1.Optimization of the in vitro culture system for chicken small intestinal organoids.
Jing LI ; Liya WANG ; Dingyun MA ; Senyang LI ; Juanfeng LI ; Qingda MENG ; Junqiang LI ; Fuchun JIAN
Chinese Journal of Biotechnology 2024;40(12):4645-4659
In order to establish a stable in vitro culture platform for chicken small intestine three-dimensional (3D) organoids, in this study, crypt cells were collected from the small intestine of 18-day-old embryos of AA broilers. On the basis of the L-WRN conditioned medium, we optimized the culture conditions of chicken small intestinal organoids by adjusting the proportions of nicotinamide, N-acetylcysteine, LY2157299, CHIR99021, Jagged-1, FGF, and other cytokines to select the medium suitable for the long-term stable growth of the organoids. The optimization results showed that the addition of 1.5 µmol/L CHIR99021 significantly improved the organoid formation efficiency and organoid diameter. When 0.5 µmol/L Jagged-1 was added, a small amount of bud-like tissue appeared in organoids. After the addition of 50 ng/mL FGF-2, the rate of organoid germination was significantly increased. The 1.5 µmol/L CHIR99021, 0.5 µmol/L Jagged-1, and 50 ng/mL FGF-2 added in the medium can cooperate with each other to improve the formation and speed up the proliferation and differentiation of organoids, while improving the stemness maintenance of cells. The morphology, cell types, and culture characteristics of chicken small intestinal organoids were studied by HE staining, transmission electron microscopy, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), indirect immunofluorescence, and immunohistochemistry. The results showed that the 3D organoids of the chicken small intestine cultured in vitro were morphologically consistent with the chicken intestinal tissue and contained differentiated epithelial cells. In summary, we successfully established an in vitro culture system for chicken small intestinal organoids, providing a new method for the subsequent research on chicken intestinal physiology, pathology, and host-pathogen interaction mechanism and the development of relevant drugs.
Animals
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Organoids/metabolism*
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Intestine, Small/drug effects*
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Chickens
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Cell Culture Techniques/methods*
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Culture Media
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Chick Embryo
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Tissue Culture Techniques/methods*
2.Bladder microenvironment actuated proteomotors with ammonia amplification for enhanced cancer treatment.
Hao TIAN ; Juanfeng OU ; Yong WANG ; Jia SUN ; Junbin GAO ; Yicheng YE ; Ruotian ZHANG ; Bin CHEN ; Fei WANG ; Weichang HUANG ; Huaan LI ; Lu LIU ; Chuxiao SHAO ; Zhili XU ; Fei PENG ; Yingfeng TU
Acta Pharmaceutica Sinica B 2023;13(9):3862-3875
Enzyme-driven micro/nanomotors consuming in situ chemical fuels have attracted lots of attention for biomedical applications. However, motor systems composed by organism-derived organics that maximize the therapeutic efficacy of enzymatic products remain challenging. Herein, swimming proteomotors based on biocompatible urease and human serum albumin are constructed for enhanced antitumor therapy via active motion and ammonia amplification. By decomposing urea into carbon dioxide and ammonia, the designed proteomotors are endowed with self-propulsive capability, which leads to improved internalization and enhanced penetration in vitro. As a glutamine synthetase inhibitor, the loaded l-methionine sulfoximine further prevents the conversion of toxic ammonia into non-toxic glutamine in both tumor and stromal cells, resulting in local ammonia amplification. After intravesical instillation, the proteomotors achieve longer bladder retention and thus significantly inhibit the growth of orthotopic bladder tumor in vivo without adverse effects. We envision that the as-developed swimming proteomotors with amplification of the product toxicity may be a potential platform for active cancer treatment.
3.Compassion fatigue of medical staff working in centralized quarantine sites and its influencing factors during COVID-19 epidemic
Shuai LIU ; Lei ZHANG ; Rong LI ; Qing ZHANG ; Zuhong ZHU ; Juanfeng GUO
Chinese Journal of General Practitioners 2022;21(1):59-65
Objective:To investigate the compassion fatigue of medical staff working in centralized quarantine sites and its influencing factors during the COVID-19 epidemic.Methods:From September 2020 to October 2020, a cross-sectional study was conducted on 213 medical staff working in 13 quarantine centers in two districts of Shanghai. A self-made general information questionnaire, professional quality of life scale, general self-efficacy scale and perceived social support scale were used in the study.Results:The score of empathy satisfaction was 33.00(28.25, 37.75), and there were 141 cases (66.20%) with supercritical value. The score of job burnout was 26.00(21.50, 30.50), and 96 cases (45.07%) exceeded the critical value. The score of secondary traumatic stress was 25.00(20.50, 29.50), and 192 cases (90.14%) exceeded the critical value. Multivariate logistic regression analysis showed that average daily working hours, job satisfaction, self-evaluation of work pressure, general self-efficacy and perceived social support were the influencing factors of compassion fatigue among medical staff. Compared with those who worked for 0 h in the polluted area, those who worked for more than 8 h had a higher degree of compassion fatigue ( OR=7.787, 95 %CI: 1.127-53.820);compared with those who were more satisfied with their work, those who were less satisfied with their work had a higher degree of compassion fatigue ( OR=6.209,95 %CI:1.474-26.157); compared with those with no or low stress, those with high stress had higher degree of compassion fatigue ( OR=2.567, 95 %CI: 1.228-5.366); compared with those with high self-confidence, those with low self-confidence or moderate self-confidence had higher degree of compassion fatigue ( OR=13.519, 95 %CI: 3.394-53.848; OR=3.462, 95 %CI: 1.038-11.548); compared with those with high perceived social support, those with low perceived social support had higher degree of compassion fatigue ( OR=2.071, 95 %CI: 1.027-4.175) (all P<0.05). Conclusion:The status of compassion fatigue of medical staff working in the centralized quarantine sites need to be improved. We should pay more attention to the those medical staff having long daily working hours in the polluted area, those with low job satisfaction, poor self-evaluation of work pressure, poor general self-efficacy and low perceived social support.
4.Regulation of vascular function by nitric oxide-related S-nitrosylation.
Jing LI ; Juan FENG ; Xian WANG
Acta Physiologica Sinica 2017;69(5):557-570
Nitric oxide (NO), well known as the endothelium-derived relaxing factor (EDRF), has become increasingly recognized as an important determinant of cardiovascular diseases. A molecular pathway by which NO accomplishes various biochemical processes is the cGMP-independent post-translational modification of protein, S-nitrosylation. Here we review the biochemistry and regulating mechanisms of NO-related S-nitrosylation of proteins, and discuss the multiple roles of S-nitrosylation in vascular functions and pathology in particular.

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