1.Role of autophagy in acute kidney injury
International Journal of Pediatrics 2017;44(7):451-455
Acute kidney injury(AKI)is a major kidney disease associated with poor clinical outcomes.The pathogenesis is characterized by tubular cell injury and death.Recent studies have demonstrated autophagy induction in proximal tubular cells during acute kidney injury.In the pharmacological or autophagy-related gene knockout AKI models,autophagy has been demonstrated to reduce the cell death and protect kidney through inhibiting the inflammatory response and removing the injury organelles.However,some studies suggest that autophagy has damage effects on the AKI.In conclusion,illuminating the roles and regulatory mechanism of autophagy in the AKI may provide the important theoretical clue for the therapy and prognosis of AKI.
2.The changes of β-arrestin 2 and microtubule-associated protein light chain 3 in renal tissue of rats with renal ischemia reperfusion injury
Manman CHU ; Yubin WU ; Yue DU
Chinese Pediatric Emergency Medicine 2017;24(2):137-143
Objective To investigate the expression of β-arrestin2 and microtubule-associated pro-tein light chain(LC)3 in renal of rat with acute renal ischemia reperfusion injury,and to analyze the relation-ship between them and renal injury. Methods Fifty-four male SD rat(3-4 weeks old) were randomly divid-ed into three groups:control group,sham group,acute ischemic reperfusion injury group. We established the acute renal ischemia reperfusion injury model through removing the right kidney and clamping the left renal for 45 minutes with noninvasive arterial clip. We obtained the kidney and blood samples respectively at 12 h, 24 h,36 h,48 h,72 h,96 h after the surgery. Expressions ofβ-arrestin2 and LC3 protein were detected by the immunohistochemistry method and Western blot method. The renal function and morphological changes were assessed. Results Compared with control group and sham group,the serum creatinine and kidney pathologi-cal grading of acute ischemia reperfusion injury group obviously rised. The kidney injury was the most serious at the 24 h after acute ischemic reperfusion injury. The expressions of β-arrestin2 and LC3 were little in the control group and sham group. However,the expressions of these two indicators were obviously higher and reached the peak at the 12 h after acute ischemia reperfusion injury. All these results suggested that the chan-ges of these two indicators were anterior to the histopathological changes. The expressions ofβ-arrestin 2 and LC3 protein were in positive correlation with the kidney injury(r=0. 821,P<0. 05;r=0. 913,P<0. 05). Conclusion In the acute renal ischemia-reperfusion injury,β-arrestin2 may be as a kind of upstream regula-tory protein involving in the kidney pathological process through the regulation of the autophagy.
3.A Soil Water Extraction Method Using Accelerated Solvent Extraction Technique for Stable Isotope Analysis
Qingzeng ZHU ; Qing SUN ; Zhiguo SU ; Manman XIE ; Junyong SONG ; Yabing SHAN ; Ning WANG ; Guoqiang CHU
Chinese Journal of Analytical Chemistry 2014;(9):1270-1275
Soil water is one of the most important components in hydrological cycle. The stable hydrogen and oxygen isotopes in soil water have been increasingly used in the ecological, environment and hydrological research. In view of different techniques for extracting soil water, there is significant difference in theδD andδ18 O composition. This paper presents a method for analyzing hydrogen and oxygen isotopes in soil water by using elemental analyzer and isotope ratio mass spectrometry with accelerated solvent extraction for sample pretreatment. The conditions are: extraction solvent: dichloromethane, temperature: 100 ℃, pressure of 10. 3 MPa, static time:10 min. The samples were extracted three times, and with cycle values of four, four and three, respectively. Comparing with the added water, the deuterium and oxygen isotope values in the extracted soil water enrich 2. 12‰-4. 58‰ and 0. 17‰-0. 93‰, respectively. The reproducibility of replicate extractions of soil water is around ±0. 89‰ for δD and ±0. 37‰ for δ18 O.
4.Enterovirus D68 protease 2A affects anti-viral interferon type Ⅰ pathway
Huiwen ZHENG ; Zhiyao YANG ; Zening YANG ; Jie SONG ; Xing HUANG ; Nan LI ; Lisha DING ; Heng LI ; Hongzhe LI ; Lei GUO ; Manman CHU ; Haijing SHI ; Longding LIU
Chinese Journal of Microbiology and Immunology 2019;39(6):401-409
Objective To analyze how enterovirus D68 (EV-D68) protease 2A affects the anti-vi-ral interferon typeⅠ(IFN-Ⅰ) pathway in 293T cells following infection. Methods Western blot was used to detect the expression of recombinant protease 2A, IFN-α and signal transducers and activators of tran-scription 1 (STAT1) at protein level. Expression of EV-D68 viral protein (VP1) and protease 2A was ana-lyzed by immunofluorescence at different time points. Cytopathic effects were recorded to calculate 50% cell culture infective dose ( CCID50 ) . Expression of the genes involved in the anti-viral IFN-Ⅰ pathway was measured by real-time PCR (RT-PCR). Results The recombinant plasmid pCLIPf-2A was successfully constructed and the expression of recombinant protease 2A could be detected by Western blot 24 h after transfection. The recombinant protease 2A promoted the proliferation of EV-D68 at the late stage of infection and induced the production of IFN-α. Expression of the genes involved in the anti-viral IFN-Ⅰ pathway at mRNA level was up- or down-regulated to different degrees with various trends in different groups following infection. Expression of STAT1 was enhanced in all groups. Conclusions EV-D68 protease 2A promoted the activation of anti-viral IFN-Ⅰpathway in response to viral infection and enhanced the proliferation of virus at the late stage of infection.