The development of tailor-made tumor vaccines targeting specific tumor antigens is crucial for improving treatment accuracy. Tumor mRNA, as an emerging and promising vaccine modality, holds distinct technical edge in addressing this challenge, thus attracting considerable attention in cancer treatment. This article reviews the design and synthesis processes of tumor mRNA vaccines, highlighting the latest advances in antigen identification, optimization of RNA sequence and structure, and its delivery. Additionally, it discusses the challenges these vaccines face and outlines potential future directions for development.

Homologous recombination is a crucial biological event during meiosis, and its initial step involves the formation of programmed DNA double-strand break (DSB). However, for a long time, how to reconstitute the process of meiotic DSB formation in vitro remained an unresolved challenge in the scientific community. Recently, scientists have achieved a breakthrough. For the first time, they successfully reconstructed the formation of meiotic DSB in vitro. The findings reveal that weak dimerization is a key characteristic of the SPO11 (sporulation protein11)-TOP6BL (TOP6B like) core complex. In vivo, the activity of SPO11 may be limited by its intrinsically weak dimerization ability. This discovery suggests that the dimerization process of SPO11 plays a vital role in regulating meiotic DSB formation. This achievement not only fills a gap in related research fields but also provides critical insights into the molecular mechanism of meiotic DSB formation. Moreover, it offers important directions and implications for future studies in this field.

Osteosarcoma is a highly aggressive primary bone tumor that predominantly affects children and adolescents. Conventional treatment modalities are often hindered by issues such as high recurrence rates and the development of drug resistance. Proteolysis-Targeting Chimeras (PROTACs) represent an innovative therapeutic strategy for osteosarcoma, which are designed to selectively degrade pathogenic proteins. This article offers a comprehensive analysis of the mechanisms underlying PROTACs, their targeted applications in osteosarcoma—including bromodomain and ultra-terminal domain targeting—and the associated challenges, such as delivery efficiency and off-target toxicity. Additionally, it suggests optimization strategies for bone-targeted delivery (such as using DNA nanoflowers as carriers) and combination therapies (like targeted therapy and chemotherapy), informed by recent research findings, thereby providing a theoretical framework for clinical translation.

Occult hepatitis B virus (HBV) infection (OBI) represents a potential reservoir for HBV transmission, capable of spreading through routes such as blood transfusion. Additionally, OBI can contribute to the chronic progression of hepatitis B-related diseases, sustaining a state of chronic HBV infection. In individuals with compromised immune function or undergoing immunosuppressive therapy, OBI may lead to HBV re-activation, potentially triggering severe liver conditions such as acute hepatitis or liver failure. As a result, OBI poses a significant public health challenge, profoundly impacting the health and well-being of affected populations and complicating HBV infection control efforts in China. Clinically diagnosing OBI remains challenging, but its hallmark is serum hepatitis B surface antigen negative and the presence of HBV covalently closed circular DNA (cccDNA) in the liver. With the increasing focus on achieving functional cure for chronic hepatitis B, both domestic and international guidelines have refined functional cure. Notably, these guidelines acknowledge that cccDNA may persist in the liver tissue of individuals who have achieved functional cure, suggesting resemblance of an occult infection state. Here, we provide a comprehensive overview of OBI, including its definition, classification, public health implications, underlying mechanisms, and clinical reactivation. By updating the understanding of OBI, we aim to raise awareness among clinicians and public health professionals regarding the significance of OBI in the current context and encourage greater attention to this population.

With the continuous increase in global obesity prevalence, the impact of obesity on reproductive physiology has garnered widespread societal attention. As a metabolic disorder, obesity is typically accompanied by multiple abnormal physiological phenomena, such as excessive adipose accumulation and exacerbated inflammatory responses, which severely compromise the reproductive health of humans and animals. Reproductive damage induced by obesity involves a series of complex biochemical reactions and in vivo metabolic pathways, manifesting as impaired male sperm quality and female fertility. To better understand the relationship between obesity and reproductive physiology, this review summarizes the reproductive injuries caused by obesity and their underlying mechanisms. In the obese state, conditions such as oxidative stress, insulin resistance, and hyperinsulinemia are induced, with adipokines (leptin, adiponectin, resistin, etc.) and inflammatory factors (TNF-α, IL-6, IL-1β, etc.) interacting synergistically to affect the reproductive system. Oxidative stress activates the MAPK and NF-κB pathways, interfering with insulin signaling, while chronic inflammation leads to adipocyte secretory disorders and disrupts the hypothalamic-pituitary-gonadal regulatory axis. Studies have shown that obese males exhibit significantly decreased testosterone levels and impaired sperm quality, whereas obese females suffer from reproductive hormone imbalance, ovulation disorders, and polycystic ovary syndrome. This review discusses how obesity-induced metabolic disorders lead to impaired reproductive physiology in both males and females, along with the underlying mechanisms, providing a theoretical basis for the prevention and treatment of obesity-related reproductive disorders in the future.

Ferroptosis, a novel form of programmed cell death driven by iron-dependent lipid peroxidation, plays a crucial role in both disease treatment and microbial control due to its multi-level regulatory mechanisms. It mainly involves iron metabolism, lipid peroxidation, and antioxidant systems. In the field of disease treatment, ferroptosis is regarded as a highly promising therapeutic target because of its key role in autoimmune diseases, cancer, and cardiovascular diseases. This review systematically summarizes the core regulatory factors of ferroptosis, such as glutathione peroxidase 4 (GPX4) and long-chain acyl-CoA synthetase 4 (ACSL4) and their interaction networks, and deeply explores the application prospects of targeted intervention strategies based on ferroptosis signaling pathways in disease treatment and microbial control. Additionally, we also summarize the current issues faced by ferroptosis in practical applications and proposes strategies, such as nanodelivery, improved drug chemical stability and enhanced water solubility to optimize therapeutic efficacy. We aim to provide a theoretical basis and practical guidelines for exploring more targeted treatments using ferroptosis.

Gastric cancer remains one of the most prevalent and lethal malignancies of the digestive tract worldwide, underscoring the urgent need for more effective targeted therapeutic strategies. Poly (ADP-ribose) polymerase (PARP) inhibitors have demonstrated remarkable efficacy in tumors with homologous recombination repair (HRR) deficiency; however, their clinical application in gastric cancer remains limited. Clinical evidence suggests that patients harboring Helicobacter pylori infection in combination with HRR gene mutations exhibit a significantly elevated risk of developing gastric cancer, thereby supporting the potential benefit of PARP inhibition in this setting. In this study, a kinase inhibitor library was screened in combination with the PARP inhibitor olaparib in gastric cancer cells. And we identify the cyclin-dependent kinase 8/19 (CDK8/19) inhibitor Senexin A as a compound that synergistically enhances the cytotoxic effect of PARP inhibition (P< 0.05). Phenotypic validation using CCK-8 and colony formation assays demonstrated that the combination treatment significantly suppressed cellular proliferation and clonogenic potential compared to either monotherapy (P< 0.0001). Mechanistically, alkaline comet assays revealed a significant increase in DNA damage in the combination treatment group relative to either single-agent group (P < 0.0001), suggesting that the synergistic effect results from the exacerbation of DNA damage via impaired DNA repair mechanisms. In addition, treatment with CDK8/19 inhibitors alone markedly increased the formation of γH2AX and 53BP1 foci in irradiated gastric cancer cells (P< 0.0001), indicating inhibition of DNA damage repair pathways. Transcriptome sequencing further revealed that CDK8/19 inhibition impacts critical cellular pathways, including DNA repair, cell cycle regulation, and RNA splicing. Co-immunoprecipitation assays confirmed that inhibition of CDK8/19 kinase activity significantly reduces the phosphorylation level of PARP1, suggesting a potential regulatory interaction. Immunohistochemical analysis of tumor and adjacent non-tumor tissues from gastric cancer patients demonstrated that CDK8 is significantly overexpressed in tumor tissues, supporting its potential as both a prognostic biomarker and a therapeutic target. Collectively, this study elucidates a mechanistic basis by which CDK8/19 inhibition enhances the sensitivity of gastric cancer cells to PARP inhibitors. These findings provide a strong rationale for the combined use of CDK8/19 and PARP inhibitors as a targeted therapeutic strategy and offer promising translational implications for advancing personalized medicine in gastric cancer treatment.

Fucoidan (FUC) is a natural seaweed-derived drug. Previously, our experiments have shown that FUC can significantly inhibit the cell viability of human osteosarcoma 143B cells and induce cell death, but the mechanism remains unclear. Ferroptosis, a novel form of cell death, has emerged as an important target for tumor therapy. This study aims to investigate whether FUC induces ferroptosis of 143B cells and elucidate its underlying molecular mechanisms. CCK-8 and LDH assays result showed that FUC (10, 100, 400 μg/mL) significantly reduced cell viability of 143B cells and induced cell death. Calcein-AM staining, FeRhoNox-1 staining, and C11 BODIPY 581/591 staining indicated that FUC obviously increased the levels of labile iron pool (LIP), Fe^2＋, and lipid reactive oxygen species (Lip ROS) in 143B cells. Chemical colorimetric analysis revealed that FUC markedly decreased intracellular Glutathione (GSH) contents. Real-time quantitative PCR showed that FUC dramatically reduced the mRNA levels of ferroptosis-related factors solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4), while increasing the mRNA levels of prostaglandin endoperoxide synthase 2 (PTGS2) and acyl-CoA synthetase long-chain family member 4 (ACSL4). Western blotting analysis demonstrated that FUC significantly reduced the protein levels of SLC7A11 and GPX4, and the ratios of p-PI3K/PI3K, p-AktSer473/Akt, and p-AktThr308/Akt, but increased the protein level of ACSL4. Immunofluorescence staining showed that FUC obviously inhibited the nuclear translocation of p-AktSer473. The ferroptosis inhibitor ferrostatin-1 (Fer-1) and iron chelator deferoxamine (DFO) remarkably suppressed cell death induced by FUC in 143B cells. Additionally, the PI3K/Akt pathway activator 740Y-P significantly inhibited FUC-induced iron overload and lipid peroxidation in 143B cells, and restored the protein levels of SLC7A11 and GPX4. In conclusion, FUC can induce ferroptosis of 143B cells by inhibiting the PI3K/Akt signaling pathway, which may be a potential target for the prevention and treatment of osteosarcoma.

Breast cancer (BRCA) remains one of the leading causes of cancer-related deaths worldwide due to its high rates of metastasis and recurrence, making it crucial to explore its underlying molecular mechanisms. Our previous study demonstrated that miR-326 inhibits BRCA progression by targetingEPH receptor B3(EPHB3). This study further explores the molecular mechanism by which long non-coding RNAs (LncRNAs) regulates BRCA progression via the competing endogenous RNA (ceRNA) mechanism, in which it competes with EPHB3 for miR-326 binding. Bioinformatics analysis identified LncRNA Small Nucleolar RNA Host Gene 12 (SNHG12) as a potential miR-326-binding molecule. SNHG12 was found to be significantly upregulated in BRCA tissues, exhibiting a negative correlation trend with miR-326 and a positive correlation trend with EPHB3, suggesting its potential involvement in the ceRNA regulatory network. Nuclear-cytoplasmic fractionation assays revealed cytoplasmic localization of SNHG12, while dual-luciferase reporter assays confirmed its direct binding to miR-326. Functional experiments demonstrated that SNHG12 knockdown significantly suppressed BRCA cell proliferation, invasion, and migration, while miR-326 inhibition reversed these effects. Furthermore, miRNA pulldown assay revealed significant enrichment of SNHG12 and EPHB3 in the miR-326 pulldown products, indicating direct binding between them. Western blotting and rescue experiments revealed that SNHG12 upregulates EPHB3 expression by sponging miR-326, thereby promoting the malignant behaviors of BRCA cells. Collectively, this study revealed that LncRNA SNHG12 promotes BRCA progression by regulating the miR-326/EPHB3 axis through a ceRNA mechanism. The SNHG12/miR-326/EPHB3 pathway may represent a promising target for the molecular diagnosis and targeted therapy of BRCA.

Diabetes-related cognitive impairment (DCI) is a major complication of type 2 diabetes mellitus (T2DM). Although exercise is essential in alleviating DCI, the underlying mechanisms remain unclear. The aim of this study is to investigate the role and mechanism of exosomal miR-126a-5p induced by exercise in ameliorating DCI. Twenty-four 16-week-old male db/db mice were randomly divided into diabetes group (n=12; DM) and exercise intervention group (n=12; DE). The control group consisted of male m/m mice of the same age group (n=12; CON). The DE group underwent 8 weeks of moderate intensity treadmill training (10 m/min, 5 days a week). In the MWM experiment, compared to the CON group, the DM group exhibited prolonged escape latency (P<0.01), reduced swimming speed and target quadrant time (P<0.001), and decreased expression of miR-126a-5p and EX-miR-126a-5p in hippocampal tissue (P<0.001). After exercise intervention, the DE group showed improved performance with decreased escape latency (P<0.05), increased swimming speed and target quadrant time (P<0.05), and elevated levels of exosomal miR-126a-5p (P<0.001). Morphological staining revealed a decrease in the expression and proportion of NeuN in hippocampal neurons and an increase in the expression and proportion of glial cells in the CA1 and CA3 regions of DM group mice compared to CON group mice (P<0.05), while DE group mice showed increased fluorescence intensity and proportion of neurons (P<0.05). Western blotting analysis revealed that the DM group also showed significant upregulation of amyloid β (Aβ), high mobility group box 1 (Hmgb1), and NF-κB in the hippocampus (P<0.05), which were reduced after exercise (P<0.05). Moreover, exosomal miR-126a-5p overexpression greatly decreased the levels of Hmgb1, NF-κB, and amyloid precursor protein (APP) in HT22 cells and TNF-α, IL-1β in supernatant exposed to HG (P<0.05), while inhibition of miR-126a-5p led to increased levels of these proteins (P<0.05). In conclusion, eight weeks of treadmill exercise improved cognitive function in db/db mice, likely through the EXs-miR-126/HMGB1/NF-κB pathway to reduce inflammation in hippocampal tissue.

Pathological cardiac hypertrophy is an early and significant cardiac structural characteristic that contributes to the onset and progression of heart failure (HF). Its mainly structural feature is the abnormally enlarged cardiomyocyte. Effective intervention targets for abnormally enlarged cardiomyocyte remain to be identified. Previous studies have shown that the cellular shape and size can be regulated by the actin related protein 2/3 (Arp2/3) complex, which is an actin-binding protein complex involved in the actin nucleation and assembly. However, the roles of the Arp2/3 complex in cardiomyocyte hypertrophy remain unknown. Here our study identifies its novel roles in the occurrence and development of cardiomyocyte hypertrophy. We found that mRNA levels of all subunits from the Arp2/3 complex are significantly upregulated (P<0.05) in the angiotensin II (Ang Ⅱ)-induced neonatal rat primary and H9c2 cardiomyocyte hypertrophy. Further studies showed that siRNA-directed ARPC2 silencing inhibits the reactivation of fetal genes and enlargement of cardiomyocyte area induced by Ang II in neonatal rat primary cardiomyocytes (NRCMs) and H9c2 cells (P<0.05). In addition, the upstream activators of the Arp2/3 complex including SH3 protein interacting with Nck, 90 kD (SPIN90) and Ras-related C3 botulinum toxin substrate 1 (Rac1)/WASp family Verprolin-homologous protein-2 (WAVE-2) are upregulated (P<0.05) in Ang Ⅱ-induced neonatal rat primary and H9c2 cardiomyocyte hypertrophy, indicating the excessive activation of the Arp2/3 complex. We further show that CK666, a specific Arp2/3 complex inhibitor, prevents the reactivation of fetal genes and the enlargement of cardiomyocyte area induced by Ang II in NRCMs and H9c2 cells (P<0.05). Our results reveal that the Arp2/3 complex plays a crucial role in Ang Ⅱ-induced cardiomyocyte hypertrophy, which is beneficial to further studies about the molecular mechanisms by which the Arp2/3 complex regulates pathological cardiac hypertrophy.

Monkeypox is a viral zoonotic disease, and there is currently a lack of safe and effective vaccines against the monkeypox virus. Therefore, screening and developing vaccine candidates is of significant practical importance. With the rapid advancement of molecular biology and plant genetic engineering, plant bioreactors offer promising potential for producing vaccine proteins due to their advantages, including safety, cost-effectiveness, and scalability. In this study, we focused on the monkeypox protein B6R. The recombinant expression plasmid pFolia40108-B6R-Fer was successfully constructed using amplification, enzyme digestion, and flexible linker tandem ferritin technology. A complete transient expression system in Nicotiana benthamiana and a purification system for the recombinant monkeypox protein were established. The optimal expression time was determined to be 12-14 days, with a final purified protein concentration of approximately 1 mg/mL and a yield of 0.85 mg/kg fresh weight. The purified B6R-Fer recombinant protein self-assembled into spherical virus-like particles (VLPs) with an average particle size of 24 nm. The B6R-Fer recombinant protein from this study shows promising potential for use in the development and screening of plant-derived monkeypox vaccine candidates.

The CDH23 gene is a pathogenic mutant gene of the USH1D subtype in Usher syndrome. In this study, two wild-type Cdh23 full-length plasmids (~16 kb) with different promoters were constructed, and fluorinated polyethylene imine (FPEI) was used as a delivery vector to transfect thehouse ear institute-organ of corti 1 (HEI-OC1) and the optimal expression plasmid was obtained by evaluating the transfection efficiency in vitro. Firstly, the results of the synthesis of FPEI were analyzed using Fourier transform infrared absorption spectroscopy to prove the successful synthesis of FPEI. After that, the plasmid encapsulation ability of FPEI and the surface potential and hydration diameter of the formed complexes were characterized by agarose gel blocking assay, Zeta potential assay, and dynamic light scattering assay. It was found that FPEI had good plasmid encapsulation ability, and the FPEI plasmid complexes were all positively charged at high mass ratio, with the distribution of particle sizes in the range of 100-300 nm. The low cytotoxicity and high transfection efficiency of FPEI in HEI-OC1 cells were verified by Cell Counting Kit-8 (CCK-8) and flow cytometry. Comparing FPEI with Lipofectamine 3000 and different quality PEI (25K, 40K) transfection reagents, the transfection efficiency of FPEI was found to be significantly better than that of the traditional transfection reagents. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot results showed that the CAG promoter was better than the CMV promoter, which could be used as the optimal expression plasmid for the subsequent in vivo experiments.In addition, it was verified by cellular immunofluorescence that CDH23 was mainly distributed in the cytoplasm after overexpression. The above results demonstrated that FPEI can be used as an efficient delivery vector for in vitro overexpression of large genes represented by Cdh23, which provides an important experimental basis for subsequent in vivo gene therapy of USH1D syndrome.

The integration of science and education is not only an important strategy for promoting social progress and technological development, but also a modern form of higher education aiming at cultivating innovative talents. Conducting scientific research training for undergraduate medical students is one of the important ways to cultivate their innovative abilities and comprehensive qualities. Our team proposed a "teaching, science, and ideology trinity" teaching model to comprehensively cultivate students' scientific research comprehensive abilities under the value orientation of ideological and political education by organically integrating molecular biology experimental teaching with the scientific research training of undergraduate medical students. In this teaching activity, taking the experiment of gene polymorphism as an example, our team selected students with research potential from the whole grade and divided them into 4 project groups that were instructed by 4 teachers. The students were trained in the whole process of scientific research, including topic selection, project writing, experimental designing, application for research ethics, and project summary. Our team has always adhered to student-contentedness of educational concepts to stimulate students' intrinsic motivation throughout the teaching process. Students are the designers and implementers of the project, and teachers are only guides and promoters of learning. After this training, students not only became familiar with the writing and implementation of scientific research projects, but also improved their literature reading, experimental designing, experimental skills, and problem-solving abilities. More importantly, this teaching activity also cultivated students' awareness of research ethics and academic moral standards.

In order to investigate the teaching effectiveness of artificial intelligence (AI) in molecular biology, this study selected students from the Animal Medicine major of the College of Animal Science and Technology at Hebei North University in 2022 and 2023 as research subjects. There was no significant difference in professional foundation, admission scores, and other aspects between the two grades, and they were taught by the same lecturer to ensure the reliability of research results. The 2022 students will adopt the traditional teaching mode, while the 2023 students will implement the AI-enabled teaching mode, which includes four stages: pre class exploration, in class assistance, post class learning support, and teaching reflection and improvement. Before class, the teaching team pushes teaching videos of various knowledge points in the course and relevant preview materials organized by the AI system to students to complete self-learning, and use AI systems to track students' learning difficulties. In class, the teacher uses various teaching methods such as case teaching, group discussions, AI animation demonstrations and virtual experiments, etc.. Thus, they provide in-depth explanations of key contents based on the feedback data from the intelligent learning companion AI system, promoting students' understanding of the knowledge. After class, the AI system generates personalized learning plans for students and provides different levels of learning resources to broaden their horizons. At the same time, the AI system provides teachers with students' learning data analysis reports, and teachers can adjust and optimize their teaching plans accordingly. Research has found that students in the 2023 AI-empowered teaching class have significantly higher satisfaction in multiple dimensions such as learning interest, understanding and mastery of knowledge points, and cultivation of scientific research thinking than those in the 2022 traditional teaching class. In terms of student participation in comprehensive activities, the proportion of 2023 students participating in subject competitions and innovation and entrepreneurship activities has significantly increased. In terms of academic performance, the mid-term, laboratory, and final grades of 2023 students are higher than those of 2022 students, with a significant increase in the excellence rate and a significant decrease in the failure rate. The results indicate that the application of AI technology in molecular biology teaching has stimulated students' interest in learning, helped them better understand and master knowledge, significantly improved their academic performance. In sum, it has a positive impact on improving teaching quality.