We have established a quantitative analysis of multi-components by single marker method (QAMS) for the simultaneous determination of apigenin-7-glucuronide, quercitrin, yuankanin, luteolin, apigenin, hydroxygenkwanin, and genkwanin in Qutan Zhike Granules. The chromatographic column used was an Agilent EC-C18(150 mm × 4.6 mm, 4 μm), the mobile phase was methanol-0.15% phosphoric acid solution (gradient elution), and the detection wavelength was 338 nm. Apigenin was chosen as the internal reference standard, the relative correction factors for the six components were determined by multi-point correction method and included apigenin-7-glucuronide, quercitrin, yuankanin, luteolin, hydroxygenkwanin and genkwanin. According to the two-point correction method combined with the relative retention time correction of the components to be tested, the peak location was determined. The contents of these seven compounds in 10 batches of Qutan Zhike Granules samples were determined with relative correction factors, and the relative error (RE) was used to compare the results to that of the external standard using the External Standard Method to verify the accuracy of QAMS. The relative correction factors for apigenin-7-glucuronide, quercitrin, yuankanin, luteolin, hydroxygenkwanin and genkwanin were 1.762 8, 2.310 4, 1.898 4, 1.282 8, 1.191 3 and 1.066 9, respectively. RSDs of the relative correction factors were all lower than 3%. The peaks for each constituent were accurately located by the two-point correction method combined with relative retention time correction, and the predicted retention time was close to the actual retention time. The relative error of the contents by QAMS and determined by ESM in 10 batches of Qutan Zhike Granules were between -5% and 5%. This content determination method can be used for the simultaneous determination of seven components in Qutan Zhike Granules.

Acute lung injure (ALI) is a severe diffused lung disease, which is caused by pathogen-induced infections, inhalation of irritates, and so on. It could lead to acute respiratory distress syndrome (ARDS). Long oil (LO) is a lipidic mixture extracted from multiple medicinal animals and plants. It has been used for clinical wound repair. Here, an O/W LO emulsions (LOE) was prepared, which was composed of LO, Tween-80, propylene glycol, xanthan gum, and water. The droplet size of LOE was 671.63 ± 7.21 nm, and the zeta potential was-17.8± 1.26 mV. The size of LOE was small and homogenous, and the stability was satisfied. The aerosols had an aerodynamic diameter of 2.25 ± 0.05 μm after atomization of LOE with a vibrating screen atomizer, where the percentage of particle sizes within 1-5 μm was 81.40%, indicating effective deep lung deposition and suitable pulmonary inhalation. The safe dose of LOE was high to 12.50 μg·mL^-1 on human embryonic lung fibroblast MRC-5 cells. In the range of 0.02-2.50 μg·mL^-1 of LOE, the proliferation and migration of mouse fibroblast L929 cells were improved. Animal experiments were approved by the Ethics Committee of Institute of Radiation Medicine, Academy of Military Medical Sciences, and the experiments were conducted by relevant guidelines and regulations. No significant toxicity was observed after intratracheal (i.t.) administration of LO (3.25 mg·kg^-1) to mice. Mouse i.t. administration of LOE remarkably attenuated lung injury induced by lipopolysaccharide with mitigations of inflammatory factors (tumor necrosis factor-α, interleukin-6) and total proteins. LOE is a promising inhaled formulation for the treatment of ALI.

A fungal strain WXF1904, was isolated from a starfish sample corrected in the South China Sea. According to its internal transcribed spacer (ITS) analysis, the strain was identified as a member of the genus Aspergillus and designated as Aspergillus sp. WXF1904. One new isocoumarin containing halogennamed 6-chloro-5, 7-dihydroxy-3, 8-dimethylisocoumarin (1), along with six known compounds pilobolusate (2), p-hydroxybenzaldehyde (3), methyl orsellinate (4), catechol (5), vanillic acid (6), and wasabidienone E (7), were isolated from the cultures of Aspergillus sp. WXF1904 by silica gel column chromatography, ODS gel column chromatography, and high performance liquid chromatography (HPLC). Their structures were elucidated by high resolution mass spectrometry (HR-MS), nuclear magnetic resonance (NMR) as well as literature comparison. The new compound 5-chloro-6, 7-dihydroxy-3, 8-dimethylisocoumarin and compound 2 showed weak acetylcholinesterase inhibitory activity.

Identification of metabolites of Danshen-Honghua herb pairs in isolated rat intestinal flora based on HPLC-Q-TOF-MS/MS technique. By incubating enterobacteria in isolated rats as well as inactivated enterobacteria in the incubation solution. The extracts of Danshen-Honghua herb pairs were added separately and co-incubated under anaerobic conditions. Animal experiments and protocols were approved by the Laboratory Animal Ethics Committee of Shaanxi University of Traditional Chinese Medicine (approval number: TCM-2020-030-E05). A total of 14 compounds, including 5 prototypes and 9 metabolites, were identified in the isolated rat intestinal incubation fluid. In contrast, no metabolites were detected in the inactivated enterobacterial fluid, except for the prototype component. The results showed that the main components of the Danshensu, salvianolic acid B, rosmarinic acid, lithospermic acid, and hydroxysafflor yellow A, could be metabolized by the intestinal flora, and these active ingredients were mainly metabolized in the rat intestinal flora in isolation by hydroxylation, decarboxylation, deoxygenation, decarboxylation and dehydration in phase Ⅰ, sulfate esterification and methylation in phase Ⅱ. This proved that the Danshen-Honghua herb pair could be transformed into various metabolites by the action of rat intestinal flora, further clarifying the role of intestinal flora in the metabolic transformation of the active ingredients of Chinese medicine and laying the foundation for perfecting the potent substances of the pair.

A new rapid, quality control method based on quantitative water tests has been established for the quality evaluation of Indigo Naturalis. The Turbiscan stability index (TSI) of 26 batches of Indigo Naturalis was measured by a stability analyzer. The parameters, including the method by which the ingredients are added, their particle size, amount, and the testing temperature, were systematically optimized and the methodological indexes such as repeatability and stability were determined. The content of indigo and indirubin in 26 batches of Indigo Naturalis was determined by high performance liquid chromatography and the total ash was measured. The correlation analysis between the active ingredients, total ash content and TSI value of Indigo Naturalis was determined by SPSS 26.0 and Origin 2021. This research shows that the best way to prepare samples for testing is to add 0.2 g of Indigo Naturalis powder which has passed through a No. 7 sieve but failed to pass through a No. 9 sieve to a glass bottle containing 20 mL pure water by a funnel and scan at 25 ℃ with a stability analyzer. Consistency analysis showed that the content ranking of indigo and indirubin is opposite to the TSI value, and the content ranking of total ash is generally consistent with the TSI value. Correlation analysis showed that the correlation coefficients of indigo and indirubin content and TSI value were -0.850 and -0.801, respectively, and R² was 0.72 and 0.64. The correlation coefficient between total ash content and TSI value was 0.724, R² was 0.52. Using the change in TSI value of Indigo Naturalis powder in water, this study establishes the range of classification of Indigo Naturalis decoction pieces and the content of relevant components, which can be used to authenticate Indigo Naturalis and evaluate its quality.

This study was designed to investigate the role of activation of the ferroptosis pathway in the inhibition of esophageal cancer proliferation and metastasis by realgar, using esophageal cancer Eca109 and KYSE150 cells as the target cells. The rate of inhibition and half-inhibitory concentration (IC₅₀) were measured by the CCK-8 method; clone formation ability was measured by clone formation assay; the changes in reactive oxygen species (ROS) were detected by flow cytometry; the ultrastructure of the cells was observed by transmission electron microscopy; the distribution of intracellular iron particles was observed by Prussian blue staining; the expression of glutathione peroxidase 4 (GPX4) was detected by immunofluorescence staining; the scratch assay was used to detect the cell migration ability; the Transwell assay was used to detect the cell invasion ability; and western blotting was used to detect E-cadherin, Slug and N-cadherin protein expression in the cells. The results show that realgar inhibited the proliferation of Eca109 and KYSE150 cells in a time- and concentration-dependent manner, and the IC₅₀ of Eca109 and KYSE150 cells was 64.297 and 51.337 μmol‧L^-1, respectively. Compared with the control group, many mitochondria in the cytoplasm of Eca109 and KYSE150 cells in the realgar 2IC₅₀ group were swollen and blue-stained particles of different sizes and amounts were found, and ROS fluorescence intensity was significantly increased while GPX4 protein expression was significantly reduced (P < 0.01). Compared with the realgar group, the proliferation, migration, membrane penetration and Slug and N-cadherin protein expression were significantly increased, and the cell inhibition rate and E-cadherin protein expression were significantly decreased in Eca109 and KYSE150 cells in the realgar+ZVAD-FMK group (P < 0.05). The proliferation, migration, membrane penetration and Slug and N-cadherin protein expression were significantly decreased, and the cell inhibition rate and E-cadherin protein expression were significantly increased of Eca109 and KYSE150 cells in the realgar +erastin group (P < 0.05). The above results show that realgar inhibited the proliferation of Eca109 and KYSE150 cells and induced partial ferroptosis in the cells, and the proliferation and metastasis effects of realgar on esophageal cancer cells may work through partial ferroptosis pathway activation.

Synthetic organoselenium compounds can be used as antioxidants, enzyme inhibitors, neuroprotective agents, antitumor agents, anti-infectious agents, and immunomodulators. Diphenyl diselenide (DPDS) is a synthetic organoselenium compound with simple and stable structure, and possesses lower toxicity than inorganic selenium and most organoselenium compounds. Moreover, DPDS has strong antioxidant properties due to its strong glutathione peroxidase-mimetic activity, involvement in thioredoxin reductase catalytic reactions, and activation of the nuclear factor E2 associated factor 2 (Nrf2)/Kelch like epichlorohydrin associated protein-1 (Keap1) signaling pathway. DPDS has been shown to possess a variety of pharmacological activities including anti-inflammatory, anti-diabetic, antifungal, neuroprotective, antitumor, hepatoprotective, renoprotective, and cardiovascular protective effects in a variety of animal and cellular models. This paper reviews the research progress on the physicochemical properties, main pharmacological effects and mechanisms of DPDS by compiling relevant research results, so as to provide reference for future research and reasonable development and application of DPDS.

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1, 5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

The liver is an important organ of the body, which has many functions, such as metabolism and detoxification. Due to the rapid change of lifestyle and the improvement of public health, the incidence rate of non-communicable diseases has increased significantly, which fundamentally changed the disease characteristics in most parts of the world. At present, the global prevalence of non-alcoholic fatty liver disease (NAFLD) is about 25%. Moreover, about 59.10% of NAFLD patients progress to non-alcoholic steatohepatitis (NASH) within 5 years, and about 41% of NASH patients progress to fibrosis. NAFLD has become one of the most important liver diseases in the world and may become the main cause of end-stage liver disease in the next few decades. In addition, NAFLD and related cirrhosis will bring huge economic burden to patients, health care system and society. Since there are currently no medications available that have been approved by Food and Drug Administration (FDA), NAFLD is still treated mainly through lifestyle changes such as exercise and diet. Oxidative stress and inflammation are the most important pathological processes in the occurrence and development of liver diseases. Nuclear factor erythroid-2-related factor 2 (Nrf2) is a key regulator of the body's antioxidant stress system, with anti-inflammatory, antioxidant and other functions. Many studies have shown that Nrf2 pathway significantly affects the progression of liver diseases. In this review, we aimed to summarize the regulatory role of the Kelch-like ECH-associating protein 1 (Keap1)-Nrf2-antioxidant response element (ARE) signaling pathway in the pathogenesis of NAFLD, and to reveal the potential of Nrf2 as a therapeutic target for NAFLD.

We used metabolomics to investigate the ability of a traditional Mongolian medicine called modified Tabusen-2 (MT-2) to improve kidney yang deficiency (KYD) in rats. All animal experiments were conducted under the guidance and standards of the Medical Ethics Committee of Inner Mongolia Medical University. SD rats were divided into 6 groups of six rats: a normal group, a model group, Jinkuishenqi pill administration group (1.26 g·kg^-1), and MT-2 administration in high-, medium- and low-dose groups (1.512, 0.756, and 0.378 g·kg^-1). KYD was established by intramuscular injection of hydrocortisone (HC) and biochemical indicators and clinical characterization was used to confirm that KYD was established. All groups received intragastrically administered drug (Jinkuishenqi pill or MT-2) or saline. Serum from each group was collected after 8 weeks and analyzed by UPLC-Q-exactive-MS to measure various biochemical indicators. The biomarkers affected by MT-2 were identified and the metabolic pathways of KYD regulated by MT-2 were analyzed by metabolomic analysis. The results show that MT-2 can decrease serum creatinine (Cr) in KYD rats and significantly increase (P < 0.05) the content of thyroid stimulating hormone (TSH) and luteinizing hormone (LH). In serum samples, 38 biomarkers such as corticosterone, L-phenylalanine, and DL-tryptophan were measured as possible indicators for disease development in KYD rats. MT-2 lowered 18 biomarkers of KYD, including corticosterone, deoxycorticosterone, and L-phenylalanine, and altered 13 related metabolic pathways including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and steroid hormone biosynthesis, resulting in an overall improvement in KYD. MT-2 appears to be important in improving KYD in rats mainly by regulating metabolites such as amino acids, steroids and lipids.