Objective:To discuss the bacteria detection before and after disinfection by using central negative pressure suction device terminal socket, reduce cross infection probability in hospitals and provide evidence for disinfection management specification. Methods:one hundred and thirty clinical central terminal devices with negative pressure were disinfected spirally with iodine swabs and among them, Department of respiration 48, Department of cerebral surgery 46, and Pediatrics 36, were sampled and sent to be cultured by clinical laboratory personnel before and after disinfection. The specimens were administered in a sterile blood AGAR plate culture medium, and the bacterial growth conditions were observed to detect the bacteria number as pollution standards. Results: The pollution rates of clinical equipment central pressure suction into the terminal socket after disinfection were: Department of respiration 8.33%; Department of cerebral surgery 6.52%; Pediatrics 5.56%, and the average pollution rate was 7.69%.The detected cases of pathogenic bacteria were gram-negative bacteria pollution, and contrasting the pollution rates before and after disinfection was statistically significant(x2=14.08, P<0.05). Conclusion: Using based iodine swabs disinfection central pressure suction mouth terminal socket can reduce the cross infection between the patients with risk and provide patients with safe medical environment.

Objective To explore the immunophenotype features and folate receptor expression of blasts in patients with myelodysplastic syndromes(MDS).Methods Four-color flow cytometry using conventional and secondary gating strategies was used into analysis the immunophenotype features and folate receptor(FR) expression of blasts and CD+34 cells in bone marrow nucleated cells with MDS.The patients with acute myeloid leukemia-M2(AML-M2) were as positive control.Results with progression of MDS from RA/RAS,RAEB to RAEB-T,using conventional gating strategy,the proportion of CD+34 cells were gradually increased(P＜0.05).Moreover,the expression of HLA-DR,CDll7,CD13,CD33 were also gradually increased and the expression of CDl5 was gradually decreased(P＜0.05).Using secondary gating strategy,the expression of HLA-DR,CD117,CD13,CD33 on blasts were higher than those by conventional gating(P＜0.05).However,there was no significant difference(P＞0.05) in the expression of above mentioned antigens on CD+34 cells among different MDS subtypes.On the other hand,there were no expression of FR on blasts and cD+34 cells with different MDS subtypes.Conclusion With progression of MDS,the antigens of blasts surface change into more immature immunophenotype of medullary system.But these antigens abnormal expression only illustrates the increase of ascendant malignant clone quantity,it can not reflect the nature of the disease.Using flow cytometry technique can not detect whether or not FR expression on the blasts with MDS.

The aim of this study was to construct the lentivirus expression vector of human FLT3-ITD mutation and to screen the AML cells for stable expression of FLT3-ITD.We screened the AML patient with FLT3-ITD mutation by peripheral blood DNA extraction and PCR,and then the ORF of FLT3-ITD was constructed using the method of homologous recombination,which used the BamH Ⅰ and Not Ⅰ double enzyme digestion of the ORF and the FIV vector.Results were confirmed by restriction enzyme identification and PCR.The constructed recombinant vector was co-transfected with VSVG and g/p into HEK293T cells to produce the lentiviral parti cles by transfection reagent.The lentiviral particles were transduced into K562 cells and the infection efficiency was measured by fluorescence microscope.The cells were sorted by the Flow Cytometer.The expression of FLT3 in the stable cell lines was detected by Western blot.There were 42 patients with FLT3-ITD mutation among the 207 patients we collected.We successfully amplified the ORF of FLT3 of the patient with highest mutant/wt ration.Then constructed the lentivirus expression vector FLT3-ITD/FIV and overexpressed it in K562 cell lines.We screened the cells expressed FLT3-ITD/FIV by fluorescence activated cell sorting,and Western Blot results proved the expression of FLT3-ITD.The over-expression 1 entiviral vector of human FLT3-ITD/FIV has been successfully constructed and cell lines for stable expression of FLT3-ITD have been successfully established.This will shed a light on the future study of FLT3 function in AML and provide a new in vitro model for AML.

The treatment of hepatobiliary malignant tumor is characterized by the coexistence of multiple treatment methods and multiple disciplines. In order to evaluate the clinical efficacy of different treatment measures or multiple treatment combinations, and to promote the standardized development of comprehensive treatment patterns for hepatobiliary malignant tumor, the Hepatobiliary Center of the First Affiliated Hospital of Nanjing Medical University constructs the registry and follow-up database in hepatobiliary tumor patients based on the information-based platform of the hospital, which will help guide clinicians to make scientific decisions and improve the level of clinical diagnosis and treatment. This study describes the framework design, function modules, data acquisition process and quality control of the database of hepatobiliary malignant tumor. Based on the observational bidirectional cohort study design, the previous clinical data can be sorted to match the current database, on the other hand, the clinical data can be prospectively collected including basic information, admission evaluation, surgical information and postoperative situation, comprehensive treatment measures, regular reexaminations and long-term follow-up, etc. The data quality control system can be improved by formulating standardized operation procedures, regularly personnel training and full-process data management plans. This database will provide high-quality real-world data for clinicians, researchers, and guideline experts, and then provide high-level medical evidence for the standardized development of comprehensive treatment patterns of hepatobiliary malignancies.

Objective:To evaluate the role of epidermal growth factor (EGF) in repair of lung tissues in mice with acute respiratory distress syndrome (ARDS).Methods:Fifty SPF male C57BL/6 mice, aged 6-8 weeks, weighing 21-23 g, were divided into 5 groups ( n=10 each) using a random number table method: control group (group C), EGF group, LPS+ PBS group, LPS+ EGF group and AG1478+ LPS+ EGF group.PBS 0.1 ml was intraperitoneally injected in group C. EGF 10 μg (0.1 ml) was intraperitoneally injected in group EGF.The equal volume of PBS and EGF 10 μg was intraperitoneally injected at 12 h after tracheal infusion of LPS in group LPS+ PBS and group LPS+ EGF, respectively.EGF receptor (EGFR) antagonist AG1478 1 mg was intraperitoneally injected, 30 min later LPS was tracheally instilled, and 12 h later EGF 10 μg was intraperitoneally injected in group AG1478+ LPS+ EGF.ARDS model was developed by endotracheal instillation of LPS 3 mg/kg.The mice were sacrificed on the 1st and 5th days after development of the model, and lung tissues were obtained for microscopic examination of the pathological changes which were scored after HE staining.Bronchoalveolar lavage was performed on 5th day after development of the model and before sacrifice, and bronchoalveolar lavage fluid (BALF) was collected to detect total protein concentration (by BCA method) and IL-6 and TNF-α concentrations (by enzyme-linked immunosorbent assay). Lung tissues were obtained for determination of the wet/dry lung weight ratio (W/D ratio), expression of lung surfactant associated protein C (SP-C) and proliferating nuclear antigen (PCNA) (by immunofluorescence method), and expression of EGFR, phosphorylated EGFR (p-EGFR), protein kinase B (Akt), and phosphorylated Akt (p-Akt) (by Western blot). Results:Compared with group C, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly increased, the number of cells co-expressing SP-C and PCNA was increased, and p-EGFR/EGFR and p-Akt/Akt ratios were increased in group LPS+ PBS ( P<0.01), and no significant change was found in the indexes mentioned above in group EGF ( P>0.05). Compared with group LPS+ PBS, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly decreased, the number of cells co-expressing SP-C and PCNA was increased, and p-EGFR/EGFR and p-Akt/Akt ratios were increased in group LPS+ EGF ( P<0.01). Compared with group LPS+ EGF, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly increased, the number of cells co-expressing SP-C and PCNA was decreased, and p-EGFR/EGFR and p-Akt/Akt ratios were decreased in group AG1478+ LPS+ EGF ( P<0.01). Conclusions:EGF can promote the repair of lung tissues in mice with ARDS, and the mechanism may be related to activation of EGFR signaling pathway and promotion of proliferation of alveolar epithelial cell type Ⅱ.