Autophagy is an important homeostatic cellular recycling mechanism responsible for degrading injured or dysfunctional subcellular organelles and proteins in all living cells. The process of autophagy can be divided into three relatively independent steps: the initiation of phagophore, the formation of autophagosome and the maturation/degradation stage. Different morphological characteristics and molecular marker changes can be observed at these stages. Morphological approaches are useful to produce novel knowledge that would not be achieved through other experimental methods. Here we summarize the morphological methods in monitoring autophagy, the principles in data interpretation and the cautions that should be considered in the study of autophagy.
Autophagy ; Homeostasis ; Humans ; Organelles ; Phagosomes

Adult ; Arsenic Poisoning ; diagnosis ; drug therapy ; Humans ; Male ; Middle Aged ; Occupational Diseases ; chemically induced ; diagnosis ; drug therapy

Inhibitor of DNA binding 1 (ID1) has an aberrantly high expression in multiple cancer tissues, including colon cancer, lung cancer, breast cancer, and so on, which is closely related to cancer aggressiveness and poor clinical outcomes in cancer patients. It has been reported that ID1 maintains colorectal cancer cells (CRCs) stemness traits and contributes to the CRC drug resistance. While, the biological molecular mechanisms have not been fully elucidated. In this research, we found that ID1 upregulates octamer binding transcription factor (OCT4) protein level as well as OCT4 signaling pathway via Western blot, gene set enrichment analysis (GSEA), dual-luciferase reporter assay, and real-time PCR. Through the in vitro sphere formation assay, we found that overexpression of OCT4 reverses the inhibitory effect of knocking down ID1 on CRC sphere formation ability. With the help of JASPAR and GEPIA database, we predicted a novel transcriptional repressor—forkhead box D3 (FOXD3) of OCT4. Finally, by using co-immunoprecipitation (Co-IP), confocal and real-time PCR, we demonstrated that ID1 interacts with FOXD3 to inhibit its transcriptional repression activity and therefore to upregulate OCT4 transcription and OCT4 signaling pathway. In conclusion, this study provides a new theoretical basis for the regulation mechanism of colon cancer stem cells, and the newly found protein-protein interaction of ID1-FOXD3 provides a potential drug target for the therapy of CRC.

Objective: To investigate the gravity center and spatial clustering of silicosis in Zhejiang Province, so as to provide the reference for improving silicosis prevention and control measures. Methods: Data on cases of silicosis in Zhejiang Province from 2013 to 2022 was collected from the National Occupational Disease and Health Risk Factors Monitoring Information System. The spatial and temporal characteristics of silicosis incidence in Zhejiang Province were analyzed using the gravity center migration and spatial autocorrelation analysis. Results: Totally 2 902 silicosis cases were reported in Zhejiang Province from 2013 to 2022. The reported cases in 2013, 2019 and 2014 ranked the top three, with 545 (18.78%), 373 (12.85%) and 372 (12.82%) cases, respectively. The silicosis cases were predominantly males, accounting for 99.04% (2 874 cases). The main occupation was mining and related industries, accounting for 50.41% (1 463 cases). Gravity migration analysis showed that the gravity center of silicosis incidence was in Shengzhou City in 2013; from 2014 to 2018, it migrated to the southwest, and was located in Dongyang City and Yiwu City; from 2019 to 2020, it migrated to the southeast, and was located in Xianju County and Tiantai County; then from 2021 to 2022, it migrated to the west, and was located in Pan'an County and Xianju County. Spatial autocorrelation analysis showed that there was spatial correlation of silicosis incidence in 2014, 2016, and 2021 (all Moran's I >0, all P<0.05). Ningbo City, Quzhou City, Wenzhou City and Taizhou City were high-high clustering sites. Conclusions The gravity center of silicosis incidence in Zhejiang Province from 2013 to 2022 had a slight migration, and mainly gathered in the central regions. The high-high clustering sites were migrated from Ningbo City and Quzhou City to Taizhou City.

Objective To evaluate the effect of off-pump coronary endarterectomy (CE) plus off-pump coronary artery bypass grafting (off-pump CABG) on patients with chronic total occlusion (CTO) combined with diffuse distal atherosclerosis. Methods From October 2006 to August 2009,65 CTO patients with 176 angiographically confirmed vascular stenosis or occlusive lesions, 70 of which were complete occlusion, underwent off-pump CABG. During the operation, diffuse intimal thickening distal to occlusive lesion was found, and blood flow of the bridges was unfavorable.Results Therefore endarterectomy was performed, followed by CABG. The blood flow in the bridges were 2-10 ml/min versus 14-37 ml/min before versus after endarterectomy. Pulsatility index (PI) was 5.1-15.6 versus less than 5 before versus after endarterectomy. Left ventricular ejection fraction was also improved significantly [before operation: (0.47±0.12)%, after operation: (0. 52±0.15)%, t=2.17, P＜0.05]. Peri-operative myocardial infarction occurred in 2 cases, but without significant cardiac homodynamic changes. And 23 patients underwent coronary angiography to evaluate graft patency 3-18 months after operation, all of them had favorable blood flow. Conclusions It is feasible to perform off-pump CABG plus coronary endarterectomy for patients of chronic coronary total occlusion combined with diffuse distal atherosclerosis. This treatment is safe and effective.

Autophagy is an active research area in the biomedical field as its role has been identified in many physiological and pathological processes. Accordingly, there is a growing demand to identify, quantify and manipulate the process accurately. Meanwhile, there is great interest in identifying compounds that modulate autophagy because they may have applications in the treatment of a variety of autophagy-related diseases. In this review, we summarize the current status of autophagy screening systems to facilitate identification of autophagy modulators.
Autophagy ; Humans

Objective To study the therapeutic efficacy of Chinese botulinum toxin type A (CBTX- A) in masticatory spasm patients.Methods 16 patients with masticatory spasm were treated with CBTX-A local injection.12 patients showed jaw clench with masseter and temporalis affected (type Ⅰ).Four showed jaw clench and deviation to one side with pterygoid muscle affected unilaterally or bilaterally (type Ⅱ).All patients showed paroxysmal clenched jaw with difficulty in opening their mouths.There were no other clinical manifestations.CT and MRI did not reveal any intracerebral abnormalities.The efficacy and adverse effects were observed.Results CBTX-A were injected into 16 patients,resulting in a significant improvement of symptoms in 13 cases (4 cases of unilateral type Ⅰ,7 of 8 cases bilateral type Ⅰ,2 of 4 type Ⅱ).The spasms ceased within 3-10 days after the injection,and the effects lasted for 8-26 weeks.Four patients were observed to have slight masseter weakness after the injections,which recovered within a few weeks.The benefit persisted after identical repeated injection.Conclusion CBTX-A injection is an effective and safe treatment for masticatory spasm.

With the development of therapeutic antibodies over the past decade, they have become the treatment options for immunity and inflammation diseases. Major limitations of mouse antibodies as therapeutic agents - immunogenicity, lack of effectors' functions and short serum half-life -- were subsequently identified and largely overcome by the advent of humanized and fully human antibody technologies. The therapeutic antibodies for immunity and inflammatory diseases are primarily utilized in the treatment of allograft rejection, autoimmune disease, autoinflammatory syndromes, allergies and other chronic inflammation. The action mechanisms of therapeutic antibody include blocking ligands or receptors, regulating receptor activity, clearing the target cells or activating receptor. Strategies for generating the antibody drugs with high efficacy and low side effects can be realized by modulation of Fc-mediated activities and optimization of antigen-binding domains.

Autophagy is an important homeostatic cellular recycling mechanism responsible for degrading injured or dysfunctional cellular organelles and proteins in all living cells. Aging is a universal phenomenon characterized by progressive deterioration of cells and organs due to accumulation of macromolecular and organelle damage. Growing evidences indicate that the rate of autophagosome formation and maturation and the efficiency of autophagosome/lysosome fusion decline with age. Dysfunctional autophagy has also been observed in age-related diseases. Autophagy disruption resulted accumulation of mutated or misfolded proteins is the essential feature of neurodegenerative disorders. However, in cancers, fibroproliferative diseases or cardiovascular diseases, autophagy can play either a protective or destructive role in different types of disease, and even in different stages of the same disease. The review will discuss the cellular and molecular mechanisms of autophagy and its important role in the pathogenesis of aging and age-related diseases, and the ongoing drug discovery strategies for therapeutic intervention.
Aging ; Autophagy ; Drug Discovery ; Humans ; Lysosomes ; metabolism ; Neurodegenerative Diseases ; Phagosomes ; metabolism ; Protein Folding