ABSTRACT
The construction of high-quality public hospitals is the key to the construction of high-quality and efficient medical service system. The authors introduced the implementation of " 1+ 5″ engine-driven high-quality transformation development strategy in Zhejiang Provincial People′s Hospital, that is, adhering to the party building to lead the development of the hospital, and formulating measures to build a high-quality hospital from the five aspects of quality and safety, operational efficiency, brand influence, innovation driven and sustainable development, so as to provide some reference for other hospitals′ high-quality development.
ABSTRACT
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.
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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.
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Objective Cryptococcosis is a potential life-threatening systemic mycosis with a heterogeneous susceptible popu-lation which is classified into three groups according to the current guidelines, including AIDS patients, organ transplantation recipients ( OTR) , and non-HIV-infected and non-transplant hosts ( NHNT) .This study aimed to discuss the influence of basic diseases on the clinical features and prognosis of NHNT cryptococcosis patients. Methods Using a retrospective cohort study design, we retrieved the clinical data about 73 NHNT cryptococcosis patients treated in Jinling Hospital.Based on the presence or absence of immunodefi-ciency or infection-increasing complications, we divided the patients into a basic disease group ( n=35) and a non-basic disease group ( n=38) and analyzed their clinical characteristics, chest radiographic features, laboratory results, and clinical outcomes. Results Compared with the non-basic disease group, the basic disease group showed a significantly higher incidence rate of disseminated disea-ses (62.9%vs 21.1%, P<0.01), more cases of patchy consolidation (47.4%vs 16.7%, P<0.05) and mixed lesion (31.6%vs 3.3%, P<0.05) in chest radiography, and a higher mortality (30.0%vs 5.3%, P=0.016). Conclusion Basic diseases have a great impact on the clinical features and prognosis of NHNT cryptococcosis.NHNT patients with basic diseases are susceptible to dis-seminated diseases with severer clinical symptoms and a higher mortality.
ABSTRACT
Objective To study the effects of linderae on alcoholic-induced acute liver injury in SD rats. Methods Forty SD rats were randomly divided into normal group, model group, biphenyl double ester group,and radix lin-derae group. The levels of serum alanine aminotransferase ( ALT) ,aspartate aminotransferase ( AST) were detected with routine laboratory methods using an autoanalyzer. Histopathological changes were assessed by HE. The activities of superoxidedismutase ( SOD) and the level of malondialdehyde ( MDA) in liver homogenates were measured by spectrophotometry. The expression of interleukin-1β( IL-1β) ,nuclear factor-κB( NF-κB) , and tumor necrosis fac-tor-α ( TNF-α) in the liver were analyzed by immunohistochemistry. Results Compared with the normal group, the levels of serum ALT, AST in rats treated with radix linderae were significantly decreased. The activity of serum and liver tissue SOD was significantly increased. Furthermore, the levels of IL-1β, NF-κB and TNF-αwere signifi-cantly decreased after treatment with radix linderae ( P<0. 05 ) . Conclusion The results of our study indicate that extracts from radix linderae alleviate alcoholic liver injury, in part maybe by improving the SOD activity of liver tis-sue and by increasing the antioxidant capacity. Meanwhile, the ability of linderae to reduce proinflammatory cyto-kine has a direct protective effect on hepatocytes.