OBJECTIVE To investigate the distribution and the drug resistance of pathogens in infective cases and to provide a foundation for reasonable application of antibacterials.METHODS The information of all bacterial cultures and susceptibility tests from the inpatients in the year of 2006 was analyzed.RESULTS A total of 336 strains of pathogens were isolated,and the pathogens causing lower respiratory tract infection rated the top(60.7%).The resistance rate of Acinetobacter baumannii was the highest in Gram-negative bacilli.It was 33.3% to imipenem,and more than 66.6% towards the other antibacterials.All Gram-positive cocci were sensitive to vancomycin.The resistance rate of Enterococcus and Staphylococcus aureus was more than 75% to the most antibacterials.CONCLUSIONS Inspecting pathogens and strengthening susceptibility tests are very important in reducing drug abuse,decreasing the resistance rate and raising the cure rate in hospital.

Objective To investigate the appropriate setting up of normal reference ranges of lymphocyte subsets in some flow cytometry laboratories and to study the effects of different flow cytometers and various reagents by different manufacturers on the analysis of peripheral blood lymphocyte subsets. Methods Three FCM labs (named A, B and C) in Beijing region were selected representing 3 commonly used flow cytometers (Beckman Coulter Epics XL, Beckman Coulter Cytomics FC500, BD FACS Calibur). 50 samples from healthy donors were distributed to 3 labs and tested according to individual lab's standard operating procedure to verify whether the normal reference ranges of peripheral blood lymphocyte subsets established were appropriate. The application of internal quality control was also investigated. Commercial blood quality control reagents were given to the 3 FCM labs and tested within 20 working days paralleled with routine samples. In addition, 20 patients' samples were prepared using 4 different combinations of reagents ( a , b , c and d). The results from combination a, which used the Beckman Coulter reagents and instrument, were compared to the results from combination b, c and d, which used reagents from different manufacturers. Then the prepared samples were tested on Beckman Coulter Epics XL to evaluate the effects of different combinations of reagents on the results of peripheral blood lymphocyte subsets analyzed by the same instrument. Furthermore, 24 patients' samples prepared by same reagents from Beckman Coulter company were tested on both Beckman Coulter Epics XL and BD FACS Calibur respectively to assess the effects of different instruments on peripheral blood lymphocyte subsets. 20 patients' samples prepared by same reagents and instruments were analyzed by Beckman Coulter Epics XL analytic system and BD FACS Calibur analytic system respectively to assess the effects of the two analytic systems on the lymphocyte subsets. Results Over 10% of the results for NK and T4/T8 in lab A as well as T4 in labs B and C fell outside of their normal reference ranges. The probabilities exceeding corresponding normal reference ranges were 16% ( 9/50 ), 24% ( 12/50 ), 22% (11/50) and 12% ( 6/50 ), respectively. The results using internal blood quality control in 3 FCM labs within 20 working days were all within the reference ranges of the quality control provided by the kit. The biases from b and c reagent combinations were substantial compared with that of reagent a combination. Among the biases from b and c reagent combinations, the lowest probability of bias exceeding 10% was T8 of combination b, which had probability of 70% (14/20). The highest probabilities of hias exceeding 10% were T3 and T4 of b and c reagent combinations, which reached 100% (20/20) . Furthermore, the biases of T3, T8 and B of d reagent combination compared with that of reagent a combination were also substantial. The probabilities of bias exceeding 10% were 35% (7/20) ,85% (17/20) and 75% (15/20), respectively. Comparing the results of samples prepared and analyzed by reagents and instruments from different manufacturers to that of samples prepared and analyzed by the same company's reagents and instruments showed that there were great discrepancies in T3, T4 , T8 , B and NK. The probabilities of bias exceeding 10% were 71% ( 17/24), 80% (19/24) ,38% (9/24), 33% (8/24) and 92% (22/24), respectively. The biases of T8, NK and B were substantial when compared the results from Beckman Coulter Epics XL analytic systems and BD FACS Calibur analytic systems. The probabilities of bias exceeding 10% were 55% (11/20 ), 70% ( 14/20 ) and 55% (11/20), respectively. Conclusions FCM labs should set up their own normal reference range for peripheral blood lymphocyte subsets. The normal reference range should be verified periodically. It is important to apply internal blood quality control regularly and accumulate the quality control results. The reagents and instrument for preparing peripheral blood samples should be from the same manufacturers.

Objective To observe the changes of the depressive behavior and amygdaloid nucleus nerve growth factor(NGF)expression in estro?gen?deprived female rats and explore the possible mechanism and targets of estradiol in depression treatment. Methods A total of 30 adult SD fe?male rats were randomly assigned to 3 groups:sham operation group(SHAM,n=10);ovariectomized group(OVX,n=10)and ovariectomized rats treated with estradiol group(OVX+E,n=10). The behavior changes were observed by tail suspension test(TST)and sucrose preference test (SPT) after 8?week estradiol treatment. Subsequently ,immunohistochemical staining detect NGF expression in amygdaloid nucleus. Results Compared with the SHAM group rats,sucrose preference ratio significantly decreased in SPT(P<0.01),immobile time prolonged in TST(P<0.01),serum estradiol level and amygdaloid NGF expression significantly decreased(all P<0.01). 8?week estradiol treatment ameliorated depres?sion?like behavior and increased serum E2 level and NGF expression in amygdaloid nucleus in OVX+E group rats when compared with the OVX group(all P<0.01). Conclusion Estradiol treatment can improve the depressive behavior of ovariectomized rats ,which may be related to the in?crease of serum estradiol level and the expression of NGF in amygdaloid nucleus.

Objective To study the effects of glycosaminoglycan extracted from Pinctada martensii on the proliferation, differentiation and mineralization of newborn rat calvarium osteoblast in vitro. Methods MTT, PNPP and ARS methods were used to measure the cell proliferation, activity of ALP and the function of mineral nodes formation of osteoblasts cultured in vitro. Results Glycosaminoglycan at concentration of 0.008～0.5g?L -1 inhibited mildly the proliferation of osteoblast cells, however, this range of concentrations of glycosaminoglycan markedly increased the ALP activity and stimulated mineral node bone formation in osteoblast. Conclusion Glycosaminoglycan extracted from the Pinctada martensii showed stimulating effects on the differentiation and mineralization of newborn rat calvarium osteoblast in vitro, but the stimulating effect on cells proliferation was not observed.

Objective To investigate the resistance rates of the Candida albicans strains isolated from patients with vulvovaginal candidiasis to 5 antifungal agents and examine the mechanism of azole resistance in these strains.Methods A total of 1 646 C.albicans strains were collected in Obstetrics and Gynecology Hospital of Fudan University from January to December 2015.The resistance rates of these isolates to five antifungal agents were analyzed.Azole-resistant (n=30),dose dependent sensitive (S-DD) (n=13),and susceptible isolates (n=10) were randomly selected from the microbiology laboratories of three obstetrics and gynecology hospitals in Shanghai.The expression levels of drug efflux pump related gene CDR1,CDR2,MDR1 and drug target enzyme gene ERG11 were analyzed by real-time fluorescence quantitative polymerase chain reaction (PCR).At the same time,the ERG11 and ERG3 genes were amplified by PCR and sequenced,and analyzed for resistance-related mutations.Results Of the 1 646 C.albicans strains,5.2％,3.2％,2.5％ and 2.1％ were resistant to itraconazole,voriconazole,fluconazole and 5-fluorocytosine,respectively.All isolates were sensitive to amphotericin B.The expression of ERG11 gene was significantly higher in S-DD group and azole-resistant group than in azole-sensitive group (P＜0.05).The expression of CDR1,CDR2 and MDR1 did not show significant difference among the three groups.There were 13 missense mutations in the ERG11 gene,of which T123I,P98S and Y286D amino acid substitutions were newly discovered.Both T123I and Y132H were identified in 26 resistant isolates,of which 16 gene mutation was detected in two pan-azole-resistant isolates.Conclusions The C.albicans strains isolated from vulvovaginal candidiasis showed higher resistance rates to azole antifumgal agents than that to 5-fluorocytosine and amphotericin B.Mutation and over-expression ofERG11 gene may be one of the prevalent molecular mechanisms underlying azole resistance in C.albicans.were pan-azole-resistant.In addition,the ERG3 heterozygous

Objective To study the effects of lactate peritoneal dialysis solution(L-PDS) with different concentrations on apoptosis of human peritoneal mesothelial cells(HPMC),the expressions of bcl-2,bax and activity of caspase-3.Methods HPMC were separated using enzyme digestion and cultivated stably in vitro.After HPMC were co-cultivated with different concentrations(1.50%,2.50%,4.25%) L-PDS,flow cytometry was used to test the apoptosis of HPMC,RT-PCR was used to observe the expressions of bcl-2 and bax,fluorometric method was used to detect the activity of caspase-3.Results Compared with control group, L-PDS could induce the apoptosis of HPMC,especially in high concentration.With the increasing of L-PDS concentration,the expression of bcl-2 mRNA decreased,the expression of bax mRNA increased,the activity of caspase3 raised.There were significant differences of the indexes mentioned above between 4.25%,2.50% L-PDS groups and control group(P0.05).Conclusion L-PDS could induce HPMC apoptosis,which may be executed by alternating of the expressions of bcl-2,bax and activating of caspase3.

ObjectiveTo explore the application value of acoustic radiation force impulse(ARFI) in the differential diagnosis of breast lesions.Methods Eighty six patients with 107 lesions were performed ARFI elastography.The stiffness of the masses in virtual touch tissue image(VTI) were scored.The pathological diagnosis as the gold standard,drawed the ROC curve to find the cut off point of area ratio to predict breast cancer.Shear wave velocity(SWV) within the breast masses were measured using virtual touch tissue quantification(VTQ) technique.ResultsThe elastography hardness score higher than grade 4 was mainly distributed in the malignancies compared to benign lesions( P ＜0.01).1.53 as the cut-off point of area ratio,sensitivity and specificity of diagnosing breast cancer were 95.7 ％,92.5 ％.SWV of malignant group higher than benign group,the difference was statistically significant (P ＜ 0.001 ).Conclusions Hardness score in VTI imaging,area ratio of benign and malignant lesions,SWV within the breast masses can help the differential diagnosis of breast lesions.

Objective To investigate the genotype of carbapenem-resistant Acinetobacter baumannii(CRAB) acquired from intensive care units (ICUs) environment in Shanghai hospitals and to evaluate the gene homology of CRAB isolated from patients and environment in ICUs in Shanghai hospital.Methods Sixty-one CRAB isolated from ICUs environment in Shanghai hospitals and 14 strains isolated from ICUs patients in S hospital were typed by REP-PCR based DiversiLab system.Results There were 7 types of the CRAB isolated from ICUs environment in Shanghai hospitals.Type G1 was the dominant type,accounting for 75.4％ of all CRAB,mainly in 13 hospitals in Shanghai.The percentage of the 7 types CRAB strains were 75.4％,13.1％,3.3％,1.6％,1.6％,3.3％ and 1.6％,respectively.Except for 1 strain of type G6 isolated in D hospital,type G1 CRAB was identified in every hospital which was isolated CRAB.There were 25 strains isolated from patients and environment in S hospital ICUs,which were divided into 4 types.Type G1 was the dominant type mainly distributed in SICU and CICU,including 9 from patients and 8 from environment.The distribution diagram of environmental and patients' strains indicated that the patients in adjacent bed isolated G1 CRAB in SICU and a patient in CICU and its surrounding environment both isolated G1 CRAB.Conclusion Type G1 is the dominant type in ICUs environment in Shanghai hospitals.There are inter-hospital transmission of CRAB in ICUs environment and highly homology of CRAB isolated from patients and environment in ICUs in Shanghai hospitals.Active surveillance of infection and colonization by CRAB,together with implementation of enhanced control strategies should be taken to eradicate CRAB in ICU environment.

Objective To evaluate the biological variations of 8 lymphocyte subsets using flow cytometric double-platform method.Methods Twenty healthy adults were recruited from Peking Union Medical College Hospital in September 2013.At 8:00 AM,12:00 PM,and 4:00 PM on days 1,3,and 5,venous blood was collected from the volunteers.The percent and absolute lymphocyte subset counts were measured using duel-platform method.The sample collection and handling techniques were standardized.Before each batch analysis,the instrument quality controls were performed using the same lots of reagents.The intra-individual coefficient of variation (CVI) and inter-individual coefficient of variation (CVG) were calculated by nested ANOVA with SPSS 13.0 software.The analytical coefficient of variation (CVA),index of individuality (Ⅱ) and reference change value (RCV) were calculated by Excel2003.A mean pairwise comparison was determined by one-way ANOVA and variance analysis.The values between groups were analyzed by independent sample t test.Results For T cells (CD3 +),helper T cells (CD3 + CD4 + CD8-),suppressor T cells (CD3 + CD4-CD8 +) and B cells (CD3-CD19 +),the intra-individual coefficient of variation (CVI) and inter-individual coefficient of variation (CVG) were 0.03,0.06,0.05,0.14 and 0.12,0.16,0.23,0.31 respectively,which were all similar to those in previous studies.However,the Ⅱ and RCV of the four lymphocyte subsets were very different from those in previous studies,which were 0.26,0.40,0.22,0.44 and 8.77,16.86,14.93,39.69,respectively.Moreover,variations in absolute count,CVI,CVG,and analysis coefficient of variation (CVA) of all 8 lymphocyte subsets were greater than those of relative count.Variations in the percent and absolute counts for the CD3 + CD4-CD8-,CD3 + CD4 + CD8 +,and CD3+ CD16+ CD56+ cell subsets were relatively high.The CVI,CVG and CVA for the cells of CD3 + CD4-CD8-were 0.12,0.49 and 0.16.The CVI,CVG and CVA for the cells of CD3+ CD4+ CD8+ were 0.40,0.93 and 0.55.The CVI,CVG and CVA for the cells of CD3 + CD16 + CD56 + were 0.28,1.11 and 0.16.Conclusions Investigation on the CVI,CVG and CVA may allow us to obtain Ⅱ and RCV,by which we can determine the utility of traditional population based reference ranges.Documentation of the RCV indices may be used as objective delta-check values in quality management and decide whether clinical significance existed in the continuously detected results.

Objective To investigate the vestibular functions ,especially the semicircular canal function in children with secretory otitis media (SOM ) .Methods A total of 24 children with SOM of median age of 8 .5 years had effusion that lasted about 3 to18 months ,compared with 12 children without effusion in the middle ear with the median age of 7 .9 years .The two groups received pure tone audiometry ,acoustic immitance and semicircular canal functions ,including the saccadic test ,the tracking test ,the gaze test and spontaneous nystagmus test .Results The 24 children with SOM had conductive hearing loss ,including 11 mild cases and 13 moderate case .The 12 children without SOM had the normal hearing .There were significant differences in semicircular canal functions between the two groups (8/24 & 0/12 , P<0 .05) .Age ,genders ,hearing -loss degrees and vertigo were not the significant factors which affected the semicircular canal function .Conclusion The function of semicircular canal was affected in children with SOM ,but the mechanisms were still not clear .