Abstract: Objective With China’s manned spaceflight project entering the medium- and long-term flight stages, the effects of the weightless environment on cognitive function during long-term stays in space has become an important challenge. This study aimed to explore the influence of a weightless environment on learning and memory ability in mice, and to investigate its potential molecular mechanism by establishing a simulated space-weightlessness model on the ground. Methods Male ICR mice were used to establish a simulated space-weightlessness model by tail-suspension method, and the intervention lasted for 4 weeks. Changes in learning and memory ability were evaluated by behavioral tests including the Y-maze, new object-recognition, Morris water maze, step-down test, and shuttle-box test. The H₂O₂ and malondialdehyde (MDA) in serum were detected by enzyme-linked immunosorbent assay, and markers of oxidative stress (nuclear factor erythroid 2-related factor 2(Nrf2) / heme oxygenase-1 (HO-1) pathway), inflammation ( interleukin ( IL)-33, cyclooxygenase-2 ( COX-2)), apoptosis ( Caspase-3, Caspase-8,Bcl2 / Bax), and mitochondrial function (mitochondrial transcription factor A (mtTFA), phospho-mechanistic target of rapamycin (P-mTOR) / mTOR, SirT1, SirT3) were detected in the hippocampus by Western blot. Results After 4 weeks of simulated weightlessness, the discrimination index of mice in the new object-recognition experiment was reduced compared with the control group (P<0. 05), markedly prolonged escape latency in the Morris water maze (P<0. 01), extremely significantly reduced time spent in the safe zone in the step-down test (P<0. 001), and notably decreased numbers of active avoidance responses in the shuttle-box test (P<0. 01, P<0. 001, P<0. 05). The H₂O₂ and MDA contents in brain tissue were increased (P<0. 05). The relative expression levels of Nrf2 and HO-1 protein in the hippocampus were decreased in the simulated-weightlessness mice ( P<0. 001, P<0. 05). The relative expression levels of the inflammatory-related proteins IL-33 and COX-2 were increased (P<0. 05, P<0. 001); the pro-apoptosis-related proteins Bax, Caspase-3, and Caspase-8 were increased (P<0. 01); the anti-apoptosis protein Bcl2 was decreased ( P<0. 01); and the relative expression levels of the mitochondrial function-related proteins mtTFA, P-mTOR/ mTOR, SirT1, and SirT3 were decreased ( P<0. 001, P<0. 05, P<0. 01, P<0. 001) in simulated-weightlessness mice. Conclusions Simulated weightlessness can lead to a significant decline in learning and memory ability in mice, and its mechanism may be related to multiple pathophysiological processes such as increased oxidative stress, activation of the inflammatory response, and increased apoptosis and mitochondrial dysfunction in the hippocampus. This study provides an experimental basis for understanding the pathogenesis of cognitive dysfunction in a space-weightlessness environment, and lays a theoretical foundation for developing targeted protective measures.

Abstract: Objective To determine if enolase 1 (ENO1) exacerbates renal injury by regulating the nuclear factor (NF)-κB signaling pathway to mediate M1 macrophage polarization. Methods An acute kidney injury (AKI) model was established in C57BL / 6 mice ( n= 12) via ischemia-reperfusion injury ( IRI). Mice were assigned randomly to Sham and IRI groups. Renal function was detected on the 3rd day after surgery by measuring serum creatinine ( SCr) and blood urea nitrogen ( BUN), and pathological changes in renal tissue were detected by hematoxylin-eosin and periodic acid-Schiff staining. Expression levels of ENO1, kidney injury molecule-1 (KIM-1),and key proteins in the NF-κB pathway ( phospho-p65, nuclear factor-κB inhibitor α ( IκBα)) were detected by immunohistochemistry or Western blot, and levels of inflammatory cytokines ( tumor necrosis factor ( TNF)-α,interleukin (IL)-1β, IL-6) were quantified by enzyme-linked immunosorbent assay. Human renal tubular epithelial cells ( HK2 ) were exposed to tert-butyl hydroperoxide in vitro to induce oxidative stress, followed by ENO1 knockdown or overexpression to assess the impact on macrophage polarization markers (inducible nitric oxide synthase (iNOS), CD206). Results ENO1 expression was markedly upregulated in AKI mice(P<0. 05), accompanied by deteriorated renal function ( SCr, BUN, P<0. 001), elevated inflammatory cytokines, and aggravated renal tissue injury(P<0. 001). Mechanistically, ENO1 activated the NF-κB pathway, evidenced by increased p-p65 levels and IκBα degradation ( P<0. 01, P<0. 001 ), and promoted macrophage polarization towards the M1 phenotype (upregulated iNOS). Knockdown of ENO1 suppressed NF-κB activation ( P<0. 05, P< 0. 001), attenuated M1 polarization, and ameliorated inflammatory injury(P<0. 01), while ENO1 overexpression further enhanced NF-κB activity and exacerbated inflammatory responses. Conclusions IRI or oxidative stress can induce the upregulation of ENO1 expression in renal tubular cells. As a novel paracrine regulatory mediator, ENO1 promotes macrophage M1 polarization via activating the NF-κB pathway (with no significant effect on the M2 phenotype), thereby aggravating the inflammatory cascade in AKI. This suggests that ENO1 may serve as a potential therapeutic target for AKI, providing a novel insight for intervening in the inflammatory response.

Abstract: Objective To establish a model to evaluate the therapeutic effect of Biejiajian pills (BJJP) on bile duct ligation (BDL)-induced liver fibrosis in rats, and to investigate the molecular mechanisms by which BJJP alleviates trimethlylamine oxide (TMAO)-aggravated liver fibrosis in BDL rats. Methods A rat model of therapeutic BJJP for BDL-induced hepatic fibrosis was established. Thirty rats were divided randomly into Sham, BDL, and BJJP low-dose (-L), medium-dose (-M), and high-dose (-H) groups. Collagen-fiber deposition was analyzed by Masson and Sirius Red staining. Expression levels of α-smooth muscle actin ( α-SMA) and type Ⅰ collagen α1 chain (COL1A1) proteins in liver tissue in each group were detected by immunohistochemical staining and Western blot.Feces were collected from rats in the Sham, BDL, the BJJP-H groups for 16S rRNA high-throughput microbial sequencing and non-targeted metabolomics analysis. Serum and feces levels of trimethylamine ( TMA) and TMAO were determined in the Sham, BDL, and BJJP-H groups, respectively, by non-targeted and targeted metabolomics technologies. To investigate the therapeutic effects of BJJP on TMAO-aggravated hepatic fibrosis and its potential molecular mechanisms, a further 20 Sprague Dawley rats were divided randomly into Sham, BDL, BDL+TMAO, and BDL+ TMAO + BJJP-H groups. Changes in histomorphology and collagen fibers in liver tissue were detected by hematoxylin-eosin, Masson, and Sirius Red staining. Expression levels of α-SMA, flavin-containing monooxygenase 3(FMO3), phosphorylated (p-) protein kinase B (AKT), and p-phosphatidylinositol 3-kinase (PI3K) in each group were analyzed quantitatively by Western blot. Results Compared with the Sham group, liver levels of collagen fiber,α-SMA and COL1A1 protein were increased in the BDL group(P<0. 000 1, P<0. 000 1, P<0. 001). Compared with the BDL group, levels of collagen fiber in the BJJP-L, BJJP-M, and BJJP-H groups were significantly decreased(P<0. 05, P<0. 05, P<0. 001), as well as the levels of α-SMA protein and COL1A1 protein ( all P<0. 05). BJJP improved the intestinal flora and metabolic disorders in BDL rats, and reduced feces levels of TMA and serum levels of TMAO (all P<0. 05). Compared with the Sham group, rats in the BDL group exhibited disorganized hepatic cord arrangement, evident hepatocyte swelling, and extensive collagen-fiber deposition, along with up-regulation of α-SMA, FMO3, p-AKT, and p-PI3K protein expression(all P<0. 05). The above indicators in liver tissues of rats were further increased in the BDL+TMAO group compared with the BDL group(P<0. 001, P<0. 05, P<0. 000 1, P<0. 05). Compared with the BDL + TMAO group, rats in the BDL + TMAO + BJJP-H group exhibited more orderly hepatocellular arrangement, reduced collagen-fiber deposition(P<0. 000 1), and decreased protein expression of α-SMA, FMO3, p-AKT, and p-PI3K (P<0. 001, P<0. 01, P<0. 001, P<0. 05). Conclusions BJJP can improve hepatic fibrosis while TMAO can aggravate hepatic fibrosis in BDL rats. BJJP may alleviate the hepatic fibrosis process in BDL rats by regulating intestinal flora TMAO.

Abstract: Objective This study aimed to preliminarily investigate the mechanism by which Sanqiang formula acts on quadriceps muscle atrophy in unstable knee osteoarthritis ( KOA), and to provide experimental evidence for its clinical application and development. Methods The KOA rat model was established by transection of the anterior cruciate ligament (ACL) of the right posterior knee joint. In the Sham operation group, only the skin was incised, and the ACL was completely preserved. The successfully modeled rats were divided into the Model group, celecoxib group, and low-, medium-, and high-dose Sanqiang formula groups, with ten rats in each group.The Sham operation and model groups were administered normal saline by gavage; the other groups were administered the corresponding doses of the drug by gavage. Behavioral tests were conducted before and after drug administration.After the end of administration, the passive range of motion of the rat knee joint was measured; the quadriceps femoris muscle weight, and serum interleukin ( IL )-1β and tumor necrosis factor ( TNF )-α levels were determined.Hematoxylin-eosin staining was used to observe quadriceps muscle tissue damage in SD rats of each group, and Masson staining was used to observe quadriceps muscle fibrosis changes. RT-qPCR was used to detect the expression of Atrogin-1 and MuRF1 mRNA in skeletal muscle tissues of each group; Western blot was used to detect the expression of PI3K, p-PI3K, AKT, p-AKT, FOXO1, p-FOXO1, Atrogin-1, and MuRF1 proteins in skeletal muscle tissues of each group. Results Sanqiang formula improved behavioral changes in rats ( P<0. 05, P<0. 001),significantly increased passive range of motion (P<0. 001), and increased quadriceps muscle weight (P<0. 05). It decreased serum IL-1β and TNF-α (P<0. 01, P<0. 001), increased muscle cell area in the medium- and high-dose groups (P<0. 001), significantly reduced interstitial connective tissue among groups, and reduced collagen volume fraction ( P<0. 001 ). Treatment improved pathological morphological changes in articular cartilage, increased p-PI3K/ PI3K, p-AKT / AKT, and p-FOXO1 / FOXO1 values in quadriceps muscle (P<0. 05, P<0. 01, P<0. 001),and inhibited Atrogin-1 and MuRF1 protein and mRNA expression in quadriceps muscle ( P<0. 01, P<0. 001). Conclusions Sanqiang formula promoted the activation of PI3K/ AKT / FOXO1 signaling in the quadriceps muscle of KOA rats, inhibited the expression of Atrogin-1 and MuRF1, and improved quadriceps muscle atrophy.

Abstract: Objective To investigate the therapeutic effect and mechanism of transcutaneous electrical acupoint stimulation ( TEAS) combined with human chorionic gonadotrophin ( hCG) on blastocyst implantation dysfunction in rats. Methods Female rats were divided randomly into control, model, and treatment (TEAS+hCG)groups (n= 6 per group). Rats in the treatment group received TEAS+ hCG injections for 3 consecutive days starting from the late diestrus phase. On the day of proestrus, females were co-housed with male rats at a 2 ∶ 1 ratio. On gestation day 3, rats in the model and treatment groups received a subcutaneous injection of mifepristone suspension (5 mg / kg) to establish a blastocyst implantation dysfunction model. On gestation day 9, implanted blastocyst numbers, endometrial morphology, and expression levels of estrogen receptor α ( ERα), progesterone receptor (PR), and leukemia inhibitory factor ( LIF) were evaluated. Results The number of implanted blastocysts was significantly lower in the model group compared with the control group (p= 0. 005). The number was significantly increased after treatment with TEAS + hCG ( p= 0. 027) compared with the model group. The endometrium was significantly thinner in the model group compared with the control group, with underdeveloped pinopodes. TEAS+hCG restored the endometrial thickness to a similar level to the control group, and improved the development of pinopodes.Expression levels of ERα mRNA ( p= 0. 028), ERα protein ( p= 0. 012), and PR protein ( p= 0. 014) were significantly lower in the model group compared with the control group. Treatment with TEAS+hCG resulted in upward trends in the protein expression levels of ERα, PR, and LIF. Conclusions TEAS+hCG treatment may regulate endometrial expression of ERα, PR, and LIF, promote endometrial and pinopode development, improve endometrial receptivity, and effectively treat blastocyst implantation dysfunction in rats.

Abstract: Objective To investigate the regulatory effect and molecular mechanism of annexin A5 (AnxA5)gene on cognitive function in mice. Methods AnxA5 gene knockout (AnxA5^{- / -}) mice were constructed by CRISPR/Cas9 technology. The regulatory effect of AnxA5 on cognitive function in mice was verified by Morris water maze, Ymaze and novel object recognition tests. Western blot, qPCR, immunofluorescence, real-time cell analysis (RTCA) and other method were used to detect the effects of AnxA5 deletion on the proliferation and morphology of mouse hippocampal neurons, and to explore its regulatory mechanism. APP / PS1 double transgenic mice were used as positive controls for Alzheimer’ s disease. Results Compared with wild-type (WT) C57BL / 6 mice, the cognitive function of AnxA5^{- / -}mice was significantly reduced(P<0. 05). After interfering with the expression of AnxA5 gene in HT-22 cells, the cell proliferation ability was significantly weakened(P<0. 05), the cell morphology was irregular,and AnxA5 deletion could activate the P65 signaling pathway. Conclusions AnxA5 gene deletion can up-regulate the expression level of inflammatory factors by activating the P65 pathway, thereby damaging mouse hippocampal neurons and ultimately leading to cognitive impairment in mice.

Abstract:Ferroptosis is an iron-dependent form of programmed cell death that plays a key role in the pathological process of intervertebral disc degeneration (IVDD). Traditional Chinese medicine, with its advantages in overall conditioning, has demonstrated unique potential in the prevention and treatment of IVDD. This review considers the pathological mechanism of IVDD and its interaction with ferroptosis, revealing ferroptosis as not only a pathological driving factor, but also a therapeutic regulatory target. The theory of ferroptosis has great potential in guiding interventions for IVDD using traditional Chinese medicine. By targeting the key pathways of ferroptosis and exploring the potential mechanisms of ferroptosis-related genes, it provides a new perspective and theoretical basis for the prevention and treatment of IVDD using traditional Chinese medicine, and also lays the foundation for the development of precise treatment strategies.

Abstract:Ischemic stroke (IS) is a cerebrovascular disease with high global mortality. Its occurrence and development are closely related to oxidative stress. Vascular recanalization is mainly used to restore cerebral blood circulation; however, it may also cause cerebral ischemia-reperfusion injury and further aggravate neurological damage. The single-target treatment strategy of modern medicine has certain limitations, making the overall intervention advantages of multicomponent, multichannel, and multi-target traditional Chinese medicine increasingly valued. This paper systematically summarizes research into oxidative stress involved in the pathological process of IS by mediating inflammatory responses, affecting the function of neurovascular units, regulating key signaling pathways,destroying the integrity of the blood-brain barrier, inducing apoptosis, and regulating autophagy. In recent years,extensive research on traditional Chinese medicine monomers and active ingredients has been performed, showing that it has good potential for improving clinical symptoms and neurological function recovery in IS. With an emphasis on oxidative stress, this paper aims to summarize the mechanism of action and clinical research progress of traditional Chinese medicine in IS treatment to provide a theoretical reference and ideas for the clinical prevention and treatment of this disease and the development of new drugs. Follow-up studies should focus on elucidating the mechanism of multicomponent synergistic anti-oxidative stress effects of traditional Chinese medicine, constructing a sequential treatment scheme of integrated traditional Chinese and western medicine, and carrying out high-quality research to establish clinical evidence. This review provides accurate prediction models, timing optimization strategies, and highlevel medical evidence for the prevention and treatment of cerebral ischemia-reperfusion injury, promotes the transformation of basic research result into clinical practice, and provides new strategies for IS prevention and treatment.

Abstract:Sj?gren’s syndrome (SS) is a chronic autoimmune disease that usually occurs because of immune system disorders; however, the specific mechanism is still unclear, and there is no cure. T lymphocytes are an important type of immune cell; their role in SS occurrence and development has been widely described, and they are an important target for anti-SS treatment. Although modern medical treatment can alleviate symptoms and delay progression to some extent, there are obvious clinical limitations. Additionally, the overall treatment effect is not satisfactory, making it important to find an effective and safe treatment method. Traditional Chinese medicine treats diseases based on the fundamental principles of syndrome differentiation and holistic concepts; it has been found to have multi-pathway, multilevel, and multi-target anti-SS effects. Active ingredients of traditional Chinese medicine,such as Fangchinoline, total glucosides of paeony, and Salidroside, as well as traditional Chinese medicine compound prescriptions such as Moisturizing and Drying Formula, Yangyin Yiqi Huoxue Formula, and Jiefu Huaxue Shengjing Formula, can regulate the changes of different T lymphocyte subsets through signaling pathways such as PI3K/ AKT /mTOR, STAT3, and NF-κB. These treatments synergistically alleviate inflammatory responses, reduce immune infiltration, and improve glandular function to achieve a therapeutic effect in SS. This article reviews and analyzes the relevant recent research on traditional Chinese medicine treatments for SS, and discusses their mechanisms in regulating T lymphocytes against SS to provide new ideas for the clinical treatment of this condition.

Abstract:Post-stroke depression (PSD) is the most common mental complication after stroke, seriously affecting recovery of neurological function and quality of life. Although antidepressants are currently the first-line treatment, they have limitations such as cardiotoxicity, gastrointestinal reactions, and relatively high costs. As a nonpharmaceutical intervention method integrating traditional Chinese medicine and modern neuroanatomy, auricular therapy has shown unique advantages in PSD treatment through a multi-target regulatory mechanism. From the perspective of neurobiology, this article systematically reviews the mechanism of auricular therapy in PSD intervention, including: ( 1 ) the regulation of monoamine and amino acid neurotransmitter homeostasis;(2) promoting neural plasticity and the reorganization of brain network functions; (3) inhibiting neuroinflammation and oxidative stress responses; (4) and regulating the neuroendocrine system. This review aims to comprehensively explain the mechanism by which auricular therapy alleviates PSD through a multidimensional neuro-immune-endocrine regulatory network to promote further research and application in PSD treatment.

Abstract:Cancer is the second leading cause of death worldwide. Despite significant recent progress in the treatment of cancer, the incidence and mortality rate are still high, and finding effective drugs to treat cancer thus remains a focus of research. The natural compound extract, isoliquiritigenin (ISL), has been widely studied in terms of its anti-tumor effects in recent years. This article reviews recent research progress on the anti-cancer effects of ISL,and discusses its role in suppressing the occurrence of cancer by controlling the cell cycle, metastasis, angiogenesis,

Abstract:Rheumatoid immune diseases (RID) are chronic autoimmune inflammatory diseases characterized by immune inflammatory cascade reactions, which are closely related to the occurrence and development of immune homeostasis imbalance. Macrophage efferocytosis plays a key regulatory role in delaying the progression of RID by clearing apoptotic cells, maintaining immune homeostasis, and promoting inflammation resolution. Recent studies have shown that active ingredients in traditional Chinese medicines such as Astragalus polysaccharide, Triptolide, Berberine, can upregulate the expression of efferocytosis-related proteins such as Mer tyrosine kinase, growth arrestspecific protein 6, scavenger receptor class A, and microtubule-associated protein 1 light chain 3 by regulating related signaling pathways, activating macrophage efferocytosis and regulating the immune microenvironment, inflammatory response, angiogenesis, epithelial mesenchymal transition, and endothelial-mesenchymal transition, delaying the progression of RID such as rheumatoid arthritis, systemic lupus erythematosus, and systemic sclerosis. This article summarizes the research result on the regulation of macrophage efferocytosis by active ingredients in Chinese medicine for the treatment of RID to provide new ideas for clinical application.