Objective: Long non-coding ribose nucleic acids (LncRNAs) are a class of RNA molecules exceeding 200 nt in length, but they lack protein-coding capacity. A growing body of research indicates that LncRNAs play an important role in the lineage-specific differentiation of embryonic stem cells (ESCs). However, the upstream regulatory mechanisms that govern LncRNAs remain unclear. Methods: The RNA-seq and ChIP-seq technologies were employed to systematically identify LncRNAs regulated by the Hippo-YAP signaling pathway in ESCs. The ROSE software was further used to identify LncRNAs regulated by YAP via super-enhancers, followed by RT-qPCR validation of Hippo-YAP-regulated LncRNAs in mouse ESCs. Results: ChIP-seq analysis identified 918 LncRNAs regulated by Hippo-YAP, with the deletion of Mst1/Mst2 significantly altering their expression and inducing histone modification remodeling at their genomic loci. Further studies revealed that LncRNAs such as Lncenc1 are subject to regulation by the Hippo-YAP pathway through the action of super-enhancers. Experiments confirmed that YAP overactivation induced by Mst1/Mst2 knockout markedly upregulated the expression of Lncenc1 and other LncRNAs, whereas YAP knockdown in Mst1/Mst2-deficient ESCs eliminated super-enhancers, thereby reversing their aberrant upregulation. Conclusion: Our study preliminarily elucidates the molecular mechanism by which the Hippo-YAP signaling pathway regulates a group of stem cell-associated LncRNAs through super-enhancers, providing new insights into the maintenance of pluripotency and differentiation regulation in ESCs.

Objective: To screen and identify the proteins interacting with Neuritin and explore the possible molecular mechanism by which Neuritin promotes neurite outgrowth. Methods: The proteins interacting with Neuritin were screened by the yeast two-hybrid technique. Recombinant plasmids pcDNA3.1-His-Neuritin and pcDNA3.1-Myc-Syap1 were constructed and then transfected into HEK 293 cells, and the interaction between Neuritin and Syap1 was determined by CO-IP. The effect of Neuritin expression levels on Syap1 expression levels was detected by Western blot analysis. Indirect immunofluorescence was used to observe the effects of Neuritin overexpression on Syap1 transport and axon morphology of SH-SY5Y cells. Results: A total of 57 proteins interacting with Neuritin were obtained by sequencing and sequence comparison analysis of the yeast two-hybrid positive clones. Syap1, which has a function that is highly consistent with Neuritin, was listed as a candidate protein. The forward and reverse CO-IP results showed that Neuritin and Syap1 had a specific interaction. Following the overexpression and underexpression of Neuritin, Western blot results showed that Neuritin was a negative regulator of Syap1. Neuritin overexpression can promote the terminal transport of the Syap1 protein in SH-SY5Y cells to the axon, and thereby promote axon elongation. Conclusion: Neuritin has specific interaction with Syap1 and is a negative regulator of Syap1. Overexpression of Neuritin can promote the terminal transport of Syap1 protein to the axon and neurite prolongation.

Objective: The demand for artificial bone substitute materials continues to grow for bone repair. Calcium phosphate bone cement exhibits good biocompatibility, self-setting ability, and plasticity. We should improve its mechanical properties and osteogenic activity, while regulating the degradation rate to enhance its therapeutic effect. Methods: This study synthesized a composite of α-tricalcium phosphate (α-TCP), α-calcium sulfate hemihydrate(α-CSH), and mineralized collagen (MC), using water and nano-silk fibroin fibers (SFF) as curing agents to prepare composite bone cement. The structure and composition of the materials were characterized, and the setting time, compressive strength, degradation properties, and osteogenic activity of the materials were studied. Results: The results indicate that as the MC content increases gradually, the solidification time of the material accelerates. The compressive strength of bone cement reaches the maximum when 5 wt% MC and 0.4 wt% SFF are used as the curing liquid. SFF can adjust the compressive strength and degradation rate of bone cement. MC and SFF can act in a synergistic manner to promote cell proliferation and osteogenic differentiation of MC3T3. Conclusion: The composite material studied enhances the osteogenic activity of pure calcium phosphate bone cement, potentially leading to broader applications in orthopedic clinical practice.

Objective: To isolate and purify the enzyme that specifically hydrolyzes the rhamnosyl group from the side chain of pulchinenoside H3 (PSGaseⅠ) from the enzymatic extract of Absidia sp. P3800r fermentation, and to characterize its enzymatic properties, thereby supporting enhanced bioavailability of pulchinenoside H3 and further pharmaceutical development.Methods: PSGaseⅠ was purified using a DEAE Sepharose Fast Flow anion-exchange column. Its molecular mass, optimal pH and pH stability, optimal temperature and thermal stability, the effects of metal ions, enzyme kinetics, and substrate specificity were systematically analyzed.Results: The molecular mass of PSGaseⅠ was approximately 53 kDa. It exhibited maximum activity at a pH of 5.0 and remained stable within pH 4.0 ~ 6.0. The optimum temperature was 40℃, with stable activity maintained between 20 and 40℃. Metal ions such as K⁺, Na⁺, Mg²⁺, and Ca²⁺ showed no significant effect on enzyme activity, whereas Cu²⁺, Zn²⁺, and Fe³⁺ strongly inhibited it. Kinetic parameters were determined as follows: K_m 1.93 mmol·L^-1, V_max 0.072 9 mmol·L^-1·h^-1, k_cat 0.038 s^-1, and k_cat/K_m 19.6 L·mol^-1·s^-1. Substrate specificity analysis revealed that PSGaseⅠ hydrolyzed pNP-α-L-rhamnopyranoside and the rhamnosidic bond of pulchinenoside H3. It showed trace hydrolytic activity toward ginsenoside Re, but did not affect the rhamnosidic bonds in flavonoids.Conclusion: PSGaseⅠ was successfully purified and characterized, providing a basis for the large-scale enzymatic preparation of secondary saponins with improved bioavailability and facilitating further pharmaceutical research and application development.

N6-methyladenosine (m⁶A) is the most prevalent internal messenger RNA modification found in animals, which can be dynamically regulated by methyltransferases and demethylases. Targeted editing of RNA m⁶A modifications is crucial for studying the function of specific m⁶A sites, elucidating the relationship between the presence of specific m⁶A and phenotypic outcomes, and developing molecular therapies targeting m⁶A. Currently, the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (CRISPR-Cas) systems have been widely used for gene editing due to its efficiency, ease of manipulation, and good target specificity. In particular, a series of CRISPR-Cas-based RNA m⁶A editing tools have been developed by fusing m⁶A methyltransferases or demethylases with deactivated Cas9/Cas13 variants. These tools have been successfully applied in mammalian cells, stem cells, and plant systems, revealing the functional roles of m⁶A in gene expression, viral infection, and developmental regulation. This paper summarizes the systematic construction of various m⁶A editors, their protospacer adjacent motif/protospacer flanking sequence (PAM/PFS) requirements, off-target effects, and editing windows, as well as potential methods for improvement. Furthermore, we discuss the advantages, limitations, and application prospects of these tools, providing references for the further improvement and application of m⁶A editing tools.

Vaccine adjuvants, as key components in enhancing vaccine immunogenicity, play a vital role in modern vaccinology. A systematic review was conducted on the historical evolution and mechanisms of action of vaccine adjuvants, with particular emphasis on the innovative advancements in aluminum-based, emulsion-based, and nucleic acid adjuvants. Specifically, aluminum-based adjuvants enhance immune responses through particle formation/antigen adsorption, inflammatory cascade activation, and T-cell stimulation. Emulsion-based adjuvants increase immunogenicity via antigen depot effects and localized inflammation, while nucleic acid adjuvants like CpG oligonucleotides directly activate B cells and dendritic cells to promote Th1-type immune responses and memory T-cell generation. The article further explores the potential applications of these novel adjuvants in combating emerging pathogens, particularly highlighting their significance in improving vaccine potency and durability. Moreover, the critical role of adjuvant development in the design of next-generation vaccines was emphasized, providing a theoretical foundation for the creation of safer and more effective vaccine adjuvants.

For pathogens that use the digestive tract as the main route of infection, oral vaccines, by virtue of their effect of stimulating mucosal immune responses, become an important immune barrier to defend against the invasion of such pathogens. With the continuous development of vaccination technology, oral vaccines have become a hot topic in the prevention and control of gastrointestinal infectious diseases due to their convenience, painless vaccination, and the ability to induce effective mucosal, humoral, and cellular immunity. However, due to the complexity of the digestive tract environment and mucosal immune mechanism, oral vaccines are easily destroyed by gastric acid and digestive enzymes in the gastrointestinal tract, resulting in antigen inactivation and reduced immune efficacy. To overcome this problem, current research focuses on the development of various delivery systems and new mucosal adjuvants to enhance the immune efficacy of oral vaccines. This article reviews the mucosal immunity and the mechanism of action of oral vaccines, delivery vectors, and mucosal adjuvants. It also discusses the challenges faced by oral vaccine development, aiming to provide a reference for the research and development of oral vaccines.

Ulcerative colitis (UC) is a subtype of inflammatory bowel disease (IBD), but the cause is not yet clear. UC is a chronic and recurrent intestinal inflammatory disease. In the treatment of UC, oral administration is an ideal way of administration because of its good compliance, high safety, convenient operation and direct action on intestinal mucosa. However, oral administration is easily disturbed by gastrointestinal physiological environment in vivo, which leads to low bioavailability and a poor therapeutic effect. A large number of studies have proved that natural polysaccharides are not degraded in human gastric or small intestinal juices, but can be degraded into monosaccharides or oligosaccharides by enzymes produced by microorganisms present in the colon. Using a natural polysaccharide as UC drug delivery carrier material not only has good biocompatibility, biodegradability and bioactivity, but also can adhere to the intestinal mucosa through non-covalent interactions such as van der Waals forces, hydrogen bonding and electrostatic attraction. This can significantly improve the targeted delivery efficiency of drugs, making it an ideal drug carrier material for UC treatment at present. Preparing natural polysaccharides in the form of hydrogels, microspheres, and nanoparticles can not only improve the bioavailability of drugs, but also achieve synergistic therapeutic effects. Among the natural polysaccharide carrier materials for UC treatment, sodium alginate has become the most commonly used natural polysaccharide carrier material for UC drug delivery because of its relatively good solubility, degradability, pH response and strong targeting properties. In this paper, the properties, modification, dosage form design, application effects, mechanisms, current challenges and future development directions of natural polysaccharide sodium alginate in UC treatment are comprehensively reviewed, in order to provide ideas for the development and application of drug delivery system for UC treatment.

Raman spectroscopy, as a non-invasive and highly sensitive process analytical technology (PAT), has demonstrated significant application potential in the biopharmaceutical field in recent years due to its strong chemical specificity and the elimination of the need for sample pretreatment. This paper systematically reviews the technical principles and classifications of Raman spectroscopy and focuses on its key applications throughout the entire biopharmaceutical process. Additionally, it analyzes the core advantages (such as real-time capability and non-destructiveness) and limitations (such as issues with signal intensity and resolution accuracy) of Raman spectroscopy. Furthermore, it proposes that the integration of Raman spectroscopy with artificial intelligence (AI) algorithms can overcome traditional analytical limitations and drive the transformation of traditional biopharmaceutical manufacturing towards intelligent manufacturing. In the future, the deep integration of Raman spectroscopy and AI will not only accelerate the development of novel PAT sensors but also provide multidimensional data support for process optimization and quality control, facilitating the biopharmaceutical industry’s transition towards digitalization and intelligentization.

Based on extracellular electron transfer (EET), microbial electrochemistry (MEC) is an emerging technology as a combination of molecular biology, microbiology, and electrochemistry. It has demonstrated a bright prospect in applications such as microbial fuel cell (MFC), and microbial electrosynthesis (MES). EET, as the chemical foundation of MEC, has great influences on the reaction rate and Colomb efficiency of MEC. Enhancing EET via synthetic biology and material science methods is the fundamental means to further enhance the rate of MEC reaction. In this review, the synthetic biological strategies are systematically summarized to improve EET through both direct and mediated electron transfer, aiming to provide a reference for increasing the EET rate and expanding the application range of the MFC technology.

Visual detection technology has attracted wide attention due to its intuitiveness and convenience, but the problems of insufficient sensitivity and limited trace analysis ability still need to be solved. Rolling circle amplification (RCA), as a new nucleic acid isothermal amplification technology, has the advantages of high sensitivity and high specificity, and has great application prospects in the fields of in vitro diagnosis and food safety detection. Therefore, the combination of visual detection and RCA technology greatly improves the detection sensitivity and has the potential of point-of-care testing (POCT). In this paper, the principles, types, and signal amplification mechanism of RCA technology, as well as various visual biosensors based on RCA, including fluorescence and colorimetry, are reviewed in order to provide new ideas for the development and application of RCA technology in the field of rapid visual detection.

Artificial intelligence-assisted new drug research and development leverages its robust data analysis, pattern recognition, and predictive capabilities to process biomedical data. Through virtual screening of novel molecules and structures, toxicity prediction, activity monitoring, and release mechanism analysis, AI can rapidly identify active compounds, thereby shortening the drug development cycle. This technology has broad applications at every critical stage of drug development. The integration of cutting-edge information technologies, particularly AI, into pharmaceutical R&D has significantly enhanced efficiency and success rates while reducing costs. Initial evidence confirms the practicality and immense potential of AI in this domain. China places significant emphasis on the convergence of biotechnology and information technology, and with policy support, AI-assisted drug development has progressed rapidly. However, challenges remain, including inadequate scientific competition and research ecosystems, insufficient high-quality data, lack of policy guidance and support for large-scale R&D models, shortage of interdisciplinary talent, and absence of key common platforms. Given its strong foundation in biomedicine and burgeoning AI technology, China should seize this opportunity to optimize scientific competition and research ecosystems, enhance the data resource integration and sharing, support the development of foundational biological models, strengthen the cultivation of high-end interdisciplinary talent, improve key common technology platforms, and foster a favorable environment for the AI pharmaceutical industry. Strategic planning and early layout of the AI pharmaceutical sector will promote accelerated innovation and transformation in the biomedical industry, enhancing quality and efficiency.