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Objective:To establish a rapid method to detect the minimum inhibitory concentration (MIC) of imipenem in Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) based on ompK36 gene′s GD mutation. Methods:This was a methodological evaluation study. A total of 258 isolates of Klebsiella pneumoniae were collected from Lishui Municipal Central Hospital from March 2011 to December 2019. Porin gene ompK36 and carbapenemase genes blaKPC, blaNDM, blaIMP and blaOXA-48 were amplified by PCR and confirmed by sequencing. The MIC was detected and confirmed by microbroth dilution susceptibility test, and the corresponding patterns of genotype and MIC were constructed. Based on the patterns, a method for rapid detection of imipenem MIC by real-time fluorescence PCR (RT-PCR) was designed and established. The 159 isolates of non-repetitive Klebsiella pneumoniae collected by Lishui Disease Prevention and Control Center (CDC) from 2017 to 2019 were used for further verification. The sensitivity and specificity were calculated by fourfold table. Kappa test was used to compare the consistency between RT-PCR and microbroth dilution susceptibility test. Results:Among 258 isolates, 109 isolates did not carry carbapenemase gene, 65 isolates carried ompK36 gene GD mutation, 127 isolates carried blaKPC, 15 isolates carried blaNDM, 9 isolates carried blaIMP, and blaOXA-48 was not detected. With mircobroth dilution susceptibility test as the standard, there were 3 corresponding patterns between the drug resistance gene and the imipenem MIC of Kp: when all the 4 carbapenemase genes were negative, MIC≤1 mg/L, the sensitivity was 100% (107/107) and the specificity was 98.4% (125/127); when blaKPC was positive and ompK36 gene GD mutation was negative, 4 mg/L≤MIC≤16 mg/L, the sensitivity was 88.2% (60/68) and the specificity was 98.8% (164/166); when blaKPC and ompK36 gene GD mutation were both positive, MIC≥32 mg/L, the sensitivity was 96.6% (57/59) and the specificity was 96.6% (169/175). RT-PCR detected blaKPC, blaNDM, blaIMP, blaOXA-48 genes accurately.The RT-PCR results of ompK36 gene GD mutation in the KPC-producing isolates were 100% consistent with the sequencing results. In the 159 isolates from Lishui CDC, the sensitivity and specificity of imipenem MIC detected by RT-PCR were higher than 95% in all 3 patterns with mircobroth dilution susceptibility test as the standard, and Kappa value was 0.971. Conclusion:The RT-PCR based on ompK36 gene GD mutation was helpful to quickly determine the MIC range of imipenem in KPC-Kp.
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Extracellular vesicles (EVs) are phospholipid bilayer vesicles actively secreted by cells, that contain a variety of functional nucleic acids, proteins, and lipids, and are important mediums of intercellular communication. Based on their natural properties, EVs can not only retain the pharmacological effects of their source cells but also serve as natural delivery carriers. Among them, plant-derived nanovesicles (PNVs) are characterized as natural disease therapeutics with many advantages such as simplicity, safety, eco-friendliness, low cost, and low toxicity due to their abundant resources, large yield, and low risk of immunogenicity in vivo. This review systematically introduces the biogenesis, isolation methods, physical characterization, and components of PNVs, and describes their administration and cellular uptake as therapeutic agents. We highlight the therapeutic potential of PNVs as therapeutic agents and drug delivery carriers, including anti-inflammatory, anticancer, wound healing, regeneration, and antiaging properties as well as their potential use in the treatment of liver disease and COVID-19. Finally, the toxicity and immunogenicity, the current clinical application, and the possible challenges in the future development of PNVs were analyzed. We expect the functions of PNVs to be further explored to promote clinical translation, thereby facilitating the development of a new framework for the treatment of human diseases.
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Bioelectrical impedance measurement technology is a non-invasive detection technology for extracting human physiological and pathological information. The analysis method of the relationship between bioimpedance and human physiological parameters is an important part of this technology. In order to calculate the internal and external liquid volume of human cells more accurately, based on the Moissl equation for calculating the internal and external fluid volume of human cells, a segmented human bioimpedance spectrum measurement model and an improved calculation method of intracellular and external fluid capacity were proposed. The measurement and calculation experiments of the intracellular and extracellular fluid volume before and after the human body's water intake were designed and compared with the Moissl calculation method. The results show that the improved calculation method can calculate the intracellular and extracellular fluid volumes more effectively, and the relative error is less than 5%, which may provide new ideas or more accurate methods for the analysis of human body components, facilitating the diagnosis and treatment of diseases.
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Humanos , Água Corporal , Impedância Elétrica , Líquido Extracelular , Líquido IntracelularRESUMO
Objective To analyze the drug resistance phenotype and genetic background of New Delhi metallo-β-lactamase (NDM)-producing bacterial strains in Lishui area of Zhejiang province.Methods The imipenem-resistant Enterobacteriaceae strains were isolated from January 2012 to December 2016 in Lishui Municipal Central Hospital of Zhejiang Province.Mrieux Vitek 2 Compact system was used to identified strains and PCR was used to screen for blaNDMgene.Susceptibility was detected by K-B method and MICs were obtained by Vitek 2 with GN13 cards.Plasmids typing was carried out by DNA sequencing of the replication initiator with the transconjugates as templates.The blaNDMgenetic contexts were detected by PCR and complete genome sequencing.Results A total of 102 strains of carbapenem-resistant enterobacteria (CRE), mainly Escherichia coli and Enterobacter cloacae, were isolated, of which 15 were positive for blaNDMwith a positive detection rate of 14.7%.The resistance rate to β-lactam antibiotics was 100%, and the resistance rates to aztreonam, compound sulfamethoxazole , tobramycin and gentamicin were all >80%;and the resistance rate to quinolones was >50%.Among the 15 NDM-producing strains , 12 strains were positive for Hodge test, and 2 strains of Enterobacter cloacae and 1 strain of Escherichia coli were negative. There were 11 strains of blaNDM-1and 4 strains of blaNDM-1.A variety of plasmid types such as IncX 3, IncFIIγ and IncA/C were detected, and 4 blaNDM-5genes were located in the IncX3 plasmid.Four blaNDMsurrounding gene structures were found, of which 8 blaNDM-1genes were located in ISAb125-hyp-blaNDM-1-bleMBL-TrpF-DsbC-IS26, while blaNDM-5was located in IS3000-IS5-blaNDM-5-bleMBL-TrpF-DsbC-IS26 structure, and the former was reported for the first time in China.Conclusion NDM-producing bacterial strains in Lishui area are prevalent at a low level and have high sensitivity to fosfomycin and polymyxin .The blaNDMgene may have multiple sources, but IncX3 plasmid is still the main source for gene transfer.Some new types of blaNDM-1 gene structure have been also found in this study.
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To today of its development,the tenacious vitalities of traditional Chinese medicine (TCM) beyond doubt.From this point of view,it has a positive meaning to analyze and understand TCM,from which to find its advantages and characteristics for better improvement and development of TCM-theory,so that better for the benefit of mankind.Therefore,they were discussed from the perspective of philosophy of science with the conclusion as follows:1.It is a major current Chinese medicine theory Features that the conceptual system of metaphysical nature made the organic combination of syndrome differentiation and treatment,and the Chinese herbal medicines and other therapy as a means of feedback to verify and correct TCM's theory.2.The theory of current Chinese medicine is different from the theory of TCM,because its theoretical deduction and its argument are different from the focus of ancient Chinese medicine.These development and innovation in current Chinese medicine needs to be studied and summarized.3.Chinese medicine and Western medicine should be parallel to the two medical theory systems,both of which enrich the prevention and treatment of diseases.4.As an independent medical theory system,Chinese medicine theory must also need to develop and even revolution.And this process depends on the overall level of development and improvement of science and technology.
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Objective To study the effects of long-term injection of corticosterone (CORT) on the hippocampal astrocytes and synaptic plasticity in mice.Methods Male C57BL/6N mice (n=40) were randomly divided into control group and model group.Mice in the model group were treated with subcutaneous injection of CORT for 4 weeks to generate chronic stress depression model.Chronic stress model was proved to be established by tail suspension test and sucrose preference test and serum level of CORT was determined by radioimmunoassay.Protein expression levels of hippocampal synaptophysin (SYP) and postsynaptic density-95 (PSD-95) were detected by Western blot.Somal volume and protrusion length of glial fibrillary acidic protein-positive astrocytes were assayed by immunocytochemistry and quantitative stereological techniques.Results Immobility time in model group ((137.95±6.22) s) was significantly extended in comparison with that in the control group ((114.05 ± 4.12) s) (P< 0.01).Sucrose preference in the model group (62.42 ± 6.75)% was lower than that in the control group (86.52±5.08)% (P<0.01).CORT levels in model group ((11.48±0.62) ng/ml) was significantly increased compared with that in the control group ((1.11±0.05) ng/ml) (P<0.01).Hippocampal SYP (0.54±0.04) and PSD-95 (0.57±0.07) expression levels in model group were lower than those in the control group (0.99±0.14),(1.03±0.10) (P<0.01).Somal volume in the model group ((132.04±9.23) μm3) and protrusion length ((1.39± 0.05) × 107 μm) was lower than that in the control group ((168.49±9.01)μm3),(1.77±0.10) × 107 μm) (P<0.05,P<0.01).SYP and PSD-95 expression level was found to be positively correlated with the reduction of the protrusion length(r=0.660,P<0.01;r=0.614,P<0.01)in astrocytes.Conclusion The results suggest that depression-like behavior-in-duced by long-term CORT treatment is possibly caused by the alternation of the synaptic plasticity associated with the reduction of the reducing the protrusion length.
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AIM:To study the effect of chronic corticosterone ( CORT) injection on the depression-like behav-iors and the brain glycogen level in mice.METHODS:Male C57BL/6N mice (n=40) were randomly divided into nor-mal control group and model group.The mice in model group were subcutaneously consecutively injected with CORT for 4 weeks.The mouse model of chronic stress depression was constructed.The forced swim test and open field experiment were conducted to prove chronic stress model.The serum level of CORT in the mice was measured by radioimmunoassay.The protein levels of hippocampal synaptophysin ( SYP) and brain-derived neurotrophic factor ( BDNF) were detected by West-ern blot.Hippocampus glycogen, glycogen synthase and glycogen phosphorylase were determined by indirect fluorescence measurement.RESULTS:Compared with normal control group, the immobility time of the forced swim test in model group was significantly lengthened (P<0.01), and the ability of spontaneous activity was reduced (P<0.01), indicating that chronic CORT injection induced depression-like behaviors in mice.The CORT level increased significantly (P<0.01) in model group.CORT injection decreased the protein expression of hippocampal SYP and BDNF (P<0.01), reduced hipp-ocampal glycogen level (P<0.05) and glycogen synthase activity (P<0.05), and increased glycogen phosphorylase ac-tivity (P<0.05).CONCLUSION:Chronic CORT injection causes hippocampal neuron damage and induces the depres-sion-like behaviors of mice, which may be associated with decreasing hippocampal glycogen level by CORT.
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AIM To explore pharmacokinetics of Berberine in YL2000 in normal and febrile rats. METHODS The levels of Berberine in plasma were measured through HPLC and secondary parameters were obtained by fitting the dose time data of Berberine making use of 3P87 programme. RESULTS In normal and febrile rats, the plasma concentration of Berberine was peaked at (3 4?0 3) h vs (0 3?2 1) h( P