Kynurenine 3-monooxygenase (KMO) is a key rate-limiting enzyme in the downstream catabolism of kynurenine pathway (KP). Under the catalysis of KMO, the intermediate product kynurenine is metabolized into various active metabolites, including 3-hydroxykynurenine (3-HK), quinolinic acid (QA) and nicotinamide adenine dinucleotide (NAD⁺). More and more studies have shown that abnormal KMO expression activity mediates KP metabolic disorders, and is involved in the occurrence and development of nervous system diseases, autoimmune diseases, infectious diseases and tumors, suggesting that KMO can be used as a potential and effective drug therapeutic target. This article focuses on the role of KMO in the pathological mechanism of various diseases, and summarizes the existing KMO inhibitors to provide methods and ideas for targeted KMO therapy.

Bacterial resistance has always been a challenge in the field of antibacterials research. As one of the most problematic pathogens causing nosocomial infections, Acinetobacter baumannii (A. baumannii) was a severe threaten to public health. In recent years, as the rate of multidrug-resistant, extensively drug-resistant, and even pan drug-resistant A. baumannii has been increasing, there is an urgent need to develop new strategies to combat this pathogen. Iron is an essential element for the survival and the infection process of A. baumannii. Bacteria have developed a series of iron uptake strategies to survive in conditions of the host iron starvation, including the secretion of siderophore for iron chelation, the uptake of ferroheme, and the transport of ferrous by Feo system. Developing antibacterial strategies to inhibit the iron uptake is an effective way to deal with A. baumannii infection. This article will review the mechanisms of iron uptake by A. baumannii and the antimicrobial strategies developed based on them.

As the second largest cofactor after ATP in body, S-adenosyl-L-methionine (SAM) is responsible for methyl donor in SAM-dependent methyltransferases (MTases). The methylation of essential ingredients (e.g., DNA, RNA, protein) plays a critical role in epigenetic regulation, cellular signal transduction and metabolic cycles, which is closely related to different kinds of diseases. Therefore, SAM-dependent methyltransferases are considered as promising drug targets. Currently, a growing number of nucleoside analogues have been developed as SAM-competitive inhibitors, blocking the downstream signaling pathways to cure diseases. In the review, we outline the design strategy and optimization process of methyltransferase inhibitors, analyze the shortcomings and solutions of developing nucleoside derivatives as MTase inhibitors, to provide guidance and broad direction to the development of nucleoside MTase inhibitors.

Traf2- and Nck-interacting kinase (TNIK) is closely related to the cancer occurrence, development, and prognosis. TNIK plays an important role in regulating Wnt/β-catenin pathway, thus becoming a potential target for cancer treatment. Various small-molecule TNIK inhibitors with diverse structures and potent in vitro activities have been developed to treat tumors. However, no TNIK inhibitor has entered clinical phase mainly due to poor selectivity, toxicity, and unsatisfied in vivo anti-tumor efficiency. This review summarizes the research progress of small molecule inhibitors targeting TNIK, with the aim of giving some guidance for future research and development of small-molecule inhibitors.

Pentacyclic triterpenoids are a class of widespread natural compounds containing six isoprene structures with a wide range of pharmacological activities, including antibacterial, anti-inflammatory, antiviral, antitumor, immune regulation, etc. The structural modifications of pentacyclic triterpenoid natural products and the drug development have always been a hot research topic. This article reviews the recent progresses in the structural modifications, pharmacological effects, and clinical studies of different kinds of pentacyclic triterpenoid natural products.

Deuterated drugs have been new strategies in drug design and development for advantages in improving pharmacokinetic properties, reducing toxic metabolites, and stabilizing chiral structures over their non-deuterated counterparts. Deuterium isotopic impurities, including isotopologues and isotopomers, are inevitably generated even with state-of-the-art deuteration techniques, which significantly affect the safety and potency of drugs through deuterium kinetic isotope effects. Therefore, deuterium isotopic purity is the most characteristic critical quality attribute of deuterated drugs compared to other drugs. However, it is challenging to separate isotopic mixtures with routine analytical methods, and regulatory agencies have not proposed guidelines for deuterated drugs in terms of analytical techniques and quality control, which presents great analytical challenges for the development of deuterated drugs. This paper reviews the analytical techniques for deuterium isotopic purity of deuterated drugs and discusses the features and scopes of different methods. It aims to provide references for the development and relevant study of deuterated drugs.

In recent years, the abuse of antibiotics has led to antibiotic tolerance in the process of bacterial treatment, the morbidity and mortality caused by drug-resistant bacterial infection have further increased significantly. Drug delivery systems can be precisely designed to achieve controlled drug release, thereby reducing the risk of antibiotic toxicity and resistance, it is urgent to seek novel drug delivery systems to address the challenges posed by bacterial infections. This review first outlines the epidemic and prevention situation of bacterial infection, and further summarizes living microorganisms and their derivatives-based drug delivery systems, focusing on their natural characteristics such as surface specific proteins, physiological signal sensing, directed movement, and secretion of antibacterial substances, which show great potential in the treatment of bacterial infectious diseases by demonstrating their antibacterial effects. This review aims to provide ideas for the development of novel drug delivery systems based on living microorganisms and their derivatives for the treatment of bacterial infectious diseases.

Metal-organic frameworks (MOFs) are crystalline materials with a multidimensional porous network structure, formed through coordination bonds with metal ions as nodes and organic ligands as connecting bridges. Due to their excellent physicochemical properties, MOFs have extensive applications in the field of biomedicine, ranging from antibacterials, drug carriers, imaging to sensors. Nanoscale metal-organic frameworks (nMOFs), commonly utilized drug carriers, can gain enhanced safety, targeted delivery, and superior therapeutic effect through endocytosis. In this review, we comprehensively summarize the factors influencing the endocytosis of nMOFs, focusing on three key physicochemical properties, particle size, morphology and surface modification. Based on different illness models, the review succinctly summarizes the latest advancements in understanding the endocytosis pathways of nMOFs while critically reflecting on the inherent limitations of current research methods. Lastly, the review offers valuable insights into future research methodologies and objectives, aiming to lay the groundwork and provide meaningful guidance for the synthesis and development of nMOFs as promising versatile drug carriers.

Antibody-drug conjugate (ADC) has become an effective method for treating various diseases, especially cancer, due to its clear target and good selectivity in clinical practice. However, the monoclonal antibodies in traditional ADC have poor tissue permeability, high modification costs, pose risks such as immunogenicity and immunotoxicity. The nanobody (Nb) which is extracted from the blood of camel animals, is the smallest antibody fragment known to have complete antigen binding ability. It has advantages such as strong tissue permeability, strong specificity, low immunogenicity, and high stability, and can replace traditional monoclonal antibodies to participate in the construction of nanobody-drug conjugate (NDC). This article reviews and discusses the advantages of Nb structure, the construction and application of NDC in the hope of providing ideas for the research and development of NDC.

Salvia miltiorrhiza, a commonly used traditional Chinese medicine, has been widely recognized for its blood-activating and stasis-removing properties in the clinical treatment of cardiovascular and cerebrovascular diseases. The synthesis and regulatory mechanism of tanshinones, the key active constituents of Salvia miltiorrhiza, have been a hot topic of research. The paper summarized the research findings on the regulation of tanshinone biosynthesis by transcription factors such as AP2/ERF, bHLH, MYB, bZIP, and WRKY in recent years. The review identifies the existing issues in the transcriptional regulation studies of Salvia miltiorrhiza and discusses the research direction of transcription factors in the regulation of tanshinone biosynthesis, providing a theoretical basis for the further discovery and utilization of functional genes involved in the regulation of tanshinone bioactive constituents.

Monosodium urate (MSU)-induced the gouty arthritis (GA) model was used to investigate the effect of Nod-like receptor protein 3 (NLRP3) inhibitor N14 in alleviating GA. Firstly, the effect of NLRP3 inhibitor N14 on the viability of mouse monocyte macrophage J774A.1 was examined by the cell counting kit-8 (CCK-8) assay. The expression of mature interleukin 1β (IL-1β) and cysteinyl aspartate specific proteinase-1 (caspase-1) p20 in the cell supernatant and the expression of NLRP3, caspase-1 and pro-IL-1β proteins in the cell lysates was detected by Western blot for the inhibitory effect of N14 on the MSU-induced NLRP3 inflammasome activation in J774A.1 cells. Animal behavioral tests were used to detect redness, swelling, heat and pain in mice with gouty arthritis. Hematoxylin-eosin (H&E) staining revealed pathologic changes and inflammatory infiltration in foot sections. Protein expression of NLRP3, caspase-1, and pro-IL-1β in mouse hind paw tissues were assessed by Western blot. The effect of N14 on the plasma levels of alanine transaminase (ALT), aspartate transaminase (AST), creatinine (CRE), urea, and uric acid (UA) was investigated by the MSU-induced gouty arthritis model in mice. All animal experiments in this paper were approved by the Scientific Ethics Review Board of Qingdao Marine Biomedical Research Institute (grant No. E-MBWNL-2024-20). The experimental results showed that N14 did not exhibit cytotoxicity in mouse monocyte macrophage J774A.1 cells at concentrations up to 100 μmol·L^-1, and N14 effectively prevented MSU-induced activation of NLRP3 inflammasome. In the mouse gouty arthritis model, N14 significantly ameliorated the redness, swelling, heat and pain caused by GA, and down-regulated the levels of NLRP3 inflammasome-associated proteins in mouse hind paw tissues. Meanwhile, N14 appeared to be well tolerated, as it did not significantly affect various biochemical indices in mouse plasma. In conclusion, N14 effectively alleviated GA in mice by inhibiting the NLRP3 inflammasome pathway, which is important for both prevention and treatment of related diseases.

Constipation is a prevalent ailment which might significantly impact the quality of people's life and rise some associated deseases risks. In this study, a chronic constipation mouse model was established using loperamide hydrochloride. Mice were gavaged an Angelica sinensis Cistanche Fiber Compound, comprised of Dang Gui [Angelica sinensis (Oliv.) Diels], Rou Cong Rong (Cistanche deserticola Y.C.Ma), wheat fiber, and low-molecular-weight xylan at high dosage (3.6 g·kg^-1, 30 times the recommended human dose) and low dosage (0.6 g·kg^-1, 5 times the recommended human dose) for 14 days. The study assessed the therapeutic efficacy of the compound by observing fecal morphology, measuring water content, and conducting small intestine motility experiments. Furthermore, enzyme-linked immunosorbent assays (ELISA) were conducted to evaluate the serum levels of gastrointestinal hormones including motilin (MTL), gastrin (GAS), the inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). Histopathological examination with H&E staining was employed to evaluate colonic tissue damage. Additionally, Western blot and immunohistochemistry experiments were conducted to examine the expression levels of aquaporin-3 (AQP3) and c-Kit in the colon. The results indicated that the Angelica sinensis Cistanche Fiber Compound could improve fecal morphology, increase fecal water content, enhance small intestinal transit rates in mice. Additionally, there was a significant elevation in the serum levels of gastrointestinal hormones and a notable reduction in the levels of inflammatory factors. The improvement in colonic histopathological damage was accompanied by a marked decrease in the expression of the colonic water channel protein AQP3 and a significant increase in c-Kit expression. These results collectively suggested the presence of a dose-response relationship. These findings indicate that the Angelica sinensis Cistanche Fiber Compound effectively alleviates constipation in mice. Its action is associated with the regulation of colonic water channel protein AQP3 and c-Kit expression, along with the modulation of serum inflammatory factors and gastrointestinal hormones. This experiment was approved by the Animal Experiment Ethics Committee of Jinan University.

The present study identified chemical constituents of Honghua Xiaoyao Tablet (HXT) and explored its biological connotation and characteristics on the premenstrual syndrome (PMS) treatment from the "disease-syndrome-symptom" association network. UHPLC-Q Exactive Orbitrap HRMS technology was applied to analyze the chemical constituents in HXT. According to the composition principles, the compatible herbs of HXT were divided into the Shugan Jieyu group, Huoxue Tiaojing group and Yiqi Jianpi group. The candidate targets of the corresponding prescriptions of HXT efficacy groups were collected from the Pharmmapper database and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP) v2.0. The gene set related to the clinical symptoms included in Traditional Chinese and Western Medicine diagnosis and treatment standards were obtained from SoFDA, GeneCards, DisGeNET, MalaCards and literature published. The "HXT candidate targets-PMS (liver depression, Qi stagnation, and blood stasis syndrome) genes" network was constructed based on the gene interaction information, and further, the core network targets were screened out by topological characteristics of calculating network, and the functional exploration was carried out based on Kyoto Encyclopedia of Genes and Genomes (KEGG) for exploring the therapeutic advantages in PMS treatment of HXT efficacy groups, which were further verified experimentally in vitro. A total of 109 components from HXT were identified, including 20 components from Shugan Jieyu group enriched in the neurological system, estrogen regulation, and "immune-inflammation" related pathways, 77 components from Huoxue Tiaojing group enriched in the blood-circulation system, "immune-inflammation" and estrogen regulation related pathways and 30 components from Yiqi Jianpi group regulating immune inflammation, digestive system, and hormone levels. The biochemical indicator detection demonstrated that both the levels of 5-HT and DA in the hypothalamus tissues and the levels of E₂, NO, VEGF and RLN in the uterine tissues of PMS model rats were lower than those in controls, which the levels of OT, PROG, IL-6, IL-1β and TNF-α in the uterine tissues were increased in PMS group, which were all reversed by the administration of HXT, indicating that this prescription may regulate the synthesis and secretion of estrogen, intervene in neurotransmitter synthesis and signal transduction, reverse the imbalance of "immune-inflammation", and regulate digestive system function through various biological pathways, exerting the liver-smoothing, Qi-regulating, blood-activating and spleen-tonifying comprehensive effects, leading to alleviating the "neuroendocrine-endocrine-immune" system and blood-circulation disorders. The relevant results may provide a reference for clarifying the advantages and efficacy of HXT in treating PMS with liver stagnation, Qi stagnation, and blood stasis syndrome, and exploring its therapeutic advantages. The animal experiment of this study was approved by the Ethics Committee of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences (approval number: 2023B248).

Inflammatory bowel disease (IBD) is characterized by chronic relapsing intestinal inflammation and encompasses ulcerative colitis (UC) and Crohn's disease (CD). IBD has emerged as a global healthcare problem. Clinically efficacious therapeutic agents are deficient. This study concentrates on models of ulcerative colitis with the objective of discovering novel therapeutic strategies. Previous investigations have established that schisandrin A demonstrates anti-inflammatory effects in vitro, concurrently enhancing the transcriptional activity of farnesoid X receptor (FXR). FXR inversely modulates the transcriptional activity of NF-κB, which has an important role in regulating inflammatory responses. Consequently, the current study was to explore the safeguarding influence of schisandrin A on ulcerative colitis and delineate the mechanism by which it regulates this effect through the FXR signaling pathway. The effect of schisandrin A on the mRNA levels of inflammatory factors was evaluated in RAW264.7 cells. The dual luciferase reporter gene assay was used to verify the relationship between schisandrin A and FXR targeting. The animal experiments were performed in accordance with the regulations of the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval No. PZSHUTCM2304250005). Acute ulcerative colitis was induced in wild-type or FXR knockout C57BL/6 mice by drinking 3% dextran sodium sulfate (DSS) for 7 days, and schisandrin A was administered via gavage for a continuous treatment period of 7 days. The body weight and faecal were monitored daily. The mRNA levels of inflammatory factors in colon tissue and FXR target genes were measured by RT-qPCR. The findings revealed that schisandrin A, in vitro, impeded the lipopolysaccharide (LPS)-induced elevation in mRNA levels of inflammatory factors and schisandrin A could augment transcriptional activity of FXR. In wild-type mice, schisandrin A significantly improved weight loss, colon shortening, loose stools and blood in stools in mice with acute ulcerative colitis, and schisandrin A significantly reduced the expression of pro-inflammatory factors genes and significantly increased the expression of FXR target genes in colon tissues. In FXR knockout mice, the administration of schisandrin A failed to yield ameliorative effect on acute ulcerative colitis in mice. In conclusion, schisandrin A can reduce intestinal inflammation through the FXR signaling pathway to alleviate acute ulcerative colitis in mice. Implications arise that Schisandra lignans could serve as lead compounds for drug development aimed at inflammatory bowel disease.

Diabetic retinopathy (DR) is a diabetic ocular complication that can lead to poor vision and blindness. This experiment aimed to investigate the ameliorative effect and its mechanism of Panax notoginseng saponins (PNS) eye drops on streptozotocin (STZ)-induced non-proliferative diabetic retinopathy (NPDR) in rats. All experiments were approved by the Animal Research Committee of Shanghai University of Traditional Chinese Medicine. Animal welfare and the animal experimental protocols were strictly consistent with related ethics regulations of Shanghai University of Traditional Chinese Medicine (No. PZSHUTCM 211115004). Diabetes mellitus was induced by a single intraperitoneal injection of STZ into rats. Two months later, PNS solution was dropped binocular twice per day at 6 h intervals at dose of 20, 40, and 80 mg·kg^-1 continuously for 1 month. The morphological structure and activation of microglia of the retina were observed by hematoxylin-eosin staining and immunohistochemical assay. The disruption of the blood-retina barrier (BRB) was conducted by the Evans blue dye leakage assay. The number of acellular capillaries in the retina was assessed by digesting and hematoxylin-eosin staining assay. The number of retinal leukocyte adhesion was observed by fluorescein isothiocyanate-coupled concanavalin A lectin labeling assay. The serum expression of inflammatory factors was measured using enzyme-linked immunosorbent assay. Western blot experiments were used to detect the expression of relevant proteins in retinal tissue and transcriptional activation of nuclear factor kappa-B (NF-κB). The results revealed that PNS eye drops significantly increased the thickness of the retina, decreased the number of acellular capillaries, and up-regulated the expression of the tight junction proteins claudin-1 and occludin, thereby alleviating BRB damage. In addition, PNS eye drops were also able to significantly reduce leukocyte adhesion and microglia activation, the expression of inflammatory factors in serum, and the nuclear translocation of retinal p65 proteins, effectively inhibiting the occurrence of retinal inflammation. The above results showed that PNS eye drops played a role in improving NPDR by inhibiting the activation of the NF-κB signaling pathway and reducing inflammation.

Coronaviruses of the genus Coronavirus contain a variety of human pathogenic viruses, and the development of anti-coronavirus drugs is of great value. The development of antiviral drugs targeting host cells is not only helpful for the development of new antiviral strategies, but also for solving problems such as drug resistance due to viral mutations. Our preliminary study identified that cell cycle-dependent protein kinases (CDKs) involved in coronavirus replication, for which they would be potential anticoronaviral targets. In this study, we found that the broad-spectrum CDK inhibitor flavopiridol significantly inhibited severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA dependent RNA polymerase (RdRp) activity. Further studies showed that flavopiridol suppressed the RNA synthesis efficiency of SARS-CoV-2 RdRp. In addition, flavopiridol effectively restricted the replication of human coronavirus OC43 (HCoV-OC43). Therefore, our study suggested that the CDK inhibitor flavopiridol may be a potential anticoronaviral drug.

Lipopolysaccharides (LPS) are major outer membrane components of Gram-negative bacteria. Unlike most Gram-negative bacteria, Acinetobacter baumannii can still survive and acquire polymyxin resistance after complete loss of LPS. Previous studies of LPS-deficient Acinetobacter baumannii mostly focused on LPS-deficient strains induced by polymyxin, whose background was complex and unstable. To investigate LPS loss-mediated polymyxin resistant-Acinetobacter baumannii, this study constructed a stable and clear background LPS-deficient strain by knocking out lpxC gene in Acinetobacter baumannii ATCC 19606 with CIRSPR/Cas9, and then studied the phenotypic changes of the lpxC-deficient strain including morphology, growth rate, antibiotic susceptibility, virulence, membrane permeability, and membrane potential. Animal experiments were approved by the Animal Care and Welfare Committee Institute of Medicinal Biotechnology, CAMS and PUMC (approval number: IMB-20240119D₉). The results indicated that the lpxC gene of Acinetobacter baumannii ATCC 19606 was successfully knocked out. After losing the lpxC, the strain underwent the morphological change from rod-shaped to spherical. Furthermore, it leads to reduced growth rate, enhanced membrane permeability, decreased membrane potential, lower virulence, and increased antibiotic susceptibility to β-lactams, quinolones, aminoglycosides, macrolides, glycopeptides. The lpxC deletion results in significant changes in membrane homeostasis and adaptability of Acinetobacter baumannii. Understanding the phenotypic changes of colistin-resistant Acinetobacter baumannii mediated by LPS loss is useful for exploring the resistance mechanism of Acinetobacter baumannii and developing new therapeutic strategies.

Kidney ischemia reperfusion injury (IRI) is a leading cause of acute kidney injury (AKI) with a poor prognosis and high mortality rate. Recent studies have reported that chrysophanol may have a renal protective effect, but its specific impact and mechanism on IRI remain unclear. This study aimed to explore the effects and mechanisms of chrysophanol on AKI induced by IRI. By utilizing a unilateral kidney IRI mouse model, histopathological changes in the kidney, serum levels of creatinine and urea nitrogen, and protein expressions of apoptosis and mitophagy in kidney tissue were examined. Additionally, a hypoxia/reoxygenation (H/R) model of human kidney-2 (HK-2) cells was established to measure mitochondrial membrane potential levels and reactive oxygen species (ROS). Functional enrichment analysis was performed to screen relevant targets of chrysophanol and AKI, and to verify key targets and pathways. The animal experiments conducted in this study were ethically approved by the Experimental Animal Ethics Committee of Peking University (No. LA2021503). The findings indicate that the IRI group exhibited elevated levels of creatinine and urea nitrogen in serum, significant renal tissue damage, and increased expression of renal injury markers (KIM1), apoptosis-related proteins (cleaved-caspase 3, caspase 3, cytochrome C), and mitochondrial autophagy protein (PINK1) compared to the sham surgery group. Chrysophanol treatment ameliorated the aforementioned pathological changes in a dose-dependent manner in an IRI model. Additionally, it exhibited significant improvements in mitochondrial membrane potential and inhibition of ROS production in HK-2 cells subjected to H/R conditions. Through network pharmacological analysis, HSP90AA1 and PIK3R1 were identified as key targets primarily enriched in the phosphoinositide 3 kinase/protein kinase B (PI3K/Akt) pathway. Real-time quantitative PCR (qPCR) validation confirmed that chrysophanol significantly decreased HSP90AA1 and PIK3R1 mRNA levels in HK-2 cells under H/R conditions, while also enhancing the protein expressions of p-PI3K, PI3K, p-Akt, and Akt. In conclusion, chrysophanol has the potential to enhance AKI by selectively modulating HSP90AA1 and PIK3R1, activating the PI3K/Akt pathway, decreasing apoptosis, regulating mitochondrial autophagy, enhancing mitochondrial membrane potential, and suppressing ROS production. These findings suggest that chrysophanol could serve as a promising therapeutic option for the treatment of AKI.

The effect of different concentrations of glycyrrhizic acid (GA) and Zn²⁺ on the self-assembly of metal complexes was investigated by forming metal complexes, and the properties and assembly mechanisms of the formed carrier-free supramolecular hydrogel were characterised. Scanning electron microscopy (SEM) and zeta potential were used to characterise the microscopic morphology and stability of the GA-Zn complex hydrogel, which had spherical-like particles of about 1 μm with good stability; the rheometer was used to detect its materialistic properties, which showed excellent stability, self-healing property and reversibility; through in vitro bacterial inhibition, it was found that the GA-Zn carrier-free supramolecular hydrogel has enhanced bacterial inhibition function after assembly. The hydrogel was also found to possess both anti-inflammatory and antioxidant efficacy when evaluated using LPS and H₂O₂ induced RAW 264.7 cell damage models, respectively. The above results suggest that GA-Zn hydrogel not only has good materialistic properties, but also possesses good antibacterial, anti-inflammatory and antioxidant activities, which has the value of clinical research as a carrier-free multi-functional antimicrobial dressing, and the present study provides a reference for the discovery of novel biomedical materials from active molecules of natural traditional Chinese medicine.

Microsporidia is a group of intracellular parasitic eukaryotic microorganisms that pose threats to livestock fish production and human health, but there is no special chemo-therapeutic drug available for the treatment of microsporidia currently. In this study, sulfonamide derivatives of spectinomycin were designed by taking into consideration the low toxicity of spectinomycin and the favorable pharmaceutical properties of sulfonamides. Through the exploration of reaction conditions, a total of 21 target molecules were synthesized with a yield of 52%-74% and their chemical structures were confirmed by ¹H NMR, ¹³C NMR, and high resolution mass spectrometry (HR MS). By screening in vitro, the host cell cytotoxicity and biological activity of target molecules to Encephalitozoon hellem, a representative of vertebrates infecting microsporidia, were studied. It was found that four target molecules each could inhibit or promote the proliferation of E. hellem. In this study, the synthesis method of sulfonamide derivatives of spectinomycin was established, and new compounds effective against E. hellem were discovered, which provided a new idea for the further derivatization of spectinomycin and the development of therapeutic drugs targeting microsporidia.

Three new compounds, including a naphthoquinone, a reduced naphthoquinone derivative naphthalenone, and a tricarboxylic acid, along with five known naphthalenone derivatives were isolated from ethyl acetate extract of rice fermentation products of the fungus Pleosporales sp. by multiple column chromatographic methods, including Sephadex LH-20 gel column chromatography, silica gel column chromatography, reversed-phase HPLC, and chiral chromatography. Their structures were elucidated by MS, NMR, and specific rotation spectroscopic analyses as well as ECD calculations. Three novel compounds were named as pleospathone A (1), pleospathone B (2), and pleosporalic acid A (3). Five known compounds were separately identified as (3S, 4R)-3,4,8-trihydroxy-6-methyl-3,4-dihydronaphthalen-1(2H)-one (4), (4R)-3,4-dihydro-4,6,8-trihydroxy-1(2H)-naphthalenone (5), (-)-scytalone (6), (3S, 4S)-3,4-dihydro-3,4,6,8-tetrahydroxy-1(2H)-naphthalenone (7), and cis-4-hydroxyscytalone (8). Compounds 4-8 were isolated from the Pleosporales fungi for the first time. Compound 1 shows inhibitory activities against human cervical carcinoma cell line HeLa and murine leukemia cell line P388 with the IC₅₀ values of 78.93 and 98.80 μmol·L^-1, respectively.

In order to study the compounds from Glechoma longituba (Nakai) Kupr. and explore the substance bases of its dissipating blood stasis, MCI, silica gel, Sephadex LH-20, ODS column chromatography and preparative thin layer chromatography were used to isolate and purify the compounds. The structures were identified by MS, 1D NMR, and 2D NMR spectra analysis, etc. Eight triterpenoids were isolated from dichloromethane fraction of ethanol extract from G. longituba and identified as 2α,3α,16β-trihydroxy-13α,27-cyclours-11-en-28-oic acid (1), 28-norurs-12-ene-3β,17β-diol (2), 28-norolean-12-ene-3β,17β-diol (3), oleanonic acid (4), 3β,13β-dihydroxyurs-11-en-28-oic acid (5), uvaol (6), oleanolic acid (7), ursolic acid (8). Compound 1 is a new hexacyclic triterpene with 13α,27-cyclopropane structure, named euscaphic acid H; compounds 2 and 3 were isolated from Lamiaceae for the first time while compounds 4 and 5 were isolated from this genus. The in vitro anticoagulant activity of triterpenoids was evaluated by four coagulation indexes (activated partial thromboplastin time, APTT; thrombin time, TT; prothrombin time, PT; fibrinogen, FIB). Among them, compounds 1 and 6 showed obvious anticoagulant effects.

Eleven compounds were isolated from the twigs and leaves of Xylopia vielana Pierre by various chromatographic techniques such as silica gel, ODS and the semi-preparative HPLC. Their chemical structures were identified by HR-ESI-MS, NMR, ECD and other spectroscopic methods as vielana A (1), vielana B (2), vielana C (3), 10-oxo-isodauc-3-en-15-al (4), 1α-hydroxyisodauc-4-en-15-al (5), mokko lactone (6), 11β,13-dihydrocostunolide (7), eurylosesquiterpenol E (8), epi-α-cadinol (9), mustakone (10), 7-epi-amiteo (11). Among them, compounds 1-3 were new compounds, and the rest compounds were isolated from this plant for the first time. Compounds 1 and 2 increased the transcriptional activity of the farnesoid X receptor (FXR) downstream target gene BSEP promoter, indicating that they have potential of FXR activation.

Eighteen compounds were isolated from the methanol extract of the fruits of Litsea cubeba by silica gel, ODS, Sephadex LH-20 column chromatography, semi-preparative RP-HPLC, chiral HPLC and recrystallization. Their structures were elucidated by spectroscopic analyses and by comparison with reported spectroscopic data and physicochemical properties, and the absolute configurations of the enantiomers were established by experimental and calculated electronic circular dichroism spectra. These compounds were identified as (+)-(R)-4-hydroxypiperitenone (1a), (-)-(S)-4-hydroxypiperitenone (1b), (3S,4S,6R)-3,6-dihydroxy-1-menthene (2), (4S,5R)-4-hydroxy-5-isopropyl-2-methylcyclohex-2-en-1-one (3), (R)-6-hydroxy-3-(2-hydroxypropan-2-yl)-6-methylcyclohex-2-enone (4), (4S,6R)-4-hydroxy-6-isopropyl-3-methylcyclohex-2-enone (5), (1R,3S,4R)-3-hydroxy isopulegol (6), subamone (7), (6S)-3,7-dimethyl-7-hydroxy-2(Z)-octen-6-olide (8), (6S)-6,7-dihydroxy-3,7-dimethyloct-2(Z)-enoate (9), holostylactone (10), sesamin (11), dimethylmatairesinol (12), p-hydroxybenzoylcarbinol (13), syringaldehyde (14), p-hydroxybenzaldehyde (15), 4-hydroxy-3-methoxybenzaldehyde (16), 5,4ʹ-dihydroxy-7-methoxyflavone (17). Among them, compounds 1a and 1b were a pair of new monoterpenoid enantiomers, 8 and 9 were new natural products, 2-7, 10, 11 and 17 were isolated from Litsea genus for the first time. The in vitro anti-inflammatory effects of compounds 1-17 were evaluated using lipopolysaccharide-stimulated RAW264.7 cells, the results showed that compound 14 exhibited significant anti-inflammatory activity with NO inhibitory rate of 66.27% at a concentration of 40 μmol·L^-1.

The abuse of novel phenylcyclohexylpyridine drugs poses a significant threat to societal safety. The novel psychoactive substance 2-methyl-deschloroketamine (2-MDCK), belonging to the phenylcyclohexylpyridine class, has recently surfaced as a new compound. However, there is a lack of understanding regarding its metabolic pathways and the identification of suitable biomarkers. In this study, a human liver microsomal model was established, and ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) technology was applied to investigate the in vitro metabolism and products of 2-MDCK. The results indicate that 2-MDCK undergoes various metabolic reactions, including dehydrogenation, deamination, hydroxylation, and demethylation, leading to the formation of eight metabolites. By conducting actual testing on hair samples from 2-MDCK abusers, the existence of six metabolites was confirmed. Comparing the metabolic products in the liver microsomal in vitro model, M3.1 and M4.1 were identified as biomarkers for 2-MDCK consumption. Five samples of abusers of 2-methyl-dechloroketamine were provided by the Anti-Drug Brigade of the Hangzhou Public Security Bureau. Negative hair samples were provided by laboratory volunteers, and all samples were obtained with the informed consent of the volunteers. These findings provide a scientific basis for the detection and identification of 2-MDCK and its metabolites, as well as crucial support for the study of the metabolic mechanisms of similar novel psychoactive substances in the phenylcyclohexylpyridine class. This research holds significant importance in addressing the issue of abuse of new psychoactive substances.

The study utilized spectral correlation analyses combined with bioactivity evaluation to examine the effective components of antidepressants in the Baihe Dihuang decoction. Firstly, the chemical fingerprints for different extraction parts in the Baihe Dihuang decoction were achieved using HPLC and UHPLC-MS technology. Then, in order to evaluate the antidepressant effect of Baihe Dihuang decoction, the animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Hunan University of Chinese Medicine (No. LLBH-202104270001), in compliance with the Institutional Animal Care Guidelines. We recorded the distance of autonomous movement of mice in open field experiment, the immobility time of tail suspension test, and the forced swimming. Additionally, we measured the content of neurotransmitters. Finally, Pearson analysis, grey correlation analysis, and orthogonal partial least squares regression analysis were utilized to establish the correlation between antidepressant efficacy indicators and fingerprinting. The spectrum-effect relationship results were confirmed through the in vitro activity verification. This study demonstrated that regaloside A, B, C, catalpol, and Isoacteoside might be the main antidepressant components in Baihe Dihuang decoction. Furthermore, it was found that using diverse mathematical models and bioactivity evaluation could enhance the accuracy of the spectral correlation analyses results.

The paper is to establish an UPLC-MS/MS method for the simultaneous determination of 19 components in Microctis Folium from different production areas. The 50% methanol was used as extraction solvent. The Agilent ZORBAX SB C18 (150 mm × 2.1 mm, 1.8 μm) column was used; mobile phase was acetonitrile - 0.1% acetic acid with gradient elution, flow rate was 0.3 mL·min^-1, colume temperature was 30 ℃, and the injection volume was 2 μL; electrospray ionizaton source was used and detected in negative ion mode. The results showed that the established UPLC-MS/MS method could well separate the 19 components, and the methodological investigation results of 19 components were good. By means of orthogonal partial least squares discriminant analysis (OPLS-DA), 28 batches of Microctis Folium samples from different production areas can be divided into three categories, Guangdong, Guangxi and Hainan are each classified into one category, and 10 signature compounds which affecting the quality differences of different production areas were screened out. The established method is accurate, reliable, sensitive and reproducible. It can provide a basis for the establishment of the quality standard of Microctis Folium, as well as for safety and quality research.

This research established a simple, rapid and sensitive ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) method to investigate the metabolic profiles of cajanonic acid A (CAA) in rats. After intragastric administration of CAA (30 mg·kg^-1) to rats, the biological samples were detected by UPLC-Q-TOF-MS/MS. Relevant data was collected and processed, the accurate mass and MS² spectra of the metabolites were compared with the parent compound. As a result, a total of 23 metabolites were detected, including 15 in urine, 11 in bile, 11 in feces, and 9 in plasma. The major metabolic pathways related to CAA included dehydrogenation, reduction, hydroxylation, methylation and glucuronide conjugation. This experiment was approved by Animal Ethics Committee of Guizhou Medical University (approval number: 1603137).

Cation exchange chromatography, as a commonly used separation and purification technique in biopharmaceutical manufacturing, is often employed for downstream processes to separate target monoclonal antibodies from their charge variants. For samples with complex and poorly resolved charge variant profiles, the collection solely based on ultraviolet detection does not provide specific compositional information for individual charge variants, making it challenging to determine the range of pooled fractions directly. Subsequent laborious fractionation analysis is then required to guide collection according to production requirements. A mechanistic model for the cation exchange chromatography process of the target monoclonal antibody's critical components was established, and it was employed to assist in product collection. The model accurately predicted the elution peak shapes of the modeled variants, with a root mean square error between predicted and actual values below 0.009. In comparison to the online ultraviolet-based collection method, the model-assisted collection method not only visualized the chromatographic process but also increased the relative productivity by fourfold while ensuring compliance rate.

An integrated evaluation model based on the combination of traditional trait identification and modern chemical analysis was used for the identification of key indexes of grade classification and the establishment of grade quality standard of Lycium barbarum fruits (LBF) from Ningxia genuine producing area. Under the guidance of herbal examination and market survey, particle size, skin color and imperfection were taken as the external trait indicators; focusing on the comprehensiveness and validity of the quality evaluation, 2-O-β-D-glucopyranosyl-L-ascorbic acid (AA-2βG), zeaxanthin dipalmitate (ZD) and volatile ether extractives were used as important parameters involving in quality evaluation in addition to the pharmacopoeia test items and determination indicators such as moisture, ash, water-soluble extractives, Lycium barbarum polysaccharide (LBP) and betaine. The quality differences between different traits and grades of LBF as well as the relevance between character and quality were further investigated by mathematical and statistical means such as t-test analysis, correlation analysis and principal component analysis (PCA). Furthermore, the activities of extractives from different grades of LBF were compared at cellular level. The results showed that LBF were mainly preferred for large and red ones; except for betaine, the other index components were relatively high (moisture and ash were low) in the first-grade and samples with bright-red skin color. Besides, there was a significant correlation between skin color and moisture, water-soluble extractives, LBP, AA-2βG and ZD contents. Considering the results of the study and the market situation, the present study took the grain size and skin color of LBF as the core of evaluation, and at the same time introduced the multi-indicator components including LBP, AA-2βG, ZD, water-soluble extractives and moisture to further guide the grading, and established the quality standard of LBF grades combining the trait indexes and the chemical composition. More importantly, in vitro cell models both preliminarily confirmed that the first-grade LBF played a more significant "nourishing liver and kidney" effect, which provided an experimental basis for the quality control of LBF and promoted the realization of high quality and good price.

The anti-inflammatory efficacy of Callicarpa nudiflora extract were evaluated upon lipopolysaccharide (LPS)-induced infective inflammation in rats. Pathological changes of lung and colon tissues observed with hematoxylin-eosin (H&E) staining, together with inflammatory factors in serum, including nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), were applied to assess the mitigating effects of C. nudiflora water extract on model rats. The experimental protocol strictly adhered to the guidelines of the Ethics Committee of Animal Research of Guangdong Pharmaceutical University (Approval: gdpulac2022132). Quality markers with anti-inflammatory activities were recognized by network pharmacology analysis. Subsequently, a reliable method for simultaneously quantifying six components was established. As a result, the pathological injury on lung and colon tissues of model rats induced by LPS were improved by C. nudiflora water extract. Compared with model group, C. nudiflora extract had decreased the release of proinflammatory factors in serum, including TNF-α and IL-6, and reduced the NO (P < 0.01). Network pharmacological analysis obtained 83 chemical components, 466 component targets, 584 disease targets and 129 action targets, which were mainly concentrated in 624 biological processes such as inflammatory response and positive regulation of nitric oxide biosynthesis, as well as 162 pathways such as phosphatidylinositol-3-hydroxykinase-protein kinase (PI3K-Akt) signal pathway and Janus kinase-signal transducer and activator of transcription 3 (JAK-STAT3) signal pathway. Upon analyzing their specificity, effectiveness, detectability, and quantity transfer rule from herb to extract, 6-hydroxyluteolin 7-glucoside, cynaroside, caffeic acid, acteoside, isoacteoside, and forsythoaside B were identified as the quality markers. Their anti-inflammatory activities were verified with LPS-induced inflammation model of RAW264.7 cells. Then contents of these 6 quality markers in 16 batches of C. nudiflora were determined. Thereby, the anti-inflammatory efficacy of C. nudiflora extract were confirmed in vivo, 6-hydroxyluteolin 7-glucoside, cynaroside, caffeic acid, acteoside, isoacteoside, and forsythoaside B were identified as anti-inflammatory ingredients, which can be used as quality control indicators for the herb and related formulation.

To investigate the pharmacokinetic characteristics and metabolites of Src homology 2 region-containing protein tyrosine phosphatase 2 (SHP2) protein inhibitor in SD rats, a triazole quinolinone based derivative NC-55-122 was utilized. Firstly, rats were randomly divided into groups and given compound NC-55-122 intragastric and intravenous administration, respectively. Blood samples were collected at different time points. Taking carbmazepine as the internal standard, ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to determine the concentration of NC-55-122 in rats, and the methodology was verified. DAS 2.0 software was used to calculate the main pharmacokinetic parameters, and GraphPad Prism 8.0.1 software was used to plot the blood concentration-time curve. At the same time, UPLC-Q-TOF/MS was established to analyze plasma samples, and UNIFI metabolite prediction software was used to analyze metabolites after oral gavage and intravenous injection. The linear range of the mass concentration of compound NC-55-122 was from 1 ng·mL^-1 to 1 600 ng·mL^-1, and the linear relationship was good. The matrix effect, extraction recovery, precision, accuracy and stability were investigated in this linear range, which met the requirements of biological analysis. Secondly, the analysis of pharmacokinetic parameters showed that the oral bioavailability of the compound was low, with F%= 3.09%, indicating that the compound was absorbed slowly in vivo. Finally, five possible metabolites were deduced by analyzing the ion flow diagram and combining UNIFI software. The detection method established in this experiment is highly sensitive, specific, rapid and efficient, which is suitable for the determination of the blood concentration of compound NC-55-122 in rats and the analysis of metabolites, and lays a foundation for the structural modification and druggability evaluation of the later anti-tumor drug NC-55-122. All animal experiments were approved by the Experimental Animal Ethics Committee of Guizhou Medical University (approval number: 2200823).

In this study, doxorubicin (DOX) was used as the model drug, new indocyanine green (IR820) as the photosensitizer, and temperature sensitive liposomes (TSL) as the carrier. H460-NCI photoheat-sensitive liposomes coated with cell membrane of human cell lung cancer (DOX-IR820-TSL@CCM) for highly effective multi-pathway tumor targeting in chemical-photothermal therapy and photodynamic therapy. DOX-IR820-TSL was prepared by reverse evaporation, cancer cell membrane (CCM) was prepared by lysis, crushing and centrifugation, and DOX-IR820-TSL@CCM was prepared by nanomembrane extrusion. The drug-loading conditions of DOX-IR820-TSL were finally determined: the ratio of organic phase to aqueous phase was 4.02, the dosage of dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was 10.04 mg, and the lipid ratio was 0.12, and the significant phase transition temperature (T_m) of DOX-IR820-TSL was 43.05 ℃. The average particle size of DOX-IR820-TSL@CCM was 153.4 nm, the PDI was 0.279, and the zeta potential was -26.2 mV. The transmission electron microscope (TEM) image shows a homogeneous spherical structure and a translucent film layer. Under near-infrared irradiation, the drug release rate reaches 63.98%, which has adjustable photothermal conversion capacity and the ability to generate reactive oxygen species. Through SDS-PAGE electrophoresis, Western blot, cytotoxicity experiments and cell uptake experiments, it was proved that the design of cell membrane coating can well retain CD47, N-cadherin, CD44, CD326 and other related functional proteins, so that DOX-IR820-TSL@CCM has good immune evasion, homologous adhesion and homologous targeting. In this paper, DOX-IR820-TSL@CCM with camouflage properties and tumor targeting properties were successfully prepared, which can be used as a promising synergistic therapeutic diagnostic platform for future lung cancer treatment. All animal research programs have been approved by the Animal Ethics Committee of Heilongjiang University of Chinese Medicine (number: 2022121011).

Antisense oligonucleotides are a type of gene therapy that targets mRNA and inhibits gene expression. They have been applied in the treatment of various diseases, but there are still problems with poor enzyme stability and high dosage in vivo, due to the shortage of appropriate delivery system. Insulin-like growth factor 1 receptor (IGF1R) is a cell surface receptor with tyrosine kinase activity. Its expression is abnormal in a variety of malignant tumors. It mediates the malignant proliferation, migration and invasion of tumor cells through a variety of ways. In this study, an antisense oligonucleotide (ASO, N04) targeting IGF1R mRNA was designed and chemically modified (PS, 2'-OMOE), then neutral cytidine lipid DNCA and cystine backbone cationic lipid CLD (Mix) were used to encapsulate ASOs. The particle size, polymer dispersity index and ζ potential of the formulations were 151 nm, 0.18 and -3.9 mV. The nanoparticles entered liver cancer cells (HepG-2, Huh-7), silenced target mRNA, arrested cell cycle in S phase, promoted apoptosis, and inhibited the proliferation efficiently. These results indicate that Mix/N04_MOE5 has great potential in tumor treatment, which provides a basis for further research on novel agents against hepatocellular carcinoma.

Combined radiation and burn injury (CRBI) is a severe syndrome, which is induced by the simultaneous or successive radiation and burn; but no appropriate clinical therapies are available. Loong oil (LO) is a traditional Chinese medicine oil composed of the oil extracts of cuttlebone, safflower, walnut oil, and rapeseed oil, which has been demonstrated to own anti-radiation and tissue healing functions. In this study, glyceryl monostearate (GMO) was used for the preparation of lyotropic liquid crystals that loaded LO to obtain Loong oil-lyotropic liquid crystals (LOL) for the treatment of skin CRBI. The hexagonal phase structure of LOL was proved by small X-ray scattering (SAXS) analysis with an approximate $ \sqrt[]{7}:\sqrt[]{9}:\sqrt[]{11} $ ratio of peaks and under the polarizing microscope with the birefringence phenomenon. LOL had a high LO loading capacity (4%) and good bio-adhesive force, favoring skin administration. Animal experiments were approved by the Ethics Committee of the Beijing Institute of Radiation Medicine, Academy of Military Medical Sciences and the experiments were conducted in accordance with relevant guidelines and regulations (approval number: IACUC-DWZX-2022-834). Compared to triethanolamine creams, LOL had higher skin permeability with total skin penetration within 1 h, benefiting the treatment of deep CRBI. A deep Ⅱ degree burn CRBI mouse model was established after whole-body irradiation with 5 Gy ⁶⁰Co γ-ray and the next 6-second 100 ℃ burn injury. LOL promoted LO skin penetration, improved epithermal cell proliferation and the formation of hair follicles, accelerated wound healing, and inhibited scarring, which benefitted the therapy of CRBI. In this study, LO application fields are expanded and LO is a promising medication for the treatment of CRBI.

MADS-box protein family are important transcriptional regulatory factors in plant growth and development. The AGAMOUS 12 (AGL12) subfamily is believed to play an important regulatory role in the process of plant flowering transition. To explore the potential mechanism of AGL12 subfamily involved in regulating the flower development of Lonicera macranthoides, quantitative real-time polymerase chain reaction (qRT-PCR), prokaryotic expression and yeast two-hybrid techniques were used to analyze the expression pattern, protein expression, and protein-protein interaction pattern of LmAGL12 based on transcriptome data. The results showed that the LmAGL12 gene contains a 603 bp open reading frame (ORF), encoding 200 amino acids, and the encoded protein was stable and hydrophilic without a transmembrane region and signal peptide. Through homologous sequence alignment and phylogenetic analysis, it was confirmed that LmAGL12 protein belongs to the MADS-box protein family and is closely related to the AGL12 protein of Heracleum sosnowskyi and Daucus carota subsp. sativus. The LmAGL12 gene was cloned into prokaryotic expression vector pET-28a and the recombinant constructs were transformed into Escherichia coli BL21 (DE3), which inducing the target protein successfully. The yeast two-hybrid results showed that LmAGL12 protein interacts with LmSVP protein, LmSOC1 protein and LmAP1 protein, respectively. The qRT-PCR results showed that LmAGL12 gene were differentially expressed in different development stages of flower bud, stem, and leaves of 'Longhua' and 'Baiyun' in L. macranthoides. The LmAGL12 gene showed the highest expression level in the middle stage of the 'Longhua' floral bud; For the 'Baiyun' variety, the relative expression level of LmAGL12 gene is the highest in the stem. This study cloned the LmAGL12 gene in L. macranthoides and analyzed it expression for the first time, enriching the research on flower organ development and providing a research basis for further exploring the molecular mechanisms of long bud stage and non-unfolded corolla in L. macranthoides, as well as for variety improvement.

Polygonum capitatum is a characteristic Miao medicine in Guizhou, commonly used in clinical practice to treat gastrointestinal and urinary tract infections. Research has found that it has good antibacterial and anti-inflammatory effects, and its main active ingredient is flavonoids. Lavonoid O-methyltransferase (FOMT) is a key enzyme for oxymethylation modification of flavonoid compounds. In order to understand the function and properties of FOMT protein in Polygonum capitatum, the transcriptome was sequenced by Illumina HiSeq 4000 high throughput sequencing technology. Then, the obtained transcripts were annotated and analyzed, and the whole genome of the FOMT gene family in Polygon capitalis was mined and identified. A total of 99 298 Unigenes were obtained, of which 71 514 were successfully annotated by the public database. In the genome of Polygonum capitatum, a total of 50 FOMT genes were identified. The phylogenetic tree showed that FOMT genes were divided into two subfamilies: caffeoyl CoA O-methyltransferase (CCoAOMT) and caffeic acid O-methyltransferase (COMT). Gene sequence analysis showed that the number of FOMT encoded amino acids ranged from 99 to 1 053 aa, the molecular weight ranged from 11 224.91 to 86 687.42 Da, and the isoelectric point ranged from 4.79 to 9.45. The 50 FOMT family members were hydrophobic proteins. Subcellular localization results showed that 54% of FOMT subfamily CCoAOMT and COMT members were located in cytoplasm and 28% were located in chloroplasts. The FOMT gene was tissue specific and highly expressed in the flowers of Polygonum capitatum, followed by the stems, and the least expressed in the roots. In this study, the FOMT gene family of Polygonum capitatum was identified and analyzed to provide theoretical basis for further study of FOMT function and biosynthesis of methylated flavonoids.

GNPS-based mass spectrum-molecular networks is an effective strategy for rapidly identifying known natural products and discovering novel structures. The chemical diversity of azaphilones from the fermentation extracts of Talaromyces sp. HK1-18 was studied by molecular network technique. Three linear tricyclic azaphilones, sequoiamonacins A-C (2a, 2b, 1), were isolated by silica gel column chromatography and high performance liquid chromatography from the extracts of the fungal strain of HK1-18, and their structures were identified by nuclear magnetic resonance and high-resolution mass spectrometry. Guided by the mass spectra of sequoiamonacins A-C (2a, 2b, 1), the cluster of sequoiamonacinoid analogues was discovered from the full molecular networking of HK1-18. By analyzing the MS/MS fragments of each parent ion in this cluster, 7 azaphilones (3-9) included 6 new ones (4-9) were predicted successfully. Then the MS/MS cracking regularity of this type of azaphilones was revealed. Compound 1 showed anti-inflammatory activity, which can inhibit the production of interleukin-1α (IL-1α) in lipopolysaccharide (LPS)-induced mouse macrophage RAW264.7, with an inhibitory rate of 29% at the concentration of 12.5 μg·mL^-1.

Salvia apiana Jepson, commonly known as white sage, is a perennial sub-shrub of the Salvia genus in the Lamiaceae family with a long medicinal history. In this study, the complete chloroplast genome of S. apiana was sequenced using PacBio HiFi third-generation sequencing technology. The physical map of the genome was constructed, and the sequence structure features, codon preference, and repetitive sequences were analyzed. Furthermore, a comparative analysis of the chloroplast genome and phylogenetic evolution with closely related species within the same genus was conducted. The chloroplast genome of S. apiana was found to have a length of 151 701 bp (GenBank accession number: OR389048), with a typical quadripartite structure and a GC content of 38.06%. A total of 132 genes were annotated, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Among them, 17 genes contained introns, and 18 genes were present in duplicate copies. Codon preference analysis revealed a preference for codons ending with A or U. Analysis of repetitive structures in the S. apiana chloroplast genome identified 170 simple sequence repeat (SSR) sites and 65 scattered repeat sequences, with the majority of SSR sites composed of A and T. Phylogenetic analysis of the complete chloroplast genomes of 21 species within the same genus showed that S. apiana is most closely related to Salvia hispanica Ettling. ex Willk. & Lange, Salvia leucantha Cav., and Salvia tiliifolia Vahl. Comparative analysis of the chloroplast genomes revealed slight contraction and expansion of the inverted repeat (IR) boundaries in S. apiana, as well as multiple highly variable regions in the chloroplast genome sequence. This study establishes a method for de novo assembling the chloroplast genome of S. apiana using third-generation sequencing data and provides a comprehensive analysis of its chloroplast genome, which can serve as a theoretical basis for studies on chloroplast genetic engineering, genetic diversity analysis, molecular breeding, and species identification.

WRKY transcription factor is a type of transcription factor unique to plants and plays an important role in various physiological processes of plants. This study is based on the transcriptome data of Atractylodes lancea, the correlation between the FPKM values of the AlWRKY gene and the AlTPS gene of Atractylodes lancea was analyzed. Combined with the analysis results, the candidate gene AlWRKY65 with a significant positive correlation with the expression levels of AlTPS1 and AlTPS6 genes and a relatively high FPKM value was screened. The candidate gene was cloned to obtain the open reading frame ORF of the AlWRKY65 gene, and the related information of its encoded protein was analyzed and the gene expression was studied. The results showed that AlWRKY65 contains a 681 bp open reading frame and encoding 226 amino acids. Through amino acid sequence homology analysis, it was found that AlWRKY65 amino acid sequence had high homology with several plants such as HaWRKY65 and LsWRKY65; AlWRKY65 protein had a typical WRKYGQK domain, belonging to IIe subgroup of WRKY transcription factor family; phylogenetic analysis indicated that AlWRKY65 protein had the higher homology with CcWRKY65 protein; the expression of AlWRKY65 gene in different tissues of two producing areas of Atractylodes lancea were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves and also has tissue differences. The expression level of AlWRKY65 gene was down-regulated within 48 h of methyl jasmonate (MeJA) induction; subcellular localization and transcriptional activation assay suggested that AlWRKY65 was located in the nucleus and had no transcriptional activation activity. This study provides a reference for further elucidating the biological function of AlWRKY65 in Atractylodes lancea.