Hepatic encephalopathy is a common metabolic neuropsychiatric syndrome in the development of end-stage liver disease. Since the concept of intestinal-liver-brain axis was proposed, the relationship between the pathogenesis of hepatic encephalopathy and the gut microbiota has been a hot research topic. In recent years, studies have confirmed that gut microbiota is involved in and affects various pathological processes of hepatic encephalopathy. This article combines the latest research progress at home and abroad to elaborate on the research status of regulating gut microbiota and thus interfering with the pathological process of hepatic encephalopathy, hoping to provide new ideas and methods for the intervention of hepatic encephalopathy based on the regulation of gut microbiota.

Depression is one of the diseases with the highest disability rate in the world. A large number of studies have shown that the intake of unsaturated fatty acids can deal with depression while chronic overconsumption of saturated fatty acids is a risk factor for depression. It was suggested that the mechanism of saturated fatty acids inducing depression is related to the following four aspects: regulating the function which links to depression in whole brain and specific brain regions, including the hippocampus, the hypothalamic-pituitary-adrenal axis, the striatum, and the prefrontal cortex; stimulating the secretion of inflammatory factors; affecting the balance and function of metabolic regulatory hormones, including leptin, adiponectin, glucocorticoid, and insulin; inducing the disturbance of intestinal flora. This article reviews the relationship between dietary fatty acids and depression, and the possible mechanisms by which saturated fatty acids induce depression from the four aspects mentioned above.

Malignant tumors seriously endanger human life and health, and their treatment has always been a research focus of scientists all over the world. Natural flavonoids and their derivatives have a variety of biological activities, especially regarding antitumor growth, with unique biological activities. They can interfere with the growth cycle of tumor cells, change the mitochondrial membrane potential, promote apoptosis, and can reduce the immune escape of tumor cells and prevent tumor metastasis by improving human immunity. In the human body, they regulate the biological signal transduction, leading to the up-regulation of pro-apoptotic protein expression. They inhibit the growth of solid tumors by regulating the growth of vascular epithelial cells and blocking the formation of blood vessels in tumor tissue. Recent studies have shown that these compounds can play an important role in the treatment of various human tumors and are expected to be developed into new antitumor drugs. This review summarizes the recent research results on the antitumor mechanism of flavonoids and their ability to inhibit tumor growth.

Epilepsy is one of the most common neurological conditions, which is characterized by recurrent unprovoked seizures. Drug treatment is still the main method for the disease. Although remarkable progress has been made in the development of antiepileptic drugs in recent years, there is still a poor curative effect on patients with refractory epilepsy. This review will focus on the current status and pathogenesis of epilepsy, as well as the antiepileptic drugs (targeting sodium channels, calcium channels, potassium channels, and the balance of γ-aminobuyric acid/glutamate system, respectively) that have been developed based on classical epileptogenic mechanisms. Further the antiepileptic drugs acting on new targets (epigenetic interferers, synaptic vesicle glycoprotein 2A modulators, mammalian target of rapamycin signal pathway blockers, carbonic anhydrase inhibitors, cannabidiol and adenosine inhibitors) have also been discussed.

Macrophages are highly plastic and heterogeneous. In different types of inflammatory diseases, or at different stages of the same disease, macrophages can undergo phenotypic transformation to elicit different functions. Hence, exploring new regulatory mechanism of macrophage polarization and seeking for new key mediators will lay the foundation for the diagnosis and treatment of macrophage-related diseases, such as inflammatory diseases, autoimmune diseases, and cancer. Interferon regulatory factors (IRFs) have been reported to play an important role in the maturation and differentiation of macrophages. In this review, we will describe the structure and modulation pattern of IRFs, and then further summarize the molecular mechanism and signal regulation network of IRFs in pathological processes of related diseases through controlling macrophage polarization. Our review will explore the new therapeutic strategy and potential drug targets for related diseases.

Anxiety disorders are one of the most common mental disorders in adults, the cause of which derives from a combination of genetics and environmental factors. A series of animal models have been established according to their pathogenesis to measure the level of anxiety or induce anxiety only, and these models have been widely applied in the non-clinical evaluation of anxiolytics. In this review, we present the current trends in the study of anxiety disorders and summarize typical non-clinical anxiety animal models, including models that both measure anxiety levels and induce anxiety, and models that induce anxiety only. This review summarizes the important issues in standardized non-clinical research of anxiety disorders and proposes criteria for the selection of an appropriate R & D model.

In recent years, with the improvement in living standards, the morbidity and mortality of cardiovascular and cerebrovascular diseases has increased markedly. Atherosclerosis is the main pathological basis for cardiovascular and cerebrovascular diseases, and there are many risk factors for atherosclerosis. The pharmacological effects of puerarin are broad, and considerable clinical data confirms that puerarin has a definite effect on cardiovascular diseases resulting from atherosclerosis. The use of puerarin for atherosclerosis has increased in recent years. This article reviews the effect and mechanism of puerarin on atherosclerosis.

Aristolochic acids (AAs) are widely distributed in Aristolochiaceae, and are important toxic components in medicinal plants of Aristolochiaceae. As one of the most powerful carcinogens in the Carcinogenic Potency Database (CPDB), AAs can induce hepatotoxicity, nephrotoxicity, carcinogenicity, mutagenicity, and other adverse reaction. AAs also can produce a series of metabolites such as AA-DNA adducts in the body, and their specific metabolites can be used as biomarkers for early diagnosis and treatment of related diseases. Thus, the current discovery for technical means that can quickly and accurately detect biomarkers possesses significant research value. AAs can be attenuated by processing, compatibility, molecular breeding, and other methods to improve the clinical safety of Chinese medicine containing AAs. In this review, we report the distribution of AAs, attenuation strategies and biomarker detection. We would like to provide a reference for the quality control of AAs-containing Chinese medicines, as well as for the prevention and control of diseases caused by AAs.

Protein-protein interactions (PPI) are involved in a variety of biological processes, including cell-to-cell interactions, metabolism and development control. The misregulation, post-translational modification and interference of PPI are related to a variety of human diseases, making the regulation of these interactions a very attractive field of drug discovery. In recent years, the interaction between MDM2 and p53 has become a research hotspot, which plays an important role in the treatment of tumors. But unfortunately there are no such inhibitors approved all over the world. In this view, recent advances of MDM2-p53 inhibitors were briefly described and its inhibitors with potential therapeutic activities in clinical studies were introduced.

Natural products are valuable resources for discovering new drugs. So far, screening bioactive compounds from organism extracts is still an important and challenging task. Traditional biometric guided method involves repeated fractionation steps and bioactivity tests, which are time-consuming, labor-consuming, and inefficient. Ligand fishing is a bioanalysis method for screening ligands from complex organism extracts based on intermolecular affinity interactions. It has the characteristics of strong specificity, high efficiency, and less requirement for sample pretreatment. In this review, we summarize the classification of ligand fishing strategy and its application in enzyme inhibitors screening. Finally, the development prospects of this technology are forecasted.

Lefamulin (BC-3781) is a semi-synthetic pleuromutilin antibiotic, approved for the treatment of community-acquired bacterial pneumonia (CABP) by Food and Drug Administration (USA) in August 2019, with the commodity name of Xenleta. It is the first pleuromutilin antibiotics used for systemic treatment of bacterial infections in human. Lefamulin binds to the peptidyl transferase center of the 50S ribosomal subunit to prevent peptide transfer, thus inhibits protein synthesis. Lefamulin displays expanded activity against gram-positive organisms, and also shows high activity against atypical microorganism like Mycoplasma pneumoniae. This review discusses the mechanism, bacterial spectrum of activity, preclinical and clinical data of Lefamulin.

Tumor microenvironment (TME) is a complex comprehensive system composed of immune cells, inflammatory cells, tumor-associated fibroblasts, microvessels, and various cytokines and chemokines. As the living environment of tumor cells, it's closely related to the occurrence, metastasis, and recurrence of tumors. The characteristics of tumor microenvironment include: weakly acidic environment, low oxygen, high concentration of reactive oxygen species (ROS) and reducing substances, immunosuppression, etc. A weakly acidic environment favors metastasis of tumor cells, hypoxia is conducive to the emergence of drug resistance, high concentration of ROS and reducing substances are beneficial for tumor treatment, and immunosuppressiveness facilitates immune escape. With the rapid development of nanotechnology, more and more nanostructures have been reported to achieve tumor treatment by regulating the tumor microenvironment. This review summarizes recent advances in the nanostructures used to regulate tumor microenvironment through changing elements, including hydrogen-ion concentration (pH), the concentration of oxygen (O₂) and reactive oxygen species, and the activity of immune cells. Moreover, research directions in the future are pointed out in this review.

In the past few decades, our understanding of platelets has made great progress. Platelets play an unexpected central role in cancer and greatly affect the behavior of cancer cells. At the same time, the physiology and phenotype of platelets are also affected by cancer cells. Therefore, platelet-based tumor targeted therapy strategies have attracted the attention of researchers, but the limitations of their application require more attention. In this paper, the strategies of platelet-based tumor targeted therapy are summarized, and the strategies of platelet mimicking nanocarrier delivery, platelet hitch riding, platelet membrane coating biomimetic and engineered platelet targeting are mainly introduced. The easy activation, hard storage and unknown functional and phenotypic changes of platelets were discussed. At the same time, the strategy of platelet-based targeted tumor therapy is reviewed from theoretical basis and practical application. The development potential of platelets in the field of tumor diagnosis and treatment is discussed, which will provide some theoretical reference for the study of platelet-related tumor diagnosis and targeted therapy.

To study the regulating effect of total phenolic acids from the stems and leaves of Salvia miltiorrhiza Bge. on the intestinal flora and short-chain fatty acids in spontaneous type 2 diabetic nephropathy mice, db/db mice were taken as the research object, and were treated with the total phenolic acid of Salvia miltiorrhiza Bge. Animal welfare and experimental procedures followed the regulations of the Animal Ethics Committee of Nanjing University of Chinese Medicine Drug Safety Evaluation Research Center. Fresh feces and cecal contents of mice were collected for analysis of intestinal flora composition and differential flora. Gas chromatography was used to detect short-chain fatty acids in fresh feces and cecal content. Then the correlation analysis of the two results was made. Compared with the normal group, the most significant decreased differential flora in the model group were g_Rikenellaceae_RC9_gut_group and g_Bacteroidales_S24-7_group, while the most significant increased were g_unclassified_f__Coriobacteriaceae and g_unclassified_p__Firmicutes. Compared with the blank group, the contents of isovaleric acid and valeric acid in fresh feces and the contents of 6 short-chain fatty acids in the cecal contents of the model group were significantly reduced (P < 0.01). After drug intervention, the intestinal flora disorder and the reduction of short-chain fatty acids were improved to varying degrees, and the effect of the total phenolic acids from the stems and leaves of Salvia miltiorrhiza Bge. was slightly better than that from the roots in regulating some flora and short-chain fatty acids. The results of correlation analysis showed that g_Rikenellaceae_RC9_gut_group was moderately positively correlated with acetic acid and isobutyric acid in the cecal contents (r > 0.4). It is suggested that the total phenolic acid from the stems and leaves of Salvia miltiorrhiza Bge. can improve the intestinal flora disorder of mice with type 2 diabetic nephropathy, and can regulate the content of short-chain fatty acids in the intestine via adjusting the content of some short-chain fatty acid-producing bacteria, thereby helping to restore normal.

Insulin resistance refers to the impaired ability of insulin to regulate glucose metabolism in peripheral organs, which is considered to be the etiology of type 2 diabetes. This study aims to explore the mechanism of improving insulin resistance by compatibility of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos (DH). Insulin resistance was conducted on C56BL/6J mice by treatment of high fat diet. The energy intake and body weight, plasma levels of triglycerides, total cholesterol, insulin and glucose, oral glucose tolerance test (OGTT) and insulin tolerance test (ITT), as well as gene transcription and protein expression levels of insulin signaling pathway in liver, heart, kidney, and skeletal muscle of insulin resistance mice were evaluated. Animal experiments and welfare were performed in compliance with the guidelines of Animal Ethics Committee of Nanjing University of Chinese Medicine. The results showed that DH treatment significantly alleviated the excessive food intake and weight gain, and significantly decreased the levels of plasma triglycerides and total cholesterol, and constantly mitigated the hyperinsulinemia in insulin resistance mice. The results of OGTT and ITT suggested that DH treatment dramatically improved the response of insulin resistance mice to insulin stimulated glucose metabolism. Furthermore, the imbalance of metabolic arm and mitogenic arm of insulin signaling pathway in insulin resistance mice was normalized after DH treatment. DH treatment regulated insulin signaling pathway and improved the ability of glucose metabolism of insulin resistance mice.

This study investigated the effects of ginkgolide B on the long-chain fatty acid metabolism-related enzyme protein peroxisome proliferators-activated receptors α (PPARα), long-chain specific acyl-CoA dehydrogenase (LCAD), carnitine palmitoyl transterase-1 (CPT-1), and acyl coenzyme A oxidase 1 (ACOX1) expression in the liver of rats with non-alcoholic fatty liver disease (NAFLD). All the animal welfare and experimental procedures are in accordance with the regulations of the Animal Ethics Committee of Yunnan University of Traditional Chinese Medicine. After successfully building the rat model of non-alcoholic abnormal liver disease, the rats were divided into the model group, the simvastatin group, and the low-dose, middle-dose, and high-dose groups of ginkgolide B according to random number method, and were given corresponding drug treatment 4 weeks. We detected liver pathological indicators and determined blood lipids, transaminase and anti-oxidation indexes. Western blot and RT-PCR assays were used to detect the protein and mRNA levels of PPARα, LCAD, CPT-1, and ACOX1 in livers. The results showed that: ① the liver histopathology showed that the liver slices of the model group had obvious structural disorder, the nucleus was squeezed, and there were obvious fat vacuoles. The treatment groups improved significantly compared with the model group; ② compared with the normal group, the liver function and blood lipid indexes of the model group increased significantly, while the anti-oxidation indexes decreased significantly. Compared with the model group, each treatment groups were significantly improved; ③ compared with the normal group, the protein and mRNA expression levels of PPARα, ACOX1, CPT-1, and LCAD in the model group were significantly reduced, compared with the model group, those indexes in the treatment groups were significantly up-regulated. This study found that ginkgolide B could regulate the expression of long-chain fatty acid metabolism-related proteins PPARα, ACOX1, CPT-1, and LCAD, meanwhile improve the body's antioxidant capacity, thereby reduce blood lipids, further improve liver function and protect the liver.

Abnormal aggregation of amyloid-β protein (Aβ) in brain plays a vital role in the occurrence of Alzheimer's disease (AD). Hence, inhibiting Aβ aggregation is one major tactic for therapy of AD. Previous studies have found that tolcapone can inhibit Aβ42 aggregation and reduce the cytotoxicity induced by Aβ42 aggregates, but clinical studies have found that tolcapone has strong liver toxicity. To reduce the liver toxicity of tolcapone, its side chain structure was modified to obtain its derivative phenethyl (E)-2-cyano-3-(3, 4 dihydroxy-5-nitrobenzene)-acrylate (PCDNA). Thioflavin T (ThT) and atomic force microscopy (AFM) assays were used to explore the inhibitory effect of PCDNA on Aβ42 fibrillogenesis. The cytotoxicity assays were used to explore the inhibitory effect of PCDNA against the cytotoxicity induced by Aβ42 aggregates. In addition, the depolymerization effect of PCDNA on mature Aβ42 fibrils was also explored. Finally, molecular docking was used to explore the interaction between PCDNA and Aβ42 pentamer. These results lay the foundation for the study of the structural analogues of tolcapone as Aβ inhibitors.

Z-VAD-FMK was combined with hypoxia-reoxygenation (H/R) injury to establish a necroptosis model of H9c2 cells to mimic the pathological changes of myocardial ischemia reperfusion injury (MIRI) in vitro and to study the effect and mechanism of tilianin against myocardial ischemia-reperfusion injury. A cell counting kit-8 (CCK-8) was used to detect cell viability, and commercial kits were used to detect lactate dehydrogenase (LDH) and superoxide dismutase (SOD) in the cell culture supernatant. Hoechst 33342/PI immunofluorescence staining was used to detect cell death. DCFH-DA, BBcellProbe^TMM61, and JC-1 probes were used to detect reactive oxygen species (ROS), mitochondrial permeability transition pore (mPTP), and mitochondrial membrane potential (MMP), respectively. An enzyme-linked immunosorbent assay (ELISA) method was used to detect the release of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6). The results show that the cell viability, SOD activity, and MMP of the model group induced by H/R injury decreased, as compared with control group, but the necroptosis rate, LDH level, and ROS release increased significantly. Furthermore, mPTP of the model group cells opened, and TNF-α, IL-1β, and IL-6 levels were significantly higher. Molecular docking modeling showed that tilianin can bind to calmodulin-dependent protein kinase Ⅱ (CaMKII), and Western blot results showed that compared with control group, the expression levels of p-CaMKII and phospho-mixed lineage kinase domain-like protein increased in the model group, and tilianin could decrease the expression level of these proteins. The above results indicate that tilianin can protect H9c2 cells by inhibiting the phosphorylation of CaMKII at threonine 287, protecting mitochondrial function, and inhibiting the opening of mPTP to prevent necroptosis. This study has value for research on new methods to treat H/R injury.

In recent years, there has been an increase in the incidence of herbal-induced liver injury due to the accidental ingestion of herbal medicines containing pyrrolizidine alkaloids (PAs) in domestic. Salvianolic acid B (Sal B), a hydrophilic component in Salvia miltiorrhiza Bge., shows activities of anticoagulation, antioxidation, and other pharmacological activities. This research aims to investigate the protective effect of Sal B on hepatotoxicity induced by senecionine (SEN) and its potential mechanism. The animal experiment was approved by the Experimental Animal Ethical Committee of Shanghai University of Traditional Chinese Medicine, and all mice have received humane care in compliance with the institutional animal care guidelines. Mice were treated with Sal B (10 mg·kg^-1) 3 days before and 1 day after SEN (50 mg·kg^-1) treatment. The animals were sacrificed 48 h after SEN administration. As a result, Sal B effectively ameliorated SEN-induced liver injury. The mice in the group treated with Sal B showed lower serum activities of alanine aminotransferase and aspartate aminotransferase, less hepatic sinusoidal hemorrhage, and reduced hepatocyte necrosis. Besides, contents of pyrrole-protein adducts, the marker for PA-induced toxicity, were also decreased in serum. The key factors related to coagulation, oxidative stress, and liver fibrosis were further analyzed. It was found that Sal B inhibited the coagulant system by reducing the expression of plasminogen activator inhibitor-1. Sal B also modulated glutathione and superoxide dismutase levels and improved the anti-oxidative defense system. In addition, Sal B decreased the excessive deposition of extracellular matrix and inhibited the progression of liver fibrosis via down-regulating several key factors related to liver fibrosis, including matrix metalloproteinase 9, transforming growth factor-β1, signal transducer and activator of transcription 3, and chemokine 1. In conclusion, Sal B ameliorated SEN-induced liver injury in mice by regulating the blood coagulation system, improving oxidative stress, and modulating liver fibrosis-related factors. Our present study pointed to the possibility of utilizing salvianolic acid B for protection against PA-induced liver injury clinically.

Bioorthogonal fluorogenic probes are becoming an ideal tool for live-cell fluorescence imaging. With the tetrazine bioorthogonal fluorogenic probe that displays fluorescence enhancement, the tetrazine plays the dual-role of a bioorthogonal reaction unit and the fluorescence quenching unit. The "off" and "on" states of the fluorescence probe are mainly controlled through inverse electron demand Diels-Alder (IEDDA) bioorthogonal reaction. We designed a series of turn-on tetrazine fluorescent probes with Donor-π-Acceptor (D-π-A) structure to achieve a high signal-to-noise ratio and specificity of fluorescence imaging. This series of probes reacted with the dienophile bicyclononyne, and then generated pyridazine structure in-situ that acted as an electron acceptor, resulting in a new D-π-A effect of fluorescent dyes, turning on the intramolecular charge transfer (ICT) effect. By adjusting the electron-donating groups and the degree of conjugation, tunable fluorescence spectra between 400-647 nm with fluorescence turn-on enhanced up to 500-fold have been achieved. This research lays the foundation for the further optimization of tetrazine bioorthogonal fluorescent probes and their applications in molecular imaging and biomedical fields.

Bidehydrocorydaline A (1), a new dimeric alkaloid of proberberine connected by a methylenedioxy group, was isolated from the vinegar-prepared Corydalis yanhusuo by various chromatographic methods, including column chromatography over macroporous adsorption resin and silica gel, reverse-phase MPLC, and semi-preparative HPLC. Its structure was determined by spectroscopic methods, including UV, IR, HR-ESI-MS, 1D and 2D NMR. Bidehydrocorydaline A (1) showed potent inhibitory activity against LPS-induced NO production in RAW 264.7 macrophages, with an IC₅₀ value of 2.33 ±0.57 μmol·L^-1.

The structure and diversity of the intestinal flora in rats exposed to high altitude hypoxia was investigated. Animal experiments strictly follow the regulations of Medical Laboratory Animal Ethics Committee of Qinghai University, School of Medicine. SD rats were randomly divided into a control group, a moderate altitude hypoxia group, and a high altitude hypoxia group. The pH value of the feces was measured and histopathological changes in the small intestine were determined by HE staining, and the intestinal flora were characterized by 16S rDNA high throughput sequencing technology on the 3rd, 7th, 15th, and 30th day of hypoxia exposure. Compared with the control group, the fecal pH value of rats in the moderate altitude hypoxia group and the high altitude hypoxia group was decreased significantly. The lamina propria and submucosa capillaries were slightly dilated and congested on the 3rd day in the moderate altitude hypoxia group. In the high altitude hypoxia group the submembrane capillaries were dilated and congested, the lamina propria of the mucosa showed mild edema, and the lymphatic vessels were dilated on the 7th day. The composition and diversity of intestinal flora in these rats changed significantly with prolonged exposure to the high altitude hypoxic environment. A total of 35 phyla, 87 classes, 205 orders, 337 families, 638 genera, and 256 species were annotated in the three groups of rats, including Firmicutes, Clostridia, Clostridiales, Ruminococcaceae, Akkermansia, and Lactobacillus_murinus. Compared with the control group, the intestinal flora of the hypoxic groups showed the most significant changes by the 15th day. There were 9 microbiota of gut microorganisms with relative abundance in the moderate altitude hypoxia group, of which Rikenellaceae_RC9_gut_group bacteria was the most common, there were 19 different microbiota of gut microorganisms with higher relative abundance in the high altitude hypoxia group, of which Ruminococcaceae bacteria was the most common. The results of this study indicate significant changes in the intestinal flora with high altitude hypoxia, and establish a foundation for further research on the initiation and development of diseases and drug metabolism in high altitude hypoxia.

Studies have found that metformin is not only the preferred drug for lowering blood sugar, but also shows lipid-lowering and weight-loss effects. The purpose of this study was to use a hyperlipidemia hamster model to investigate the lipid-lowering effect of metformin and its effect on important metabolic pathways in lipid metabolism disorders. Fifty golden hamsters were divided into a control group, a model group, metformin high- and low-dose groups, and a simvastatin group. A high-fat diet was fed for 1 week to create the model, and then drug was administered for 11 weeks with the high-fat diet. Serum was taken for measurement of blood lipid and blood glucose at 2, 6, and 9 weeks after administration, and at weeks 3, 5, and 9 feces and urine were collected for ¹H NMR metabolomics tests. After 11 weeks of intravenous injection of[U-¹³C₆] glucose, serum was collected for a ¹³C NMR metabolic flux test. The results showed that the administration of metformin can significantly reduce blood lipids and glucose levels and can significantly affect metabolic pathways such as sugar metabolism, lipid metabolism, ketone metabolism, amino acid metabolism, and intestinal flora metabolism. The results of the metabolic flux analysis showed that the high-fat diet reduced the metabolism of tricarboxylic acids by 37.48%. After administration of low and high doses of metformin the metabolism of tricarboxylic acid increased by 98.14% and 143.10%, respectively. After administration of simvastatin tricarboxylic acid metabolism increased by 33.18%. The results indicate that metformin has a significant effect on promoting energy metabolism. This study used a combination of metabolomics and metabolic flow to explore the effect of metformin on lipid metabolism disorders and quantifies changes in the key pathway of energy metabolism-the tricarboxylic acid cycle. This study provides useful information for the study of the efficacy and mechanism of metformin, as well as a practical technical method for the screening of lipid-lowering drugs based on a hamster model.

Qualitative and quantitative methods were used to establish the quality of different medicinal parts of Poria cocos (Poriae Cutis, rubra Poria, white Poria, Poria cum Radix Pini) by using ultra-performance convergence chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry (UPC²-PDA-Q-TOF/MS^E). A total of 18 chromatographic peaks were detected from Poria cocos by UPC²-PDA. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used to compare the four medicinal parts. The results showed that there were significant differences in the components of different medicinal parts, and the main triterpenoic acids were poricoic acid A, poricoic acid B, dehydroeburicoic acid, and dehydrotrametenolic acid. When combined with the common active component polyporenic acid C, a method for determination of five triterpenoic acids in different parts of Poria cocos was established. These components could be separated within 15 min, and the amount of methanol was 3.63% of that of HPLC method. Taking the five triterpenoid acids as an index, the content of triterpenoid acids in different parts of Poria cocos from high to low were Poriae Cutis, rubra Poria, white Poria and Poria cum Radix Pini. The method is simple, rapid, and uses minimal solvent. The mobile phase of environment-friendly gas carbon dioxide has unique advantages in reducing environmental pollution, which can provide a basis for the development and standard formulation of Poria cocos and its related products.

Metabolomics based on liquid chromatography coupled with mass spectrometry (LC-MS) was used to study the initiation and development of diabetes in rats, and the ability of Ginkgo biloba extract (GBE) to ameliorate this pathology. Diabetes mellitus (DM) was induced by intra-peritoneal injection of streptozotocin. The rats were randomly divided into a normal control group treated with drug-free solution (NC), a normal control group treated with GBE (N-GBE), a DM group treated with drug-free solution (DM), and a DM group treated with GBE (D-GBE); rats were maintained on this protocol for 9 weeks. Rat plasma was collected from the sixth week to the ninth week and then analyzed with LC-MS. Animal experimentation was approved by the Committee on the Ethics of Animal Experiments of Xuzhou Medical University. Twelve plasma metabolites with continuous differentiation were monitored to indicate dysfunction of metabolic pathways including fatty acid metabolism, phospholipid metabolism, amino acid metabolism, tricarboxylic acid cycle activity, bile acid metabolism, and purine metabolism to confirm the occurrence and development of DM. Treatment with GBE partially reversed the changes seen in five metabolites in DM rats, indicating that GBE could prevent the occurrence and development of DM by acting on fatty acid metabolism, phospholipid metabolism, amino acid metabolism, and the tricarboxylic acid cycle.

Using a H₂O₂-induced BRL cell senescence model, we investigated the anti-aging effects of drug-containing serums of Erzhi Wan (EZW) and various polar extracts (petroleum ether, ethyl acetate, n-butanol, water, and iridoid glycoside-enriched fractions). Cell viability was detected by MTT assay. Cell senescence was evaluated with β-galactosidase staining assay. Intracellular reactive oxygen species (ROS) were measured by flow cytometry. UFLC-Q-TOF-MS/MS was used to identify chemical components in EZW and the extracts, and molecular docking technology was used to predict the anti-aging components of EZW. Results showed that treatment of cells with 600 μmol·L^-1 H₂O₂ for 72 h markedly induced cell senescence, inhibited cell proliferation and increased intracellular β-galactosidase activity and ROS levels. If cells were pretreated with drug-containing serum of EZW this induction of senescence was decreased. A total of 49 chemical compounds were identified in EZW by liquid chromatography-mass spectrometry, 14 of these were identified by molecular docking as potential active ingredients. Based on these analyses, and the in vitro experiments with polar extracts, we conclude that the anti-aging components of EZW are triterpenes, flavonoids and phenyl alcohols, providing a basis for further elucidation of the active agents and mechanism of the anti-aging effect of EZW.

Acetaminophen (APAP, also known as paracetamol)-induced liver injury is the leading cause of drug-induced liver injury in the world. Wuzhi Tablet (WZ, an ethanol extract of Schisandra sphenanthera) is widely used in clinical practice to protect liver function. Our previous studies have shown that pretreatment with WZ for 3 days can significantly protect against APAP-induced liver injury; however, the effect of different intervals between APAP and WZ treatment on APAP-induced liver injury remains unclear. In this study, the change in liver injury indexes, APAP metabolites, and the activity of cytochrome P450 (CYP450) enzymes after treatment with WZ and APAP at different intervals were determined. The animal experiment was reviewed and approved by the Animal Ethics Committee of Sun Yat-sen University. The results show that 0 h, 0.5 h, and 2 h pretreatment with WZ significantly protected against APAP-induced liver injury in mice, as evidenced by a significant decrease in biochemical parameters such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and malonaldehyde (MDA). WZ inhibited the metabolic activation of APAP mediated by CYP450 enzymes and reduced the formation of APAP metabolites. This study further demonstrates that pretreatment with WZ at different intervals (0 h, 0.5 h, and 2 h before APAP dosing) exerts a significant hepatoprotective effect against APAP-induced liver injury, and a single-dose of WZ inhibits the activity of CYP450 enzymes related to APAP metabolic activation, thereby protecting against APAP-induced hepatotoxicity.

This study aims to establish the design space of the key processes for drop-on-powder 3D printing based on design of experiment (DoE). By utilizing Minitab, an experimental scheme with three factors, two levels and three center points was designed to analyze the factors that significantly affected the tablet quality attributes. Furthermore, the factor interactions were analyzed using Minitab. subsequently, the computer aided drafting (CAD) software was used to adjust the model volume with fixed radius/height ratio (r/h=1.25) and establish a linear regression equation between model volume and dose. As a result, the drug dose could be controlled in a flexible manner. The finally determined process parameters were: ink-jet level is 12, layer thickness is 150 μm, and the X-axis printing head speed of 635 mm·s^-1. Regression equation between drug content (y) and model volume (x) was y=0.062 x-0.582 7 (R²=0.999 9) showing good linear relationship. This indicated that robust and feasible process parameters were obtained through DoE, and the preparation of personalized-dose tablets was realized with good reproducibility.

To investigate the influence factors and effects of metal or dissolving microneedles on the formation and healing of skin microchannels, the pencil-shaped or conical stainless steel microneedles with different lengths, tip to tip space and base area, and the pencil-shaped dissolving microneedles with different tip to tip space were used. The microneedles were applied to the skin of epilatory mice and rats, and the effects of various microneedle parameters, application parameters, and animals on the microchannels were explored by the transepidermal water loss (TEWL) and methylene blue staining. Visually observe the skin local irritation caused by the microneedles. The animal experiments were approved by the Animal Experiments Welfare and Ethical Committee of Zhejiang University of Technology. The application time of the microneedle should be maintained at 30 s or more. When the insertion forces were 2, 4, 8 N, and the TEWL initial values of the formed microchannels were 12.9, 33.0, 40.4 g·m^-2·h^-1, respectively. When the length of metal microneedle were 400, 600, 800 μm, and the TEWL initial values of the formed microchannels were 37.1, 40.4, 49.5 g·m^-2·h^-1, respectively. When the tip to tip space of metal microneedle were 400, 600, 800 μm, and the TEWL initial values of the formed microchannels were 33.2, 40.4, 55.8 g·m^-2·h^-1, respectively. When the base area of metal microneedle were 0.16, 0.35, 0.62 cm², and the TEWL initial values of the formed microchannels were 35.1, 40.4, 67.1 g·m^-2·h^-1, respectively. The effects of conical and pencil-shaped microneedles are similar. When the tip to tip space of pencil shaped dissolving microneedle were 400, 600, 800 μm, and the TEWL initial values of the formed microchannels were 49.8, 60.5, 70.5 g·m^-2·h^-1, respectively. The TEWL baseline values of animal skins of different genders and series are different, but the tendency of microchannels formation and closure is similar. Visual inspection revealed that the slight erythema caused by the microneedles subsided within 24 h. Microneedles of different parameters have different effects on microchannels. The microchannels closed within 24 or 48 h, and the skin local irritation caused by microneedle was mild.

MYB transcription factors play many important regulatory roles in plant growth and development, secondary metabolism, and stress adaptation processes. In this work, an MYB gene containing a complete open reading frame (ORF) was selected from the transcriptome database of R. palmatum L. RpMYB4 ORF and cloned, encoding a polypeptide of 245 amino acids with a molecular weight of 26.99 kDa. RpMYB4 lacks a signal peptide or transmembrane domain but contains two conserved DNA binding domains (HTH-MYB) of the R2R3-MYB subfamily at the N-terminus. Multiple-sequence alignment demonstrated that RpMYB4 shared as high as 61% identity with many MYB proteins from other species. Phylogenetic analysis showed that RpMYB4 had the closest relationship with FtMYB8 and was clustered in the S4 subfamily. Subcellular localization by confocal microscopy showed that an RpMYB4-GFP-fusion protein localized to the nucleus in tobacco. Real-time fluorescence quantitative PCR analyses revealed that RpMYB4 was differentially expressed in various tissues, with the highest expression in leaves, followed by petioles, rhizome, and roots, and with the lowest level in mature seeds. After treatment of R. palmatum L. seedlings with 200 μmol·L^-1 MeJA, the expression of RpMYB4 in leaves was down-regulated within 24 h, and significantly up-regulated after 200 μmol·L^-1 SA treatment at 12 h and 24 h. However, gene expression did not change with 200 μmol·L^-1 ABA treatment. The transcripts of RpMYB4 under drought, high temperature, and mechanical injury stresses reached a peak at 24 h, 24 h, and at 3 h, respectively, while RpMYB4 expression was inhibited by low temperature stress, reaching its lowest value at 6 h. The gene showed no significant response to salt stress. Overall, RpMYB4 was cloned from R. palmatum L. for the first time, showed high expression in leaves, and was responsive to SA and various abiotic stress treatments including drought, high temperature, and mechanical injury. The results will be useful for further analysis of secondary metabolism and stress adaptations in R. palmatum L.

To explore the characteristics of soil microbial communities of Cistanche deserticola and Cynomorium songaricum, two typical parasitic medicinal plants that live in an extreme saline alkali environment, 16S PCR was used to sequence the soil microbial communities of C. deserticola and C. songaricum in Ebinur Lake, Xinjiang. Redundancy analysis and correlation analysis were carried out based on the abundance of core microbiome and ecoclimatic factors. The results show that the diversity of the soil microbial community of C. deserticola was significantly higher than that of C. songaricum. The core microbial groups of C. deserticola and C. songaricum were Marinomona, Halomonadaceae, Rhizobiales, Halomonas, and Acidimicrobiales. Six specific biomarkers were identified as Micrococcacea, Echinicola, Glutamicibacter, Galbibacter, Pseudoalteromonas, and Marinobacterium_rhizophilum. The results of redundancy analysis and correlation analysis show that the average temperature in the driest season and the average temperature in the coldest season, and the clay content and soil texture classification were the main ecological factors affecting the composition of these soil microbial communities. This study provides a theoretical basis for finding molecular markers of C. deserticola and C. songaricum and promoting the quality of C. deserticola and C. songaricum.

In this study we isolated and cloned four flowering-related genes from Bupleurum chinense DC., named BcSVP, BcPAF1, BcCO, and BcFT. Actin and EF-1α were used as double internal standards to analyze the spatial and temporal differences in the expression of four genes in different tissues and flowering stages of Bupleurum chinense DC. qRT-PCR analysis showed that the BcSVP gene was mainly expressed in root, and its relatively expression level was low. BcPAF1 and BcCO were both highly expressed in different parts, and their relative expression level showed an increasing trend and then slowly decreasing with the flowering development process. BcFT was mainly expressed in the stem, and the relatively expression level was increased sharply in the flowering stage. In summary, four genes related to flowering of Bupleurum chinense DC. were cloned and their relative expression levels were characterized, laying a foundation for elucidating the molecular mechanisms that regulate the flowering stage of Bupleurum chinense.