conclusion Compared with NCPAP,early application of BiPAP can obviously reduce the invasive respiratory support rate of children with NRDS with intubation.It is worthy of promotion.

OBJECTIVE To understand flora distribution and four antifungal drugs′in vitro antifungal activity of deep fungi in nosocomial infection in order to provide help to clinics.METHODS Fungi were cultured and isolated by routine procedure which identified by VITEK microbe automatic system.Drug susceptibility test used Rosco paper disk diffusion and broth dilution method with NCCLS M27-A.RESULTS Totally 156 strains with 9 species of deep fungi that main fungi were Candida albicans,and C.tropicalis with 57.69%,and 31.41%,respectively,were isolated from nosocomial infection.The major isolating rates of clinical infection specimens were from respiratory,cardiovascular surgery,and neurological departments with 26.28%,12.18%,and 9.62%,respectively.The main infection specimens were from respiratory tract and urinary tract with 71.15% and 16.67%,respectively.Drug resistance rates to fluconazole,amphotericin B,itraconazole,and ketoconazole with Rosco paper disk diffusion were 23.08%,2.56%,12.18%,and 17.36%,MIC90 were 64.0,2.0,8.0,and 16.0mg/L,respectively.CONCLUSIONS The main deep fungi are C.albicans and C.tropicalis.Antifungal activity of amphotericin B is the highest than others.The drug resistance rate to fluconazole is more and more higher.Clinics should use antifungal drug rationally in accordance with drug susceptibility test results.

OBJECTIVE To enhance the positive rate of the blood culture in order to make pathogenic diagnosis actually and quickly and conducting usage of antimicrobial agents in clinic.To value the clinical applied circumstance of BacT/Alert 3D automated blood culture system.METHODS The 3728 blood cultures were detected by BacT/Alert 3D automated blood culture system,and bacterial susceptibility test was conducted on all isolates using Kirby-Bauer methods with CLSI standards.To statistically analyze the examined time,the positive rate and variety and drug resistance for all kinds of pathogens.RESULTS The blood culture positive rate was 6.0% in the 3728 blood cultures with 222 strains of bacteria.The false positive rate was 0.2% and the false negative rate was 0.4%.The positive rate of blood culture was 0.89% in 12 hours,2.01% in 18 hours,and 3.51% in 24 hours.Among all the 222 isolates,29.7% were Enterobacteriaceae,the drug resistant rates to amikacin,ceftazidime,ciprofloxacin,and imipenem were 13.6%,36.3%,42.4%,and 3.0%,respectively;20.3% were Staphylococcus,the drug resistant rates to erythromycin,levofloxacin,cefoxitin,and vancomycin were 75.6%,33.3%,68.9%,and 0,respectively;11.7% were Enterococcus,the drug resistant rates to errythromycin,levofloxacin,fosfomycin,and vancomycin were 96.2%,80.8%,23.1%,and 0,respectively;11.3% were non-fermented bacilli,the drug resistant rates to amikacin,ceftazidime,ciprofloxacin,and imipenem were 32.0%,52.0%,32.0%,and 32.0%,respectively;12.6% were fungi.CONCLUSIONS The pathogens in the blood specimens are more wider in distribution and more higher in drug resistance rates than before.BacT/Alert 3D automated blood culture system can be an important tool for the blood culture,it can provide the diagnostic help quickly for the clinic and increase the positive rates in blood culture.It can not only shorten the check-up time,but also be more quick and more exact.

OBJECTIVE To understand the distribution or change and drug resistance of the bacteria flora in nosocomial infection,in order to provide laboratory evidence for controlling nosocomial infection and to indicate clinical antimicrobial agents usage.METHODS All isolates were identified by routine procedure and VITEK microbe automatic system and API identified system.Drug susceptibility test was used K-B paper disk diffusion method in accordance with the CLSI/NCCLS standards.RESULTS There were 11 464 strains of bacteria which were obtained from 2002 to 2006.Sputum,secretion and middle urine were the major specimens.The major bacteria floras were Pseudomonas aeruginosa,Escherichia coli,Acinetobacter,Enterococcus,Staphylococcus and Candida albicans,whose isolating rates were 57.02% in 2002,64.46% in 2003,57.83% in 2004,57.49% in 2005,and 60.73% in 2006.E.coli and Klebsiella pneumoniae produced ESBLs rates were from 39.87% to 61.98% that drug resistance rates to cefoperazone/sulbactam and imipenem were 14.06% and 0.64%.The drug resistance rates of nonferment Gram-negtive bacteria to cefoperazone/sulbactam were below 33.22%,Gram-positive cocci to vancomycin was below 0.34%.CONCLUSIONS Now,the important bacteria flora is nonferment Gram-negative bacteria in nosocomial infection.The P.aeruginosa,E.coli,Acinetobacter baumannii,Enterococcus faecalis,C.albicans,and meticillin-resistant staphylococcus(MRS) were prevalent important bacteria in nosocomial infection.The situation of producing ESBLs in Enterobacteriaceae is very severe.Glycopeptides are the best choice to MRS.To zymogenic bacteria and the nonferment Gram-negtive bacteria that cause severe infection,combining-drug treatment can be chosen according to drug susceptibility test results.

BACKGROUND: The transplantation of allogeneic cartilage has local immunological rejection, and it is necessary to further reduce the rejection to promote its application in clinic, thus it is significant to perform a series of experiments to induce local immune privilege.OBJECTIVE: To observe the in vivo growth of tissue engineered allogeneic cartilage reconstructed by chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL.DESIGN: A randomized controlled observation.SETTING: Shanghai Jiao Tong University.MATERIALS: Thirty-six allogeneic New Zealand rabbits as recipients and 45 1-week-old chinchillas as donors, either sex,were purchased by the experimental animal center of Chinese Academy of Sciences. Amphotropic recombinant retrovirus coated cell line PT67 was purchased from Clontech Company; Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), G418 and Polybrene were bought from GIBCO BRL.METHODS: The experiment was carried out in original Shanghai Second Medical University from January 2000 to July 2005. The New Zealand rabbits were randomly divided into three groups: FasL-transfected group (n =12), untransfected group (n =12) and blank control group (n =12). The rabbit allogeneic cartilages were constructed by the compound of pLNCX2-FasL transfected chondrocytes and tissue engineered material of pluronic F-127. ① Gross observation and mass changes of the grafts: Corresponding materials were infused subcutaneously, the grafts were removed at 1, 2 and 3 months after transplantation for gross observation and the mass changes. ② Staining observation: The grafts were removed at 1, 2 and 3 months after transplantation, then prepared into sections, and observed by hematoxylin and eosin (HE), Safranin'O and Masson's trichrome stainings. ③ Antibody detection: Blood samples (1 mL) were collected at 1 and 2 months after transplantation, the chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, and separated by electrophoresis in agarose medium, then acted with serum of recipient to observe whether corresponding antibody generated. ④ Complement dependent cytotoxicity (CDC) test: The chondrocytes of chinchillas were prepared into cell suspension (2×109/L), and then seeded into 96-well plate, attached grew for 24 hours, then recipient serum was added for the CDC test, and the percentage of apoptotic cells was counted under microscope.MAIN OUTCOME MEASURES: ① Gross observation and mass changes of the grafts;② Histological changes; ③ Results of the antibody detection; ④ Percentage of apoptotic cells.RESULTS: All the 81 rabbits were involved in the analysis of results. ① Gross observation and mass changes of the grafts: Two weeks after inoculation, there were obvious nod formations at the inoculated sites, but no nod formed in the blank control group. The new cartilage tissues became smaller gradually and completely disappeared at 4 months in the untransfected group, whereas those in the FasL-transfected group became smaller, but still existed after 7 months. The masses of grafts in the FasL-transfected group were higher than those in the untransfected group (P ＜ 0.05). ②Histological observation: Plenty of lymphocytic infiltrations around cartilage tissue could be observed in the untransfected group, and obviously decreased in the FasL-transfected group. No lymphocyte was observed inside the chondrocytes.Masson's trichrome staining was performed, and it was observed under light microscope that the small white parts in the middle were immature chondrocytes, and there were green collagen around most of the mature chondrocytes. Safranin O staining showed strong positive reaction, suggested that there were rich glycosaminoglycan in matrix. ③ Antibody detection: The chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, then acted with serum of recipient, and the results showed that no corresponding antibody generated. ④ Percentage of apoptotic cells: The percentages of serum CDC apoptotic cells in the FasL can ransfected group, untransfected group and blank control group were 5%, 6% and 1%, which were all negative.CONCLUSION: Rabbit allogeneic chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL can reconstruct tissue engineered cartilage, and can postpone the degeneration by 3 months.

BACKGROUND: At present,enhanced green fluorescent protein(EGFP)is proved to be the best labeled molecule,with unique advantages,such as high fluorescence specificity,easy to be detected,and so on.Recombined retroviral vector EGFP-pLNCX2,which can stably express EGFP,can be construct using gene-engineering technique.Transfecting allogenic chondrocytes is indeed very useful to investigate the target gene expression and process of constructing tissue engineered cartilage in vivo.OBJECTIVE:To construct recombined retroviral vector EGFP-pLNCX2 which can stably express EGFP,and investigate optimal conditions for retrovirus transfection of chondrocytes.DESIGN:Randomized controlled observation.SETTING:Shanghai Jiao Tong University.MATERIALS:Thirty New Zealand rabbits of either gender,1 week of age,were purchased from Experimental Animal Center of Chinese Academy of Sciences.Amphotropic retrovirus package cell line PT67 pLNCX2 and pEGEP-C1 were purchased from Clontech Company;NIH 3T3 cell line was purchased from ATCC Company;DH5a Bacterium coli was preserved by the laboratory of Shanghai Jiao Tong University;Retroviral vector pLNCX2 was purchased from Clontech Company;pEGFP-C1 plasmid with EGFP were donated by professor Cong Xiao-qian from Chinese Academy of Sciences.METHODS:This experiment was carried out in the Shanghai Jiao Tong University in August 2005.Chondrocytes of New Zealand rabbits were isolated and cultured.The recombinant retroviral vector EGFP-pLNCX2,which can stably express EGFP,was constructed to transfect cultured chondrocytes from New Zealand rabbits by using gene engineering technique.Transfection results were observed under fluorescence microscope.Altogether 6×105 chondrocytes incubated in 10 cm-diameter flat plate were used to transfect retrovirus-EGFP immediately and at 12,24 and 48 hours after inoculation.One week later,EGFP expression efficiency was measured with flow cytometer,and best occasion for retrovirus transfecting primary chondrocytes.Enzyme-digested chondrocytes were inoculated for 24 hours to transfect retrovirus.After 250 mg/L G418 was added,chondrocytes were screened on the 2nd,3rd,4th,5th and 6th days separately.After phosphate buffer solution(PBS)washings,the transfection efficiency of chondrocytes was detected by flow cytometer,and the best occasion for G418 screening was observed.MAIN OUTCOME MEASURES:①EGFP-pLNCX2 transfection efficiency.②The best occasions for retrovirus transfecting primary chondrocytes and G418 screening.RESULTS: ①Retroviral vector EGFP-pLNCX2 transfected primary chondrocytes of rabbits,and high expression of transfected chondrocytes could be obtained through preliminary screening of G418.After being screened and expressing EGFP,chondrocytes kept normal morphology,with pseudopod adhering to the wall and matrix secreting vigorously.②The best occasion for retrovirus transfecting primary chondrocytes was at 24 hours after cell inoculation.The transfection efficiency determined with flow cytometer was 19.14% on the 5th day.The best occasion for G418 screening was on the 5th day after culture.Transfection efficiency of G418 screening was 55.75% on the 7th day.CONCLUSION:Recombinant retroviral vector EGFP-pLNCX2 can effectively transfect chondrocytes.The best occasion for retrovirus transfecting primary chondrocytes is at 24 hours after inoculation, and the best occasion for G418 screening is on the 5th day after culture.

OBJECTIVE:To prepare the gene recombined human interferon-alpha poly(lactic acid-co-glycolic)microspheres and study its physicochemical properties.METHODS:Interferon-? microspheres were prepared by W/O/W solvent extraction technique;the in vitro release of microspheres were examined;the rudimental organic solvent was determined with GC-MS.RESULTS:The IFN-?-MS were regular in their morphology,and the particle size distribution was narrow.The average diameter was 45.54?m.The encapsulation rate and the loading efficiency were 83.49% and 8.03%,and the in vitro accumulative release rate was 80.32% in 30 days.The in vitro interferon-? release from the microspheres was best described using Higuchi diffusion model.The rudimental of dichloromethane was less than 0.05%.CONCLUSIONS:The prepared IFN-?-MS is very good in all aspects of its physicochemical properties.

Objective To investigate antifungal activities of AMB, ICZ, VRC, CBF against 72 strains of filamentous fungi in vitro. Methods Based on CLSI M38-P and M38-A scheme, MIC of antifungal drugs were determined. The growing inhibitory concentration of 100%, 100%,≥80%, for AMB, VRC ,ICZ act as respective MIC. For caspofungin, the minimal effective concentration (MEC) was determined as the lowest drug concentration showing morphology change of filaments. The fractional inhibitory concentration (FIC) was used to evaluate the effect of combination therapy. FIC was calculated by the following equation: FIC = MICcombination/MICA drug alone+ MICcombination/MICB drug alone. Results MIC90 of AMB, ICZ, CBF, VRC against 72 isolates of filamentous fungi were 8 μg/ml, 4 μg/ml, 2 μg/ml, 8 μg/ml, respectively. MICs range of combined AMB + ICZ, AMB + VRC, ICZ + VRC were 0. 125-16. 97, 0. 2452-1.25, and 0.0625-8. 25 μg/ml respectively. The percent of synergistic interaction of AMB + VRC against filamentous fungi (20.0%-88.9% ) was higher than those of AMB + ICZ ( 10.0% -62.5% ) and ICZ + VRC ( 20.0% - 44.4% ) ( P=0.007 <0.05 ). Conclusions The antifungal activities of four kinds antifungal drugs against 72 strains of filamentous fungi vary in vitro. The therapy of AMB combined with VRC is maybe better than AMB + ICZ and ICZ + VRC for severe fungi infection.

Exosomes are vesicular bodies secreted by living cells containing proteins and RNA, and play an important role in the process of physiology and pathology.Pancreatic cancer is one of the common gastrointestinal tumor with high morbidity and mortality.As a hotspot in oncology, exosomes have potential values in the research of development, diagnosis and treatment of pancreatic cancer.This article reviewed the role of exosomes in pancreatic cancer.