Abstract: Objective To explore the mechanism of gout attacks in hyperuricemic rats under ice-water swimming conditions based on the purinergic P2X7 receptor (P2X7R). Methods Male Sprague-Dawley rats were divided randomly into Control group (n= 6), HPD+IWS group (n= 12), HPD group (n= 6), and IWS group (n=6). Hyperuricemia was induced by gavage with potassium oxonate combined with yeast extract. Ice-water swimming was performed by 5-minute endurance swimming in an ice-water mixture. The experiment lasted for 21 days. Serum uric acid levels were detected by the uricase method, serum and liver xanthine oxidase (XOD) levels were detected by colorimetry, and serum ATP levels were detected by phosphomolybdic acid colorimetry. Uric acid crystals in joint fluid were observed by polarized light microscopy. During the experiment, the right ankle joint diameter and foot thickness were measured every 2 days, and the swelling index was calculated. The pathological changes of ankle joint and synovial tissue were observed by hematoxylin-eosin (HE) staining. The serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were detected by ELISA. Neutrophil count in venous blood was detected by full self-help blood cell analyzer. The expression level of P2X7R protein in synovial tissue was detected by Western blot. The expression level of nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) protein in synovial tissue was detected by immunohistochemistry. Results Compared with Control group, on the 7th, 14 th and 21 st days of the experiment, the serum uric acid levels of HPD+IWS group and HPD group were significantly increased (P<0. 01 or P<0. 05). Compared with HPD+IWS group, the serum uric acid level of HPD group was significantly decreased on the 7th and 21st days of the experiment (P<0. 01). And the serum uric acid level of IWS group was significantly decreased on the 7th, 14th and 21st days of the experiment (P<0. 01). Compared with Control group, the serum and liver XOD levels in HPD+IWS group were significantly increased (P<0. 01). Compared with HPD+IWS group, the serum XOD levels of HPD group and IWS group were significantly reduced (P<0. 01). And the liver XOD level of HPD group was significantly reduced (P<0. 05). Compared with HPD+IWS group, the serum ATP of HPD group was significantly increased on the 7th, 14th and 21st days (P<0. 01 or P<0. 05). And the serum ATP level of IWS group was significantly increased on the 14th day (P<0. 01). Urate crystals were observed in HPD+IWS group. Compared with Control group, the swelling index of the right ankle joint in HPD+IWS group increased significantly on the 5th, 7th, 9th, 11th and 13th days ( P<0. 01 or P<0. 05). Compared with Control group, the foot thickness swelling index of HPD+IWS group was significantly increased on the 5th, 7th, 9th, 11th, 13th, 15th, 17th, 19th and 21st days (P<0. 01 or P<0. 05). The result of HE staining showed that synovial hyperplasia and inflammatory cell infiltration appeared in the ankle joint cavity of HPD+IWS group. Compared with Control group, the levels of serum IL-1β, IL-6 and TNF-α in HPD+IWS group were significantly increased (P<0. 01). Compared with HPD+IWS group, the levels of serum IL-1β, IL-6 and TNF-α in HPD group and IWS group were significantly decreased (P<0. 01). Compared with Control group, the neutrophil count in HPD+IWS group was significantly increased (P<0. 01). Compared with HPD+IWS group, the neutrophil count in IWS group was significantly reduced (P<0. 01).Compared with Control group, the expression of P2X7R and NLRP3 protein in the synovial tissue of HPD+IWS group was significantly increased (P<0. 01 or P<0. 05). Conclusions Ice-water swimming under hyperuricemic conditions can lead to ATP metabolic imbalance, increase uric acid levels and urate deposition, and activate P2X7R, leading to gouty arthritis attacks via the ATP-P2X7R-NLRP3 pathway.

Abstract: Objective To explore the expression and potential clinical value of Discs large homologyassociated protein 5 (DLGAP5) in hypoxic pulmonary hypertension (HPH). Methods The transcriptome data of non-HPH patients and HPH patients were downloaded from the GEO database, and bioinformatics analysis was performed to explore the expression of DLGAP5 in the non-HPH patients and HPH patients. Then, lung tissue samples from 9 HPH cases and 4 non-HPH cases were collected, and the expression levels of DLGAP5 in different lung tissues were verified using Western blot experiments. Rat models of HPH were created using Sugen5416 combined with chronic hypoxia ( SuHx) and the mean pulmonary artery pressure (mPAP) was detected. The rats were divided into HPH rat model group and normal group, with 6 rats in each group. Pulmonary vascular remodeling and DLGAP5 expression in HPH were detected by hematoxylin-eosin ( HE), immunohistochemistry ( IHC), and immunofluorescence (IF) staining. Finally, differential gene enrichment analysis was used to explore the downstream signaling pathway of DLGAP5 in regulating HPH. Results DLGAP5 was significantly upregulated in lung tissues from HPH patients (P<0. 01). Compared with the normal group, the mPAP was significantly increased in SuHxinduced HPH rat model group (P<0. 0001). HE and IHC staining showed a thickened pulmonary artery media and obvious vascular remodeling in HPH rat model group. IF staining showed increased DLGAP5 expression in pulmonary artery media smooth muscle cells in HPH rat model group compared with the normal group (P<0. 0001), mainly distributed in the cytoplasm. Finally, differential gene enrichment analysis suggested that DLGAP5 may regulate HPH by influencing the cell cycle, especially the G2 / M phase (P<0. 001). Conclusions DLGAP5 may be a potential new target for HPH, indicating new ideas for the diagnosis and treatment of HPH.

Abstract: Objective The 4D-FastDIA proteomic method was employed to explore the mechanism of LiqiTongfu massage in treating functional dyspepsia (FD), focusing on its effects on protein expression. Methods 30 rats were categorized into CTL group, MOD group, TN group ( n=10 rats per group), using a random number table approach. Rats in CTL group were fed normally. Rats in the MOD and TN groups were treated with tail clips,ice water, and a non-standard diet. After successful modeling, TN group was treated with LiqiTongfu manipulation for 14 days. Overall health and digestive movement were then assessed, pathological changes in the stomach tissues were monitored, and protein expression levels were measured in each group. Results Rats in CTL group exhibited pure white, thick, and lustrous fur, well-formed stools without odor, and were in good spirits with high activity levels. Rats in MOD group rats displayed dull, disheveled, sparse, and lackluster fur, loose stools with an acidic odor, lethargy, reduced activity levels, and decreased gastric emptying and small intestinal propulsion rates (P<0. 01). Rats in TN group exhibited restored dense, glossy fur, well-formed feces without an acidic odor, and improvements in mental state, activity levels, gastric emptying rate, and small intestinal propulsion rate ( P<0. 01). Hematoxylin-eosin staining showed no clear infiltration of inflammatory cells in the gastric mucosa, and no difference in gastric histopathology across the three groups. Proteomic analysis showed changes in 263 proteins in MOD group compared with CTL group, and changes in 237 proteins in TN group compared with MOD group. The protein expression profiles differed between MOD group and CTL group, but massage treatment reversed the differences in 95 proteins, of which 16 were up-regulated and 79 were down-regulated. Conclusions LiqiTongfu massage therapy can effectively improve FD. This may be related to the regulation of structural proteins, smooth muscle and cell motility-associated proteins, protein activity-regulating proteins, metabolism-related proteins, signal transduction and regulation-related proteins,as well as immune and stress-related proteins.

Abstract: Objective To investigate the pathological role of circadian rhythm disorders in insomnia by destroying the molecular oscillation mechanism of hypothalamus-amygdala core circadian clock genes, and to provides dynamic molecular evidence for the Ying-Wei theory of insomnia circadian clock gene disorder. Methods 104 sixweek-old female ICR mice were divided randomly into normal and PCPA groups (n= 52 per group). Mice in PCPA group received intraperitoneal injections of PCPA for 3 days and normal mice received equivalent volumes of saline.Behavioral tests were conducted to evaluate sleep latency, total sleep duration, and anxiety-depression-like behaviors. Nuclear translocation of period circadian regulator 1 (PER1) and cryptochrome circadian regulator 1 (CRY1) in the hypothalamus and amygdala were detected by immunofluorescence at different time point (6:00,12:00,18:00,24:00), and mRNA and relative protein expression levels of circadian clock genes were detected by quantitative reverse transcription-polymerase chain reaction and Western blot, respectively. Results Compared with normal group, mice in PCPA group exhibited prolonged sleep latency, reduced sleep duration (P<0. 01), decreased sucrose preference (P<0. 01), increased grooming frequency, and extended central exploration distance in the open field test ( P<0. 01). Compared with normal group, mRNA and protein levels of core clock genes (PER1, CRY1, CLOCK, and BMAL1) differed at different time points, with inconsistent expression patterns between the hypothalamus and amygdala. Peak nuclear co-localization of PER1 and CRY1 proteins in the hypothalamus was advanced by 4~6 hours in PCPA group, while their nuclear co-localization peaks in the amygdala were phase-shifted by approximately 12 hours compared with normal group. Conclusions Circadian rhythm disturbance caused by an imbalance of Camp and Defense may be related to the circadian rhythm oscillation and phase shift of core clock genes in different brain regions, potentially manifested as phase shifts of PER1 and CRY1 into the nucleus to form complexes.

Abstract: Objective To investigate the role of the inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) /glucose-regulated protein 75 (GRP75) / voltage-dependent anion channel 1 (VDAC1) axis in regulating mitochondrial calcium (Ca²⁺) transport in apoptosis in an in vitro hypoxia-induced injury model of pulmonary artery smooth muscle cells (PASMCs). Methods Rat PASMCs were cultured in vitro and divided randomly into three groups: normal control ( NC) group, hypoxia group, and 4-phenylbutyric acid ( 4-PBA) group. The Ca²⁺concentrations in the endoplasmic reticulum ( ER ), cytoplasm, and mitochondria of PASMCs were measured using Ca²⁺immunofluorescence probes. Mitochondrial structure and morphological changes were observed by transmission electron microscopy and cell apoptosis was detected by flow cytometry. The messenger RNA (mRNA) and protein expression levels of key factors in the Ca²⁺pathway and mitochondrial fission and fusion were determined by quantitative reverse transcription-polymerase chain reaction ( RT-qPCR) and Western blot, respectively. Results Compared with NC group, Ca²⁺concentrations in the ER and cytoplasm of PASMCs were increased, whereas mitochondrial Ca²⁺concentration was decreased in hypoxia group (P<0. 01). Mitochondria showed reduced volume, swelling, disordered cristae structure, and increased rupture, and PASMCs apoptosis was decreased (P<0. 01). The mRNA and protein levels of the key Ca²⁺transport pathway factors IP3R2, GRP75, and VDAC1, and expression of the mitochondrial fission factor dynamin-related protein 1 (DRP1) were all upregulated (P<0. 01), while expression of the mitochondrial fusion factor mitofusin-2 (MFN2) was downregulated (P<0. 01). Compared with hypoxia group,Ca²⁺concentrations in the ER and cytoplasm of PASMCs were decreased, whereas mitochondrial Ca²⁺concentration was increased in 4-PBA group ( P<0. 01). Mitochondrial structure was restored, and expression levels of IP3R2,GRP75, VDAC1, and DRP1 decreased(P<0. 01), while MFN2 expression increased(P<0. 05, P<0. 01), with an increase in PASMCs apoptosis(P<0. 01). Conclusions The IP3R2 / GRP75 / VDAC1 axis enhances mitochondrial Ca²⁺transport in hypoxia-induced PASMCs, promoting PASMCs apoptosis. These result provide a theoretical basis for the prevention and treatment of hypoxic pulmonary hypertension.

Abstract: Objective To explore the expression and clinical significance of inhibitor of DNA binding 1 (ID-1) in breast cancer and its molecular mechanism in human breast cancer MCF-7 cells by targeting the nuclear factor (NF)-κB / Src homology 2 domain-containing protein tyrosine phosphatase ( SHP2) / SMAD/ Src signaling pathway. Methods The expression of ID-1 in breast cancer tissues and adjacent normal tissues was detected using immunohistochemistry, and its clinical significance was analyzed. Bioinformatics analysis was employed to examine the correlation between ID-1 and key proteins. In the in vivo experiment, 45 female mice were selected to establish a breast cancer model and divided into five groups with 9 mice each: Vivo-control group, Vivo-BMP2 group, Vivo-ID-1 mimic + BMP2 group, Vivo-BMP2 + PHPS1 group, and Vivo-ID-1 mimic + PHPS1 group. Tumor tissues from each group were dissected, observed, and weighed. Human breast cancer MCF-7 cells were used for in vitro experiments and divided into NC group, BMP2 group, ID-1 mimic+BMP2 group, sulfasalazine+BMP2 group, ID-1 mimic+sulfasalazine+BMP2 group, BMP2+PHPS1 group, and ID-1 mimic+BMP2+PHPS1 group. Protein expression levels were determined by Western blot, and cell migration and cell invasion were evaluated by scratch and Transwell assays, respectively. Results Immunohistochemical result showed that the expression of ID-1 in breast cancer tissues was significantly higher than that in adjacent normal tissues, the difference was statistically significant (P<0. 001). The expression status of ID-1 was closely associated with histological grade, TNM stage, lymph node metastasis, and distant metastasis ( P<0. 05). Bioinformatics analysis indicated that ID-1 was correlated with BMP2, NF-κB,SMAD1 / 8, and Src in breast cancer(P<0. 05). ID-1 promoted the progression of breast cancer tumors in vivo, while inhibition of SHP2 slowed tumor progression ( P<0. 05, P<0. 01). Inhibition of SHP2 significantly decreased expression levels of ID-1, NF-κB, phospho ( p)-SHP2, p-SMAD1 / 5 / 8, and p-Src proteins in vitro( P<0. 05,P<0. 01). Similarly, inhibition of NF-κB reduced expression levels of ID-1, NF-κB, p-SHP2, p-SMAD1 / 5 / 8, and pSrc proteins(P<0. 05,P<0. 01). Both ID-1 and BMP2 promoted MCF-7 cell migration, while inhibition of SHP2 or NF-κB significantly reduced cell migration(P<0. 05,P<0. 01). ID-1 and BMP2 also enhanced MCF-7 cell invasion,while inhibition of SHP2 or NF-κB reduced cell invasion( P<0. 05,P<0. 01). Conclusions ID-1 may promote breast cancer invasion and migration by activating the NF-κB/ SHP2 / SMAD/ Src signaling pathway.

Abstract:To compare the application value of rabbit brain infusion ( TF) and lipopolysaccharide ( LPS)-induced disseminated intravascular coagulation (DIC) models in medical laboratory teaching, this study systematically contrasts these two classical models across four dimensions: pathogenic mechanisms, modeling techniques, detection systems, and teaching application design. Results indicate that the TF model simulates acute traumatic DIC through exogenous coagulation activation, while the LPS model mimics chronic infectious DIC via endogenous inflammatorycoagulation cascades. Both models exhibit complementary characteristics in pathogenesis timing, indicator profiles,and experimental skill training. Teaching optimization strategies grounded in mechanism-oriented approaches, skilltiered training, and clinical mapping can effectively enhance students’ understanding of DIC’s complex pathological processes and practical competencies. This provides theoretical justification and a feasible teaching design framework for selecting and integrating DIC models in medical experimental education.

Abstract:Macaques are widely used as experimental animals in biomedical research. Monkey B virus (BV) is a zoonotic pathogen that naturally infects macaques, with an infection rate as high as 70% to 100%. Although BV infection in macaques is usually asymptomatic or has only mild symptoms, once transmitted to humans, it can cause severe encephalomyelitis and neurological sequelae, with a fatality rate of over 40%. BV can also remain latent in the human body for a long time and then be reactivated under specific circumstances, leading to serious neurological diseases. Most people infected with BV are animal caregivers, veterinarians, and laboratory researchers. The severity of human BV infection makes it the main zoonotic disease of concern among those in contact with macaques. There is currently no vaccine to prevent human infection with BV and no specific antiviral drugs. Implementing preventive measures through occupational health and safety management, conducting standardized emergency responses to exposure incidents, and promptly blocking or reducing the invasion of BV are thus key strategies for lowering the risk of human BV infection and mortality. This article reviews the characteristics of human BV infection, the preventive measures for occupational health and safety management related to BV, emergency treatment measures after exposure,exposure risk assessment, post-exposure prophylaxis, and the key points of post-exposure follow-up, with the aim of providing useful references for laboratory animal institutions and medical institutions in the practice of BV prevention and control.

Abstract:Growth arrest-specific protein 7 (Gas7) is rich in α-helix structures and is highly expressed in the central nervous system, particularly in the cortex, hippocampus, and cerebellum. The abnormal expression of Gas7 is closely related to the occurrence of various nervous system diseases and drug addiction; however, the molecular regulation and underlying mechanisms of Gas7 remain to be fully elucidated. This review comprehensively examines the diverse functions of Gas7 in the nervous system, including its critical regulatory roles in nervous system development, neuronal differentiation and maturation, microtubule dynamics, cell cycle regulation, neuronal migration, axon guidance, neuronal mitophagy and repair, and synaptic plasticity. Further investigations may offer novel insights into the pathogenesis of neurological diseases and provide a theoretical foundation for the development of innovative therapeutic strategies.

Abstract:Cerebral ischemia-reperfusion injury (CIRI) is a key pathological link in the deterioration of neurological function after stroke. Its mechanism is closely related to oxidative stress and the inflammatory response,and ultimately leads to severe neuronal damage. The nuclear factor E2-related factor 2 / heme oxygenase-1 (Nrf2 / HO-1) signaling pathway, which mediates endogenous protective effects, has recently received extensive attention. This pathway plays a central role in maintaining cellular redox homeostasis by regulating antioxidant, anti-inflammatory,and anti-apoptotic processes. This review explores the activation mechanism and multi-directional protective effect of the Nrf2 / HO-1 signaling pathway in CIRI, and analyzes the therapeutic potential of targeting this pathway, to provide a theoretical basis for the development of new neuroprotective drugs to optimize clinical treatment strategies and improve the prognosis of patients with CIRI.

Abstract:Heart failure has high incidence and mortality rates worldwide. Artificial intelligence (AI) is increasingly used in the cardiovascular field. This article summarizes progress in combining AI technology with clinical examination method for the prediction of heart failure. AI can evaluate the structure of the heart, and predict heart failure by learning parameters from echocardiography. The electrocardiogram model established by AI has high agility in predicting heart failure. New biomarkers and gene involvement discovered by AI can guide the prediction of heart failure and the screening of high-risk populations. AI can also identify the risk of heart failure by learning other disease characteristics. AI technology has the advantages of convenience, reliability, and high efficiency for the prediction of heart failure, presented new possibilities and challenges for this field and provided valuable references for the clinical decision-making of medical specialists.

Abstract:Sj?gren ’ s syndrome ( SS ) is a chronic autoimmune disease characterized by a complex pathogenesis and suboptimal treatment outcomes. Its onset is closely linked to dysregulation of the immune system. As key components of this system, immune cells influence the development of SS by modulating lymphocyte infiltration,inflammatory responses, and epithelial cell proliferation. miRNAs play a crucial role in regulating gene expression,thereby impacting immune responses, metabolic processes, and cellular activities, and are widely implicated in the pathogenesis, progression, and prognosis of SS. Specific miRNAs, including miR-31-5p, miR-100-5p, and miR-125b, have been shown to target signaling pathways including the p38 mitogen-activated protein kinase and Janus kinase / signal transducer and activator of transcription pathways. Via these pathways, miRNAs can modulate the function of macrophages, dendritic cells, T lymphocytes, and B lymphocytes, ultimately reducing inflammation, improving glandular function, and exerting anti-SS effects. This article reviews the current literature regarding the specific mechanisms by which miRNAs regulate immune cells in the context of SS treatment, aiming to provide insights and references for future therapeutic strategies.

Abstract:The cyclic GMP-AMP synthase ( cGAS)-stimulator of interferon genes ( STING) signaling pathway is a crucial DNA recognition and signaling mechanism in the innate immune system, which is widely involved in the occurrence and development of immune-related diseases. However, excessive activation of this pathway in inflammatory lung diseases may lead to chronic inflammation and tissue damage, thereby promoting disease progression. Research has indicated that cGAS converts ATP and GTP into cGAMP, activating the STING pathway, which triggers the production of type I interferons ( IFN) and participates in the immune response, cellular senescence, and inflammation processes. This review systematically discusses the role of the cGAS-STING signaling pathway in inflammatory pulmonary diseases such as chronic obstructive pulmonary disease (COPD), asthma, and pulmonary fibrosis, as well as the latest advancements in related drug research. The latest progress in the study of cGAS-STING-related inhibitors is reviewed. This review provides a reference for further studies of the pathogenesis of inflammatory pulmonary diseases and targeting therapeutic strategies, offering new perspectives for future research.

Abstract:Breast cancer is one of the most common cancers in women. Chemotherapy, as an important part of its comprehensive treatment, often induces a variety of adverse reactions, among which cancer-related fatigue (CRF) is one of the most common ones. At present, the pathogenesis of CRF remains unclear, and there is a lack of reliable diagnosis and treatment method. Intestinal barrier dysfunction is considered to be one of the important pathogenic mechanisms of CRF, while probiotics can alleviate CRF symptoms by repairing the intestinal barrier,regulating immunity and inflammatory responses. This review summarizes the research progress of probiotic intervention in CRF of breast cancer patients undergoing postoperative chemotherapy, aiming to provide new ideas for clinical adjuvant therapy.