Against the backdrop of rising multidrug-resistant Acinetobacter baumannii (MDR AB) threats, this study explores the in vitro antibacterial activity and mechanism of Senecio scandens (a Miao ethnic medicinal herb) crude extract. Using 10 clinical MDR AB strains, we reassessed antibiotic sensitivity and then applied microbroth dilution to determine MIC/MBC, time-kill curves for bactericidal kinetics, and SEM/TEM for structural changes. Proteomics identified downregulated proteins, cross-referenced with VFDB/CARD to target membrane-related proteins (msbA, lptD), while molecular docking validated the strong binding of linarin/hyperoside to these targets. qPCR confirmed lptD/msbA mRNA downregulation (p < 0.05) by linarin/hyperoside (MIC = 312.5 μmol/L). The extract showed concentration-dependent bactericidal effects (MIC = 640 μg/mL), disrupting cell wall/membrane integrity. This study first reveals that linarin and hyperoside inhibit MDR AB by downregulating lptD/msbA, compromising outer membrane integrity, offering novel therapeutic candidates.

Infections caused by carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-HVKP) pose a critical challenge in clinical management due to limited treatment options and high mortality. Elucidating their virulence determinants and resistance mechanisms is essential for optimizing antimicrobial strategies and improving patient outcomes. We conducted a retrospective analysis of 102 carbapenem-resistant CRKP strains. Antimicrobial susceptibility testing was performed using the broth microdilution method, and resistance genotypes were characterized via PCR and Sanger sequencing. Virulence factors were identified through genomic sequencing and validated in a murine survival model (n = 6 mice per strain). Conjugation assays were performed to assess the transferability of resistance and virulence genes, with recipient strain E. coli J53, and transfer events were confirmed by PCR. CRKP strains were predominantly derived from sputum-, urine- and blood-specimens. Patients with CRKP infections predominantly had pulmonary infections. CRKP exhibited high resistance to other β-lactam antibiotics, mainly due to the presence of blaKPC-2, high resistance to quinolones mediated by the carriage of aac (6”-Ib-cr), QnrS and QnrB, and high resistance to aminoglycosides mediated by the carriage of rmtB, ant(3”)-I and armA. CR-HVKP is mainly composed of ST11. Aerobactin (iucA, iutA), Ent siderophore (fepA, entB), Salmochelin (iroN), Yersiniabactin (ybtS), Type 3 fimbriae (mrkD), Type I fimbriae (fimH), and Regulation (prmpA) were detected in the CR-HVKP isolates. The CR-HVKP strains had a median lethal dose (LD50) of 2 × 103 to 5 × 103 CFU in the mice, which was similar to that of the positive control NTUH-K2044. Conjugation assays revealed that the genes iucA, iutA, iroN, prmpA, blaKPC-2, blaNDM-1, blaVIM, blaSHV-12, blaTEM-1b, QnrA, QnrB, QnrS, and rmtB can be transferred to E. coli J53. Compared with those of the recipient E. coli J53, the MICs of meropenem, imipenem, levofloxacin and amikacin in the transconjugants increased by 4–128 times. These findings reveal that carbapenems in combination with aminoglycosides and quinolones may not be an effective option for the treatment of CRKP infections. Furthermore, the virulence and resistance genes may spread rapidly, which posing a public health risk and a significant threat to clinical care. Therefore, it is necessary to further strengthen hospital infection monitoring, prevention and control measures and to provide strict management and training on the rational use of antimicrobial agents in intensive care units.

Numerous studies have confirmed that Tongmai Yangxin pill (TMYXP) is effective in the treatment of coronary heart disease (CHD) with a deficiency of both Qi and Yin in traditional Chinese medicine theory, but whether it works through the regulation of gut microflora (GM) for the treatment of the disease is unclear. The network pharmacology and molecular docking were used to explore the potential mechanism of TMYXP in treating CHD by regulating GM. Based on network pharmacology, the potential target of TMYXP in the treatment of CHD by GM was identified. The network diagram of protein-protein interaction was established to screen the key targets. The biological function and enrichment pathways of TMYXP were analyzed using the DAVID database, and the biological pathway of TMYXP in treating CHD by regulating GM was explored. The key active ingredients and proteins were analyzed through molecular docking simulation. TMYXP may regulate 27 targets and related 34 types of intestinal bacteria to exert cardiovascular protection, with the active substance basis including 145 active ingredients represented by Quercetin and Luteolin. GM plays an important role in the treatment of CHD with TMYXP, whose therapeutic mechanism relates to multi-target and multi-pathway.

Multidrug-resistant (MDR) Pseudomonas aeruginosa (PSA), recently reclassified by the World Health Organization (WHO) as a high-priority antimicrobial-resistant pathogen, continues to impose a substantial global health burden due to escalating resistance and stagnant therapeutic innovation. Colistin retains critical clinical utility against MDR P. aeruginosa infections; however, its dose-limiting nephrotoxicity and neurotoxicity necessitate strategies to optimise therapeutic indices. This study investigated the molecular mechanism underlying the synergistic activity of aspirin in potentiating colistin efficacy against MDR P. aeruginosa. In vitro analyses revealed marked synergistic bactericidal activity (FIC index ≤0.5), with metabolomic profiling demonstrating suppression of key metabolic pathways integral to bacterial membrane biogenesis, including glycerophospholipid metabolism and fatty acid biosynthesis. Ultrastructural imaging confirmed irreversible disruption of membrane integrity via combined treatment. In a rat model of P. aeruginosa-induced pneumonia, colistin-aspirin co-administration demonstrated superior efficacy to monotherapy, significantly reducing pulmonary bacterial load (3 to 4-log CFU/g reduction vs colistin alone; p < 0.01), attenuating histopathological injury, and suppressing pro-inflammatory cytokine levels (IL-6, IL-8, TNF-α) by 30-47%. Critically, this synergy enabled a reduction of colistin dosing to one-sixteenth while maintaining bactericidal potency. These findings provide mechanistic insights into aspirin-mediated colistin sensitisation and evidence supporting combinatorial regimens to circumvent colistin toxicity barriers. This work establishes a rational foundation for clinical translation of repurposed aspirin-colistin therapy against MDR P. aeruginosa infections.

ETHNOPHARMACOLOGICAL RELEVANCE:Relinqing Granules (RLQ), a single-herb formulation derived from Polygonum capitatum Buch.-Ham. ex D. Don (THL), possesses diverse pharmacological properties-including antimicrobial, anti-inflammatory, and antioxidant activities-which suggest their potential utility in treating pneumonia. This study seeks to elucidate the underlying mechanisms of RLQ to support its future clinical application in managing inflammatory lung diseases. AIM OF THE STUDY:This study aimed to investigate the therapeutic effects of RLQ on pneumonia and elucidate its underlying mechanisms, with a particular focus on the phosphoinositide 3-kinase/protein kinase B/mechanistic target of rapamycin (PI3K/AKT/mTOR) signaling pathway. MATERIALS AND METHODS:Pneumonia was induced in mice via lipopolysaccharide (LPS) administration, while A549 cells were similarly stimulated with LPS to establish an in vitro inflammation model. Mice received RLQ treatment by oral gavage for seven consecutive days. Subsequently, lung tissues were collected for hematoxylin-eosin (H&E) staining, enzyme-linked immunosorbent assay (ELISA), Western blotting, and quantitative real-time polymerase chain reaction (qPCR). In vitro, RLQ-treated A549 cells were evaluated through ELISA, Cell Counting Kit-8 (CCK-8), flow cytometry, Western blotting, and qPCR. To further elucidate the underlying mechanism, A549 cell models with mTOR knockdown and overexpression (OE) were constructed to assess the regulatory role of mTOR in inflammation and apoptosis. RESULTS:RLQ alleviated LPS-induced lung inflammation by shifting the regulatory T cell/T helper 17 cell (Treg/Th17) balance toward an immunoregulatory state and modulating pro-inflammatory cytokine expression, potentially through the PI3K/AKT/mTOR signaling pathway. In vitro, RLQ treatment reduced cytokine levels, improved cell viability, and inhibited apoptosis in LPS-injured A549 cells by regulating this pathway and its downstream apoptotic proteins. Furthermore, in mTOR knockdown and OE models, RLQ significantly mitigated LPS-induced cellular damage, enhanced cell viability, and suppressed apoptosis, which correlated with the downregulation of mTOR pathway activation. CONCLUSION:RLQ effectively protected against LPS-induced lung and alveolar epithelial cell injury by inhibiting PI3K/AKT/mTOR pathway phosphorylation, restoring the Treg/Th17 immune balance, and suppressing apoptosis. These findings provide new mechanistic insights into the therapeutic potential of RLQ in the treatment of pneumonia.

Radix pseudostellariae (RP) has been used for the treatment of various diseases, including gastric disorders such as Chronic gastritis (CG), However, the multi-targeting mechanism of RP on CG still needs to be clarified. Drug targets were established through TCMSP, PubChem and Swiss Target Prediction online database. GeneCards database was used to search for chronic gastritis related targets. Cytoscape was utilized to construct the "drug-active ingredient-target" network relationship diagram. Venny plots of drug targets and disease targets were obtained from online software to get the intersecting genes, and the coincidence targets were imported into STRING database to construct a PPI network. The intersecting targets were subjected to pathway enrichment analysis KEGG and GO functional enrichment analysis with the help of the DAVID database. Molecular docking was performed by Autodocktools software and then visualized by PYMOL. 8 active ingredients and 117 action targets were obtained for Radix Pseudostellariae, 758 action targets for chronic gastritis disease, 31 intersecting targets for Radix Pseudostellariae in chronic gastritis were obtained. PPI network analysis showed that the core targets of the drug affecting chronic gastritis were TNF, MMP9, AKT1, etc. The molecular docking validation results showed good binding of core components and core targets.

This study aims to analyze the effective components of Polygonum capitatum (PC) inhibiting Escherichia coli based on network pharmacology methods and predict its molecular mechanism of action. PC compounds and targets were collected from the TCMSP database, Swiss Target Prediction, and the literature. E coli targets were searched using the GeneCards database. The targets of E coli and the targets of the active ingredients of PC were taken as intersections to obtain the intersecting targets. The resulting overlapping targets were uploaded to the STRING database to construct the protein interaction network diagram of E coli target inhibition. The key targets for the inhibitory effect of PC on E coli were obtained. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed by uploading key targets into the DAVID database. The results showed that there were 50 targets for PC to inhibit E coli. Among them, there are 5 core targets, mainly including AKT1, TNF, EGFR, JUN, and ESR1. A total of 196 gene ontology functional analysis results and 126 Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis results were obtained. These include cellular response to cadmium-ion, cellular response to reactive oxygen species, pathways in cancer, prostate cancer, and PI3K-Akt signaling pathway. Molecular docking results indicate that Lutedin, Hirsutin, Flazin, and Ellagic acid in PC have high affinity for the target genes AKT1, TNF, MAPK3 and EGFR. PC exerts its inhibitory effect on E coli through multi-targets and multi-pathways, which provides a new basis for the new use of PC as an old medicine.

Background: Radix Wikstroemia indica (RWI) is one of the most commonly used drugs in Miao medicine. However, RWI is characterized by high toxicity. Objectives: In this study, we aimed to observe the effect of the “sweat soaking method” processed on reducing the hepatotoxicity of RWI and on cytochrome P450 (CYP) protein expression. Materials and Methods: The study focused on investigating the impact of both RWI raw products and RWI processed products on the proliferation of L-02 cells. To assess this, a cell counting kit-8 was employed. After the administration of RWI through oral gavage for a duration of 15 days in rats, the corresponding kits were utilized to determine the serum levels of the liver index factor. Furthermore, both hematoxylin and eosin (H&E) staining and western blot analysis were conducted to analyze the liver tissues. Results: Both the raw RWI and the processed RWI at high doses inhibited the proliferation of L-02 cells, but the cell viability of the concoction group was higher than that of the raw group. The liver tissues of rats in the control and processed product groups were normal, while those in the raw product groups showed different degrees of liver damage. Additionally, compared with the control group, the activity of alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine transaminase (ALT) in the raw product groups was notably increased in a dose-dependent manner; the three enzyme activities of the processed group were lower than those of the raw product group. In addition, the expression levels of CYP3A4, 2C19, 2C9, 1A2, and 2E1 in the liver tissue of rats in the processed group were lower than those in the raw group, while the protein expression trend of CYP2D6 was irregular. Conclusion: The detoxication mechanism of RWI, after undergoing processing, appears to be associated with reduced protein expression of CYP1A2, CYP2E1, CYP2C9, CYP2C19, and CYP3A4, ultimately leading to decreased liver injury.

Background Radix Wikstroemia indica (RWI) is one of the most commonly used drugs in Miao medicine. However, RWI is characterized by high toxicity. Objectives In this study, we aimed to observe the effect of the “sweat soaking method” processed on reducing the hepatotoxicity of RWI and on cytochrome P450 (CYP) protein expression. Materials and Methods The study focused on investigating the impact of both RWI raw products and RWI processed products on the proliferation of L-02 cells. To assess this, a cell counting kit-8 was employed. After the administration of RWI through oral gavage for a duration of 15 days in rats, the corresponding kits were utilized to determine the serum levels of the liver index factor. Furthermore, both hematoxylin and eosin (H&E) staining and western blot analysis were conducted to analyze the liver tissues. Results Both the raw RWI and the processed RWI at high doses inhibited the proliferation of L-02 cells, but the cell viability of the concoction group was higher than that of the raw group. The liver tissues of rats in the control and processed product groups were normal, while those in the raw product groups showed different degrees of liver damage. Additionally, compared with the control group, the activity of alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine transaminase (ALT) in the raw product groups was notably increased in a dose-dependent manner; the three enzyme activities of the processed group were lower than those of the raw product group. In addition, the expression levels of CYP3A4, 2C19, 2C9, 1A2, and 2E1 in the liver tissue of rats in the processed group were lower than those in the raw group, while the protein expression trend of CYP2D6 was irregular. Conclusion The detoxication mechanism of RWI, after undergoing processing, appears to be associated with reduced protein expression of CYP1A2, CYP2E1, CYP2C9, CYP2C19, and CYP3A4, ultimately leading to decreased liver injury. Keywords Radix , Wikstroemia indica , sweat soaking method , cytochrome P450 , liver toxicity , processing mechanism

Background: Infections caused by carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-HVKP) are a great challenge for physicians in the administration of clinical therapy, and clarification of virulence factors and resistance mechanisms can aid in the selection of appropriate antimicrobial agents. Methods: We analysed the clinical characteristics of patients with CRKP infection, CRKP susceptibility to clinical first-line antimicrobial agents and resistance genotypes. The lethality and associated virulence factors of CR-HVKP are described. The possibility of the transfer of virulence and resistance genes was also explored. The susceptibility of CRKP and CR-HVKP to last-resort antimicrobial agents was also tested. Results: CRKP strains were predominantly derived from sputum specimens, urine specimens and blood specimens. Patients with CRKP infections predominantly had pulmonary infections and were predominantly elderly males in the intensive care unit (ICU). CRKP exhibited high resistance to other β-lactam antibiotics, mainly due to the presence of blaKPC, high resistance to quinolones mediated by the carriage of aac (6''-Ib-cr), QnrS and QnrB, and high resistance to aminoglycosides mediated by the carriage of rmtB, ant(3'')-I and armA, respectively. CR-HVKP is mainly composed of ST11 and capsule K1. Aerobactin (iucA, iutA), Ent siderophore (fepA, entB), Salmochelin (iroN), Yersiniabactin (ybtS), Type 3 fimbriae (mrkD), Type I fimbriae (fimH), and Regulation (prmpA) were detected in the CR-HVKP isolates. The CR-HVKP strains had a median lethal dose (LD50) of 2 × 103 to 5 × 103 CFU in the mice, which was similar to that of the positive control NTUH-K2044. Conjugation assays revealed that the genes iucA, iutA, iroN, prmpA, blaKPC, blaNDM, blaVIM, blaSHV, blaTEM, QnrA, QnrB, QnrS, and rmtB can be transferred to E. coli J53. Compared with those of the recipient E. coli J53, the MICs of meropenem, imipenem, levofloxacin and amikacin in the transconjugants increased by 4-128 times. Conclusions: These findings suggest that virulence and resistance genes may spread rapidly in Enterobacteriaceae, posing a public health risk and a significant threat to clinical care. Therefore, it is necessary to further strengthen hospital infection monitoring, prevention and control measures and to provide strict management and training on the rational use of antimicrobial agents in intensive care units.

The present study utilizes network pharmacology and molecular docking methodologies to investigate the mechanism of action behind the intervention of Polygonum capitatum Buch.-Ham.ex D. Don (THL) in treating pulmonary nodules (PN). This research aims to provide a theoretical foundation for broadening the clinical application of THL. Active components of THL were identified and screened through an extensive literature review and the PharmMapper database, followed by an analysis of their target interactions. Relevant targets associated with PN were selected using databases such as OMIM and GeneCards, with an intersection of the two sets being determined. STRING11.5 facilitated the acquisition of protein-protein interaction data, which was then imported into Cytoscape 3.7.2 to establish a protein interaction network topology. This enabled the identification of pivotal targets affected by THL intervention in PN. The study further employed the Metascape database to conduct GO and KEGG bioinformatics enrichment analyses, which illuminated core pathways involved in THL's therapeutic effects on PN. A comprehensive component-target-pathway diagram was constructed utilizing Cytoscape 3.7.2 software, with molecular docking validations carried out via Maestro software. A total of 49 active THL ingredients were discerned, implicating 67 PN-relevant targets. Subsequent software analysis pinpointed 10 key targets, including ALB, EGFR, and SRC. Molecular docking studies indicated strong binding affinities for most protein-compound pairs, with 44 out of 60 docking results exhibiting binding energies below -5 kcal/mol. Enrichment analysis highlights that key targets are mainly involved in pathways such as cancer, lipid metabolism and atherosclerosis, estrogen signaling, IL-17 signaling, complement and coagulation cascades, and chemical carcinogenesis through receptor activation. Through comprehensive network pharmacological approaches, this research delineates the synergy of THL's multiple components, targets, and pathways in mitigating PN. It posits that primary active ingredients of THL - quercetin, salidroside, and oleanolic acid - may exert effects on targets like ALB, EGFR, SRC, potentially modulating pathways associated with cancer, lipid and atherosclerosis, and IL-17 signaling in the context of PN intervention.

头花蓼为贵州传统苗药,具有清热利湿、利水通淋等功效.其主要化学成分为黄酮类、木脂素类及挥发油类,还含有萜类、鞣质类等成分.近年来,因头花蓼具有抗炎、抗菌、抗氧化及降血糖等多种药理作用而成为研究热点.研究头花蓼相关国内外文献,综述其化学成分及药理作用机制,以期对头花蓼的药理研究及临床应用提供参考.

Ethnopharmacological relevance: DBD is a traditional Chinese medicine formula that is used to cure 'deficiency of Qi and Blood' by benefit Qi and generate Blood. It is frequently used to treat female menstrual disorders, amenorrhea, infertility, and perimenopausal syndrome in contemporary therapeutic practice.Objective: Currently, there is uncertainty about how DBD affects ovarian function in women with premature ovarian failure (POF). In this work, we investigated the molecular basis of the effect of DBD on ovarian function in cisplatin-induced POF rats.Materials and methods: Six groups of 90 female SD rats were created: control, model, vitamin D (VD) (0.9 g/kg), H-DBD (7.2 g/kg), M-DBD (3.6 g/kg), and L-DBD (1.8 g/kg). The cisplatin intraperitoneal injection and 35-day treatment with VD and DBD were used to create the POF rat model. Rats' vaginal epithelial cells' morphology and percentage were studied using methylene blue staining. Rat ovarian histological structure and follicular development were examined using hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate the levels of serum estradiol (E2), estrogen receptor alpha (ERα), anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), and 25-hydroxyvitamin D3 [25(OH)D3]. By using a quantitative PCR test and Western blot to determine the levels of the vitamin D receptor (VDR), 25-hydroxyvitamin D3-1-hydroxylase (CYP27B1), vitamin D3-24-hydroxylase (CYP24A1), P53, and Bcl-2 mRNA and protein expression in rat ovaries. Immunofluorescence was used to measure the amounts of the BMSCs-specific proteins CD29 and CD90, as well as the proteins VDR, CYP27B1, CYP24A1, P53, and Bcl-2 in the bone marrow and the BMSCs-specific proteins CD29 and CD90 in the ovaries of rats.Results: In POF rats, DBD treatment can significantly increase the proportion of vaginal epithelial cells, control the motility cycle, encourage the development of mature follicles and prevent the formation of atretic follicles, as well as regulate serum sex hormone and vitamin D levels to enhance ovarian function. Moreover, DBD controls the P53/Bcl-2 signaling pathway, enhances bone marrow mesenchymal stem cells (BMSC) proliferation, differentiation, and homing function via the vitamin D system, inhibits ovarian apoptosis and boosts follicular reserve capacity, and delays the pathological development of POF.Conclusions: These results imply that DBD may reduce ovarian cell apoptosis and modulate the P53/Bcl-2 signaling pathway via the VD system. Moreover, DBD may help prevent and cure POF by enhancing follicular reserve capacity and boosting the proliferation, differentiation, and homing function of BMSCs via the VD system.

Background: Radix Wikstroemia indica is a traditional Chinese medicine (TCM) used as anti-inflammatory and anti-tumor drug. However, it has serious hepatotoxicity, "Sweat soaking method" processed could effectively decrease its hepatotoxicity. Objective: The objective of this study is to study the effects of Radix Wikstroemia indica on six kinds of cytochrome P450(CYP450) isozymes of rat liver microsomes before and after processing, and to study the mechanism of Radix Wikstroemia indica processed by the "Sweat soaking method" to reduce liver toxicity in rats. Methods: In this study, the effects of Radix Wikstroemia indica and processed Radix Wikstroemia indica on the six main CYP450 isoforms (2E1, 1A2, 2C6, 2D1, 2C11, and 3A1) were investigated in vitro. Using a cocktail probe of CYP450 isoform-specific substrates and their metabolites, we carried out in vitro enzymatic studies in liver microsomal incubation systems via UPLC-MS/MS. Results: The results showed that the established UPLC-MS/MS method was precise and reliable. Compared with the blank group, the activities of six enzymes in the RWI and PRWI groups were higher than those in the blank group. At the same dose, the enzyme activities of CYP2E1, CYP1A2, CYP2C6, CYP2C11, and CYP3A1 increased with the increase in dose, and the enzyme activities of the RWI group were higher than those of the PRWI group. The enzyme activities of CYP2E1 and CYP1A2 in the Radix Wikstroemia indica group were significantly increased compared with the blank group, CYP3A1 in the RWI high-dose group was higher than that in the blank group and PRWI group with statistical differences (p<0.05 or p<0.01). Conclusion: The processed Radix Wikstroemia indica could reduce liver injury, and its detoxication mechanism might be related to the decrease in enzyme activity of CYP1A2, CYP2E1 and CYP3A1.

目的 探讨热淋清颗粒对脓毒症急性肺损伤模型大鼠的保护作用及其机制.方法 将30只SD大鼠随机分为6组:假手术组、模型组、热淋清颗粒高剂量组[4.8 g/(kg·d)]、热淋清颗粒中剂量组[2.4 g/(kg·d)]、热淋清颗粒低剂量组[1.2 g/(kg·d)]、阳性对照组[地塞米松1.5 mg/(kg·d)],每组5只.各组均采用灌胃给药,1次/d,连续给药7 d,末次给药后各组大鼠均采用盲肠结扎穿孔法制备脓毒症急性肺损伤模型(假手术组不结扎和穿孔),造模24 h后取材.取各组大鼠右肺中叶计算肺组织湿/干重比;采用HE染色法观察各组大鼠肺组织病理学改变并进行损伤评分;采用ELISA法检测各组大鼠血清中IL-1α、TNF-α、PGE2水平含量;采用RT-PCR法检测各组大鼠肺组织中TLR4、MyD88、NF-κB p65 mRNA相对表达量;采用Western-Blot检测各组大鼠肺组织中TLR4、MyD88、NF-κB p65蛋白相对表达量.结果 与假手术组比较,模型组大鼠肺组织湿/干重比、肺组织病理损伤评分显著升高(P＜0.01);与模型组比较,热淋清颗粒高剂量组和阳性对照组可显著降低大鼠肺组织湿/干重比、肺组织病理损伤评分(P＜0.05);与假手术组比较,模型组大鼠血清TNF-α、IL-1α、PGE2水平及肺组织TLR4、MyD88、NF-KB p65 mRNA和蛋白表达量均显著升高(P＜0.01);与模型组比较,热淋清颗粒高、中剂量组和阳性对照组大鼠血清TNF-α、IL-1α、PGE2水平及肺组织TLR4、MyD88、NF-κB p65 mRNA和蛋白表达量显著降低(P＜0.05);热淋清颗粒高剂量组上述各指标改善情况优于阳性对照组.结论 热淋清颗粒能减轻脓毒症大鼠急性肺损伤,其机制可能与调节TLR4/MyD88/NF-κB p65通路有关.

Background: Daphnoretin, as a known bicoumarin compound that contained various pharmacological activities, was isolated from Wikstroemia indica C.A. Mey (RWI). Objective: The study aims to investigate the pharmacokinetic characteristics of daphnoretin from RWI ethanol extracts in rat plasma and to determine daphnetin in rat plasma and various tissues by a rapid, reliable and sensitive ultra high performance liquid chromatography with tandem mass spectrometry method. Methods: The UPLC-MS/MS method was established. Daphnoretin and IS (buspirone) were chromatographed on an agilent Zorbax XDB-C18 column (2.1 mm × 50 mm, 3.5 μm), and Gradient elution of acetonitrile-0.15% formic acid in aqueous solution. Quantification was performed using electrospray ionization in positive ion multiple reaction monitoring mode of the transitions m/z 353.1→179.1 for daphnoretin and m/z 386.3→122.3 for IS. Results: Good linearity between 5-10000 ng/mL for cyperidin in plasma and tissue samples (r ≥ 0.99) was resulted. The accuracies of plasma and tissue homogenates ranged from-3.31% to 9.00%, and the precision was less than 5.78%. After that, the validated method was successfully applied to the pharmacokinetics and tissue distribution study of daphnoretin after oral administration of ethanol extract from the roots of RWI to rats. Conclusion: Daphnoretin was well absorbed in the systemic circulation after oral administration and was widely distributed in tissues, with the highest concentration in lung tissue. This study is beneficial to the development and utilization of RWI and provides a reasonable reference for its clinical administration.

Pulmonary fibrosis (PF) is a serious interstitial disease that includes diffuse collagen deposition of lung tissue. Polygonum capitatum Buch.-Ham. ex D. Don (THL) is a traditional vaccine that has antibacterial and anti-inflammatory effects. In this research, to investigate the mechanism of action of THL in the intervention of pulmonary fibrosis by network pharmacology and molecular docking related research methods, in order to provide a theoretical basis for expanding the scope of THL medication. A total of 49 active ingredients were analyzed and screened in Cephalus cephalusis , including 35 pulmonary fibrosis targets, and 10 key targets such as ALB, EGFR were screened after software analysis. The molecular docking results showed that there were 44 binding energies less than –3 kcal·mol −1 in the 60 docking results, indicating that most proteins had strong binding energies with compounds. The key targets of KEGG enrichment analysis were mainly enriched in 20 core action pathways, such as hemostasis-related pathway, regulation of kinase activity. This study shows that based on network pharmacology, the multicomponent–multitarget–multipathway effect of THL intervention in pulmonary fibrosis is discussed.

The etiology of premature ovarian failure (POF) is mainly related to inflammatory diseases, autoimmune diseases, and tumor radiotherapy and chemotherapy; however, its specific pathogenesis has not been clarified. Vitamin D (VD), a fat-soluble vitamin, is an essential steroid hormone in the human body. Neutrophil extracellular traps (NETs) are meshwork structures that are formed when neutrophils are stimulated by inflammation and other factors and are closely associated with autoimmune and inflammatory diseases. Notably, VD inhibits NET formation and intervenes in the development of POF in terms of inflammatory and immune responses, oxidative stress, and tissue fibrosis. Therefore, this study aimed to theorize the relationship between NETs, VD, and POF and provide new ideas and targets for the pathogenesis and clinical treatment of POF.

Based on metabolomics, to study the mechanism of Radix Wikstroemia indica (RWI) "Sweat soaking method" processing detoxification. The raw drug group and processed products was given raw RWI and processed RWI respectively by gavage. The control group was given the same amount of 1% sodium carboxy methyl cellulose distilled water by gavage. After 7 days of continuous gavage, blood samples were collected. The blood samples of rats in each group were analyzed by 1H-NMR technology to explore the changes of endogenous metabolism and the possible metabolic pathways to rats before and after processing. Compared with the control group, the raw RWI could significantly reduce 16 metabolites and increase 10 metabolites. The processed RWI can increase the levels of most metabolites that decrease to the raw RWI, such as 13 metabolites such as alanine, L-glutamine, L-valine, L-serine, betaine and glutamic acid; At the same time, the metabolites that increased in the level of crude products were down-regulated, such as asparagine, lactic acid, 2hydroxyisobutyric acid, sucrose, glucose and D-glucose. Compared with raw products, RWI treated with "Sweat soaking method" can reversely regulate or reduce amino acid, choline metabolism, energy and carbohydrate metabolism, thereby reducing hepatotoxicity and nephrotoxicity.

Radix Wikstroemia indica (L.) C.A. Mey. (RWI) is a toxic medicinal species primarily present in the Miao area of China. The toxicity of RWI is effectively reduced whilst maintaining the therapeutic effect when processed using the 'sweat-soaking method', which is a common method of Traditional Chinese Medicine preparation. However, there is a lack of scientific and medical evidence to explain the potential mechanisms by which the toxicity of RWI is reduced after preparation using this method, and the endogenous systemic metabolic effect of RWI remains uncertain. The aim of the present study was to explore the endogetnous metabolic alterations caused by RWI and to examine the possibility of reducing the toxicity of RWI using the sweat-soaking method using proton nuclear magnetic resonance (NMR) metabolomic analysis in rats. Principal Component Analysis, Partial Least Squares-Discriminant Analysis (PLS-DA) and Orthogonal PLS-DA were used to assess individual proton NMR spectra. A total of 34 metabolic products were altered after delivering raw RWI, and 32 endogenous metabolites were induced by processed RWI. The metabolic pathways that lead to a significant impact on energy and carbohydrate, amino acid, organic acids and lipid metabolism following raw and processed RWI use were identified. The mitochondria of hepatic and renal tubules of rats were injured in the raw RWI group, whereas the processed product reduced or interfered with energy substrate, carbohydrate and amino acid metabolism, whilst reducing the levels of metabolic markers of hepatotoxicity and nephrotoxicity, without causing damage to the mitochondria. Our previous study showed that the median lethal dose (LD50) value of raw RWI was 4.05 g/kg in rats after oral administration; however, the LD50 value of the processed RWI could not be measured. The maximum tolerated dose and minimum lethal dose were 20 and 30 g/kg for the processed RWI, respectively, corresponding to 109 and 164 times the clinical daily dose (0.029 g/kg). Thus, the sweat-soaking method reduced the toxicity of RWI. Moreover, after processing, the toxic component YH-10 was converted into a YH-10 + OH compound, reducing the content of the toxic YH-10 by 48%, whilst also reducing the contents of the toxic components YH-12 and YH-15 by 44 and 65%, respectively. In conclusion, the present study showed that the sweat-soaking method reduced the toxicity of RWI, as evidenced by the reduction of the levels of metabolic markers and the activity of metabolic pathways, thus providing a basis for processing of RWI for clinical use.