Compared with systemic administration such as oral delivery or injection, inhaled medicines directly locate in the respiratory tract to exert therapeutic effects, offering obvious advantages in the treatment of respiratory diseases. Marketed inhaled medicines are yet difficult to meet the clinical demands, and there are considerable challenges in the discovery and development of novel inhaled medicines due to the lack of experiences- and property-based rules for inhaled compounds. Personalized modification of candidate drugs through prodrug technology to meet the requirements of inhalation therapy is the current alternative approach for inhaled drug development. In this review, we intend to summary the applications of prodrug technology in the research of inhaled medicines over the past 20 years. These studies have shown that esterified prodrugs and macromolecule conjugates could effectively prolong lung retention; mannose modification or acid-sensitive bond connection can achieve targeted drug release in alveolar macrophages; personalized modified prodrugs can obtain suitable physicochemical properties for pulmonary delivery and reduce drug toxicity. In general, the application of prodrug technology can modify the physicochemical and biopharmaceutical properties of drugs and may promote the discovery and development of novel inhaled medicines.

Pulmonary drug delivery agents deliver drugs directly to the lung tissue to obtain higher local concentration, which is beneficial to the treatment of lung diseases, and may reduce the systemic side effects. Therefore, it has become the preferred drug for the treatment of many lung diseases. However, the drugs entering the lungs are easily cleared by the lung tissues, which reduce the retention time of the drugs in the lungs and affects the efficacy. Although particles with a smaller particle size (1-5 µm) are easily inhaled into the lungs, they are also easily swallowed and cleared by lung macrophages. While the porous polymer microspheres (PPMS) with larger geometric diameter (Dg > 5 µm) and low density (ρ < 0.4 g·cm^-3) can not only effectively avoid the phagocytosis of alveolar macrophages, but also have a high effective deposition rate in the lungs due to the appropriate aerodynamic diameter, moreover, the polymer is biodegradable and non-toxic, so it has become a research hotspot for pulmonary drug delivery carriers. This article combined with the lung clearance mechanism of granules, summarized the preparation materials and methods of PPMS, as well as its quality control, etc. Furthermore, opinions are also put forward for the development of PPMS, to provide a reference for the in-depth study of PPMS.

The movement and deposition of inhaled particles in the lung tissue have an important influence on the respiratory physiology of human body. The corresponding particle dynamics model plays a vital role in exploring the pathogenic factors and treatment methods of lung diseases. Such as evaluating the optimal design of pulmonary atomization drug delivery and the impact of particulate air pollutants on the lung. According to the different knowledge and modeling methods, this paper classifies, combs and analyzes several representative particle dynamics models in lung airways in recent years. We divided the mechanism models into the biological model, physical model and numerical simulation model. The biological models include in vivo imaging and pharmacokinetic methods; physical models include bionic and microfluidic chip models; the numerical models include respiratory tract model, airflow model and particle model. Moreover, the numerical solution and fluid-structure-interaction models were also reviewed, especially the application prospect of the lattice Boltzmann method.

This article systematically reviews the background and regulatory requirements of bioequivalence of orally inhaled and nasal drug products (OINDPs), as well as the basic regulatory requirements for the assessment by the guidelines and guidance issued in China, the United States, and the European Union. Detailed statistical evaluation method considerations and calculations of the US FDA population bioequivalence (PBE) method were presented for the evaluation of in vitro bioequivalence (IVBE) for OINDPs. Using the example described in the FDA Draft Guidance for budesonide inhalation suspension, the PBE analysis statistical parameters were calculated via the R programming, and the results were compared with that in the guidelines. Moreover, pseudo-code for the PBE calculation program was provided. This paper aims to provide guidance and references for the research and development of new drug, as well as pharmaceutical quality control, and development of generic medicinal products for OINDPs.

Bacterial pneumonia is a common clinical disease and the abuse of antibacterial leads to the production and prevalence of resistant bacteria. Novel antibacterial active ingredients can be found from natural products for the treatment of bacterial pneumonia. Pulmonary delivery systems can directly deliver drugs to infected lung tissues, favoring the treatment of bacterial pneumonia. Cinnamon oil (CO) is a volatile oil extracted from cinnamon, which has antibacterial and anti-inflammation functions. However, the preparation of its pulmonary delivery systems is difficult because of its oil and volatile properties. Here, CO-β-cyclodextrin inclusion complex (CO-β-CD) was prepared. Its dry powder inhalers (DPIs) were obtained by lyophilization. The DPIs had the aerodynamic diameter of 1.34 μm and the fine particle fraction (FPF) of 30.90%, which was suitable for pulmonary delivery. The minimal bactericidal concentration of CO-β-CD was 2 mg·mL^-1 against Staphylococcus aureus (SA). The animal experiments were approved by the Ethics Committee of Academy of Military Medical Sciences and the experiments were conducted in accordance with relevant guidelines and regulations. A high intratracheal (i.t.) dose (100 mg·kg^-1) of CO-β-CD did not show significant toxicity to the mice. Bacterial pneumonia mice models were established after intraperitoneal injection of dexamethasone and then i.t. administration of SA. CO-β-CD and penicillin were separately i.t. administered to the mice. Both of them alleviated the lung injury of mice and remarkably increased the survival rates. Compared to penicillin, CO-β-CD significantly reduced the infiltration of inflammatory cells and the numbers of leukocytes and neutrophils in the blood, and the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the lung tissues were similar to those of the healthy mice, indicating that CO-β-CD had the capability of anti-inflammation besides antibacterial effect in vivo. CO-β-CD dry powder inhalers are a potential pulmonary delivery system for the treatment of bacterial pneumonia.

Conjugated linoleic acid (CLA) is a nutrient substance that exists in humans and animals. It has anti-tumor, anti-atherosclerosis, and immune-regulating functions, but its oral bioavailability is low. Conjugated linoleic acid dry powder inhalers (CDPIs) were prepared and intratracheally administered to the rats that suffered from primary lung cancer. Conjugated linoleic acid nanoemulsions were prepared first and CDPIs were with 10% mannitol after lyophilization. CDPIs are loose white powders with the aerodynamic median diameter (D_a) of 3.10 μm, which were suitable for pulmonary delivery. Rats lung cancer models were established after 45 days by instilling 3-methylcholanthrene (MCA) and N, N-dimethylnitrosamine (DEN) into the rats lung once. The animal experiments were approved by the Ethics Committee of Academy of Military Medical Sciences and conducted in accordance with the relevant guidelines and regulations. The CDPIs, gefitinib suspension and blank DPIs were sprayed into the lungs of rats with lung cancer through the trachea. Compared with the model group, both the gefitinib suspension group and the CDPIs group showed significantly fewer tumor nodules and inflammatory cells, and the CDPIs group was better than the gefitinib suspension group. The inhibition efficiency of CDPIs on CD31 and NF-κB p65 was better than that of the gefitinib suspension group. The vascular endothelial growth factor (VEGF) level in the CDPIs group was significantly reduced, which was equivalent to that of the gefitinib suspension group. The apoptosis in the CDPIs group by Tunel tests showed a significant increase, which was significantly better than the gefitinib suspension group. Therefore, CDPIs had excellent pharmacological activity on lung cancer, which provided a model for the efficient delivery of oil therapeutic agents.

High altitude pulmonary edema (HAPE) is a result of leaking of the fluids from blood vessel to the lung tissue due to pulmonary artery hypertension in the high altitude place, which happens very quickly and shows high mortality. Sildenafil (SIL) can prevent HAPE by expanding pulmonary vessels; however, only oral tablets and injections of SIL are currently available. The formulations have the disadvantages of high doses, inconvenient use, and high systemic side effects. Here, liposomal sildenafil (LS) was prepared for pulmonary delivery and the prevention of HAPE was explored. The ammonium sulfate gradient method was used for the preparation of LS with a high entrapment efficiency of nearly 100%, the particle size of 116.97 nm, and the zeta potential of -30.93 mV. All animal experiments had been approved by the Ethics Committee of Academy of Military Medicine, Academy of Military Sciences, and carried out in accordance with relevant guidelines and regulations. The dose (3 mg·kg^-1 SIL) of sildenafil suspensions, LS or blank liposomes was intragastrically or intratracheally administered to the lungs of mice. The mice with or without treatment were put in the hypobaric oxygen chamber of 5 000 m altitude for 1 h. In the open field text the model mice had the shorter total distances and longer dead time than the healthy mice and they showed the lower percutaneous oxygen saturation (SpO₂), the higher tumor necrosis factor-α (TNF-α) levels of and the lower reduced glutathione (GSH) in the lung tissues. By contrast, oral LS remarkably modified the moving ability of the mice and they had the higher SpO₂ than the model mice and the similar TNF-α and GSH levels to the healthy mice. Sildenafil suspensions and LS with the same dose (3 mg·kg^-1 SIL) were intratracheally administered to mouse lungs, respectively, and the mice of the model group and the treatment groups were put in a hypobaric oxygen chamber for predetermined time. In the rotating rod experiments, the mice in the model group showed shorter drop latency and more drop times than those in the healthy group, indicating that the physical activity of the mice was improved due to treatment. The inflammatory cytokines, TNF-α and interleukin-1β (IL-1β) in the blood of model mice were higher than those from the healthy group and they decreased after treatment, where the LS group maintained the lowest level close to the normal level. IL-1β in the blood of mice in the LS group was lower than that in the SIL group (P < 0.05) 1 hour and 48 hours post-hypoxia. Hypoxia inducible factor-1α (HIF-1α) in the lung tissues of model mice increased but decreased to the normal range after treatment. Moreover, HIF-1α in the LS group was lower than that in the SIL group, where the values were significantly different 48 hours post-hypoxia (P < 0.01). Inhaled liposomal sildenafil is a promising medication for the prevention of HAPE.

We established in vitro evaluation methods of the nebulization characteristics of budesonide suspension for inhalation and analyzed the influence factors. The delivery rate and total drug substance delivered (TDD) of two manufacturers were determined by using the breath simulator with different nebulizers. The aerodynamic particle size distribution was investigated by next generation impactor (NGI) and HPLC as well. The fine particle dose (FPD), the mass median aerodynamic diameter (MMAD), the delivery rate and TDD results of the same sample with different nebulizers were significantly different (P < 0.01), mainly due to the different design parameters of the nebulizers. The FPD of two samples were significantly different (P < 0.01) by the same nebulizers, probably due to differences in physical and chemical properties differences such as suspension particle size. The analysis method of nebulization characteristics provided in this paper can be used to select the nebulization device with appropriate delivery dose and aerodynamic particle size distribution (APSD) for different inhalation liquid preparations in clinical practice, and can also be used as the guidance for the selection of nebulization device and analysis method in the research and development of generic inhalation liquid preparations and quality consistency evaluation.

The objective of the study was to investigate the effects of nebulizers and add-on spacers on nebulization performances of Re-Du-Ning injection. Chlorogenic acid and geniposide were taken as the chemical markers of the injection. In vitro nebulization performances including real-time particle size distribution, aerodynamic particle size distribution (APSD), respirable drug delivery rate (RDDR) and percentage of respirable delivered dose (RDD%), residual rate and nebulization time were determined after nebulization of Re-Du-Ning injection via jet or mesh nebulizers. The delivery efficiency was the ratio of delivery dose to the drug content of the injection. The aerosol delivery efficiency of Re-Du-Ning injection for the jet nebulizer was about 30% of the nominal dose whereas those for mesh nebulizers varied from 24% to 35% in adult breathing pattern. Upon the inclusion of spacers, the delivery efficiencies of mesh nebulizers increased to 37%-68% whereas the presence of a spacer decreased the delivery efficiency of the jet nebulizer. When the aerosols of mesh nebulizer exhibited comparative fine particle fraction (FPF) and mass median aerodynamic diameter (MMAD) to those of the jet nebulizer, the RDD% of the former nebulizer with a spacer was more than two folds of the latter. These results indicated that the addition of spacer was demonstrated to significantly improve the delivery efficiency of Re-Du-Ning injection by mesh nebulizers.

Most small molecule drugs bind to enzymes, receptors or ion channels, which possess binding pocket for drug occupation. However, the study of drugs that interfere with protein-protein interactions has always been a challenging subject. The discovery of molecular glues and degraders has opened an avenue to tackle this issue. With the structural features of bifunctional ligand molecular glues mediate the recognition and binding of two proteins. As a useful tool for chemical biology molecular glue can not only help to find probes to undruggable targets, but also can be developed into drugs through structure optimization in medicinal chemistry. This minireview concisely describes the features of molecular glue using a few existing drugs or active compounds.

Hypoxia is one of the most significant characteristics of solid tumors. Hypoxia microenvironment can lead to the overexpression of hypoxia inducible factor-1α (HIF-1α). As the most critical transcription factor in the hypoxia response, HIF-1α activates downstream gene expression resulting in abnormal tumor cell proliferation, tumor angiogenesis, unusual energy metabolism, increased drug resistance, invasion, and metastasis. Down-regulation of HIF-1α expression is considered as a promising approach for the treatment of solid tumors, whereas the clinical efficacy of most existing HIF-1α inhibitors is restricted in low efficacy and high toxicity. Therefore, it is particularly important to develop powerful and safe novel drugs against the overexpression of HIF-1α. In recent years, numbers of studies have proved that a variety of chemical components of traditional Chinese medicine can directly or indirectly inhibit the activation of HIF-1α, which has a broad prospect in the fight against hypoxia-induced tumor progression. In this review, we summarized various anti-tumor active components of traditional Chinese medicines responsible for inhibiting the expression of HIF-1α in last ten years and analyzed the corresponding mechanism, with a view to further research as a reference.

Transient receptor potential vanilloid ion channel subtypes 2 (TRPV2) is a calcium permeable nonselective cation channel. TRPV2 is expressed in nerves, blood vessels, spleen, and other organs, and participates in the regulation of a variety of important physiological functions, such as nociception, neural development, and immune response. Here we reviewed the research progress on the structure, modulators, and physiological functions of TRPV2 channel to provide references for future research.

Growth differentiation factor 15 (GDF15) is an endocrine hormone that belongs to the transforming growth factor β (TGFβ) superfamily, which plays a role by binding to the glial cell line derived neurotrophic factor (GDNF)-family receptor α-like (GFRAL)-rearranged during transfection (RET) heterodimer receptor. A number of studies have confirmed that GDF15, as a biomarker for obesity, diabetes, tumors, non-alcoholic fatty liver disease, ischemic diseases, and so on, has become a new target for new drug discovery. At present, pharmaceutical companies around the world are carrying out drug discovery with GDF15 as a new therapeutic target involving many therapeutic areas such as anti-obesity, anti-tumor, and treatment of anorexia syndrome. Comprehensive analysis of the advantages and disadvantages of the GDF15 target, discussion, and objective evaluation of research and development of new drugs based on GDF15 will provide a scientific basis for the discovery of innovative drugs that could relieve the human suffering from illness.

Protein disulfide isomerase (PDI), also named PDIA1, is an archetype member of protein disulfide isomerase family, which mainly exists in endoplasmic reticulum and plays an important role in pathophysiological progress via enzyme activity and molecular chaperone function helping protein folding correctly. Amyotrophic lateral sclerosis (ALS) is a kind of neurodegenerative diseases characterized by abnormal protein aggregation. Recently, PDI mutants have been found in ALS patients and the important role of PDI protein in the development of ALS has also been confirmed. Here, we focus on the recent studies of PDI in ALS hoping to provide a novel target for the treatment of ALS.

The structural modification of natural product represents a powerful tool for anti-cancer drug discovery. Noscapine, as a phthalideisoquinoline alkaloid from opium, has been used as an over-the-counter antitussive drug with excellent oral bioavailability and low toxicity. Recently, the potential of this compound as a particularly attractive lead for anti-cancer drug discovery has been demonstrated. Multiple mechanisms, especially the interference of tubulin polymerization, might be involved. Thereafter, various structural modifications based on semisynthetic routes, which aims to improve the anti-cancer activity and pharmacokinetic properties, as well as to probe the mechanism, has been performed. Several analogues are emerging as possible candidates as novel anticancer therapies. This perspective mainly discusses the advancing noscapine and related analogues in the fight against malignant disease in recent years. Furthermore, the future directions of this evolving field were also preliminary prospected.

In recent years, the research on myocardial ischemia disease is increasingly in-depth, and there are more and more drugs related to the anti-myocardial ischemia, among which natural medicines play an important role. In order to deal with myocardial ischemia diseases caused by a variety of pathogenic causes, the research on natural medicines is more and more deep. At present, natural medicines containing saponins, flavonoids, alkaloids and other compounds are the research objects. At the same time, great progress has been made in the research on the clinical application of natural drugs against myocardial ischemia. This article reviews the research progress of natural medicines against myocardial ischemia at home and abroad in the past ten years.

Cancer is a malignant disease threatening human health and life. Early diagnosis of carcinoma plays an important role in the treatment of carcinoma, so tumor imaging agents have been the focus of research in this field. Mannose receptor is a C-type lectin receptor, which expressed in various tumor tissues, sentinel lymph nodes and has become an important tumor marker. In recent years, a variety of imaging agents targeting mannose receptors have been designed and developed. This review summarizes the representative achievements.

As one of the most critical post-translational modifications, glycosylation of therapeutic proteins has a profound impact on their safety, efficacy and consistent. However, glycosylation is not a template-driven process, therefore variability in the glycosylation pattern of a protein can arise. This makes challenges of glycan analysis and control. Here, we review the overall control strategy, basic requirements for standardized protocols and the novel technologies of glycosylation analysis to accelerate the development of therapeutic glycoproteins.

The activity and expression of drug metabolizing enzymes and transporters changes significantly under high altitude hypoxia. The gut microbiota is an important factor affecting the metabolism of drugs through direct and indirect actions, changing the bioavailability, biological activity or toxicity of drugs and affecting the efficacy and safety of drugs. High altitude hypoxia significantly changes the structure and diversity of the gut microbiota, which may play a role in drug metabolism. This article reviews the effects of high-altitude hypoxia on the gut microbiota and the effects these changes on drug metabolism.

We provide a new preclinical pharmacodynamic evaluation method of cerebral thrombolytics using beagle dogs with acute thrombotic cerebrovascular occlusion. An intracranial large vessel occlusion animal model was generated by pushing an autologous thrombus to the left carotid artery under X-ray angiography. The experiments were approved by the Animal Care and Welfare Committee Institute of Materia Medica, China Academy of Medical Sciences (CAMS), Peking Union Medical College. According to the degrees of cerebral blood flow reperfusion, the new method has 0, Ⅰ, Ⅱ (Ⅱa and Ⅱb), Ⅲ levels. Respectively, 0 level represents no blood flow through the area of vascular occlusion; Ⅰ level has low perfusion; Ⅱa level is less than half the infusion; Ⅱb level is more than half perfusion; Ⅲ level is completely recanalization perfusion. Alteplase was used as a positive drug to verify the accuracy of the method. Our results suggest that 100% of dogs reached the level of grade I and below in the model group, while 83% reached the level of grade IIa and above in the alteplase group. Results of a rank sum test showed that alteplase significantly improved the reperfusion of occluded cerebral vessels. We propose a method to evaluate the efficacy of thrombolytic drugs on dogs with middle cerebral artery occlusion.

In recent years, the prevalence of non-alcoholic fatty liver disease (NAFLD) has been dramatically increased in China and specific targeted therapy is still unavailable. The purpose of this research was to investigate whether schisandrin B (SchB) improves NAFLD and the potential mechanisms. Wildtype mice with C57BL/6J background were treated with special high fat diet (containing 40% fat, 22% fructose, 10% sucrose, and 2% cholesterol) for 16 weeks to induce NAFLD. Then SchB (120 mg·kg^-1) were used to treat NAFLD mice for 6 weeks. Body weight, food intake, glucose tolerance, insulin resistance, serum level of total cholesterol (TC), triglycerides (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were assayed and histopathological analysis were performed to evaluate the improvement of NAFLD induced by SchB. Furthermore, the level of lipopolysaccharide (LPS) in serum, intestinal permeability, and the expression of genes and proteins associated with mucosal defense were evaluated, intestinal flora composition in fresh cecal contents were analyzed and differential flora were identified to explore the potential mechanism. All animal experiments were approved by the Animal Research Committee of Shanghai University of Traditional Chinese Medicine. These results showed that SchB significantly reduced the body weight of NAFLD mice without changing food intake, and effectively reduced serum level of TC, ALT, and AST. SchB also significantly altered the composition of the microflora in NAFLD mice, increased the abundance of the Akkermansia muciniphila (A. Muciniphila) and elevated the expression of genes and proteins associated with the mucosal defense in ileum and colon, restored the permeability of intestinal barrier. In summary, SchB improves NAFLD by regulating the composition of the microflora and enhancing the function of intestinal barrier to further reduce the excessive lipids accumulation and hepatic inflammation.

To explore the preventive effects and potential mechanism of mulberry leaf water extract (MLWE) on type 2 diabetes mellitus (T2DM), this study observed the influence of MLWE on the metabolism of arachidonic acid (AA). T2DM mice were induced by high fat and high sucrose (HFHS) diet and intra-peritoneal injection of streptozotocin. The mice were randomly divided into a normal control group treated with drug-free solution (NC group), a normal control group treated with MLWE (NCML group), a diabetes mellitus (DM) group treated with drug-free solution (HFHS group), and a DM group treated with MLWE (HSML group); mice were maintained on this protocol for 10 weeks. Animal experimentation was approved by the Committee on the Ethics of Animal Experiments of Xuzhou Medical University. Mice livers and plasma were collected at the end of experiment. Fasting blood glucose (FBG) and fasting blood insulin (FBI) were detected by kits. The mRNA and protein expression levels of relative metabolic enzymes related to AA in mice livers were respectively detected by quantitative real time PCR (qPCR) and Western blot. The contents of AA and its relative metabolites in mice plasma were determined by liquid phase chromatography coupled with tandem mass spectrometry. The results showed that there was no significant variation for the relative expression of metabolic enzymes, contents of FBG, FBI, AA, and its relative metabolites between NCML group and NC group. Compared with NC group, the relative expression of fatty acid synthase (Fasn), phospholipase A2 (PLA2), cyclooxygenase-2 (COX-2), and lipoxygenase-5 (LOX-5), and the levels of FBG, FBI, AA, and its relative metabolites in DM group were obviously increased, while that of carnitine palmitoyltransferase 1A (CPT1A) and cytochrome P450 family 4A (CYP4A) were significantly decreased. After intervention with MLWE, those changes could be improved, indicating that MLWE could prevent T2DM by acting on AA metabolism.

Blocking the binding of programmed death 1 (PD-1) on the T cells and programmed death ligand 1 (PD-L1) on the tumor cells has become a hotspot in the field of tumor immunotherapy. Small-molecule checkpoint inhibitor targeting PD-1/PD-L1 axis is the new direction of tumor immunotherapy. In the present study, we investigated the anti-tumor role of hyperoside by regulating the PD-L1 level in non-small cell lung cancer (NSCLC). Changes of total PD-L1 and membrane PD-L1 levels were determined by Western blot, flow cytometry, and PD-1/PD-L1 interaction assays. The expression of mRNA level of PD-L1 was detected by real-time PCR. The cytotoxicity of activated human T cells toward co-cultured tumor cells was measured by cell impedance assay and crystal violet experiment. The antitumor effect of hyperoside in vivo was examined by C57BL/6 mice bearing Lewis xenograft tumor. Western blot and flow cytometry assay showed that hyperoside significantly downregulated the abundance of PD-L1 in H1975 and HCC827 cells in dose- and time-dependent manner. PD-1/PD-L1 binding assay revealed that hyperoside reduced the binding of tumor cells to recombinant PD-1 protein. In addition, hyperoside decreased the abundance of c-Myc, a key transcriptional regulator of PD-L1, in H1975 and HCC827 cells. Cell impedance and crystal violet staining indicated that hyperoside enhanced the killing activity of co-cultured T cells toward tumor cells. Animal experiments (all animal experiments were conducted in accordance with the Animal Ethics Committee of the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences) revealed that hyperoside treatment displayed significant suppression in the growth of Lewis tumor xenografts in C57BL/6 mice with an inhibition rate of 48.3% at 25 mg·kg^-1. Our results demonstrate that hyperoside exerts its anti-NSCLC activity by reducing the PD-L1 level. Our study provides an important material basis and scientific basis for developing hyperoside into a new small molecule drug for tumor immunotherapy.

Twenty-five quinolizidine alkaloids (including matrine-type 1-14, sparteine-type 15-17, cytisine-type 18-23, other types 24 and 25) were isolated from the roots and rhizomes of Sophora tonkinensis by various chromatographic methods. Their structures were elucidated by physicochemical properties, NMR and MS spectral data. Among them, 12-(1-acetoxyethyl)-cytisine (23) is a new alkaloid derivative, and compounds 13, 16, 17, 24 were isolated from the roots and rhizomes of S. tonkinensis for the first time. Compounds 1, 6, 19 and 20 showed potent inhibitory activity against LPS-induced NO production in RAW 264.7 macrophages, with IC₅₀ values of 39.86 ±0.65, 23.66 ±0.37, 34.56 ±0.45, 47.68 ±0.58 μmol·L^-1, respectively.

Five compounds were isolated from an ethanol extract of Artemisia deversa Diels by solvent extraction, Diaion HP-20, CHP20/P120 MCI, Sephadex LH-20, silica gel and preparative high performance liquid chromatography. Their structures were elucidated by MS, NMR and X-ray as artemideversal (1), 7-hydroxy-6-methoxycoumarin (2), 6, 7-dimethoxycoumarin (3), caffeic acid (4) and 4', 5, 7-trihydroxy flavone (5). Compound 1 is a new eudesmane-type sesquiterpene. In vitro cytotoxic activities of the five compounds were explored by MTT testing with HepG2, A549, HeLa and MRC-5 cell lines. Results show that compounds 1 and 5 significantly inhibited cellular proliferation. The IC₅₀ of compound 1 in A549 and HepG2 cells was 8.36 and 16.51 μmol·L^-1; the IC₅₀ of compound 5 in A549 and HepG2 cells was 17.06 and 7.95 μmol·L^-1.

An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for the simultaneous determination of seven components (chlorogenic acid, rutin, narirutin, hesperidin, sinensetin, nobiletin, tangeretin) in Citri Exocarpium Rubrum. The separation was performed over 12.0 min on a Shim-pack GIST C18 column (100 mm×2.1 mm, 2.0 μm) with a mobile phase consisting of acetonitrile (A) and 0.1% formic acid water solution (B) with gradient elution at a flow rate of 0.4 mL·min^-1; the column temperature was 30℃ and the injection volume was 1 μL. Detection was performed on a QTRAP 4500 mass spectrometer equipped with electrospray ionization. Positive and negative electrospray ionization was performed in multiple reaction monitoring mode. All of the analytes showed good linearity (r ≥ 0.997 3) in the tested ranges, with good precision, repeatability and stability for all analytes. The average recoveries were in the range of 99.32%-107.75% with relative standard deviations RSD ≤ 3.94%. The established method is accurate, stable and reproducible, and can provide a research method for the quality control of Citri Exocarpium Rubrum.

Type 2 diabetes is a common form of diabetes and can have serious consequences for diabetics when their wounds do not heal properly due to impaired function. Glycosaminoglycans (GAGs) are widely found in skin tissue. Due to the isomerization of C6 in uronic acid and different sulfuric acid substitutions, the structure of GAGs is relatively complex. The fine structure of GAGs in the skin of diabetic patients has not been studied. In this study, the structure of GAGs in the skin of streptozotocin (STZ)-induced diabetic mice was characterized by liquid chromatographytandem mass spectrometry (LC-MS/MS). All animal experiments were carried out with approval of the Animal Ethics Committee of Ocean University of China. The results indicate that the content of hyaluronic acid (HA) in STZ-induced diabetic mouse skin was significantly lower than that of non-diabetic mice. Although there was no significant difference in the content of chondroitin sulfate (CS) and heparin sulfate (HS) between diabetic mouse and normal mouse skin, the disaccharide compositions were different. The expression of CS-4S6S, HS-0S, HS-NS, and HS-6S in STZ-induced diabetic mouse skin was higher than in non-diabetic mice. While the content of HS-NS6S was lower. In addition, the degree of sulfation of HS in STZ-induced mouse skin was lower than that of normal mouse skin. These results provide a basis for the pathogenesis of diabetes and the development of wound healing dressings for diabetic patients.

The metabonomics method was used to explore the processing and synergistic mechanism of epimedium fried with suet oil in warming the kidney and enhancing yang. The kidney-yang deficiency rat model was established by injection of hydrocortisone. Then the UPLC-Q-TOF-MS (ultra-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry) metabolomics method was combined with multivariate statistical analysis methods and univariate statistical analysis to screen and identify kidney-yang deficiency potential biomarkers in plasma and urine samples. Finally the metabolic regulation mechanism of suet oil group, the epimedium raw product group, the epimedium heating product group, and the epimedium fried with suet oil group improved kidney-yang deficiency was analyzed. The results showed that the plasma and urine metabolism of rats with kidney-yang deficiency induced by hydrocortisone showed obvious trajectory changes. 15 biomarkers related to kidney-yang deficiency were identified in plasma and urine, involving 5 metabolic pathways, namely glycerophospholipid metabolism, sphingolipid metabolism, sulfur metabolism, glyoxylate acid and dicarboxylate metabolism, and cysteine and methionine metabolism. The metabolic pathway of epimedium fried with suet oil warming kidney and promoting yang involved glycerophospholipid metabolism, cysteine and methionine metabolism, and the two processing factors of epimedium fried with suet oil "heating" and "suet oil" enhanced its function of warming the kidney and promoting yang by regulating glycerophospholipid metabolism, cysteine and methionine metabolism, respectively. In this way, the processing and synergistic mechanism of epimedium fried with suet oil was clarified. The animal experiments involved in this article comply with ethical standards and have been approved by the Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine (approval number: AEWC-20200702-119).

Flavanone 3-hydroxylase (F3H), flavonol synthase (FLS) and anthocyanin synthase (ANS) are members of the 2-oxoglutarate-dependent oxygenase (2-ODD) superfamily, which play important functions in plant flavonoid biosynthesis. In this study, 16 2-ODD genes involved in flavonoid biosynthesis pathway were screened and identified from safflower genome database using bioinformatics method. The length of the proteins encoded by these genes ranged from 283 (CtFLS2) to 442 aa (CtANS6), and the molecular weight ranged from 32 062.05 (CtFLS2) to 48 245.49 Da (CtANS6). The proteins encoded by these genes were hydrophilic. Phylogeny divided it into three subfamilies, two conserved residues, H-x-D-xn-H and R-x-S, were found in this gene family using conserved motifs analysis, the analysis of cis-acting elements in the upstream 2 000 bp region showed that these genes might be regulated by environmental factors such as light, temperature and plant hormones such as salicylic acid, methyl jasmonate and so on. The members of the safflower 2-ODD gene family involved in flavonoid biosynthesis pathway were analyzed by qRT-PCR, the result showed that the expression patterns of these members were different in diverse flowering period and leaf. This study can lay a foundation for further exploring the function of 2-ODD genes involved in flavonoid biosynthesis pathway in safflower.

Twenty-five candidate MLO genes of Lonicera japonica were obtained by whole genome sequencing. Bioinformatics analysis showed that the number of amino acids in the protein family ranged from 137 to 846, the theoretical isoelectric point ranged from 5.02 to 9.50, and it was rich in basic amino acids. One protein did not contain transmembrane domain, and the other proteins ranged from 3 to 10. The results of subcellular localization showed that 21 proteins were located on the cell membrane and 1 protein was located on the chloroplast. Phylogenetic trees were constructed from 133 MLO proteins of Lonicera japonica, Triticum aestivum, Arabidopsis thaliana, Solanum lycopersicum, Nicotiana tabacum, Nicotiana sylvestris and Nicotiana tomentosiformis. The results indicated that MLO family proteins of Lonicera japonica could be divided into five subgroups. Tissue specific analysis showed that the expression of MLO genes in Lonicera japonica had obvious tissue specificity. Among them, 8 genes were highly expressed in leaves, 2 genes in stems and 2 genes in flowers. Four genes were significantly upregulated after inoculation with powdery mildew, among which MLO14 increased the most, which was more than 2 000 times higher than that of the control. In this study, the composition and expression of the MLO gene related to the occurrence of powdery mildew of Lonicera japonica were preliminarily analyzed, which laid a foundation for the further use of MLO as a target gene to develop new germplasm resistant to powdery mildew of Lonicera japonica.