Autophagy is an important physiological process that can degrade cellular components and maintain cellular homeostasis. In the process of cancer development, autophagy plays a dual role in promoting or inhibiting autophagy, and targeting autophagy is considered to be an important means of cancer treatment. According to Chinese medicine theory, autophagy has the function of regulating the balance of Yin and Yang, and the balance of good and evil in the organism, and based on the theory of "supporting the positive and dispelling the evil", the use of active ingredients of traditional Chinese medicine (TCM) to target autophagy has been proven to be effective in the treatment of cancer. In addition, autophagic cell death, as a type Ⅱ programmed cell death, is often accompanied by autophagic features, and the regulation of autophagic cell death is an important way for autophagy to achieve anti-tumor effects. In recent years, more and more studies have found that the active ingredients of TCM have good effects in cancer treatment, among which, targeting autophagic cell death is an important way for TCM active ingredients to achieve anti-tumor effects. This paper outlines the understanding of cancer and autophagy in Chinese medicine theory, and summarizes the current Chinese medicine small molecule compounds targeting autophagic cell death and their mechanisms of action based on the classification of natural medicines. Finally, the development of Chinese medicine-derived compounds targeting autophagic cell death for the treatment of diseases is summarized and prospected, which hopefully can provide clues for subsequent exploration and research.

Artificial intelligence (AI) technology is increasingly applied across various fields, particularly in handling and analyzing large volumes of data, providing breakthroughs for numerous scientific studies. In Chinese medicine research, AI demonstrates significant advantages by enhancing the systematic, efficient, and accurate nature of studies through its exceptional learning and data processing capabilities. As a discipline with a long-standing history and rich theoretical framework, Chinese medicine research requires the integration of complex information, for which AI provides crucial support. AI technologies, especially machine learning and deep learning, can decipher complex biological and chemical data, advancing new discoveries in Chinese medicine pharmacology. Researchers can systematically analyze the multi-target mechanisms of Chinese medicine components and optimize formulation efficacy through these technologies. The combination of AI with multi-omics data and its application in cell phenotype analysis aids in accurately identifying drug targets and exploring new mechanisms. Additionally, AI-integrated network pharmacology combines experimental, computational, and clinical data to analyze multi-target drug mechanisms, enhancing the efficacy of TCM formule. AI accelerates the target identification of active compounds as well as dissecting the pharmacological effects. The development of large language models also plays a crucial role in constructing Chinese medicine knowledge graphs and literature analysis, extracting valuable information from extensive literature using natural language processing to build a systematic knowledge structure. The introduction of AI technology has propelled the modernization of Chinese medicine research and has a pivotal role in the development of internationalization and precision medicine. AI not only enhances the overall level of Chinese medicine research but also provides a solid foundation for interdisciplinary collaboration and innovation. With the continuous advancement of AI technology, Chinese medicine is anticipated to have a greater influence and role globally. This process is not only a significant marker of the modernization of Chinese medicine but also a reflection of the integration of science and traditional wisdom, which will undoubtedly drive progress and development in the entire medical field.

Colorectal cancer is a major cancer threatening the life and health of people worldwide. With the transformation of modern medical models, treatment strategies represented by traditional Chinese medicine (TCM) with multiple targets have become a new research direction in the treatment of colorectal cancer. In the study of TCM, in addition to deconstructing the components and targets of TCM based on reductionism, more attention should be paid to the holistic concept in the basic theories of TCM, analyzing and understanding the mechanism of action of TCM treatment from a holistic perspective. TCM compound prescriptions, as a collection of single herbs, are composed of various monomers. Viewing TCM from different levels and considering different herbs and monomers as a whole helps us better understand TCM. At the same time, TCM has the characteristic of multiple targets. In addition to directly acting on tumor cells themselves, it can also affect the intestinal flora to improve the tumor microenvironment, treating tumors in an all-round and multi-dimensional way, regulating the physical functions of tumor patients from a holistic perspective, and comprehensively improving the health status of the individual. This review aims to systematically elaborate on the efficacy and multi-target mechanism of TCM in the treatment of colorectal cancer from three levels: TCM compound prescriptions, single herbs, and TCM monomers, based on clinical trials and preclinical research results. Meanwhile, this article will combine systems biology with the holistic view of TCM to conduct an in-depth analysis of related research on TCM treatment of colorectal cancer.

Prenylated aromatic natural products (PANPs) are widely distributed in nature. PANPs exhibit a great structural diversity due to their various core scaffolds (coumarin, lignan, benzoic acid/benzyl alcohol, flavonoid, xanthone, anthraquinone, and aromatic alkaloid, etc.) and the distinct types and substitution sites of isoprenoid moieties which may possess either linear or cyclic structures. The structural diversity of PANPs endow them with various bioactivities including anti-bacterial, anti-oxidation, anti-cancer, anti-inflammatory and analgesic effects, which makes them a group of highly promising molecules for drug development. Notably, isoprenoid moieties are often the indispensable pharmacophores in these active PANPs. Aromatic prenyltransferases (aPTs) are responsible for prenylation in the biosynthesis of PANPs. aPTs can be divided into three classes according to their evolutionary relationships and structural features, i.e. membrane-bound aPTs (UbiA type), soluble aPTs with a PT barrel structure (ABBA type and DMATS type) and terpene synthase-like aPTs. Herein, we summarize 94 aPTs belonging to the different classes which were characterized in the past ten years, in particular introduce their substrate selectivity/tolerance, regioselectivity, evolutionary relationships and structural features. This would provide cues for discovery and engineering of new aPTs, and modification and bio-production of active PANPs.

Usenamine A (UA) is a dibenzofuran compound isolated from Usnea longissima Arch. Previous studies have demonstrated that UA exhibited significant in vitro and in vivo anti-hepatocellular carcinoma activity, inducing both apoptosis and autophagy in human hepatocellular carcinoma cells. This study aims to further elucidate the molecular mechanism underlying its anti-hepatocellular carcinoma effect. Based on data obtained from gene chip assay, real-time quantitative PCR, and Western blot analysis, we found that UA effectively inhibited the expression of polo-like kinase 1 (PLK1) in human hepatocellular carcinoma HepG2 and SK-HEP-1 cells. Furthermore, the administration of BI 6727, a PLK1 inhibitor, significantly diminished the inhibitory effect of UA on the viability of human hepatocellular carcinoma cells. Additionally, the knockdown of PLK1 expression via RNA interference markedly inhibited the proliferation of human hepatocellular carcinoma cells, and the inhibitory effect of UA on cell viability was attenuated upon PLK1 knockdown. The knockdown of PLK1 expression significantly upregulated the apoptosis rate of human hepatocellular carcinoma cells and notably diminished the apoptosis-inducing effect of UA on these cells. Additionally, the inhibition of autophagy using the autophagy inhibitor 3-MA reduced the proliferation-inhibitory effect of UA on human hepatocellular carcinoma cells. Utilizing the PLK1 inhibitor BI 6727 or RNA interference, we further demonstrated that PLK1 negatively regulated autophagy in human hepatocellular carcinoma cells. Consequently, the inhibition of PLK1 attenuated the autophagy induction by UA on these cells. Thus, PLK1 plays a crucial role in the inhibition of human hepatocellular carcinoma cell proliferation and the induction of apoptosis by UA. Moreover, UA activates autophagy through the inhibition of PLK1, which subsequently exerts an inhibitory effect on the growth of human hepatocellular carcinoma cells.

Arjunic acid (AR), a main bioactive triterpenoid isolated from acorns, has been reported to exert pronounced anti-inflammatory activities. However, its anti-inflammatory mechanisms have not been elucidated. In this study, the model of lipopolysaccharide (LPS)-induced inflammation in RAW264.7 cells were established to investigate the anti-inflammatory activity of AR. The potent targets and signaling pathway of AR for the treatment of inflammation-related disease were predicted based on network pharmacology. Furthermore, the expression of pro-inflammatory cytokines and mediators was determined by Griess assay, enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and Western blot. The protein expression of NF-κB, MAPK, Nrf2/HO-1, PI3K/Akt/mTOR, and autophagy signaling pathways were gauged by Western blot. As the result, in the inflammatory model of LPS-induced RAW264.7 cells, AR could significantly inhibit the expression of pro-inflammatory cytokines and mediators, suppress the phosphorylation and translocation of NF-κB, and downregulate the phosphorylation of JNK/ERK signaling pathways. AR also inhibited ROS production and activated the Nrf2/HO-1 signaling pathway by degrading Keap1. Furthermore, AR activated autophagic flux by inhibiting the PI3K/Akt/mTOR signaling pathway. Collectively, AR was a potential natural product for the treatment of inflammation-related diseases.

Acute kidney injury (AKI) is a prevalent clinical syndrome characterized by a rapid deterioration in renal function. Naringin, a flavonoid abundant in the Rutaceae family, has been reported to provide protective effects against kidney injury. However, the mechanisms responsible for these effects remain inadequately elucidated. In the present study, the AKI mouse model was established in vivo through a single intraperitoneal injection of 20 mg·kg^-1 of cisplatin, and human kidney-2 (HK-2) cell injury was induced by cisplatin in vitro. Blood urea nitrogen (BUN) and creatinine (CRE) levels were measured using an ELISA kit. Hematoxylin and eosin (H&E) staining, along with periodic acid-Schiff (PAS) staining, were employed to evaluate changes in renal histopathology. Serum concentrations of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the mice were assessed using ELISA, and the levels of these cytokines were further analyzed. Morphological alterations in HK-2 cells were examined microscopically. The levels of reactive oxygen species (ROS) in HK-2 cells were quantified using an ELISA kit. The protein expression levels of p-MAPK/MAPK, p-JNK/JNK, p-ERK/ERK, p-NF-κB/NF-κB, p-IκBα/IκBα, IL-6, and TNF-α in renal tissue and HK-2 cells were evaluated through Western blot. The results showed that the serum BUN, Cre, IL-6, and TNF-α levels in the model group mice increased and the renal tissue structure was damaged when compared to the control group, whereas naringin could significantly ameliorate the above pathological changes. Moreover, naringin significantly reduced cisplatin-induced ROS production in HK-2 cells. Western blot results confirmed that naringin notably attenuated the expression of p-MAPK, p-JNK, p-ERK, p-NF-κB, p-IκBα, IL-6, and TNF-α proteins in vitro and in vivo. In conclusion, naringin may ameliorate cisplatin-induced AKI by inhibiting the activation of the MAPK/NF-κB pathway and reducing the production of ROS. All animal experiments were approved by the Institutional Animal Care and Use Committee of Hubei University of Chinese Medicine (approval No: HUCMS202204001).

This study aimed to clarify the mechanism and active components of Buyang Huanwu Decoction (BYHWD) in alleviating cerebral ischemia reperfusion injury (CIRI) by inhibiting pyroptosis. The key components and targets of BYHWD for CIRI were identified via network pharmacological analysis, followed by molecular docking performed with Autodock and Pymol software. The effects of BYHWD and its active components were validated in vivo and in vitro. A middle cerebral artery occlusion (MCAO) model was established in mouse to assess neural function alterations in mice under various conditions. Concurrently, an oxygen-glucose deprivation/reperfusion (OGD/R) model was developed utilizing mouse brain tissue astrocytes in vitro. Molecular biology experiments were used to verify the predicted key targets. We have determined that the principal components of BYHWD are baicalein and β-sitosterol. By analyzing genes associated with CIRI pathology alongside those linked to pyroptosis, 20 intersecting genes were identified. In conjunction with molecular docking binding energy assessment, TP53 and TNF emerged as pivotal core targets for subsequent validation. Molecular biology experiments confirmed that BYHWD effectively alleviates injury while reducing the expression level of P53. These findings indicate that the primary bioactive constituents of BYHWD were baicalein and β-sitosterol. In addition, BYHWD may inhibit pyroptosis via TNF and TP53 in protecting CIRI. The experiment has been approved by the Experimental Animal Welfare Ethics Committee of Zhejiang Academy of Traditional Chinese Medicine, approval number (KTSC2020037, KTSC2023030).

The study investigates the therapeutic effects and mechanisms of Buyang Huanwu Decoction (BHD) in treating ischemic stroke (IS). Using a middle cerebral artery occlusion/reperfusion (MCAO/R) rat model, we evaluated the neuroprotective effects of BHD, demonstrating significant improvements in neurological function scores, prolonged rotarod retention time, and reductions in both infarct volume and brain water content. An unsupervised clustering algorithm was employed to identify active components of BHD by clustering them with FDA-approved drugs for ischemic stroke treatment. Combined with network pharmacology analysis, the mechanisms of these active components were predicted to be associated with anti-inflammatory pathways. Further validation using a lipopolysaccharide (LPS)-induced BV-2 cell model demonstrated the anti-inflammatory efficacy of seven key active components, with their effects on anti-inflammatory activity and cell viability assessed via the Griess and MTT assays. Additionally, the content of these active components in BHD was quantified using liquid chromatography-mass spectrometry (LC-MS). In conclusion, this study elucidates the critical active components of BHD and their potential pharmacological mechanisms, providing valuable insights for the modernization of traditional Chinese medicine and its application in ischemic stroke therapy. All animal experiments were approved by the Animal and Medical Ethics Committee of Northeastern University (approval No.: NEU-EC-2023A052S).

The purpose of this study is not only to investigate the effects of Angelica sinensis polysaccharide (ASP) as a potential vaccine adjuvant on immune activation and cytokine release in RAW264.7 macrophages, but also to elucidate its underlying involved signaling mechanisms. Cell viability was evaluated by the CCK-8 assay. Flow cytometry was used to analyze the influence of ASP at five distinct concentration gradients on the expression of cluster of differentiation (CD) 80, CD86, and major histocompatibility complex Ⅱ (MHC Ⅱ) on RAW264.7 cell surfaces. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cell culture supernatant were determined by enzyme-linked immunosorbent assay (ELISA) method. Molecular techniques, including quantitative polymerase chain reaction (qPCR) were utilized to assess the mRNA expression levels of Toll-like receptor 4 (TLR4)-myeloid differentiation factor 88 (MyD88)-TNF receptor associated factor 6 (TRAF6)-nuclear factor kappa-B (NF-κB) signaling pathway. The levels of TRAF6, and the phosphorylation levels of IκB kinase (IKK) and p65 proteins were detected by Western blot. The results show that ASP at varying concentrations promote the proliferation of RAW264.7 cells without cytotoxicity. Surface molecules CD80, CD86, and MHC Ⅱ on RAW264.7 cells showed statistically significant up-regulation in response to ASP compared to the blank control (P < 0.05), with a dose-dependent effect within an optimal range. Furthermore, ASP also elevated cytokines IL-6 and TNF-α secretion levels by RAW264.7 cells compared to the normal control (P < 0.05), exhibiting a dose-response relationship within a specific concentration span. The qPCR results indicated that ASP groups at different concentrations all led to upregulation of mRNA expression levels of TLR4, MyD88, TRAF6, and NF-κB signaling pathway. The expression levels of TRAF6, p-IKK and p-p65 were increased by different concentrations of ASP. The TLR4 inhibitor TAK-242 significantly reduced the secretion of cytokines induced by APS (P < 0.05). This study highlights the immunostimulatory properties of ASP, emphasizing its potential as a vaccine adjuvant. By significantly enhancing the expression of co-stimulatory molecules and cytokines via the TLR4-MyD88-TRAF6-NF-κB signaling pathway, ASP offers a promising approach for modulating immune responses.

Twenty compounds were isolated from the twigs and leaves of the 95% ethanol extract of Miliusa tenuistipitata W. T. Wang by various chromatographic techniques including silica gel column chromatography, sephadex LH-20 and preparative HPLC. Their structures were determined by analysis of spectral data as tenuistone L (1), tenuistone K (2), sakuranetin (3), pachypodol (4), retusin (5), penduletin (6), casticine (7), 3, 7-dimetoxi-5, 3′, 4′-tri-hidroxiflavona (8), eupatin (9), (+)-miliusol (10), miliusane XIX (11), miliusane XVIII (12), miliusolide (13), 19, 20-dihydromiliusolide (14), debilisone C (15), goniothalamusin (16), 2, 10-dihydroxy-3, 9-dimethoxy-8-oxo-protoberberine (17), consanguine B (18), 6β-hydroxystigmasta-4, 22-dien-3-one (19) and 6β-hydroxystigmast-4-en-3-one (20), respectively. Among them, compounds 1 and 2 are new compounds, compounds 6, 9, 18 and 20 are isolated from Annonaceae plants for the first time and compounds 5, 7, 17 and 19 are isolated from the genus Miliusa for the first time. Human triple negative breast cancer cells (MDA-MB-231 and BT-549) were used to evaluate the antitumor activity of these isolates by MTT assay. Compounds 10 and 11 showed significant anti-proliferation activity against the test cells with half inhibition concentration (IC₅₀) of 0.98-4.85 μmol·L^-1.

In the study, we employed an untargeted metabolomic approach in conjunction with 16S rRNA high-throughput sequencing to identify potential antidepressant active effectors mediated by gut-flora among geniposides. Firstly, the rat depressed model was constructed using chronic unpredictable mild stress stimulation (CUMS) combined with orphaned model. Then the cecal contents of the rats were analyzed by untargeted metabolomics using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) in order to identify differential metabolites. The intestinal genera exhibiting significant differences in the cecal contents were identified through 16S rRNA high-throughput sequencing, and correlation analysis was subsequently conducted between the differential metabolites and the intestinal genera. The comparison of the metabolic profiles between the normal control group, the depression model group, and the geniposide treatment group revealed that 147 metabolites were down-regulated and 381 metabolites were up-regulated in the model group compared with the normal group. Furthermore, 212 metabolites were up-regulated and 288 metabolites were down-regulated in the geniposide treatment group compared with the model group. Subsequently, the combination of the screening conditions of FC (fold change) > 2, P < 0.05 and VIP (variable importance in the projection) > 1, secondary mapping and database comparison led to the identification of 55 metabolites with significant differences among the three groups. The results of the metabolic pathway enrichment analysis indicated that the identified differential metabolites were primarily involved in five metabolic pathways, namely tryptophan metabolism, arginine biosynthesis, phenylalanine biosynthesis with tyrosine and tryptophan, phenylalanine metabolism, and arginine metabolism with proline. The 16S rRNA sequencing results further indicated that Gardenia jasminoides extract may exert its antidepressant effects, thereby providing a new basis for the study of the gut-brain axis in the therapeutic strategy of depression. All animal experiments were conducted with the approval of the Biomedical Research Ethics Committee of the Naval Medical University.

Platinum-based complexes, particularly divalent platinum [platinum(Ⅱ), Pt(Ⅱ)] compounds, have become classic chemotherapy agents for the treatment of malignant tumors. However, their widespread clinical use is limited due to issues such as insufficient stability, the induction of acquired resistance and strong cytotoxicity. Although antibodies that interfere with the interaction between programmed cell death protein 1 (PD-1) and programmed cell death ligand 1 (PD-L1), in combination with platinum-based compounds, have shown significant clinical progress in cancer treatment, their high efficacy is often accompanied by substantial toxicity and immune-related side effects, which limit their long-term use. In contrast, platinum(Ⅳ) [Pt(Ⅳ)] complexes, with their unique octahedral geometry, have demonstrated promising anticancer potential. By modifying the axial ligands, Pt(Ⅳ)-based complexes not only show higher inertness and improved tumor selectivity, but also enable the targeted release of active ligands in the tumor microenvironment. The mechanism allows Pt(Ⅳ)-based complexes to overcome drug resistance, reduce toxicity and enhance immune system activation, making them a research hotspot in the current cancer field. More importantly, Pt(Ⅳ)-based complexes can exert anticancer effects through multiple pathways including causing DNA damage to trigger apoptosis, autophagy and ferroptosis in tumor cells. This multifaceted action mechanism not only enhances antitumor efficacy but also significantly reduces side effects associated with traditional platinum-based compounds. This review summarizes whole anticancer mechanisms of platinum-based complexes, particularly Pt(Ⅳ) complexes in chemo-immunetherapy combination therapies, discussing their potential in cancer treatment and providing theoretical support for the development of efficient, low-toxicity and highly selective Pt(Ⅳ)-based complexes.

Transient receptor potential vanilloid (TRPV), an important class of non-selective cation channels, is closely associated with a variety of physiological and pathological processes. Pungent flavour is one of the important flavours in the doctrine of five flavours in traditional Chinese medicine, which has the functions of relieving the epidermis, moving the Qi, activating the blood and so on, which has a variety of main treatments. Modern research has shown that the active ingredients in numerous pungent Chinese medicine have the ability to modulate TRPV channels. The purpose of paper is to explore the biological characteristics of TRPV channels, the modern research process of pungent Chinese medicine and its mechanism in inflammation, pain and other pathological processes. This paper discusses the research progress of the pharmacological action of pungent Chinese medicine based on TRPV channel, in order to reveal the scientific connotation of spicy traditional Chinese medicine, and also provide new research ideas and models for the modern research of Chinese medicine.

Colorectal cancer (CRC), a prevalent and deadly digestive malignancy, primarily causes death through recurrence and metastasis. Current treatments include surgery, radiotherapy, chemotherapy, and immunotherapy, but they have significant side effects and high recurrence rates. Research shows traditional Chinese medicine (TCM) can alleviate symptoms, reduce adverse reactions, enhance quality of life, prevent recurrence and metastasis post-surgery, and improve survival rates. This review explores TCM's theoretical framework, advantages in CRC management, and potential in modulating intestinal flora for CRC prevention and treatment.

Resiquimod (R848) belongs to the class of weak base organic compounds that are derivatives of imidazole quinoline and is a potent agonist of Toll-like receptor (TLR) 7/8. R848 can be considered an effective immune adjuvant as it has the ability to activate different immune cells and regulate innate and adaptive immunity. A number of researches done recently have reported R848 potent anti-tumor capacity, especially when combined with other cytotoxic therapies. Nonetheless, some hurdles remain with the clinical use of R848 such as its limited ability to reach the tumor and likelihood of inflammation and autoimmune responses which might result from repeated delivery of high dosages of the drug. Nanoscale drug delivery systems may overcome some of these challenges. Various nanoformulations, including liposomes, polymeric nanoparticles, biocarriers, inclusion compound, metal-organic frameworks and inorganic nanoparticles, as well as several R848 prodrugs or derivatives-baesd nanoparticles, have been incorporated into present R848 delivery platforms. This article reviews the physicochemical properties, immune regulatory mechanisms and nano drug delivery systems of R848, in order to provide reference and support for anti-tumor research and new drug development.

Wound healing caused by skin trauma and chronic diseases is often complicated and difficult due to infection. In such cases, the wound healing process is not only a long and continuous one, but also prone to scar repair. Conventional dressings and antibiotic treatments suffer from issues such as low drug delivery efficiency and systemic toxicity. In recent years, microneedle technology has been widely used to reduce infection and promote wound healing. Microneedles can not only penetrate the skin stratum corneum and biofilm to enhance drug delivery efficiency and reduce drug side effects, but also can be combined with a variety of materials to achieve multiple treatments and monitoring of wounds. This article will discuss the healing mechanism of infected wounds, the classification of microneedles, and their applications in the treatment of infected wounds.

Plant-derived extracellular vesicles (PDEVs) are lipid bilayer-structured nanovesicles secreted by plant cells, recognized as excellent drug carriers due to their high stability, safety, and modifiability. As an emerging drug delivery system, PDEVs are gaining increasing attention. This review systematically summarizes the latest research progress on the preparation, characterization, engineering transformation, drug loading methods, and the applications and advantages of PDEVs as drug carriers. Finally, the future development directions of PDEVs in drug delivery are discussed.

Non-small cell lung cancer (NSCLC) is the primary pathological type of lung cancer. Osimertinib, as a third-generation EGFR-TKI (epidermal growth factor receptor-tyrosine kinase inhibitor) targeted drug, effectively prolong the progression-free survival of patients with EGFR-mutated NSCLC. However, drug resistance limits the efficacy of osimertinib. Traditional Chinese medicine can effectively delay the resistance to EGFR-TKIs. In this study, Cucurbitacin B (CuB) was investigated to analyze its pharmacological effects in osimertinib-resistant NSCLC cells through cell viability, migration, and invasion experiments. Public databases were used to screen potential targets of CuB in osimertinib-resistant NSCLC cells, and the interactions between CuB and potential targets were verified through molecular docking, cellular thermal shift assay (CETSA), and microscale thermophoresis (MST). Western blot was used to detect the effects of CuB on downstream pathways of the targets. The results showed that CuB significantly reduced the proliferation activity of osimertinib-resistant NSCLC cells and inhibited the migration and invasion abilities of tumor cells. Mechanistically, CuB inhibited the expression of ERK and AKT molecules by binding to the tyrosine kinase receptor AXL. In summary, CuB exhibits resistance to osimertinib-resistant NSCLC by inhibiting the migration and invasion of resistant cells through regulating the abnormal activation of the AXL-ERK/AKT axis. This study provides a basis for the pre-clinical in vitro efficacy of CuB in the treatment of osimertinib targeted resistance in NSCLC and theoretical support for the development of clinical drug combinations.

This study mainly explored the anti-tumor effects of programmed death ligand 1/ transforming growth factor-β (PD-L1/TGF-β) bispecific antibody drugs in the huHSC-NCG mouse brain orthotopic transplantation model of human glioma cell U251-luciferase (all animal experiments in this study were approved by the Experimental Animal Ethics Committee of GemPharmatech, and the ethics review number is GPTAP20230706-3). The antibody-antigen binding activity of the PD-L1/TGF-β bispecific antibody drugs to PD-L1 target on U251 cells was detected by flow cytometry in vitro. The antitumor activity of PD-L1/TGF-β bispecific antibody drugs was evaluated using a brain glioma in orthotopic implantation model with humanized immune system. Live animal imaging and brain tissue (including tumor tissue) weight results showed that the PD-L1/TGF-β dual antibody drug had an inhibitory effect on the growth of glioma in situ. The results of drug content (ELISA assay) and drug distribution (immunohistochemistry assay) in brain tissue showed that PD-L1/TGF-β dual antibody could enter the brain glioma and play the function of immune checkpoint, thus playing an inhibitory role in brain glioma. Immunofluorescence staining of mouse brain tissue showed that the drug activated the immune system and had synergistic anti-tumor effect with the immune system. These results showed that the PD-L1/TGF-β bispecific antibody drugs have broken through the blood-tumor barrier in orthotopic implantation model of mouse brain glioma and demonstrated a strong anti-tumor effect. In conclusion, the inhibitory effects of PD-L1/TGF-β bispecific antibody drugs on human brain glioma provides a new strategy for the treatment of human brain glioma.

Tumor immunotherapy represented by blocking programmed cell death protein 1/programmed cell death ligand 1 (PD-1/PD-L1) has gained better therapeutic effect in lung cancer treatment, and the development of small molecule drugs to block PD-1/PD-L1 provides a new strategy for lung cancer immunotherapy. In this study, we applied an in vitro cell model to investigate the regulation of PD-L1 expression and the mechanism of action of baicalin on A549 lung cancer cells. The effects of baicalin on the viability of A549 cells were detected by CCK8 assay; the expression of PD-L1 protein in A549 cells was detected by Western blot; the mechanism of baicalin cytotoxicity and its correlation with PD-L1 protein expression were investigated by using small-molecule inhibitors of apoptosis and autophagy; the formation of autophagic vesicles in A549 cells treated with baicalin was observed under transmission electron microscope. And the alterations of acid autophagic vesicles and autophagic lysosomes were observed under fluorescence microscope. The results of CCK8 experiments showed that baicalin (12.5-400 μmol·L^-1) inhibited the proliferation of A549 cells in a dose-dependent manner; at the same time, the elevation of PD-L1 protein expression after interferon-γ (IFN-γ) interference could be reduced with the increase of baicalin concentration; on the other hand, treatment with autophagy inhibitors, wortmannin, and chloroquine, could call back the baicalin-induced cytotoxicity, transmission electron microscopy and fluorescence microscopy showed that the number of autophagic vesicles increased after baicalin treatment of A549 cells, and Western blot results showed that baicalin promoted the expression of autophagy-related proteins LC3-Ⅱ/Ⅰ and beclin-1; the number of baicalin-induced acidic autophagic vesicles in A549 cells was attenuated after the intervention of wortmannin, and at the same time the LC3-Ⅱ/Ⅰ expression was inhibited and the inhibitory effect on PD-L1 was regulated. The above results suggest that baicalin may exert antitumor effects by activating the autophagy protein LC3 in A549 cells to reduce the expression of PD-L1. This study lays the foundation for baicalin to have the ability to be developed as a small molecule blocker of the PD-1/PD-L1 signaling pathway.

The chemical constituents of the fruits of Forsythia suspensa (Thunb.) Vahl were investigated by using chromatographic techniques (silica gel, MCI, and sephadex LH-20 gel column chromatography, etc). Fourteen compounds were isolated from the 95% ethanol extract of F. suspensa, and their structures were identified by HR-ESI-MS, NMR, and calculated electronic circular dichroism (ECD) methods. The compounds included labdane diterpenes (1-6), clerodane diterpene (7), norlabdane diterpene (8), norclerodane diterpene (9), oleanane triterpenoid (10), ursane triterpenoids (11, 12), lupane triterpenoids (13, 14). Among them, compound 1 was a new compound and compounds 3-9, 11, and 12 were obtained from this plant for the first time. The inhibitory effect of these compounds on lipopolysaccharide-induced nitric oxide production in mouse macrophage RAW 264.7 cells also was evaluated. Unfortunately, none of these compounds exhibited significant inhibitory activity.

In this thesis, we propose to establish a phage display method for screening adalimumab (ADM)-specific binding peptide to provide a particular element of recognition for clinical studies of ADM, and to successfully establish an indirect enzyme-linked immunosorbent assay (i-ELISA) based on this peptide for clinical detection of ADM. With ADM as the target molecule, five rounds of peptide elution were performed using phage display technology by putting it into the M13 linear dodecapeptide library. By measuring the titer of each round of eluate, it was found that the recovery rate increased from 7.28×10^-6 to 1.55×10^-3, indicating that the peptides binding to ADM in the eluate were continuously enriched, and the enrichment effect was better; the fifth round of screening amplicons were sequenced, and it was found that five peptide sequences appeared with high frequency, which were considered to be potentially able to bind specifically to ADM and were synthesised; the specific binding ability of the five peptide sequences to ADM was verified and identified by surface plasmon resonance (SPR) and ELISA experiments, respectively, and the results showed that the first peptide sequence (Pep1) and the second peptide sequence (Pep2) showed specific binding to ADM, with the affinity constants (K_D) of 7.91×10^-5 mol·L^-1 and 1.67×10^-5 mol·L^-1, respectively, and of which the affinity constant of Pep1 with ADM isotype antibody IgG was 1.35×10^-4 mol·L^-1, which was one order of magnitude lower than that with ADM, and thus Pep1 was determined to be a specific binding peptide for ADM. Based on Pep1, an indirect ELISA method for the analysis of the antibody-drug ADM was successfully established. With the increasing concentration of ADM the OD₄₅₀ value showed a regular change, and the method can be used for clinical ADM blood concentration monitoring. In this study, we obtained a peptide that can specifically bind to ADM, and this peptide sequence can be used as a specific recognition element for ADM, which not only enables the monitoring of ADM blood concentration, but also provides an effective tool for the accurate qualitative and quantitative quantification of ADM in vivo and ex vivo, and at the same time provides a new strategy and idea for the discovery of recognition elements of other monoclonal antibody drugs.

The main chemical components of Gandoufumu Decoction (GDFMD) were analyzed and identified using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS^E). We utilized a ZORBAX RRHD Eclipse Plus C₁₈ column (2.1 mm × 100 mm, 1.8 μm) with a gradient elution of 0.1% formic acid in water and acetonitrile as mobile phase at a flow rate of 0.2 mL·min^-1 and a column temperature of 35 ℃. The mass spectrometry was analyzed using an electrospray ionization source (ESI) in positive and negative ion modes. The UPLC-Q-TOF/MS^E was used to detect and analyze the complex mass spectra and the chemical composition of the GDFMD. The results show that 102 compounds were identified in GDFMD, 26 flavonoids, 22 terpenoids, 19 saponins, 10 phenylpropanoids, and 25 other components. The established qualitative method can rapidly and efficiently identify the chemical components of GDFMD, providing a scientific basis for the quality evaluation and clinical application of GDFMD.

To explore the standardization of the in vitro Nb2-11 cell proliferation bioassay for recombinant human growth hormone (rhGH), we first tested the biological activity of 2 batches of rhGH drug substances, 3 batches of rhGH for injection, and 24 batches of rhGH injections, based on which we proposed the experimental effective criteria. Furthermore, we conducted methodological validation in accordance with General Rule 9401 of the 2020 edition of the Pharmacopoeia of the People's Republic of China. Subsequently, the inter-laboratory consistency, intra-laboratory and inter-laboratory precision of seven labs were studied by determining the in vitro bioactivity of six batches of rhGH products and two batches of rhGH drug substances from four different manufacturers using this method. The consistency of the in vitro and in vivo bioassays was assessed by evaluating 28 batches of rhGH drug substances with both methods. Finally, multiple batches of rhGH samples with different expiration dates were tested to determine the standard limits of the bioassay. The results of this collaborative study indicate that the Nb2-11 cell bioassay exhibits good intra- and inter-laboratory reproducibility, with a 100% pass rate for the experimental effective criteria. The Nb2-11 cell bioassay had high consistency with in vivo animal bioassay. Animal experiments were approved by the Experimental Animal Welfare Ethics Committee of China National Institute of Food and Drug Control (approval number: NIFDC (Fu) No. 2024 (B) 004). This method is applicable to rhGH drug substances and products from different manufacturers. Collectively, the Nb2-11 cell bioassay can be used as a standardized method for determining the biological activity of rhGH products, replacing in vivo animal methods for quality control and release testing.

A detection method using UHPLC-MS/MS was established for the determination of 53 prohibited veterinary drug residues in goat horns, including 16 tranquilizers, 14 β-agonists, chloramphenicol, 4 fluoroquinolones, 7 nitroimidazoles, 3 quinoxalines, and 8 other compounds. Samples were extracted using salting-out assisted liquid-liquid extraction (SALLE). A Zorbax Eclipse Plus C18 column (1.8 μm, 3.0 mm × 150 mm, Agilent) was used with 0.1% formic acid solution-acetonitrile as the mobile phase for gradient elution. Detection was performed in positive and negative electrospray ionization modes using multiple reaction monitoring (MRM). All 53 analytes showed good linearity within their respective concentration ranges, with correlation coefficients above 0.99. The average recoveries at three spiked levels ranged from 70.4% to 118.7%, with relative standard deviations (RSDs) ranging from 0.69% to 12.07%. The limits of detection (LODs) ranged from 0.1 to 50 μg·kg^-1, and the limits of quantitation (LOQs) ranged from 0.2 to 100 μg·kg^-1. This method has been applied to the determination of real samples.

The critical relative humidity (CRH) is an important indicator for measuring the hygroscopicity of solid drugs. The intrinsic mechanism of rapid moisture absorption of water-soluble drugs under CRH is not fully understood, and we conducted preliminary exploration on this topic. Firstly, the newly developed model tablet moisture absorption method and the traditional powder moisture absorption method were employed to determine the CRH values, an equivalence test between the two methods was conducted by a two-tailed t test. Then a conductivity meter was employed to measure the conductivity values of drug model tablets under different relative humidity (RH) conditions. As we know, anhydrous copper sulfate is a grayish white powder, which is easy to turn blue after absorbing water, based on this, this salt was selected as a model drug, and its CRH value was measured. Furthermore, anhydrous copper sulfate was compressed into tablets, then placed under different RH conditions. A series of photos were taken to record the formation process of the liquid film. The reason for the inflection point of the moisture absorption equilibrium curve was postulated based on changes in conductivity and color. The results showed that the CRH values of sucrose powders and tablets were 85.05% and 85.01%, respectively; and the CRH values of sodium citrate powders and tablets were 77.14% and 76.93% individually; the results of the two-tailed t test indicated that the two methods are equivalent. At RH of 75%, the conductivity of sodium citrate changed slightly, while at RH of 79% greater than the CRH, the conductivity underwent a sudden change. Sucrose and anhydrous copper sulfate showed a similar trend of change. In addition, the process of blue dots of anhydrous copper sulfate changing from discrete to continuous was successfully captured during the observation of moisture absorption. In summary, the aforementioned results suggest that the mechanism of the rising rapidly in moisture absorption curve of water-soluble drugs at CRH is probably attributed to the formation of a continuous aqueous "liquid film" on the drug surface during the process of moisture absorption.

To prepare a progesterone pressure-sensitive gel patch combined with a microneedle to enhance drug release, HPLC was used to determine the preparation's progesterone content. One-way and orthogonal experiments were used to optimize the patch's prescription. Adhesion, sensory evaluation, cumulative release, and cumulative penetration were used as evaluation indices. Three microneedles with varying needle heights were made using 3D printing, and the cumulative penetration of the patch and microneedles was calculated and compared with the patch alone. The orthogonal experiments showed that the optimal prescription for the patches was Duro-Tak 87-2677 pressure-sensitive adhesive (87.5%), tributyl citrate (2%), isopropyl myristate (5%), dibutylated hydroxytoluene (0.5%), and drug (5%). The patches were prepared according to the optimized prescription, resulting in good patch formability and adhesion. In the transdermal penetration test, the cumulative penetration of the patch was 52.35 ± 7.88 μg·cm^-2 at 24 h, and the cumulative penetration of the patch in combination with 500, 750, and 1 000 μm microneedles was 226.01 ± 7.46, 278.78 ± 6.59, 422.95 ± 16.81 μg·cm^-2, respectively. The experiment was approved by the Experimental Animal Ethics Committee of Xinjiang Medical University (IACUC-20220725-8). The optimal patch prescription was screened through one-way and orthogonal experiments, and the transdermal penetration effect of patch and microneedle combination preparation was better than that of single use, which can effectively increase the in vitro transdermal penetration of the drug, and the above study provides a theoretical basis for the application of transdermal patches of progesterone.

The HIV-1 capsid protein (CA) is an effective target for antiviral drug discovery. Targeting the capsid assembly inhibitor peptide (CAI) binding pocket in the C-terminal domain of HIV-1 CA (CA CTD) can be used to screen for CA protein inhibitors. In this study, we found that the small molecule suramin targets HIV-1 CA by binding to this pocket, with an IC₅₀ value of 2.1 μmol·L^-1 for inhibiting the CA CTD-CAI interaction, based on homogeneous time-resolved fluorescence (HTRF) technology. Bio-layer interferometry (BLI) experiments demonstrated that suramin binds directly to CA, with a binding affinity to CA hexamer (K_D = 248 nmol·L^-1) higher than to CA monomer (K_D = 227 μmol·L^-1). In addition, in vitro CA protein assembly assays showed that suramin promotes disordered multimerization of CA protein. Binding model analysis revealed that suramin binds to the interface between adjacent CA monomers in the CA hexamer through the N57, Q63, Q67 and Y169 residues, thereby enhancing CA multimerization. In summary, suramin is a CA protein inhibitor that targets the CA hexamer and can serve as a starting point for the discovery of new CA inhibitors.

Root rot represents a significant disease affecting the cultivation of Platycodon grandiflorum. The screening of biocontrol strains with antagonistic properties against this disease can provide valuable microbial resources for the environmentally friendly prevention and control of Platycodon grandiflorum diseases. In this investigation, high-throughput bacterial isolation techniques were utilized to isolate and purify endophytic bacteria from the roots of Platycodon grandiflorum. An antagonistic bacterial strain R34B7, was identified through the plate confrontation culture method, exhibiting a notable inhibitory rate of 52.18% against the pathogen causing root rot. Morphological characteristics, physiological and biochemical properties, and molecular identification results collectively confirmed that the antagonistic endophytic bacterium R34B7 belonged to the species Serratia plymuthica. The control efficacy of strain R34B7 against Platycodon grandiflorum root rot was assessed using tissue-cultured seedlings of Platycodon grandiflorum, revealing a disease control efficacy of 44.44%. Furthermore, the fermented supernatant of strain R34B7 demonstrated considerable inhibitory effects on both the mycelial growth and spore germination of the pathogen. By examining the extracellular hydrolytic enzymes and growth-promoting factors of strain R34B7, it was discovered that this strain possesses the abilities to produce protease, chitinase, fix nitrogen, solubilize phosphorus, synthesize siderophores, and produce indole-3-acetic acid, indicating its potential for growth promotion. The antagonistic bacterium R34B7 identified in this study exhibits promising biocontrol activity against root rot and holds considerable potential for further development and utilization.