LncRNA LOC103694972 promotes fibrosis of NRK-49F cells by regulating STAT3-dependent Smad/CTGF pathway via targeting miR-29c-3p

Background:Renalfibrosis is an important process in the development of chronic kidney disease.Understanding the pathogenesis andfinding effective treatments for renalfibrosis is crucial.This study aims to investigate whether a newly discovered long non-coding RNA(lncRNA)called LOC103694972 could be a potential target for treatingfibrosis of NRK-49F cells.Methods:LncRNA Chip was used to identify differentially expressed lncRNAs between TGF-β1-induced NRK-49F cells and normal cells.The dual-luciferase assay confirmed the binding between miR-29c-3p and signal transducer and activator of transcription(STAT3),as well as between miR-29c-3p and lncRNA LOC103694972.Si-LOC103694972 and miR-29c-3p mimic were then transfected into TGF-β1-induced NRK-49F cells.Results:The study found that LOC103694972 was highly expressed in TGF-β1-induced NRK-49F cells.These cells exhibited increased cell length and activity compared to the control group.The expression levels of Collagen I,α-Smooth muscle actin(α-SMA),and tissue inhibitor of metalloproteinase(TIMP-1)were increased,while matrix Metalloproteinase 2(MMP2)and matrix Metalloproteinase 9(MMP9)expression was decreased.However,transfection with si-LOC103694972 and miR-29c-3p mimics restored cell morphology and reduced cell viability.This led to a decrease in the levels of Collagen I,α-SMA,and TIMP-1,as well as an increase in MMP2 and MMP9 expression.Additionally,TGF-β1-induced NRK-49F cells transfected with miR-29c-3p mimics activated the STAT3-Smad3/CTGF pathway.Conclusion:Based on thesefindings,lncRNA LOC103694972 shows promise as a target for treating renalfibrosis.It negatively regulates miR-29c-3p and activates the STAT3-Smad3/CTGF pathway.

Investigation on the Mode of Action of the Traditional Chinese Medical Prescription-Yiqihuoxue Formula, an Effective Extravasation Treatment for Cerebral Vascular Microemboli in ApoE-/- mice

Objective: The objective of this study was to investigate the mechanisms underlying anti-embolism and extravasational effects of traditional Chinese medical prescription YiqiHuoxue(YQHX) formula in ApoE-/-mice with cerebral vascular microemboli. Materials and Methods: An ApoE-/-mice model with microemboli was developed by infusing fluorescently labeled heterologous fibrin-rich microparticles into the internal carotid artery of ApoE -/-gene knockout male mice through the common carotid artery. Before microemboli injection, the animals were randomly divided into four groups of 10 animals, treated daily for 6 weeks by intragastric administration: The ApoE-/-control group(physiological saline, 0.2 mL/10 g/d), YQHX group(0.2 ml/10 g/d), clopidogrel group(3 mg/kg/d), and atorvastatin group(3 mg/kg/d);a further group was constituted of normal male C57 BL/6 J mice(with the same genetic background as ApoE-/-mice;normal control group;no treatment;microemboli injection). The mice in each microemboli group were divided into three subgroups, the 2-h, 24-h, and 72-h subgroups, corresponding to the time after microemboli injection. Two hours(or 24 h or 72 h) after microemboli injection, the changes in aortic intima and brain tissue were analyzed by histopathology, the amounts of fluorescent emboli being measured by fluorescence microscopy image analysis. Comparison points included the microemboli induced loss of aorta functions and pathological changes, atherosclerotic plaque, brain ultrastructure and functions, and embolus extravasation. Results: Loss of aorta functions and adverse pathological changes, atherosclerotic plaque, serious damage in brain ultrastructure and functions, and reduced thrombus elimination were obviously serious in microemboli injected ApoE-/-mice. These symptoms were significantly relieved by the YQHX pretreatment:(i) the ratio of thrombus accumulation was increased with a significant decrease in thrombus extravasation in ApoE-/-mice, while YQHX induced an increased thrombus extravasation;(ii) the degree of aortic intimal thickening and brain tissue structural disorders were significantly increased in ApoE-/-mice, but overtly inhibited in the YQHX group;(iii) YQHX restored cell viability and homeostasis in the brain;(iv) YQHX regulated the expression of pro-and anti-inflammatory cytokines in the aorta;and(v) YQHX reduced cortical nerve nuclei pyknosis, edema, liquefaction, and necrosis induced by brain hypoxia, especially in the 24 h and 72 h groups. Conclusions: These findings indicate that the protective effects of YQHX on the brain against microemboli-induced injury may be attributed to the activation of extravasation mechanisms, which are involved in the cerebrovascular injury pathway and constitutively important in the progression of ischemic stroke.

A mechanism to improve early diabetic nephropathy by modulating autophagy-related mechanisms with Yuye Decotion

Objective: To investigate the molecular mechanisms of Yuye Decotion in the treatment of diabetic kidney disease using network pharmacology methods and molecular docking techniques. Methods: Obtain the transcriptome gene expression of diabetic nephropathy through GEO database, and extract genes related to autophagy. Screen the active ingredients and corresponding targets of Yuye Decoction through the TCMSP database, and map the drug prediction targets and disease targets to obtain the autophagy-related Yuye treatment targets for diabetic nephropathy point. Use String database combined with Cytoscape 3.7.2 software to construct the "drug-active ingredient-target" network and protein interaction network of Yuyetang for the treatment of diabetic nephropathy. The target point of liquid soup in the treatment of diabetic nephropathy was analyzed by GO biological process enrichment analysis and KEGG pathway enrichment analysis, and finally used Pymol and other software to analyze the core active components of Yuye Decotion and The core target protein undergoes molecular docking verification. Results: (i)100 eligible diabetic nephropathy and autophagy related genes were screened, and the potential targets of Yuye Decoction were 1,428. The acquired genes related to diabetic nephropathy and autophagy were mapped to potential targets of Yuye Decoction, and 22 therapeutic targets were obtained. GO biological process enrichment analysis and KEGG pathway enrichment analysis found that the pathways related to autophagy in the treatment of diabetic nephropathy by Yuye Decoction may include mTOR signaling pathway, phospholipase D signaling pathway, insulin resistance, EGFR tyrosine kinase inhibitor resistance, Apoptosis, PI3K /Akt signaling pathway, NF-κB signaling pathway, etc. (ii)The protein interaction network shows that VEGFA, ERBB2, GASP3, MAPK8, MYC, CDKN1A, EGFR, BCL2L1 may be the key targets of Yuye Decoction in the treatment of diabetic nephropathy. Molecular docking realizes the binding of 4 core active ingredients to 8 core target proteins. Conclusions: The research results show that Yuye Decoction treats diabetic nephropathy through multi-component, multi-target, and multi-pathway action, and provides new theoretical basis for the study of pharmacological effects and clinical application of Yuye Decoction in the treatment of diabetic nephropathy in autophagy-related aspects.

Different manual manipulations and electrical parameters exert different therapeutic effects of acupuncture

Acupuncture is an important component part of Traditional Chinese Medicine(TCM). The therapeutic effects may be influenced by a variety of factors.Stimulation quantity is one of the factors for achieving good therapeutic effects in acupuncture practice. With the development of science and technology, besides the influence of manual manipulations, the parameters adopted for electroacupuncture have become the benchmark of stimulation quantity. This study, by referring to the related literatures, is designed to explore the influence of manual manipulations and electrical parameters on therapeutic effects of acupuncture. The results from the present study show that different manualmanipulations and electrical parameters may exert different therapeutic effects of acupuncture, which are closely related to the characteristics of diseases.Different manual manipulations and electrical parameters should be adopted according to syndrome differentiation of TCM. This is very important in acupuncture treatment.

Effect of Electro-acupuncture on Vasomotor Symptoms in Rats with Acute Cerebral Infarction Based on Phosphatidylinositol System

Objective:To investigate the effect of electro-acupuncture(EA)on vasomotor symptoms in rats with acute cerebral infarction,by observing the changes in the expression of factors related to the phosphatidylinositol(PI)system.Methods:Forty-two Wistar rats were randomly divided into 3 groups by a random number table:the control group(n=6),the model group(n=18)and the EA group(n=18).The EA group was given EA treatment at Shuigou(GV 26)instantly after modeling with middle cerebral artery occlusion(MCAO)method,while the model and control groups were not given any treatment.The degrees of neurological deficiency were evaluated using neurological severity scores(NSS)and the brain blood flow was evaluated by a laser scanning confocal microscope.Western blot analysis was conducted to detect the expression levels of G-protein subtype(Gq)and calmodulin(CaM).Competition for protein binding was conducted to detect the expression level of inositol triphosphate(IP3).Thin layer quantitative analysis was conducted to detect the expression level of diacylglycerol(DAG).The expression level of intracellular concentration of free calcium ion([Ca^(2+)]i)was detected by flow cytometry.Results:The NSS of the model group was significantly higher than the control group at 3 and 6 h after MCAO(P<0.01),while the EA group was significantly lower than the model group at 6 h(P<0.01).The cerebral blood flow in the model group was significantly lower than the control group at 1,3 and 6 h after MCAO(P<0.01),while for the EA group it was remarkably higher than the model group at the same time points(P<0.01).The expressions of Gq,CaM,IP3,DAG and[Ca^(2+)]i in the model group were significantly higher than the control group(P<0.05 or P<0.01),and those in the EA group were significantly lower than the model group at the same time points(P<0.05 or P<0.01).Conclusion:EA treatment at GV 26 can effectively decrease the over-expression of related factors of PI system in rats with acute cerebral infarction,improve cerebral autonomy movement,and alleviate cerebral vascular spasm.

七氟烷麻醉诱导的认知功能损伤与内测前额叶皮层中PSD95表达量降低有关（英文）

目的:临床上接受七氟烷麻醉的病人,术后会出现认知功能损伤。本研究旨在挖掘七氟烷麻醉导致的内侧前额叶皮层中基因表达谱的改变,并探讨术后认知功能改变的机制。创新点:本研究深入探讨七氟烷麻醉导致认知功能损伤的分子机制,充分利用表达谱和动物行为学实验来揭示可能的分子机制。方法:采用十字迷宫、O迷宫和水迷宫分析接受不同时间七氟烷处理的Wistar大鼠的行为学特征;同时提取实验大鼠内侧前额叶皮层的mRNA进行表达谱分析(图3),采用实时聚合酶链反应(real-time PCR)对差异表达mRNA进行验证。结论:内侧前额叶皮层中PSD95表达量降低与七氟烷麻醉诱导的认知功能损伤有关。