In the present work, we propose drug-target co-screening strategy, a novel technique for Chinese medicine development and evaluation, which leverages whole-genome pan-receptors and genetically encoded natural compound libraries from medicinal plants and animals. This paper outlines the core components of the strategy, focusing on the identification of novel disease and syndrome-related targets based on clinical efficacy of Chinese medicine, multi-component target screening technologies for herbal medicine, technologies for multi-component identification and target screening and artificial intelligence-powered approaches for discovering active ingredients and elucidating their mechanisms. The strategy demonstrates broad applicability in areas such as the discovery of disease (syndrome) targets, the development of new Chinese medicine drugs, the analysis of multi-target, multi-pathway mechanism of action of Chinese medicine, and the study of drug properties including the five flavors. Centered on whole-genome pan-receptors, drug-target co-screening strategy provides a powerful framework to address key challenges in Chinese medicine research—namely, the ambiguity of therapeutic targets and unclear mechanisms of action. This approach offers strong support for mechanistic studies and paves the way for a new paradigm in modernizing and innovating Chinese medicine drug development.

Gut microbiota, the largest microbial community in the body, plays an important role in maintaining the immune homeostasis of the intestinal system and even the extraintestinal organs. Recently, increasing evidence has highlighted the substantial impact of gut microbiota on the cancer progression and therapy. Intervention of gut microbiota is considered as a promising anti-tumor strategy. Though the molecular mechanisms remain mostly unknown, it is widely appreciated that microbiota-derived metabolites critically mediate the interplay between host and microbiota, triggering enthusiasm to understand the mechanism behind. This article summarizes the latest progress in understanding mechanisms of microbial metabolites in regulating host immune responses, in particular in antitumor immunity. With this, we hope to provide insights for new therapeutics from the perspective of microbiota intervention.

Steatotic liver disease (SLD) is a disease characterized by abnormal accumulation of lipids in hepatocytes, which includes metabolic dysfunction-associated steatotic liver disease, alcohol-associated liver disease, and metabolic dysfunction and alcohol-related liver disease. Most patients only have simple liver steatosis at an early stage, which will gradually develop into hepatitis, liver fibrosis, and cirrhosis as the progression of the disease. Mitochondria is a highly dynamic organelle, which changes its morphology, size, and location by continuous division and fusion. This process is known as mitochondrial dynamics. The imbalanced mitochondrial dynamics can cause the dysfunction of mitochondria, leading to increased lipid deposition in liver cells, as well as antioxidant capacity deficient associated reactive oxygen species accumulation and lipid peroxidation, resulting in increased lipotoxicity and liver injury. Therefore, mitochondrial dynamics play an important role in the development of steatotic liver disease. In this review, the relationship between mitochondrial dynamics and mitophagy, mitochondrial-associated endoplasmic reticulum membranes, and mitochondrial-dependent apoptosis in steatotic liver disease were discussed. In addition, the mechanism role of natural compounds in improving steatotic liver disease via regulating mitochondrial dynamics-related cell functions was also systematically reviewed, aiming to provide the basis and clues for the subsequent investigations and further clinical development and application of natural compounds in the treatment of steatotic liver disease.

Transmembrane protein 16A (TMEM16A) is a Ca²⁺-activated Cl^- channel that is overexpressed in various malignant tumors. The gene is located in chromosome 11q13 region, and its amplification is one of the main reasons for the high expression of this protein. In addition, transcriptional regulation, epigenetic regulation and microRNAs (miRNAs) can promote TMEM16A expression in many cancer cells. TMEM16A not only activates a variety of signaling pathways that promote tumor cell proliferation, migration and invasion, but also plays an important role in inhibiting tumor cell apoptosis. Therefore, the development of anticancer drugs targeting TMEM16A is of great clinical significance. This article reviews the mechanism of TMEM16A in malignant tumors, summarizes the signaling pathways of TMEM16A in promoting tumor development, and analyzes the key role of TMEM16A as a potential biomarker in tumor development. TMEM16A inhibitors may become a new treatment for cancers with high expression of TMEM16A.

Cardiovascular diseases pose a serious threat to human life and health, with their incidence and mortality rates rapidly increasing in China in recent years. Inflammation resolution has become a key focus in cardiovascular research, and in-depth studies of this process have provided new insights into the potential mechanisms and treatments for cardiovascular diseases. Current research on inflammation resolution mechanisms primarily involves pro-resolution mediators, endogenous gases, and immune cells such as macrophages. Based on the important role of inflammation resolution in cardiovascular diseases and the potential therapeutic targets it may offer, this paper reviews the mechanisms of inflammation resolution, the research progress of inflammation resolution in cardiovascular diseases, and the current status of therapeutic strategies and drug research targeting inflammation resolution, with the aim of providing new treatment approaches for cardiovascular diseases.

Viruses infect humans and cause diseases through respiratory transmission, which is the most common pathogenic route for viruses. Viruses with respiratory transmission capabilities typically exhibit periodic and seasonal epidemic characteristics. Due to the wide variety of viruses and significant differences in their structures and life cycles, effective prevention and treatment drugs for various viruses are relatively scarce and have limited efficacy. In recent years, significant progress has been made in the repositioning of antiviral drugs, high-throughput screening and ligand-based drug design. This article systematically summarizes the approved drugs for several major viruses that primarily cause respiratory infections and reviews the latest research progress of antiviral drugs currently in clinical trials, aiming to provide a reference basis for the research and further development of antiviral drugs.

Ubiquitin-specific protease 7 (USP7) is one of the more widely studied members of the deubiquitinating enzyme family, which regulates the activity and function of a variety of proteins in cells. USP7 is closely associated with the tumor progression, which makes USP7 a highly promising target for drug therapy. However, no drug targeting USP7 has yet been approved for clinical trials, thus the development of drug targeting USP7 still requires in-depth research. This article reviews the structure and function of USP7 and the progress of drug research targeting USP7.

Seasonal influenza epidemics caused by influenza viruses continue to pose persistent threats to global public health. The core replication component RNA-dependent RNA polymerase (RdRp), distinguished by its highly conserved structure and functionally irreplaceable characteristics, has emerged as a critical target for antiviral drug development. This study systematically analyzes the structural and functional features of RdRp and the mechanisms of action of its inhibitors. It comprehensively reviews current screening technologies and emerging strategies for RdRp inhibitors across three levels: molecular, cellular, and animal model levels. By systematically evaluating the technical advantages and limitations of existing approaches, we propose future research directions to advance the development of novel high-efficiency RdRp inhibitors, thereby providing both theoretical foundations and technical frameworks for this crucial antiviral strategy.

4D printing technology represents a revolutionary breakthrough in the field of additive manufacturing. It not only inherits the spatial construction capabilities of 3D printing but also introduces the dimension of time, endowing printed objects with dynamic "life". This technology uses smart responsive materials that allow printed objects to change their shape, properties, or functions in response to specific environmental stimuli, such as temperature, magnetic fields, or light changes, as programmed in advance. This has shown great potential and application prospects in the fields of medical research and clinical applications. In this article, we first explore the smart responsive materials that enable 4D printing and introduce their unique physical and chemical properties and functions. Secondly, we review the innovative applications of these smart materials in the medical fields of tissue engineering, regenerative medicine, and drug delivery. Finally, we discuss the challenges faced by 4D printing technology in practical applications and look forward to the future development direction of the technology and its broader application potential in various fields.

Central nervous system (CNS) diseases are a global public health concern, exacerbated by the aging population. Nucleic acid drugs have shown great potential in this field. They target specific gene sequences through the principle of complementary base pairing, allowing intervention before protein expression. These drugs offer high specificity, good safety profiles, and relatively short development cycles. However, the presence of the blood-brain barrier (BBB) and the complexity of CNS pathologies restrict drug delivery, making effective delivery systems crucial for the therapeutic effect of nucleic acid drugs. This review introduces nucleic acid drugs developed for CNS disease treatment in recent years, along with their mechanisms of action. It analyzes the obstacles encountered in brain-targeted delivery of nucleic acid drugs, focusing on the physiological structure and function of the BBB, as well as potential pathways and associated mechanisms for achieving delivery. It details the mechanisms of utilizing viral vectors, nucleic acid drug modification techniques, lipid nanoparticles, polymeric nanoparticles, and exosomes to penetrate the blood-brain barrier, as well as their applications in the delivery of nucleic acid drugs. The aim of this review is to compile and analyze current strategies for enabling nucleic acid drugs to cross the BBB while improving targeting and stability. It explores methods to enhance delivery efficiency and provides guidance for future research and clinical application, ultimately promoting the development of nucleic acid therapeutics for CNS disease treatment.

This article primarily investigates the role of the neural circuit from lateral hypothalamic (LH) glutamatergic neurons to the rostromedial tegmental nucleus (RMTg) in defensive behaviors in mice. Adult male vesicular glutamate transporter 2 (Vglut2)-Cre mice (8-10 weeks old) were randomly divided into a mCherry control group and a channelrhodopsin-2 (ChR2) experimental group, who received injections of AAV2/9-EF1α-DIO-mCherry and AAV2/9-EF1α-DIO-ChR2-mCherry virus into the LH, respectively. Optical fibers were implanted in the RMTg of both groups. After three weeks of recovery from surgery and sufficient viral expression, behavioral experiments were conducted, including the open field test, real-time place preference (RTPP) test, and hiding box test. After the experiment, ascertain the extent of viral infection and optical fiber positions in the mice. Immunohistochemistry was carried out to detect c-Fos expression in the RMTg to validate the activation of the pathway from LH glutamatergic neurons to the RMTg upon 473 nm blue laser stimulation. The results showed that c-Fos expression in the RMTg was elevated in ChR2-expressing mice following blue laser illumination, confirming successful pathway activation. During a 473 nm blue laser stimulation, compared to the mCherry group, mice in the ChR2 group exhibited a notable increase in velocity in the open field test, indicating an escape behavior. In the RTPP test, the time spent on the preferred side reduced, indicating avoidance behavior. Additionally, the latency to enter the hiding box significantly shortened, reflecting hiding behavior in the hiding box test. This research indicates that the activation of the projections from LH glutamatergic neurons to the RMTg promotes defensive behaviors in mice. All experimental procedures were approved by the Committee on the Ethics of Animal Experiments of the School of Basic Medical Sciences, Fudan University (identification number: 20250427-001).

Delayed fracture healing and nonunion continue to pose substantial challenges in orthopedic clinical practice. Existing medications for fracture treatment can, to a certain degree, promote fracture healing. However, they are plagued by problems such as unstable efficacy, slow onset, and frequent adverse effects. In this study, leveraging BanGel patented technology, we developed a novel Eucommia ulmoides-based hydrogel. This was achieved by integrating Eucommia ulmoides, a traditional Chinese medicine renowned for its liver and kidney tonifying and bone-strengthening properties, with a hydrogel delivery system. A transverse mid-shaft tibial fracture rat model was established. Behavioral assessments, serological analysis, radiological and histopathological examinations were employed to observe the dynamic changes during the fracture and subsequent healing processes. The efficacy of the Eucommia ulmoides hydrogel in promoting fracture healing was objectively evaluated. Moreover, the therapeutic advantages of the hydrogel were compared with those of oral and direct topical administrations containing the same dosage of Eucommia ulmoides. All animal experiments were approved by the Laboratory Animal Welfare Ethics Committee of the Laboratory Animal Center of the Institute of Basic Theory of Chinese Medicine, China Academy of Traditional Chinese Medicine, and were conducted in accordance with the relevant guidelines and regulations (approval No. IBTCMCACMS21-2406-03). The results showed that Eucommia hydrogel can significantly alleviate the early symptoms of fractured, promote callus formation, improve trabecular parameters and cortical bone continuity, and regulate the dynamic balance of bone formation and absorption. Compared with oral and topical applications of the same dose of Eucommia ulmoides, the Eucommia ulmoides hydrogel exhibited a faster onset and stronger permeability. Further mechanism exploration and verification results showed that Eucommia ulmoides can promote fracture healing by activating the serine/threonine protein kinase 1/β-catenin (AKT1/β-catenin) signaling axis to regulate the expression of Runt-related transcription factor 2 (RUNX2) and receptor activator of nuclear factor κB ligand/osteoprotegerin (RANKL/OPG). In summary, the novel Eucommia ulmoides hydrogel shows distinct advantages in promoting fracture healing and has potential clinical application value.

This study investigated the antitumor activity and molecular mechanisms of penfluridol (PF), an antipsychotic drug, in gastric cancer treatment. In vitro assays including CCK-8, colony formation, wound healing, Transwell migration, Hoechst 33342 staining, and flow cytometry demonstrated that penfluridol significantly inhibited proliferation and migration of SGC-7901 and HGC-27 gastric cancer cells in a concentration-dependent manner, while inducing apoptosis. Transcriptomic profiling combined with molecular docking revealed stable binding conformations between penfluridol and heat shock protein 90 (HSP90) or mouse double minute 2 (MDM2). Western blot analysis further confirmed that penfluridol suppressed the HSP90/protein kinase B/MDM2/tumor protein p53 (HSP90/Akt/MDM2/p53) pathway, characterized by decreased HSP90 expression and concomitant upregulation of p53 protein. These findings collectively indicate that penfluridol exerts multifaceted antitumor effects by targeting key oncogenic pathways, providing experimental evidence for repurposing this antipsychotic agent as a potential therapeutic strategy against gastric cancer.

Voltage-gated sodium channels (VGSCs, Na_V) are essential for the generation and conduction of action potentials in excitable cells. The Na_V1.8 subtype is predominantly expressed in sensory neurons, particularly in small-diameter neurons of the dorsal root ganglia (DRG), and plays a critical role in the signal transduction of chronic, neuropathic, and inflammatory pain. Due to its tetrodotoxin-resistant properties and electrophysiological characteristics of slow activation and inactivation, Na_V1.8 is a key target for analgesic drug development, facilitating repeated neuronal firing. Consequently, establishing an in vitro drug screening model based on Na_V1.8 is crucial for identifying lead compounds with analgesic potential. However, heterologous expression of Na_V1.8 remains challenging, primarily due to low expression levels in non-neuronal cell lines, which hinders effective drug screening. In this study, we established an in vitro expression model of rat Na_V1.8 using Xenopus laevis oocytes as the expression system. Channel property was characterized using the two electrode voltage clamp (TEVC) technique, providing a platform for screening active compounds. Quantitative polymerase chain reaction confirmed that rat Na_V1.8 is predominantly expressed in the DRG and trigeminal ganglia (Guangxi University Ethics Committee Review and approval number: GXU-2022-159). We cloned the full-length Na_V1.8 gene and constructed two chimeric variants, Na_V1.8-N and Na_V1.8-C. Following in vitro transcription of the corresponding capped RNA, it was injected into Xenopus laevis oocytes for heterologous expression. TEVC recordings were used to detect Na_V1.8 and chimera expression, and channel activity was evaluated through I-V curves, steady-state activation curves, and the specific Na_V1.8 antagonists A-803467 and VX-548. The chimera Na_V1.8-C resolved the problem in stable expression of Na_V1.8 in vitro and exhibited similar pharmacological properties with native Na_V1.8, offering a valuable tool for future screening of compounds targeting voltage-gated sodium channel.

Non-nucleoside reverse transcriptase inhibitors (NNRTIs) play an important role in AIDS treatment. Diarylpyrimidine derivatives (DAPYs) represent one of the most important classes of NNRTIs. The previously reported ZK-4a showed high cytotoxicity (CC₅₀ = 5.98 μmol·L^-1), significantly limiting its therapeutic potential. To resolve this issue, we adopted a two-pronged strategy: replacing the NH fragment with an O atom and modifying the C-5 position of the pyrimidine core. These modifications enabled the design and synthesis of 27 novel deuterated-biphenyl-DAPY analogues. Fortunately, all deuterated-biphenyl-DAPYs exhibited broad and highly efficient antiviral activity, as well as obviously decreased cytotoxicity (CC₅₀ = 18.02- > 297.27 μmol·L^-1). Among them, compound 8s achieved an EC₅₀ value of 8.88 nmol·L^-1 while showing negligible cytotoxicity (CC₅₀ > 287.68 μmol·L^-1). Compound 8s also displayed exceptional resistance profiles with EC₅₀ values of 7.54 nmol·L^-1 (L100I), 8.57 nmol·L^-1 (K103N), 24.28 nmol·L^-1 (Y181C), 62.71 nmol·L^-1 (Y188L) and 14.38 nmol·L^-1 (E138K). Additionally, the target of deuterated-biphenyl-DAPYs was verified by reverse transcriptase enzyme inhibition assays.

The 95% ethanol extract of llicium micranthum was separated and purified by silica gel, Sephadex LH-20, MCI-gel and semi-preparative HPLC. The chemical structures of the isolated compounds were identified by NMR and MS. Twelve compounds were isolated and identified as illimicranol A (1), vanilloloside (2), 4-O-β-D-glucosyl-benzoic acid (3), gastrodin (4), icariside D₂ (5), caryolane-1,9β-diol (6), clovane-2β,9α-diol (7), methyl shikimate (8), protocatechuic acid methyl ester (9), p-hydroxybenzyl alcohol (10), p-hydroxyphenylethanol (11), methyl p-hydroxyphenylacetate (12). Among them, compound 1 is a unique lignan with new carbon skeleton, named illimicranol A. Compounds 2-12 are firstly isolated from this plant. The effects of compound 1 on the differentiation of PC12 cells induced by NGF and the damage of PC12 cells induced by MPP⁺ were evaluated. The results showed that compound 1 did not exhibit obvious neurotrophic and neuroprotective activities.

Five compounds were isolated from the dichloromethane fraction of a 70% ethanol extract of Epimedium sagittatum Maxim using a combination of chromatography techniques, including silica gel column chromatography, ODS column chromatography, and semipreparative high-performance liquid chromatography techniques. The structures of these five compounds were identified as 5,4′-dihydroxy-3′-prenyl-7-prenyloxyflavanone (1), 5,4′-dihydroxy-7-prenyloxyflavanone (2), 5,7,4′-trihydroxy-3′,8-diprenylflavanone (3), sebacic acid (4), and aurantiamide acetate (5), respectively, based on a comprehensive analysis of their physicochemical properties and spectroscopic data, such as NMR and MS. Notably, compound 1 was found to be a new flavonone compound, while compounds 2-5 were isolated from the Epimedium genus for the first time. Additionally, preliminary investigations indicated that compounds 1 and 3 exhibited potential anti-pulmonary fibrosis activities.

Nineteen compounds were isolated from the ethyl acetate fraction of the stems and leaves of Salvia miltiorrhiza Bge by using various column chromatographic methods, such as silica gel, Sephadex LH-20 and semi-preparative HPLC. According to their physiochemical properties and spectral data, the structures were further identified as 3′,4′-dihydroxy-4-(3-methoxy-3-oxopropyl)-[1,1′-biphenyl]-3-carboxylic acid (1), 2-hydroxy-5-(2′-hydroxybenzyl) benzoic acid (2), dehydrozingerone (3), 3,4-dihydroxy benzalacetone (4), benzyl alcohol-β-D-glucopyranoside (5), dehydrololiolide (6), rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]-oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (7), 6-hydroxy-3-(2-hydroxyethyl)-2,4,4-trimethylcy clohexa-2,5-dien-1-one (8), scopoletin (9), esculetin (10), dihydrotanshinone Ⅰ (11), 2′-deoxythymidine (12), quercetin 3-glucoside (13), sutchuenmedin A (14), 2′′-O-rhamnosylicariside-Ⅱ (15), 2,2′-oxybis(1,4-di-tert-butylbenzene) (16), 1H-indole-3-carbaldehyde (17), 3-(2-oxopropyl)-3-hydroxyind-olin-2-one (18), dysodensiol E (19). Compound 1 was a new biphenyl lignan, named as 3′,4′-dihydroxy-4-(3-methoxy-3-oxopropyl)-[1,1′-biphenyl]-3-carboxylic acid. Compound 2 was a new natural product and compounds 3-8 and 4-19 were firstly isolated from S. miltiorrhiza. MTT method was used to screen the anti-hepatoma activity of all the compounds, and none of them showed obvious effect.

The chemical profiling of Dengzhan Shengmai capsules were characterized using ultra high performance liquid chromatography/tandem quadrupole electrostatic field orbitrap (UHPLC-Q-Exactive-Orbitrap) high resolution mass spectrometry analysis. The post-acquisition strategy integrating precursor ion targeting recognition and diagnostic ion filtering was proposed to realize the identification of components in Dengzhan Shengmai capsules. The mass spectrometric data were acquired in both positive and negative ion modes using a full-scan MS1 coupled with data-dependent automatic triggering of MS/MS. By employing the similarity analysis function within the Access database software, precursor ions were promptly identified by comparing determined accurate masses to the in-house mass spectrometry database specific to Dengzhan Shengmai capsules. A diagnostic ion database encompassing a variety of compounds found in Dengzhan Shengmai capsules was compiled based on the mass spectrometry fragmentation patterns of 16 reference components. Ultimately, 248 compounds were characterized utilizing the diagnostic ion filtering method, comprising 51 caffeoyl esters, 38 flavonoids, 63 ginsenosides, 19 steroidal saponins, 11 nortriterpenoids, 21 homoisoflavones, 39 lignans and 6 other class constituents. The developed qualitative analysis approach enables swift and effective identification of chemical components present in Dengzhan Shengmai capsules, serving as a scientific basis for quality assessment and clinical utilization. This innovative strategy on high-resolution mass spectrometry data analysis, integrating precursor ion targeting recognition together with diagnostic ion filtering, introduced a novel methodology for the comprehensive analysis of complex substance systems in traditional Chinese medicine.

This study aimed to quantitatively evaluate the contents and in vivo exposure levels of 11 primary components in six commercially available Angong Niuhuang Pill (ANP) products. A chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous quantification of deoxycholic acid (DCA), cholic acid (CA), hyodeoxycholic acid (HDCA), glycocholic acid (GCA), taurocholic acid (TCA), curcumin (CC), baicalin (BC), berberine (BBR), and crocetin (CRT). A gas chromatography-mass spectrometry (GC-MS) method was established to determine muscone (MCN) and borneol (BN). The analytical methods were validated and applied to assess the content of the 11 components in six ANP products and their plasma exposure levels following intragastric administration in rats (All animal experiments had been approved by the Experimental Animal Ethics Committee of China Pharmaceutical University, ethics number: 2024-07-131). The quantitative methods utilizing LC-MS/MS and GC-MS were successfully established and validated for the simultaneous quantification of DCA, CA, HDCA, GCA, TCA, CC, BC, BBR, CRT, MCN and BN both in vivo and in vitro samples, respectively. The contents of HDCA, DCA, CA, GCA, TCA, CC, BC, CRT, BBR, BN and MCN varied over the ranges between 0.22 ± 0.03-1.47 ± 0.41, 0.22 ± 0.03-1.48 ± 0.39, 0.65 ± 0.01-4.28 ± 0.30, 0.44 ± 0.01-0.80 ± 0.02, 0.49 ± 0.05-1.36 ± 0.04, 0.001 2 ± 0.000 1-0.007 1 ± 0.001 0, 3.49 ± 0.80-4.47 ± 0.13, 0.003 6 ± 0.001 1-0.013 7 ± 0.014 8, 1.77 ± 0.05-4.84 ± 0.41, 4.70 ± 2.09-10.91 ± 3.08, 0.20 ± 0.03-0.44 ± 0.04 mg·g^-1, respectively. Similarly, the in vivo plasma exposure of HDCA, DCA, CA, GCA, TCA, BC, CRT covered the ranges over 5 088.63 ± 2 701.16-11 819.10 ± 7 585.61, 3 639.42 ± 1 970.63-18 853.46 ± 18 309.75, 19 397.89 ± 9 464.21-182 870.72 ± 236 816.06, 11 011.97 ± 4 236.99-57 331.79 ± 76 180.09, 53 607.57 ± 43 279.12-109 961.78 ± 82 288.37, 590.17 ± 175.72-2 424.90 ± 589.22, 74.95 ± 19.38-586.84 ± 194.49 h·ng·mL^-1, respectively. The plasma concentrations of the other four components (CC, BBR, BN, MCN) were below the detection limit. Three robust quantitative analytical methods were established and applied to evaluate variations of the content and in vivo exposure levels of 11 components in six ANP commercial products. The results demonstrated that the contents and in vivo exposure of some of the 11 components are consistent in the 6 products, particularly in the products containing the same natural or artificial bezoar and musk, or those manufactured by the same company. However, the inconsistency of the contents and in vivo exposure of some other ingredients suggests variations in the selection of raw herb medicines or processing technology. It suggests the crucial importance to monitor the source and quality of the raw herb medicines, and unify the processing technology. Furthermore, this study provides the data and reference methods for the supervision and consistency evaluation of quality amongst the products.

Lysosomes are intracellular organelles with lysosomal or digestive functions which contain various acid-dependent enzymes that are responsible for the recycling and degradation of intracellular substances. pH changes in lysosomes can reduce the activity of various enzymes, impairing the cell′s regulatory capacity and ultimately leading to apoptosis or pyroptosis. Therefore, monitoring lysosomal pH values is valuable for investigating alterations in intracellular physiological and pathological processes. This study developed a novel pH-responsive near-infrared (NIR) fluorescent probe targeting lysosomes, named piperazine-croconic acid-piperazine (PCP). This probe was formed by connecting the electron-donating group piperazine to both ends of croconic acid. The π-conjugated system, featuring a five-membered ring structure, endowed the probe with excellent stability and resistance to photobleaching. The tertiary amine moiety of piperazine, serving as a lysosome-targeting group, produced reversible protonation/deprotonation under varying pH conditions as result of exhibiting pH-responsive characteristics. The synthesized PCP demonstrated excellent NIR fluorescence performance, making it suitable for fluorescence imaging of lysosomal pH values in living cells. Both solution and intracellular experiments confirmed that PCP exhibited good biocompatibility and pH-responsive characteristics. Additionally, its symmetrical structure and molecular rigidity contributed to its excellent resistance to photobleaching. This study employed PCP probes to detect the pH values of the lysosomes, offering potential avenues for the treatment and prevention of diseases associated with lysosomal dysfunction.

New dressings represent a hot topic at the intersection of pharmacy, materials science, and medicine. The development of dressings with excellent biocompatibility, pharmacological activity, and mechanical properties is a new trend in this field. This study utilizes wood-based materials with inherent biocompatibility as the substrate. The ice crystal-induced phase separation method is employed to remove the tough lignin components, resulting in a three-dimensional micro-nano network structure of cellulose-hemicellulose matrix. Molecular injection is used to embed natural bioactive molecules tannin and auxiliary molecules polyvinyl alcohol in the matrix. Through multiple dynamic hydrogen bonding interactions, a novel dressing (de-lignin gel with PVA added, PLG) with both mechanical strength and biological activity is prepared. Various technical methods are employed to characterize the structure of PLG and investigate its bioactive functions. The results indicate that the designed PLG synthesis route is reasonable. The successfully obtained PLG exhibits excellent anisotropic mechanical strength and fatigue resistance (energy dissipation of 22.8 kPa·mm^-1). It also features a low linear expansion rate, outstanding adhesion capability, and anti-swelling properties. Antimicrobial experiments show that PLG exhibits antibacterial rates of 74.3% and 77.6% against Staphylococcus aureus and Escherichia coli, respectively. This indicates its broad-spectrum contact antibacterial effect. The radical scavenging rate of PLG reached 95.88%. Combined with mouse wound healing experiments, it was confirmed that PLG effectively reduces inflammatory responses through a dual antioxidant-antibacterial mechanism. The multi-layered structural design and cross-scale performance exploration of the dressing provide meaningful references for developing novel dressings that integrate mechanical adaptability, bioactivity, and multi-functional properties. The animal experiments were approved by the Laboratory Animal Ethics Committee of Zhejiang A & F University (No. ZAFUAC202514). All experiments were carried out strictly in accordance with relevant guiding principles and regulations.

Ranked second and fourth respectively in incidence rate and mortality rate among various cancers, breast cancer is characterized by strong invasion, high recurrence, poor prognosis and a lack of effective treatment, rendering the quality of life and the survival time of patients unsatisfactory. Herein, we designed a dual-response shape-transformable nanomicelle consisted of two amphipathy chains of different length, within which the chemotherapeutic agent paclitaxel (PTX) is encapsulated in the hydrophobic cavity. Based on it a combined therapy of photodynamic therapy, immunotherapy and chemotherapy was developed to augment the accumulation of therapeutic drugs in breast cancer tissues and treating efficacy. Entered in tumour microenvironment (TME), the shape transformation of micelles would be triggered by reactive oxygen species (ROS) and matrix metalloproteinase-2 (MMP-2), so as to increase the retention of the photosensitizer pyropheophorbide-a (Ppa) in tumours and release PTX and D-type programmed cell death ligand 1 (PD-L1) peptide antagonist (dPPA) concurrently, thus addressing the incompatibility of long blood circulation time, tumor penetration and retention. Under the irradiation of near infrared light, ROS produced by Ppa brought about stronger killing of cancer cells, activation of host immunity and promotion of ROS response. The dual-responsive group PTX@DTPD-PPP, along with two single-responsive groups PTX@DTPD-PRP and PTX@DPD-PPP and non-responsive group PTX@DPD-PRP were initially synthesized and characterized. Through cellular uptake experiment, light-responsive ROS generation experiment, cytotoxicity test, migration and invasion test and so forth, we proved the enhancement in both drug delivery and treating effect of dual-responsive formation. These findings demonstrate that PTX@DTPD-PPP is fulfilled with an uniform particle size (144 ± 1.3 nm) and a negative surface (-6.8 ± 0.9 mV), as well as the transformation responsiveness in 4 h within in vitro experiments. Both the drug accumulation and cellular toxicity of dual-responsive group are prominently increased in contrast with free Ppa, free PTX as well as other formations, laying a foundation for the sequent in-vivo studies.

Aloesone synthase (ALS), as a type Ⅲ polyketide synthase, plays an important role during the biosynthesis of diverse and active polyketide basic structural frameworks. In order to explore the expression pattern and function of RoALS2 in Rheum officinale, real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR), prokaryotic expression, and other techniques were used to analyze the expression pattern, in vitro protein expression, etc. of RoALS2 based on the previous transcriptome data. The overall results showed that the total length of the RoALS2 gene was 1 176 bp in size, encoding 391 amino acids, without signal peptides or transmembrane domains. Through evolutionary analysis and comparison, it was found that the RoALS2 is closely related to the ALS protein of Polygonum cuspidatum and R. palmatum. The RT-qPCR results showed that the relative expression level of RoALS2 in the roots and rhizomes of R. officinale increased first and then decreased with the change of growth years. The relative expression level was highest in the roots and rhizomes of two-year-old samples. The expression patterns of RoALS2 under different hormones' treatment were also different, and the overall trend was first upregulated, then downregulated, and then upregulated again. RoALS2 could respond to be upregulated by MeJA and SA treatments, but down-regulated by ABA and ethephon significantly. RoALS2 was then constructed into prokaryotic expression vectors pET-28a(+) and pGEX6p-1 respectively, and transformed into BL21 (DE3) competent, both of which showed successfully induction of the target fusion proteins. However, the pET-28a(+)-RoALS2 strain could be detected of the target band in the supernatant and the soluble protein was also obtained, whereas pGEX6P-1-RoALS2 recombinant protein existed in the form of inclusion bodies in the precipitate. The research results lay a solid foundation for further function characterization of the gene and elucidation of the molecular mechanisms of quality development of rhubarb from R. officinale.