Salidroside regulates tumor microenvironment of non-small cell lung cancer via Hsp70/Stub1/Foxp3 pathway in Tregs

Abstract Background: The treatment of non-small cell lung cancer (NSCLC) is challenging due to immune tolerance and evasion. Salidroside (SAL) is an extract in traditional Chinese medicine and has a potential antitumor effect. However, the mechanism of SAL in regulating the immunological microenvironment of NSCLC is yet to be clarified. Methods: The mouse model with Lewis lung cancer cell line (3LL) in C57BL/6 mice was established. And then, the percentage of tumor-infiltrating T cell subsets including Treg was detected in tumor-bearing mice with or without SAL treatment. In vitro, the effect of SAL on the expression of IL-10, Foxp3 and Stub1 and the function of Treg were detected by flow cytometry. Network pharmacology prediction and molecular docking software were used to predict the target of SAL and intermolecular interaction. Furthermore, the effect of SAL on the expression of Hsp70 and the co-localization of Stub1-Foxp3 in Treg was confirmed by flow cytometry and confocal laser microscopy. Finally, Hsp70 inhibitor was used to verify the above molecular expression. Results: We discovered that SAL treatment inhibits the growth of tumor cells by decreasing the percentage of tumor-infiltrated CD4+Foxp3+T cells. SAL treatment downregulates the expression of Foxp3 in Tregs, but increases the expression of Stub1, an E3 ubiquitination ligase upstream of Foxp3, and the expression of Hsp70. Inhibiting the expression of Hsp70 reverses the inhibition of SAL on Foxp3 and disrupts the colocalization of Stub1 and Foxp3 in the nucleus of Tregs. Conclusions: SAL inhibits tumor growth by regulating the Hsp70/stub1/Foxp3 pathway in Treg to suppress the function of Treg. It is a new mechanism of SAL for antitumor therapy.

In vitro Reconstitution of Phase-separated p62 Bodies on the Arp2/3-derived Actin Network

Microvascular Changes After Conbercept Intravitreal Injection of PDR With or Without Center-Involved Diabetic Macular Edema Analyzed by OCTA

PurposeTo investigate the intravitreal injection of conbercept as a treatment strategy for proliferative diabetic retinopathy (PDR) with or without center-involved diabetic macular edema (CI-DME) and evaluate its effect on the microvascular changes in the eyes.MethodsIn this prospective study, 43 patients including 29 cases (56 eyes) in CI-DME with PDR patients, and 14 cases (26 eyes) in the non-center involving diabetic macular edema (NCI-DME) with PDR patients were involved in this study. The best corrected visual acuity (BCVA), central retinal thickness (CRT), foveolar avascular zone (FAZ), and macular capillary vessel density (VD) of the superficial retinal capillary plexus (SCP) and deep retinal capillary plexus (DCP) were assessed before and after conbercept treatments for 1, 3, or 6 months.ResultsThe BCVA was significantly increased after conbercept treatment in the eyes of CI-DME patients. After 6 months of treatment with the conbercept, microvascular density of the inferior area in SCP and the central fovea area in DCP increased significantly, regardless of the central fovea involvement. The effect of the conbercept treatment on the VD of NCI-DME was higher than that of CI-DME. Then, after 6 months of treatment, the CRT of patients with CI-DME and NCI-DME were decreased significantly.ConclusionsIn this study, an intravitreal injection of conbercept significantly improved vision, alleviated macular edema in patients with DME. Conbercept treatment also altered the microvascular density in the retina.

Comparison of the Characteristics of Cytokine Storm and Immune Response Induced by SARS-CoV, MERS-CoV, and SARS-CoV-2 Infections

Association Between Interleukin 35 Gene Single Nucleotide Polymorphisms and the Uveitis Immune Status in a Chinese Han Population

Autoimmune uveitis is characterized by immune disorders of the eyes and the whole body and is often recurrent in young adults, but its pathogenesis is still unclear. IL-35 is an essential regulatory factor in many autoimmune diseases, which is produced by Breg cells and can induce Breg cells to regulate the immune response. The relationship between the expression and gene polymorphism of IL-35 and the immune status of patients with autoimmune uveitis has not been reported. The peripheral blood of the subjects was collected from patients with Behçet’s Disease (BD) and those with Vogt–Koyanagi–Harada (VKH) syndrome. The percentage of immune cell subsets including B cells, DC, and T cells, and the expression of IL-35 in serum of these two kinds of disease were analyzed. And then, the associations between seven IL-35 single nucleotide polymorphism (SNP) sites and disease susceptibility, the immune status, the clinical characteristics, and the serum IL-35 levels were analyzed. Our results showed that the percentage of Breg cells was significantly decreased in the blood of patients with VKH syndrome compared to that of healthy controls. The levels of IL-35 in the serum of patients with VKH syndrome or BD patients were not changed significantly, compared to that of healthy controls. Furthermore, the associations between two subunits of IL-35 (IL-12p35 and EBI3) and BD or VKH patients were analyzed. We found that there was an association between the EBI3 rs428253 and the occurrence of BD. There was an association between the IL-12p35 rs2243131 and the low level of Breg cell of VKH patients. In addition, there were associations between the polymorphisms of EBI3 rs4740 and the occurrence of headache and tinnitus of VKH patients, respectively. And the genotype frequency of IL-12p35 rs2243115 was related to the concentration of serum IL-35 in patients with VKH syndrome. Thus, the specific SNP sites change of IL-35 were correlated to the immune disorders in uveitis. And they may also play a guiding role in the occurrence of clinical symptoms in patients with uveitis, especially for VKH syndrome.