Abstract: Objective　To investigate the effect of KH-type splicing regulatory protein (KHSRP) on the malignant biological behavior of lung adenocarcinoma (LUAD) by targeting the Janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3) signaling axis. Methods　Clinical data were collected for 64 patients with LUAD, diagnosed at Huaihe Hospital from January 2017 to December 2018. Expression levels of KHSRP were detected in LUAD tissues and adjacent tissues by immunohistochemical staining. KHSRP gene expression was also detected in LUAD cell lines (SPC-A1, H1975, CL1-5, PC-9, Calu-3, H446) and normal human bronchial epithelial cells using quantitative reverse transcription-polymerase chain reaction. KHSRP expression in SPC-A1, H1975, PC-9, and Calu-3 cells was manipulated by lentivirus transfection. The effects of KHSRP on the proliferation, migration, and invasion of LUAD cells were detected by Cell Counting Kit-8 and Transwell assays. The effects of KHSRP overexpression and knockdown were also investigated in a mouse xenograft tumor model, and JAK/ STAT signaling pathway proteins were detected by Western blot. Rescue experiments were conducted to verify if KHSRP promoted the malignant progression of LUAD cells by regulating the JAK1/STAT3 signaling pathway. Results　KHSRP expression was significantly higher in LUAD tissues compared with adjacent tissues (P<0.05). Overexpression of KHSRP significantly promoted the proliferation, migration, and invasion of LUAD cells in vitro (P<0.05). KHSRP also promoted LUAD cell xenograft tumor growth and lung nodule metastasis in nude mice in vivo (P<0.01). KHSRP knockdown significantly decreased the levels of JAK1, phospho-JAK1, and STAT3 in the JAK/ STAT signaling pathway, while the situation was reversed following KHSRP overexpression (P<0.05). Rescue experiments showed that KHSRP reversed the inhibitory effect of knockdown (P<0.05). Conclusions KHSRP targets the JAK1/STAT3 signaling pathway and acts as an oncogene in LUAD.

Abstract: Objective　To establish an efficient and stable model of pelvic inflammatory disease in rats via a non-surgical method, and to evaluate its application in pharmacodynamic testing. Methods　Female Sprague-Dawley rats were divided randomly into the following groups: control group; model group with phenol for 7 d; model group with phenol for 10 d; treatment group modeled with phenol; model group with low concentration of bacteria; model group with high concentration of bacteria; and treatment group modeled with bacteria. Rats in the model and treatment groups were injected with 25% phenol gel and 2×10⁷or 2×10⁸ Escherichia coli and Staphylococcus aureus mixture via a non-surgical method, to construct a rat model of pelvic inflammatory disease. Rats in the treatment groups received the Chinese patent medicine Jingangteng capsules by gavage, and rats in the control group received the same volume of solvent solution. The health status, weight changes, and uterine appearance were monitored and the uterine coefficient was calculated. Pathological changes in the uterus and fallopian tubes, endometrial thickness, and number of glands were detected by hematoxylin and eosin staining. Serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay. Protein expression of the macrophage marker CD68 was detected by immunofluorescence. Expression of Toll-like receptor 4 (TLR4)/nuclear factor (NF)-κB pathway-related proteins in the uterus was detected by Western blot. Results　The mortality rate in the model group was only 5%. Compared with the control group, model rats showed decreased body weight, increased uterine coefficient, pathological changes in the uterus and Fallopian tubes, thinner endometrium, fewer glands, significantly higher serum levels of IL-1β, IL-6, and TNF-α and more macrophages in the uterine tissue, and activation of the TLR4/NF-κB signaling pathway. The 7 d phenol and low-concentration bacterial solution models were judged to be mild pelvic inflammatory disease models, and the 10 d phenol and high-concentration bacterial solution models were considered severe pelvic inflammatory disease models. Treatment with Jingangteng capsules relieved the pathological symptoms in the uterus and fallopian tubes, in line with the efficacy evaluation of clinical pelvic inflammatory disease. Conclusions　We established rat models of pelvic inflammatory disease using phenol and a mixed bacterial solution via a non-surgical method, to simulate the different pathological states of pelvic inflammatory disease caused by different factors. These models will be suitable for evaluating drug efficacy and elucidating the pathological mechanism of pelvic inflammatory disease.

Abstract: Objective　To establish an animal model of hand, foot, and mouth disease (HFMD) in Syrian hamsters coinfected with coxsackievirus A6 (CVA6) and coxsackievirus B1 (CVB1). Methods　42 Syrian hamsters were divided into a CVA6 infection group, CVB1 infection group, CVA6 and CVB1 coinfection group and control group. A HFMD model was established by nasal instillation of virus solution and phosphate-buffered saline. Clinical and physiological indicators and detoxification status were monitored and recorded for 15 d, and animals were selected on day 7(D7) after infection for histopathology and viral antigen and nucleic acid testing. Results　Hamsters in the single-infection and coinfection groups showed clinical symptoms similar to human HFMD. White blood cell, neutrophil, and lymphocyte result were characteristic of viral infection. Both viral nucleic acids were detected in throat swabs, feces, blood, and tissues and both viruses were isolated from fecal samples. Pathological damage and positive co-localization of CVA6 and CVB1 viral antigen proteins and nucleic acids were found in brain and other tissues. Conclusions　Nasal instillation of a CVA6 and CVB1 mixture can successfully coinfect Syrian hamsters, replicate herpes infection similar to human HFMD, and cause pathological viral myocarditis and encephalitis damage. The result showed that the coinfection group was more seriously affected than the single-infection group, with worse clinical symptoms, increased viral replication, and obvious tissue pathological damage. This study provides a reference for further basic and clinical research into human enterovirus coinfection.

Abstract: Objective　To explore the effect and mechanism of lentinan (LNT)on hepatic tissue ferroptosis in mice exposed to sodium arsenite (SA). Methods　C57BL/6N male mice were exposed to SA low-dose, SA highdose, and LNT intervention combined with SA high-dose, then, hematoxylin and eosin(HE)staining was applied to assess pathological liver tissue damage; Enzyme-linked immunosorbent and Western blot were used to detect the content or expression of tumor necrosis factor α (TNFα), interleukin-6 (IL-6), ferritinophagy or ferroptosis biomarkers. Results　Compared with the control group, SA exposure induced the elevated levels of TNFα, IL-6, ferritinophagy biomarker ferritin heavy chain 1(FTH1)and microtubule-associated protein 1 light chain 3B (MAP1LC3B)in mice liver tissue, while levels the ferroptosis biomarker GPX4 decreased(P<0.05). Compared with SA high-dose groups, LNT intervention showed the reduced pathological liver damage and the downregulated levels of TNFα, IL-6, FTH1, and MAP1LC3B, while the level of GPX4 upregulated(P<0.05). Western blot experiment showed that LNT intervention antagonized the upregulated levels of FTH1, and autophagy biomarker LC3B/A, and antagonized the increased co-expressions of FTH1 with LC3B or Ub protein in SA high-dose group(P<0.05). Conclusions　LNT antagonizes SA-exposed hepatic pathological injury and ferroptosis in mice, possibly associated with inhibition of TNFα-ferritinophagy signaling.

Abstract: Objective　To investigate the effects of Yishen Zhuyu Tongluo Decoction on chondrocyte pyroptosis and ferroptosis in rats with knee osteoarthritis (KOA). Methods　Forty SD were divided randomly into a sham operation group (n=10) and a modeling group (n=30). A KOA model was established in the modeling group by anterior cruciate ligament transection of the right hind limb knee joint. The drawer test was used to confirm the successful establishment of the model. Post-surgery, the rats were subjected to 30 min of forced activity daily to induce KOA. Lameness and hopping movements were observed after 4 weeks, and hematoxylin and eosin staining confirmed cartilage surface damage, deformation, and inflammatory cell infiltration, indicating successful modeling. The model rats were then assigned randomly to a model group (n=8), celecoxib group (n=8), and Yishen Zhuyu Tongluo Decoction group (n=8). The sham operation and model groups received 10 mL/kg of saline by gavage, the celecoxib group received 12 mg/kg of celecoxib solution, and the Yishen Zhuyu Tongluo Decoction group received 5.4 g/kg of the herbal decoction, once daily for 8 weeks. After the interventions, the rats were anesthetized, blood was collected from the abdominal aorta and serum was separated, and the knee joints were isolated. Three samples were fixed in paraformaldehyde, while the remaining cartilage tissue was reserved. Pathological changes in joint cartilage were observed by hematoxylin and eosin and Safranin O-Fast Green staining. Bone microstructure was analyzed using micro-computed tomography. Serum interleukin (IL)-1β and IL-18 protein levels were detected by enzyme-linked immunosorbent assay. Relative mRNA expression levels ofNLRP3, ASC, Caspase-1, GSDMD, IL-1β, IL-18, ACSL4, FTH-1, GPX-4, and cyclooxygenase (COX)-2 were detected by reverse transcription quantitative real-time PCR, and protein expression of NLRP3, Caspase-1, and COX-2 were detected by immunohistochemistry. Results Compared with the sham operation group, model rats showed surface damage and deformation of cartilage tissue, disordered cell arrangement in all layers, significant loss of Safranin O-Fast Green staining, and sparse and irregular trabecular bone distribution. Serum levels of the inflammatory factors IL-1β and IL-18 were elevated (P<0.01). mRNA expression levels of NLRP3, ASC, Caspase-1, GSDMD, IL-1β, IL-18, ACSL-4, and COX-2 were also significantly increased (all P<0.01), while FTH-1 and GPX-4 mRNA expression levels were decreased (P<0.01). Compared with the model group, rats in the celecoxib and Yishen Zhuyu Tongluo Decoction groups showed smoother and more intact cartilage surfaces, significantly increased cell counts, less loss of Safranin O-Fast Green staining, denser trabecular bone, and thicker cortical bone, with improved bone microstructure. Serum IL-1β and IL-18 levels were reduced (P<0.01), NLRP3 (P<0.01, P<0.05), ASC, Capase-1, GSDMD, IL-1β, IL-18, ACSL-4, and COX-2 mRNA levels were decreased (all P<0.01), and relative expression levels of FTH-1 and GPX-4 mRNA were significantly increased (P<0.01). Conclusions　Yishen Zhuyu Tongluo Decoction can treat KOA by inhibiting chondroptosis and ferroptosis in chondrocytes, reducing serum IL-1β and IL-18 levels, and improving bone microstructure.

Abstract: Objective　To explore the molecular mechanism of autophagy mediated by the protein kinase B (AKT)/mammalian target protein of rapamycin (mTOR) pathway in the rehabilitation of muscle atrophy associated with rotator cuff tears (RCTs). Methods　Forty male C57BL/6J mice were randomly assigned to the following four groups: sham group, RCTs group, RCTs + exercise group, and RCTs + exercise + rapamycin group, with 10 mice in each group. On the eighth week after grouping, healing of the bone-tendon interface and muscle cell atrophy were analyzed by histology. The mRNA expression levels of muscle-atrophy-related genes (Atrogin-1, Bnip 3, MuRF-1) in supraspinatus muscle tissue were measured by real-time quantitative reverse transcription polymerase chain reaction. The expression of LC3 and AKT/mTOR signal pathway proteins in the supraspinatus muscle tissue of the groups was detected by Western blot, and the degree of autophagy in each group was analyzed by transmission electron microscope. Results　Compared with the sham operation group, in RCTs group’s maturity score for the bone-tendon interface at the supraspinatus tendon anchorage and the cross-sectional area of the supraspinatus muscle fibers decreased significantly (P<0.001), while muscle loss and the expression of Atrogin-1, Bnip 3, and MuRF-1 increased significantly (P<0.001). Compared with the RCTs group, the RCTs + exercise group showed a significant increase in bone-tendon interface maturity score and cross-sectional area of the supraspinatus muscle fibers (P<0.01) and a decrease in muscle loss and the expression of Atrogin-1, Bnip 3, and MuRF-1 (P<0.01). Compared with the sham group, the RCTs group’s LC3Ⅰ/LC3Ⅱ and degree of autophagy in the supraspinatus muscle increased significantly (P<0.001), while p-AKT/AKT and p-mTOR/mTOR expression decreased significantly (P<0.01). Compared with RCTs group, the RCTs + exercise group’s LC3Ⅰ/LC3Ⅱ and degree of autophagy decreased significantly (P<0.01) and p-AKT/AKT and p-mTOR/mTOR expression increased significantly (P<0.001). The addition of rapamycin significantly reversed the rehabilitation effect of exercise in the RCTs group. Conclusions　This study confirmed the anti-atrophy effect of exercise rehabilitation in RCT diseases and showed that its mechanism is related to AKT/mTOR signal activation, which inhibits autophagy.

Abstract: Objective　“Lie flat” has recently become a buzz word describing a specific psychological state in some individuals. However, its psychological meaning and mechanisms remain unclear. Lack of motivation undercertain conditions may be one of the key psychological characteristics of this condition. Methods　Sixteen male SD rats were randomly assigned to two food-restriction levels (90% and 80% of baseline weight, respectively), and subjected to a sucrose-pellet self-administration task.The establishment of the model is divided into three stages. The rats were trained to self-administer sucrose pellets in a high-reward learning stage and in a low-reward learning stage. We then assessed “lie flat” behavior in a preference-test stage, in which rats could choose between high-reward- and low-reward-paired nose-pokes in a discrete choice procedure. Results　Combining self-feeding training with Western blot, the Results showed that: (1)rats showed significantly decreased nose-poke behavior for low-reward, i.e. “lie flat” behavior, but not for high-reward, when ≥90% of the maximal reward was obtained in the high-reward task and the effort (X) in the low-reward task increased to the breaking point. (2)Dopamine D1R expression in the NAc was significantly higher in the “lie flat” group compared with the “not lie flat” rats, while D2R expression was similar in both groups. Conclusions　Rats can show “lie flat” behavior , and up-regulation of D1R expression in the NAc may be a key part of the molecular basis responsible for this motivation deficiency in “lie flat” behavior. These Results extend our understanding of “lie flat” behavior, and provide a new paradigm for the study of its mechanism.

Abstract: Objective　To examine the effect of the demethylating agent 5-aza-2’-deoxycytidine (5-AzaCdR)on pericyte-myofibroblast transition (PMT)in primary rat renal myofibroblasts. Methods　Rat primary renal myofibroblasts were treated with 5-Aza-CdR 250 ng/mL for 72 h, and the degree of Epo promoter methylation was detected by pyrosequencing. Protein expression levels of α-smooth muscle actin (α-SMA), platelet-derived growth factor receptor-β (PDGFRβ)and DNA methyltransferase (Dnmt3a)were detected by immunofluorescence and Western blot, and erythropoietin (EPO)levels in the supernatant were detected. Results　Compared with the control group, 5-Aza-CdR treatment significantly decreased the expression of Dnmt3a and hypermethylation of the Epo promoter, and subsequently decreased the expression of α-SMA and the expression ratio of α-SMA to PDGFRβ in myofibroblasts. Meanwhile, 5-Aza-CdR treatment increased the level of EPO in the cell supernatant. Conclusions 5-Aza-CdR can reverse PMT by inhibiting Epo promoter hypermethylation in primary renal myofibroblasts.

Abstract: Objective　To establish an efficient DNA extraction method for the batch identification of genotypes in model mice. Methods　We extracted total DNA from transgenic mouse tails using four Methods: alkaline simplified group, alkaline routine group, protease K cleavage group and DNA extraction kit group. The purity and concentration of DNA obtained by the four Methods were measured, the effects of gel electrophoresis were evaluated, and the time and experimental costs of the four Methods were compared. Results　The protease K cleavage method produced the highest concentration of DNA, followed by the simplified alkaline boiling and routine alkaline boiling method. The reagent kit produced the highest DNA purity, followed by the simplified alkaline boiling method. The DNA templates obtained by the four method could be amplified by polymerase chain reaction and gel electrophoresis to obtain clear DNA target bands. In addition, the DNA template extracted by the simplified alkaline boiling method could be used for gene identification after storage at -20℃ for 1 month, as well as requiring the least time and lowest costs. Conclusions　The simplified alkaline boiling method is currently the simplest, fastest, and most economical DNA template-extraction method for batch identification of genotypes in model mice.

Abstract:Pollen allergen-induced allergic rhinitis (AR), also known as seasonal allergic rhinitis(SAR), typically manifests during the period of pollen dissemination by anemophilous plants. The prevalence of SAR has more than doubled over the past three decades. The etiology of SAR is multifaceted, involving factors such as pollen allergens, environmental and climatic conditions, genetic predispositions, and the immunological status of the individual. Animal models provide a critical tool for elucidating the mechanisms underlying AR and advancing the development of effective preventive and therapeutic strategies. This review synthesizes the recent pertinent domestic and international literature on pollen-sensitized AR animal experiments. It systematically delineates the factors influencing the efficacy of these models, including the selection of animal strains, the production and associated challenges of sensitizing agents, specifically pollen antigens, the utilization and limitations of adjuvants, the procedural steps involved in model creation, and the method ologies for evaluating model effectiveness. The insights provided are intended to offer guidance and support for the development of appropriate animal models of pollen-induced AR, thereby facilitating both fundamental and applied research in this area.

Abstract:A ketogenic diet(KD) refers to an eating pattern designed to achieve a low-calorie content, minimum carbohydrate intake, high-fat consumption, and standard protein levels. A ketogenic diet is used in clinical practice to treat conditions including heart disease, diabetes, obesity, autism, glioblastoma, and other cancers. Although a ketogenic diet has not been recommended for any neurological disorders except epilepsy, extensive recent research suggests that such a diet may have a neuroprotective effect and may thus represent a new dietary therapy for the treatment of Parkinson’s disease(PD). In this review, we discuss in detail the mechanisms responsible for the neuroprotective effects of a ketogenic diet in Parkinson’s disease, with the aim of providing references for future clinical and experimental studies.

Abstract:Mitochondria act as the main energy supply station for cardiomyocytes and are thus crucial for maintaining normal cardiac function. Mitochondrial autophagy plays an important positive role in maintaining cardiomyocyte homeostasis and coping with stress. The progressive exacerbation of cardiovascular diseases presents a challenge to the homeostasis of mitochondrial autophagy through as-yet-unidentified pathological mechanisms, leading to mitochondrial damage, which may in turn trigger damage to cardiomyocytes. In addition, when mitochondrial autophagy fails to meet the physiological needs of the body, mitochondrial dysfunction may be triggered, which may in turn accelerate the progression of heart failure. In this review, we explore the specific roles of mitochondrial autophagy and mitochondrial dynamics in the heart, and discuss the mechanisms in the context of major cardiovascular diseases, focusing on the latest advances and important discoveries in this field.

Abstract:In line with the rising global incidence of cancer, tumor metastasis has become the leading cause of mortality among cancer patients. Formation of the pre-metastatic niche (PMN) creates an optimal microenvironment for tumor cell colonization and metastasis, thus facilitating tumor dissemination. In this context, cancer-associated fibroblasts (CAFs)have garnered significant attention as a result of their multifaceted roles in tumor progression. CAFs enhance the formation of the PMN and promote the invasive behavior of tumor cells by remodeling the extracellular matrix, inducing epithelial-mesenchymal transition, stimulating angiogenesis and modulating the tumor immune microenvironment. These processes not only expedite metastasis but also enable tumor immune evasion. This paper reviews the fundamental mechanisms and functions of CAFs in tumor metastasis, examines their potential applications in cancer research and therapy, and offers new perspectives and directions for the development of antitumor metastasis strategies.

Abstract:Endometriosis presents a common challenge in global women’s health. Ferroptosis, as an emerging form of regulated cell death, has recently attracted attention in relation to endometriosis. In terms of disease mechanisms, ferroptosis drives ovarian endometrial fibrosis via the induction of iron overload, and promotes disease progression by regulating multiple signaling pathways, such as p38 mitogen-activated protein kinase/signal transducer and activator of transcription 6(p38MAPK/STAT6), promote angiogenesis,autophagy,etc,fueling disease progression. In terms of infertility, ferroptosis affects embryo development and ovarian function, thereby reducing fertility. From a diagnostic perspective, high expression of ferroptosis-related genes provides potential biomarkers for the early diagnosis of endometriosis, effectively distinguishing patients from healthy individuals, and showing important clinical value. In terms of treatment strategies, non-natural compounds such as Erastin, as well as natural compounds such as resveratrol, ursolic acid, and baicalein, have shown potential therapeutic effects in relieving the pathological process and related symptoms of endometriosis. This review comprehensively elucidates the key role of ferroptosis in endometriosis in terms of the disease mechanisms, infertility, diagnosis, and treatment, and explores its potential as a diagnostic biomarker and therapeutic target, thus providing new theoretical support and treatment strategies for the management of endometriosis.

Abstract:Pulmonary hypertension (PH) is a fatal disease characterized by pulmonary vascular remodeling, ultimately leading to right heart failure and death. Current treatments for PH are suboptimal, with no substantial improvement in overall survival among patients with advanced PH. Despite some progress in understanding the pathogenesis of PH, further studies at the molecular level are needed to develop more effective treatments for PH. Recent research has shown that long non-coding RNAs (lncRNAs) have an important regulatory function in the pathophysiological process of PH, and may thus be potential disease biomarkers and therapeutic targets. In this paper, we review recent progress in our understanding of the molecular mechanisms of lncRNAs in PH.

Abstract:Bone defect repair is an urgent problem in the field of orthopedics, and numerous researchers are working to develop more effective treatment plans. The accurate evaluation of bone repair after surgery is a crucial step. In line with the development of computed tomography (CT) imaging, dual-energy CT imaging has shown significant advantages in analyzing bone composition and reducing metal artifacts. This article reviews the application of dual-energy CT imaging for the evaluation of bone repair in animals.

Abstract:Colorectal cancer (CRC) is one of the most common malignant life-threatening tumors, with serious impacts on patient quality of life. Src homology 2 domain-containing protein tyrosine phosphatase (SHP2) has recently become a hot topic in the field of cancer research, and has demonstrated a close relationship with CRC. SHP2, encoded by the PTPN11 gene, is a non-receptor tyrosine kinase commonly present in various tissues and cells of the human body. Existing research shows that SHP2 plays a crucial role in regulating CRC and colitis-associated colon cancer (CAC), and the emergence of SHP2 allosteric inhibitors has identified SHP2 as a potential new therapeutic target for patients with CRC. Here we review the structure of SHP2 and its roles in CRC and CAC.

Abstract:Macrophages are important immune cells involved in innate immunity, with significant heterogeneity and polarization. Macrophages can polarize into different phenotypes (mainly M1 and M2) under stimulation by various factors in the microenvironment, leading to different roles and functions. Macrophages play an important role in the pathophysiological mechanism of renal ischemia-reperfusion injury (RIRI). An excessive immune response will inevitably lead to tissue damage. M1 macrophages are pro-inflammatory cells involved in the clearance of pathogens, while M2-type macrophages have anti-inflammatory effects and participate in the repair and remodeling of renal tissue after RIRI. The balance between these macrophage phenotypes is thus an important factor affecting the outcome and treatment of RIRI. This review considers the pathophysiologic mechanism of macrophages in RIRI and the latest treatments from the perspective of macrophage polarization, with the aim of supporting further studies of the function of macrophage polarization in RIRI and adjusting the macrophage polarization process to improve RIRI treatment strategies.