Bacteria-blocking effect of surgical gowns with new material / 中国感染控制杂志

Objective To study bacteria-blocking effect of surgical gowns with new material.Methods Semi-quantitative and qualitative testing methods were used to detect bacteria-blocking rates of key sites of surgical gowns(chest and forearm), the detected samples included sample A (composite material, unused), sample B (composite material, after washing 100 times), and sample C (monolayer material, unused).Results In semi-quantitative testing,the average bacteria-blocking rates of three samples were 75.47%, 70.78%, and 73.73% respectively.In qualitative testing,three samples could effectively block the penetration of Staphylococcus aureus under wet condition and Bacillus subtilis var.niger spores under dry condition.Conclusion In semi-quantitative testing, all three kinds of samples had bacteria-blocking effect, and the average bacteria-blocking rate was> 70%;in qualitative testing, three samples all meet requirements of bacteria-blocking effect under wet and dry condition.

Characteristics of domestic master and doctorial papers on rheumatoid arthritis:A visualization analysis / 中华医学图书情报杂志

The external and internal characteristics of domestic master and doctorial papers on rheumatoid arthritis were analyzed by visualization analysis, which revealed the publication of and hotspots and frontiers in domestic master and doctorial papers on rheumatoid arthritis, and can thus provide reference for the development of medical information resources and recent advances in rheumatoid arthritis for the broad masses of scientific workers.

Construction and identification of recombinant adenovirus vectors carrying the human transcription factor PU.1 gene / 医学研究生学报

Objective PU.1 plays a key role in innate immune function in the alveolar macrophage.This study was to con-struct and identify recombinant adenovirus vectors carrying the human transcription factor PU.1 gene. Methods The recombinant shut-tle plasmid was obtained from the PU.1 gene ( SPI1) and eukaryotic expression vector that digested by restriction enzymes and connected by T4 DNA ligase.The target fragment SPI1-IRES-EGFP was amplified by PCR.The product was cloned into the intermediate pDONR221 and then recombined with the adenovirus backbone plasmid pAd/CMV/V5-DEST to form a recombinant adenovirus vector. The recombinant adenovirus vector was linearized by PacI and then transfected into human embryonic kidney (HEK293) cells to obtain the recombinant adenovirus pAD-SPI1-IRES-EGFP, which was then propagated in HEK293 cells, filtered and purified to obtain high-con-centration adenoviruses.The adenovirus titer was determined by TCID 50 assay.The PU.1 gene expression in the HEK293 cells was con-firmed by fluorescence microscopy and real-time qPCR. Results PCR amplification, restriction digestion and sequencing analysis showed the recombinant adenovirus carried the correct PU.1 gene.The final virus titer, calculated by TCID 50, was 8 ×1011 IU/mL. Green fluorescence was observed under the fluorescence microscope. Real-time qPCR confirmed that the expression of PU.1 mRNA was increased by 2189.93 folds. Conclusion The recombinant adenovirus vector carrying the PU.1 gene was constructed and obtained successfully, which could contribute to further studies of the influence of PU.1 overex-pression on the innate defense against Aspergillus fumigatus.

Construction and identification of a recombinant adenoviral vector expressing murine dendritic cell-associ-atedC-type lectin-1 / 医学研究生学报

Objective Dendritic cell-associatedC-type lectin-1 ( Dectin-1) is one of the most important receptors in antifungal innate immune response.This study was to construct a recombinant adenovirus vector expressing themurine Dectin-1gene and acquire a high-concentration adenovirus by amplification and purification. Methods The PCR amplification product CLEC7A-pIRES2-EGFP was cloned into the intermediate vector pDONR221, and then recom-bined with the backbone vector pAD/CMV/V5-DEST to produce a re-combinant plasmid pAD-CLEC7A-pIRE2S -EGFP.The recombinant plasmid was linearized with Pac I and transfected into human embryon-ic kidney ( HEK293) cells to produce recombinant adenovirus pAD-CLEC7Ap-IRES 2-EGFP. The adenovirus was propagated in the HEK293 cells and purified by filtering through the cellulose acetate membrane and concentrating column.Fluorescence microscopy and re-al-time PCR were used to determine the expression of the Dectin-1 gene. Results PCR identification, enzyme digestion, and sequen-cing results manifested theDectin-1 gene in the vector, with the final adenovirus titer of 5×1011 IU/mL.Fluorescence microscopy revealed green fluorescence and real-time PCR assay confirmed that the expression of Dectin-1 was improved by 8677.25 times. Conclusion A relatively high-titer adenovirus expressing Dectin-1 was acquired,which may help to further study the high expression of Dectin-1 in anti-fungal innate immunity in vitro and in vivo.

Construction of microRNA Let-7b-mediated recombinant influenza A (H1N1) virus with mutated PB1 / 医学研究生学报

Objective Vaccination is a most effective method for the prevention of severe diseases caused by pandemic influenza and microRNA ( miRNA) mediated gene silencing has offered a novel approach to the construction of new vaccines.Our study aimed to construct a recombinant influenza A ( H1 N1 ) virus with the PB1 gene that carries the target fragment of miRNA Let-7b. Methods After comparing the sequence of the A/Nanjing/108/2009 H1N1 viral fragments with that of Let-7b, we selected PB1 as the optimal gene sequence, inserted the Let-7b binding target gene into PB1, ligated the modified fragments with pDP 2000, and named the recombinant plasmids pDP-mu-PB1 and pDP-sclb-PB1, respectively.We co-transfected the MDCK and 293T cells with the recombinant and other seven plasmids and injected the supernatant into the allantoic cavity of the chickenembryo for virus propagation, followed by detection of the virus by hemagglutination ( HA) assay and measurement of the viral titer by TCID50 .We amplified the viral cRNA by RT-PCR and identified the viruses by agarose gel electrophoresis and nucleotide sequence analysis. Results PB1 was the optimal sequence ( 83 bp -107bp) for the attenuation of viruses.The HA-titers of miRT-H1N1 and scbl-H1N1 were 1∶32 and 1∶64, and their viral loads were 4.68 ×105 and 7.94 ×104 TCID50/mL, respectively.Nucleotide sequence analysis showed the expected fragment in the rescued virus. Conclusion A recombinant strain vaccine was successfully constructed, which has laid the foundation for fur-ther assessment of virulence.

Clinical effects of recombinant mutant human tumor necrosis factor on the malignant pleural effusion in-duced by lung adenocarcinoma / 医学研究生学报

Objective The study aimed to observe the clinical effects and the adverse reactions of recombinant mutant human tumor necrosis factor-alpha ( rhu-TNF) on the treatment of malignant pleural effusion ( MPE) induced by lung adenocarcinoma . Methods 70 patients with MPE caused by lung adenocarcinoma hospitalized in our department were chosen as the research objects . After conventional drainage of pleural effusion , the patients were divided into two groups according to the weight , respectively receiving intra pleural injection of 2 000 000 units and 3 000 000 units of rhu-TNF, followed by the observation of clinical effects and adverse reac-tions.A further retrospective analysis were made on the effects of dexamethasone injected before operation on clinical effects and ad -verse events. Results The effective rates of 2 000 000 unit group and 3 000 000 unit group were respectively 75.68%and 87.88%(P=0.23), with no statistical difference.The adverse reactions in the group of patients being injected dexamethasone before operation significantly reduced(P=0.021) and the use of dexamethasone had no influence on the efficacy of rhu-TNF ( P=0 .486 ) . Conclusion Rhu-TNF is a safe drug with high efficiency in the treatment of MPE induced by lung adenocarcinoma , and the efficacy and safety of re-peated application in clinic still need more supporting data .

Comparison of susceptibility testing by EUCAST and CLSI broth microdilution methods against Aspergillus isolates / 中国感染与化疗杂志

Objective To compare the results of susceptibility testing by European Committee on Antimicrobial Susceptibility Testing (EUCAST)and Clinical and Laboratory Standards Institute (CLSI)broth microdilution methods against Aspergillus isolates.Methods The susceptibilities of 116 Aspergillus isolates were determined for amphotericin B, voriconazole, itraconazole,caspofungin and micafungin according to EUCAST (E.DEF 9.1 )and CLSI (M38-A2)methods.The essential agreement (EA),categorical agreement (CA),very major errors (VME)and major errors (ME)of the two methods were compared.Results The EA was 96.3%-100% between the two methods.The CA ,ME,and VME were 98.8%,0-1.2% and 0 respectively for the susceptibility of Aspergillus fumigatus to voriconazole.The CA,ME and VME was 1 00%,0 and 0 respectively for the susceptibility of Aspergillus fumigatus and Aspergillus niger to amphotericin B,or the susceptibility of Aspergillus fumigatus and Aspergillus flavus to itraconazole.Conclusions The results of susceptibility testing by EUCAST and CLSI broth microdilution methods are well consistent against Aspergillus isolates.

Invasive pulmonary aspergillosis: 20 cases / 中国感染与化疗杂志

Objective To analyze the clinical, radiological, pathological and microbiological features of invasive pulmonary aspergillosis (IPA) to improve clinical management.Methods Retrospective analysis of 20 pathologically and/or microbiologically confirmed IPA cases in our hospital from January 2005 to August 2008. Results Group A (with underlying diseases) included 13 patients (underlying malignancy in 9 patients, including 5 cases of hematological malignancy, COPD in 2 patients, pulmonary tuberculosis and bronchiectasis in 1 each). Group B (without underlying disease) included 7 patients (2 patients with a long time of fluffy toy contact, another 1 had exposure to moldy rice, and 3 had exposure to polluted water). All these 20 patients had pulmonary invasion revealed by CT imaging. Multiple changes were identified in 16 patients. Bilateral pulmonary infiltrates and/or consolidation were revealed in 7 patients. Multiple nodules were seen in 9 patients. Four patients had solitary lesions, including isolated nodules in 2 patients and segment consolidation in the other 2 patients. Pulmonary cavity without fluid level was found in 8 patients (40.0%). Eighteen cases received antifungal therapy. The overall efficacy rate was 55.6%. The efficacy rate in group A and B was 45.5% and 5/7 respectively. The average time to symptomatic relief was (12.0±2.8) days. The time to lung lesion improvement on CT was (17.4±2.9) days. The time to significant CT improvement was (34.3±9.9) days. The time to the resolution of active lesion was (56.4±6.2) days.Conclusions IPA may occur in immunocompetent patients without underlying disease. Most IPA patients have bilateral multiple pulmonary nodules and cavities on CT. The time to the resolution of active pulmonary lesions is about 6 weeks.