Objective To investigate the distribution and drug resistance of major pathogens for urinary tract infections in the First Affiliated Hospital of Nanjing Medical University.Methods Strains from midstream urine culture of patients with urinary tract infections were collected during January 1 and December 31,2011.All strains were identified by API system,and disk diffusion method was used for drug sensitivity test.Results Totally 1129 strains were isolated,in which 667 (59.1％ ) were Gram-negative strains,266 (23.5％) were Gram-positive strains,and 196 (17.4) were Candida.Among Gram-negative strains,Escherichia coli,Klebsiella pneumoniae and Proteus mirabilis were highly sensitive to carbapenem antibiotics; while Acinetobacter baumannii and Pseudomonas aeruginosa were highly resistant to most antibiotics including cephalosporins and penicillinase inhibitor,and the resistance rates were over 50％.Among Gram-positive strains,the major strains Enterococcus avium and Enterococcusfaecalis were completely sensitive to vancomycin and teicoplanin,and highly sensitive to linezolid (resistance rate below 10％ ).Candida albicans and Candida glabrata were highly resistant to voriconazole and fluconazole (with the resistance rates of 47.2％ - 60.0％ ), but were completely sensitive to amphotericin and nystatin.Conclusion Gram-negative strains account for most urinary tract infections in the First Affiliated Hospital of Nanjing Medical University with high drug resistance rates.

ObjectiveTo investigate the serum concentration of 25-hydroxy vitamin D [25(OH)D] in the second-trimester women in winter and explore its correlation with age and blood hemoglobin level. Methods The blood samples of 78 second-trimester women were collected during the 24-28 gestational weeks. Serum 25 (OH) D and blood hemoglobin levels were measured. The correlations of serum 25 (OH) D with age and blood hemoglobin levels were analyzed by Pearson correlation. ResultsOf the 78 pregnant women, the rates of vitamin D deficiency ( ≤25.0 nmol/L), insufficiency [25.0 nmol/L ＜ 25(OH) D≤50.0 nmol/L], and sufficiency were 65.38％, 30.77％, and 3.85％, respectively. Serum concentration of 25 (OH) D was positively correlated with blood hemoglobin level ( r =0.2746, P =0.015 ). ConclusionVitamin D deficiency or insufficiency is common among the second-trimester women in winter, especially among those with low hemoglobin level.

Objective To establish a semi-quantitative method for measurement of HCV RNA by use of primers and probe, which was sensitive and designed by ourselves, a new europium fluorescent chelate BHHCT. Methods 44 serum samples of HCV infected patients and 20 samples of the healthy people were collected. HCV RNA in serum sample was extracted by HCV fluorescence PCR diagnostic kit produced by Zhongshan University DAAN Gene Co Ltd, and amplified by RT-PCR in which one PCR primer was pre-labeled with biotin. The amplified products were hybridized with capture probes pre-fixed on the microplate. The biotin in the amplified products was conjugated with europium labeled streptavidin. So europium was linked to the target DNA. Then the fluorescent of europium was measured. Results The linear range of this assay was 10 - 10 copies/ml. Both sensitivity and specificity were 100%. Conclusion Europium labeled RT-PCR assay is a sensitive, specific, fast and non-radioactive contaminant method for the measurement of HCV RNA.

n positive blood culture, and they are resistant to a variety of antimicrobial agents, which should be called attention.

BACKGROUND:Many in vivo and in vitro experiments indicate that hypoxic co-cultures promote stem cells differentiate into chondrocytes.
OBJECTIVE:To evaluate the influence of hypoxia on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes.
METHODS:Adipose-derived stem cells and articular chondrocytes were mixed at the ratio of 3:1, then the mixed cells were seeded onto poly(lactic-co-glycolic acid)-gelatin scaffold at the ultimate concentration of 5.0×1010/L. The cells were cultured in normoxia (20%O 2 ) and hypoxic (5%O 2 ) conditions for 6 weeks. After culture, hematoxylin and eosin staining was performed for histological structure analysis, and alcian blue staining was used to evaluate glycosaminoglycan synthesis. Type II col agen expression was detected by immunohistochemistry staining. The content of DNA, glycosaminoglycan and hydroxyproline in the scaffold-cellcomplex was measured.
RESULTS AND CONCLUSION:In the hypoxia group, hematoxylin-eosin staining showed the formation of massive cells and extracellular matrix;alcian blue staining showed massive glycosaminoglycan formation;immunohistochemistry staining detected strongly positive expression of col agen type II, the content of DNA, glycosaminoglycan and hydroxyproline was higher than the normoxia group. Hypoxia promotes in vitro chondrogenic differentiation of co-cultured adipose-derived stem cells and articular chondrocytes. .

Objective To investigate the resistance status of different integrons of Shigella sonnei (S.sonnei) and to analyze the distribution of resistant genes in integrons in Jiangsu Province.Methods A total of 32 strains of S.sonnei isolated from six cities of Jiangsu Province in 2011 were collected.The antibiotic susceptibility was tested by disk diffusion method.The molecular homology was analyzed by pulsed field gel electrophoresis (PFGE).The detection and classification of integrons were achieved by analyzing the positive polymerase chain reaction (PCR) products using restriction fragment length polymorphism (RFLP).RFLP and DNA sequencing were used to analyze the resistance genes in integrons.Results Multi-drug resistance (MDR) was detected in 28 (87.5％) S.sonnei strains.The resistant rates to ampicillin,nalidixic acid and tetracycline were highest (87.5％,respectively).However,it was sensitive to norfloxacin.PFGE analysis showed that there were 3 kinds of homologous clones involving 31 strains of the 32 S.sonnei strains.Among them,2,5 and 24 strains had the same clones,respectively.Accordingly,they spread within one,two and five different cities.The detection rates of class 1,class 2 and the atypical class 1 integrons in S.sonnei were 62.5％ (20/32),81.3％ (26/32) and 21.9％ (7/32),respectively,and no class 3 integron was detected.Sequence analysis of class 1 integron variable area revealed that it contained multiple resistant genes (aacA4-cmlA1 and dfrA1-aadA 1) ; dfrA1-sat 1-aadA 1 from class 2 integron and blara-30-aadA 1 from atypical class 1 integron were also identified.Conclusions In 2011,homologous S.sonnei strains spread among different cities in Jiangsu Province.MDR strains are prevalent and integrons are widespread which mediated the emergence of MDR strains.

Objective To establish a proper internal quality control(QC) system for the automated hematology analyzer accord‐ing to the corresponding stipulation by ISO15189 :2012 .Methods The high ,normal and low levels of controls offered by the instru‐ment were used to detect 22 items in whole blood cells routine analysis ,draw the Levey‐Jennings chart ,analyze and evaluate the quality control data for each month or each batch .Results A proper QC system was established ,including quality goals setting , new QC target value and standard deviation calculating and the QC rules choosing .In setting the target value for new batch of con‐trol ,MCV ,HCT and MCHC should adopt the mean values of at least 20 values ,while other items could adopt the mean values of 10 values ;the difference of CV values for some items existed in different QC levels ,different brands or different instruments in the same brand .Therefore ,the quality goals and standard deviation for new batch should be set according to different levels and differ‐ent instruments .Due to the QC characteristics and existence of QC chart drift phenomenon ,therefore MCV ,HCT and MCHC should adopt the 13 s and 22 s rules .For the other items ,the 13 s ,22 s and 10x rules were adoted .Conclusion Under the corresponding stipulations of ISO15189 and by combining with the laboratory practical situation ,a set of applicable internal quality control system is established .

[ABSTRACT]AIM:TostudytheeffectofsmallinterferingRNA(siRNA)ontheexpressionofbeta2-microglo-bulin (β2M) in pre-differentiated bone marrow mesenchymal stem cells (BMSCs).METHODS: The β2M siRNA was transfected into the pre-differentiated BMSCs with Lipofectamine 2000.BMSCs were divided into transfection group , blank control group and negative control group .The expression of β2 M at mRNA and protein levels was determined by real-time qPCR, Western blotting and laser confocal microscopy .The productions of aggrecan and type II collagen in pre-differentia-ted BMSCs were determined by toluidine blue staining and type Ⅱcollagen immunofluorescence .RESULTS:The results of real-time qPCR, Western blotting and laser confocal microscopy showed that siRNA successfully inhibited the expression ofβ2 M at mRNA and protein levels in the pre-differentiated BMSCs .The results of toluidine blue and type Ⅱcollagen im-munofluorescence staining showed that siRNA does not affect the productions of aggrecan and type Ⅱ collagen in the pre-differentiated BMSCs .CONCLUSION:siRNA targeting β2 M reduces the expression of β2 M in the pre-differentiated BM-SCs and does not affect the chondrocyte characteristics of pre -differentiated BMSCs .

[ABSTRACT]AIM:Toinvestigatetheinhibitoryeffectsofresveratrolonchondrosarcomaandtherelationwith mitochondrial and PI3K/Akt pathways.METHODS:Chondrosarcoma SW1353 cells were treated with resveratrol at con-centrations of 25, 50 and 100 μmol/L for the time intervals of 24 h, 48 h and 72 h.The viability and apoptosis of the SW1353 cells in the presence or absence of resveratrol were analyzed by CCK 8 assay and Hoechst 33258 staining , respec-tively.The protein levels of Bcl-2, Bax, activated caspase-3, Akt and p-Akt were detected by Western blotting .The cell migration ability was determined by wound scratch assay .RESULTS:Exposure of the cells to resveratrol resulted in a de-crease in the cell viability in a dose-and time-dependent manner (P<0.05).visible nuclei with apoptotic characteristics in resveratrol group were observed .The protein levels of activated caspase-3 and Bax were increased , and Bcl-2 and p-Akt were decreased compared with control group .The total Akt were not significantly changed .Resveratrol also significantly re-duced the migration of tumor cells .CONCLUSION:Resveratrol induces apoptosis of chondrosarcoma , which plays a role of part through mitochondrial and PI 3K/Akt signaling pathways .

Objective To quantifying the urine human cytomegalovirus(HCMV)DNA from the HCMV infection infants and its corresponding liver function indications,and investigate the relationship between their concentrations.Methods The u-rine samples were collected from HCMV infection infants.HCMV DNA was measured by fluorescence quantitative polymer-ase chain reaction (FQ-PCR).Serum ALT,AST,ALP,GGT,T-Bil and D-Bil liver function indications were detected and the positive rate was analyzed,simultaneously.The correlation between the logarithm urine HCMV DNA (log HCMV DNA) concentration and ALT,AST,ALP,GGT,T-Bil and D-Bil were analyzed by Spearman correlation analysis.Results The dis-tribution range ofurine log HCMV DNA in 444 HCMV infection infants was <2.70~7.90;the positive rate of serum ALT, AST,ALP,GGT,T-Bil and D-Bil were 24.8%,59.0%,95.7%,31.1%,16.7% and 16.3%,respectively.The urine log HC-MV DNA was associated with GGT and the correlation coefficient was 0.099 (P < 0.05),but no associated with ALT, AST,ALP,T-Bil and D-Bil.Conclusion The positive rate of liver function indications will rise in HCMV infection infants, the urine log HCMV DNA was associated with GGT,but not associated with other liver function indications.