ISP1-anchored Polarization of GCβ/CDC50A Complex Initiates Malaria Ookinete Gliding Motility

该工作历时四年，由博士毕业生高涵（第一作者）、博士生杨振科和王旭（共同第一作者）、硕士生钱鹏戈和洪仁杰完成；袁晶教授为通讯作者；厦门大学为第一完成单位。 该工作揭示了疟疾病原疟原虫通过媒介按蚊传播过程中的关键步骤---控制动合子运动的环磷酸鸟苷cGMP信号的激活机制。 疟原虫属于顶复体亚门原生动物，每年导致数亿人口感染疟疾和超过40万病人死亡。疟原虫通过雌性按蚊在哺乳动物宿主间传播。疟疾病人被按蚊叮咬吸血，疟原虫雌雄配子在按蚊消化道中受精形成合子，进一步变形发育为具有运动能力的新月形动合子。只有获得运动能力的动合子，才能穿越按蚊消化道单层上皮细胞，成功感染按蚊媒介。在本研究中，通过大量基因修饰模型，发现GCβ缺失导致动合子运动完全丢失，进而失去按蚊感染和传播能力。研究还发现，GCβ由细胞质均匀分布改变为在动合子顶体突出一侧聚集，并且GCβ聚集和动合子成熟完全同步，显示GCβ聚集可能直接激活cGMP信号。 本工作还发现GCβ结合蛋白CDC50A，后者承担分子伴侣功能，在动合子转化和成熟动合子中，稳定GCβ。此外，进一步筛选发现动合子内膜复合物蛋白ISP1，能够结合和锚定GCβ/CDC50A复合物，在成熟动合子中维持复合物的聚集。本研究为深入开展寄生性原生动物的发育转化和信号调控提供了线索和借鉴。【Abstract】Ookinete gliding motility is essential for penetration of the mosquito midgut wall and transmission of malaria parasites. Cyclic guanosine monophosphate (cGMP) signaling has been implicated in ookinete gliding. However, the upstream mechanism of how the parasites activate cGMP signaling and thus initiate ookinete gliding remains unknown. Using real-time imaging to visualize Plasmodium yoelii guanylate cyclase β (GCβ), we show that cytoplasmic GCβ translocates and polarizes to the parasite plasma membrane at “ookinete extrados site” (OES) during zygote to ookinete differentiation. The polarization of enzymatic active GCβ at OES initiates gliding of matured ookinete. Both the P4-ATPase-like domain and guanylate cyclase domain are required for GCβ polarization and ookinete gliding. CDC50A, a co-factor of P4-ATPase, binds to and stabilizes GCβ during ookinete development. Screening of inner membrane complex proteins identifies ISP1 as a key molecule that anchors GCβ/CDC50A complex at the OES of mature ookinetes. This study defines a spatial-temporal mechanism for the initiation of ookinete gliding, where GCβ polarization likely elevates local cGMP levels and activates cGMP-dependent protein kinase signaling.We thank Dr. David Baker (London School of Hygiene and Tropical Medicine) for his comments on this manuscript. This work was supported by the National Natural Science Foundation of China (81522027, 31772443, and 31501912), the Fundamental Research Funds for the Central Universities (20720160069, 20720150165, and 2013121033), the China's 1000 Young Talents Program, the “111” Project of the Ministration of Education of China (B06016), and the Division of Intramural Research, National Institute of Allergy and Infectious Diseases (NIAID), NIH (X.S.). The authors thank Cindy Clark, NIH Library Writing Center, for manuscript editing assistance. 该研究得到国家自然科学基金委、中组部“青年千人计划”和厦门大学校长基金的资助

Exploring the mechanism by which aqueous Gynura divaricata inhibits diabetic foot based on network pharmacology, molecular docking and experimental verification

Abstract Background To predict and validate the potential mechanism by which Gynura divaricata (GD) functions in the treatment of diabetic foot (DF). Methods The main chemical constituents of GD were identified by reviewing the literature, the traditional Chinese medicine database platform (TCMIP) and the BATMAN-TCM platform. DF disease targets were identified with the GeneCards database, and the compound-target network was constructed by using the intersection of drugs and disease. The STRING platform was used to construct the protein–protein interaction (PPI) network, and Cytoscape 3.7.2 software was used to visualize the results. Moreover, the Metascape database was used for Gene Ontology (GO) enrichment analyses and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Molecular docking of the active ingredients of GD and core protein targets of DF was performed using AutoDock software. Finally, the predicted results were preliminarily verified with experiments. Results A total of 140 potential targets of GD were identified and associated with DF. According to the PPI network analysis, GD accelerated DF wound healing, and the mechanism may be related to proteins such as AKT1, TP53, IL6, CASP3, TNF, and VEGFA. GO and KEGG enrichment analyses indicated that GD may play a role in the treatment of diabetic foot by affecting various signaling pathways. Molecular docking results showed that the proteins AKT1, TP53, IL6, CASP3, TNF, and VEGFA were closely associated with the components of GD. The animal experiments showed that GD reduced the levels of IL-6 and TNF-α and increased the mRNA and protein expression of VEGFA in rats with DF. Conclusions GD regulates multiple targets and multiple pathways to promote wound healing in DF

A New Method on Calibrating PSF of Remote Sensing Space Plane Arraycamera

Point spread function (PSF) of an imaging system is an important parameter to a remote sensing imaging system, and it is the important basis for image restoration [1]. The knife-edges method has been widely used to measure PSF because it provides a relatively easy way to get the point spread function. However, the location of the edge points in edge spread function (ESF) is difficult to determine of the knife-edges method, resulting in the inaccuracy of the point spread function. In this paper, we propose a new method to solve the problem of phase influence for calibrating PSF of space plane array camera. This method include: the design of target, the process of calibration and data processing. Then this method was used on calibrating PSF of GF-4 satellite panchromatic camera. The results showed that the new method could determine the PSF correctly

Role of multiple dual-phase 18F-FDG PET/CT metabolic parameters in differentiating adenocarcinomas from squamous cell carcinomas of the lung

Purpose: To evaluate the ability of multiple dual-phase 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) metabolic parameters to distinguish the histological subtypes of non-small cell lung cancer (NSCLC). Methods: Data from 127 patients with non-small cell lung cancer who underwent preoperative dual-phase 18F-FDG PET/CT scanning at the PET-CT center of our hospital from December 2020 to October 2021 were collected, and the metabolic parameters of their primary lesions were measured and analyzed retrospectively. Intraclass correlation coefficients (ICC) were calculated for consistency between readers. Metabolic parameters in the early (SUVpeak, SUVmean, SUVmin, SUVmax, MTV, and TLG) and delayed phases (dpSUVpeak, dpSUVmean, dpSUVmin, dpSUVmax, dpMTV, and dpTLG) were calculated. We drew receiver operating characteristic (ROC) curves to compare the differences in different metabolic parameters between the adenocarcinoma (AC) and squamous cell carcinoma (SCC) groups and evaluated the ability of different metabolic parameters to distinguish AC from SCC. Results: Inter-reader agreement, as assessed by the intraclass correlation coefficient (ICC), was good (ICC = 0.71, 95% CI:0.60–0.79). The mean MTV, SUVmax, TLG, SUVpeak, SUVmean, dpSUVmax, dpTLG, dpSUVpeak, dpSUVmean, and dpSUVmin of the tumors were significantly higher in SCC lesions than in AC lesions (P = 0.049, < 0.001, 0.016, < 0.001, 0.001, < 0.001, 0.018, < 0.001, 0.001, and 0.001, respectively). The diagnostic efficacy of the metabolic parameters in 18F-FDG PET/CT for differentiating adenocarcinoma from squamous cell carcinoma ranged from high to low as follows: SUVpeak (AUC = 0.727), SUVmax (AUC = 0.708), dpSUVmax (AUC = 0.699), dpSUVpeak (AUC = 0.698), TLG (AUC = 0.695), and dpTLG (AUC = 0.692), SUVmean (AUC = 0.690), dpSUVmean (AUC = 0.687), dpSUVmin (AUC = 0.680), SUVmin (AUC = 0.676), and MTV (AUC = 0.657). Conclusions: Squamous cell carcinoma of the lung had higher mean MTV, SUVmax, TLG, SUVpeak, SUVmean, SUVmin, dpSUVpeak, dpSUVmean, dpSUVmin, dpSUVmax, and dpTLG than AC, which can be helpful tools in differentiating between the two. The metabolic parameters of the delayed phase (2 h after injection) 18F-FDG PET/CT did not improve the diagnostic efficacy in distinguishing lung AC from SCC. Conventional dual-phase 18F-FDG PET/CT is not recommended

A New Method on Calibrating PSF of Remote Sensing Space Plane Arraycamera

Point spread function (PSF) of an imaging system is an important parameter to a remote sensing imaging system, and it is the important basis for image restoration [1]. The knife-edges method has been widely used to measure PSF because it provides a relatively easy way to get the point spread function. However, the location of the edge points in edge spread function (ESF) is difficult to determine of the knife-edges method, resulting in the inaccuracy of the point spread function. In this paper, we propose a new method to solve the problem of phase influence for calibrating PSF of space plane array camera. This method include: the design of target, the process of calibration and data processing. Then this method was used on calibrating PSF of GF-4 satellite panchromatic camera. The results showed that the new method could determine the PSF correctly

Preparation of sodium alginate-polyvinyl alcohol@PVDF hybrid membrane for efficient oil-water separation

Oily sewage is widespread in petrochemical and machinery manufacturing industries. Direct discharge not only wastes water and oil resources, pollutes the ecological environment, but also affects the survival and health of humans and other organisms. The traditional oil-water separation method has strong limitations, poor economy and low separation efficiency. Sodium alginate (SA) and polyvinyl alcohol (PVA) were uniformly coated on the surface of PVDF membrane, and superhydrophilic SA-PVA@PVDF hybrid membrane was prepared by multiple cyclic self-assembly. The oil-water separation experiment was carried out by using the membrane. The initial separation efficiency was 99.8%, and the overall separation efficiency remained above 98.3% after 10 cycles of regeneration experiments. This study provides a new idea for the low-cost, green and simple preparation of superhydrophilic-underwater superoleophobic separation materials