1.Investigation and analysis of prevention and nursing of deep vein thrombosis in Department of Orthopedics of China hospitals
Juan CAI ; Liuhua QIN ; Fengli GAO ; Litao HUO ; Songhua ZHAO ; Yu JIA
Chinese Journal of Modern Nursing 2021;27(4):443-446
Objective:To explore the current status of deep vein thrombosis (DVT) prevention and nursing in Department of Orthopedics of China hospitals, and provide a basis for hospitals and related departments to promote systematic DVT prevention and nursing management.Methods:This study was a cross-sectional study, and convenience sampling method was used. In September 2019, the self-designed Domestic Orthopedic Deep Vein Thrombosis Prevention and Nursing Questionnaire was used to investigate the members of the Orthopedic Nursing Professional Committee who signed up for the China Health Promotion Foundation Orthopedic Venous Thromboembolism (VTE) Prevention and Nursing Management Forum. The survey subject came from 125 hospitals in 29 provinces, municipalities and autonomous regions across the country. Respondents completed the electronic questionnaire survey by scanning the quick response code, and 118 valid questionnaires were finally recovered.Results:The proportion of 118 hospitals carrying out in-hospital DVT special training was 82.2% (97/118) . Among the ways for nurses to acquire knowledge, the exchanges outside the hospital, literature review, and international exchange accounted for 70.3% (83/118) , 60.2% (71/118) and 14.4% (17/118) respectively. The proportion of establishing hospital DVT risk early warning information system was 57.6% (68/118) . The hospital-level professional group formulated a complete DVT prevention and nursing system process accounting for 44.1% (52/118) , and the department-level professional group formulated a complete DVT prevention and nursing system process accounting for 36.4% (43/118) . In the comprehensive evaluation of DVT prevention and nursing management, 65.3% (77/118) of hospitals rated it as excellent, and 34.7% (41/118) of hospitals rated it as fair and lacking.Conclusions:The Department of Orthopedics of China hospitals attaches great importance to DVT prevention, but in the construction of DVT prevention and nursing management system for orthopedic patients, the popularization of DVT risk early warning information system, the application of DVT preventive equipment measures, the improvement of DVT prevention and nursing related systems, and the implementation of professional management specifications needs to be further strengthened.
2.Construction of seamless genome editing system for fast-growing Vibrio natriegens.
Fengli WU ; Yanxia LIANG ; Yuanyuan ZHANG ; Yanan HUO ; Qinhong WANG
Chinese Journal of Biotechnology 2020;36(11):2387-2397
Recently, fast-growing Vibrio natriegens, as the great potential chassis, has shown a wide application in synthetic biology. Genome editing is an indispensable tool for genetic modification in synthetic biology. However, genome editing tools with high efficiency and fidelity are still to be developed for V. natriegens synthetic biology. To deal with this problem, the physiological characteristics of 6 V. natriegens strains were evaluated, and CICC 10908 strain with fast and stable growth was selected as the host strain for genome editing study. Then, the natural transformation system of V. natriegens was established and optimized. The efficiencies of optimized natural transformation that integrates antibiotic resistance marker cat-sacB or Kan(R) onto the chromosome of V. natriegens could reach 4×10⁻⁵ and 4×10⁻⁴, respectively. Based on the optimized natural transformation, a double-selection cassette was used to achieve seamless genome editing with high efficiency and fidelity. The positive rates of four different types of genetic manipulation, including gene deletion, complementation, insertion and substitution, were 93.8%, 100%, 95.7% and 100%, respectively. Finally, transformation and elimination of the recombinant plasmid could be easily achieved in V. natriegens. This work provides a seamless genome editing system with high efficiency and fidelity for V. natriegens synthetic biology.
Gene Editing
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Plasmids/genetics*
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Synthetic Biology
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Vibrio/genetics*
3.Directed evolution of tyrosine ammonia-lyase to improve the production of p-coumaric acid in Escherichia coli.
Yanan HUO ; Fengli WU ; Guotian SONG ; Ran TU ; Wujiu CHEN ; Erbing HUA ; Qinhong WANG
Chinese Journal of Biotechnology 2020;36(11):2367-2376
p-coumaric acid is an important natural phenolic compound with a variety of pharmacological activities, and also a precursor for the biosynthesis of many natural compounds. It is widely used in foods, cosmetics and medicines. Compared with the chemical synthesis and plant extraction, microbial production of p-coumaric acid has many advantages, such as energy saving and emission reduction. However, the yield of p-coumaric acid by microbial synthesis is too low to meet the requirements of large-scale industrial production. Here, to further improve p-coumaric acid production, the directed evolution of tyrosine ammonia lyase (TAL) encoded by Rhodotorula glutinis tal gene was conducted, and a high-throughput screening method was established to screen the mutant library for improve the property of TAL. A mutant with a doubled TAL catalytic activity was screened from about 10,000 colonies of the mutant library. There were three mutational amino acid sites in this TAL, namely S9Y, A11N, and E518A. It was further verified by a single point saturation mutation. When S9 was mutated to Y, I or N, or A11 was mutated to N, T or Y, the catalytic activity of TAL increased by more than 1-fold. Through combinatorial mutation of three types of mutations at the S9 and A11, the TAL catalytic activity of S9Y/A11N or S9N/A11Y mutants were significantly higher than that of other mutants. Then, the plasmid containing S9N/A11Y mutant was transformed into CP032, a tyrosine-producing E. coli strain. The engineered strain produced 394.2 mg/L p-coumaric acid, which is 2.2-fold higher than that of the control strain, via shake flask fermentation at 48 h. This work provides a new insight for the biosynthesis study of p-coumaric acid.
Ammonia-Lyases/genetics*
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Escherichia coli/genetics*
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Propionates
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Rhodotorula
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Tyrosine/genetics*
4.Aberrant NF-κB activation in odontoblasts orchestrates inflammatory matrix degradation and mineral resorption.
Fanyuan YU ; Fengli HUO ; Feifei LI ; Yanqin ZUO ; Chenglin WANG ; Ling YE
International Journal of Oral Science 2022;14(1):6-6
Inflammation-associated proteinase functions are key determinants of inflammatory stromal tissues deconstruction. As a specialized inflammatory pathological process, dental internal resorption (IR) includes both soft and hard tissues deconstruction within the dentin-pulp complex, which has been one of the main reasons for inflammatory tooth loss. Mechanisms of inflammatory matrix degradation and tissue resorption in IR are largely unclear. In this study, we used a combination of Cre-loxP reporter, flow cytometry, cell transplantation, and enzyme activities assay to mechanistically investigate the role of regenerative cells, odontoblasts (ODs), in inflammatory mineral resorption and matrices degradation. We report that inflamed ODs have strong capabilities of matrix degradation and tissue resorption. Traditionally, ODs are regarded as hard-tissue regenerative cells; however, our data unexpectedly present ODs as a crucial population that participates in IR-associated tissue deconstruction. Specifically, we uncovered that nuclear factor-kappa b (NF-κB) signaling orchestrated Tumor necrosis factor α (TNF-α)-induced matrix metalloproteinases (Mmps) and Cathepsin K (Ctsk) functions in ODs to enhance matrix degradation and tissue resorption. Furthermore, TNF-α increases Rankl/Opg ratio in ODs via NF-κB signaling by impairing Opg expression but increasing Rankl level, which utterly makes ODs cell line 17IIA11 (A11) become Trap+ and Ctsk+ multinucleated cells to perform resorptive actions. Blocking of NF-κB signaling significantly rescues matrix degradation and resorptive functions of inflamed ODs via repressing vital inflammatory proteinases Mmps and Ctsk. Utterly, via utilizing NF-κB specific small molecule inhibitors we satisfactorily attenuated inflammatory ODs-associated human dental IR in vivo. Our data reveal the underlying mechanisms of inflammatory matrix degradation and resorption via proteinase activities in IR-related pathological conditions.
Humans
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Matrix Metalloproteinases/metabolism*
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Minerals/metabolism*
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NF-kappa B/metabolism*
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Odontoblasts/metabolism*
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Osteoclasts/metabolism*
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RANK Ligand/metabolism*
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Tumor Necrosis Factor-alpha/metabolism*