Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.

OBJECTIVE: In Pakistan, traditional medicines are an important component of the medical system, with numerous varieties and great demands. However, due to the scattered resources and the lack of systematic collection and collation, adulteration of traditional Pakistani medicine (TPM) is common, which severely affects the safety of their medicinal use and the import and export trades. Therefore, it is urgent to systematically organize and unify the management of TPM and establish a set of standards and operable methods for the identification of TPM. METHODS: We collected and organized the information on 128 TPMs with regard to their medicinal parts, efficacy, usage, and genetic material, based on Pakistan Hamdard Pharmacopoeia of Eastern Medicine: Pharmaceutical Codex. The genetic information of TPM is summarized from national center for biotechnology information (NCBI) and global pharmacopoeia genome database (GPGD). Furthermore, we utilized bioinformatics technology to supplement the chloroplast genome (cp-genome) data of 12 TPMs. To build the web server, we used the Linux + Apache + MySQL + PHP (LAMP) system and constructed the webpage on a PHP: Hypertext Preprocessor (PHP) model view controller (MVC) framework. RESULTS: We constructed a new genomic database, the traditional Pakistani medicine genomic database (TPMGD). This database comprises five entries, namely homepage, medicinal species, species identification, basic local alignment search tool (BLAST), and download. Currently, TPMGD contains basic profiles of 128 TPMs and genetic information of 102 TPMs, including 140 cytochrome c oxidase subunit I (COI) sequences and 119 mitochondrial genome sequences from Bombyx mori, 1 396 internal transcribed spacer 2 (ITS2) sequences and 1 074 intergenic region (psbA-trnH) sequences specific to 92 and 83 plant species, respectively. Additionally, TPMGD includes 199 cp-genome sequences of 82 TPMs. CONCLUSION TPMGD is a multifunctional database that integrates species description, functional information inquiry, genetic information storage, molecular identification of TPM, etc. The database not only provides convenience for TPM information queries but also establishes the scientific basis for the medication safety, species identification, and resource protection of TPM.

Objective:To explore the feasibility and adaptability of laparoscopy in the surgical treatment of splenic aneurysm.Methods:The data of 28 patients with splenic aneurysms who underwent laparoscopic surgery in the Department of Hepatobiliary and Pancreatic Surgery of the Third Affiliated Hospital of Soochow University from January 2010 to December 2023 were retrospectively collected and analyzed. Among them, there were 13 males and 15 females, with the age of (57.3±7.7) years. All patients underwent laparoscopic splenic aneurysm resection, and whether to perform splenic artery anastomosis or splenectomy was determined based on the intraoperative situation. Collect the long diameter and location of splenic aneurysms, intraoperative blood loss, operation time, postoperative complications (bleeding, pancreatic fistula, splenic infarction, and splenic vein and portal vein thrombosis), postoperative hospital stay and hospitalization expenses of the patients.Results:All patients successfully underwentlaparoscopic surgery for splenic aneurysms. Aneurysms were located at the origin of the splenic artery in 3 cases (10.7%), in the middle in 8 cases (28.6%), and at the tail in 17 cases (60.7%). The long diameter of the aneurysms was (3.1±1.7) cm. Among the 28 patients, 18 cases (64.3%) underwent splenic aneurysm resection alone, 5 cases (17.9%) underwent splenic aneurysm resection combined with end-to-end anastomosis of the splenic artery, and 5 cases (17.9%) underwent splenic aneurysm resection combined with splenectomy. The operation time of 28 patients was (124.3±55.1) min, the intraoperative blood loss was 100.0 (50.0, 162.5) ml, the postoperative hospital stay was (10.9±3.8) days, and the hospitalization cost was (3.7±1.2) wanyuan. Among the 28 patients, 5 cases (17.9%) developed pancreatic fistula, 1 case (3.6%) had partial splenic infarction, and 1 case (3.6%) had portal vein and splenic vein thrombosis after the operation.Conclusion:Laparoscopic surgery for splenic aneurysm is safe and feasible, with less surgical trauma and quick postoperative recovery.

Lingguizhugan Decoction (LGZG) demonstrates significant efficacy in treating various cardiovascular diseases clinically, yet its precise mechanism of action remains elusive. This study aimed to elucidate the potential mechanisms and effects of LGZG on isoproterenol (ISO) continuous stimulation-induced chronic heart failure (CHF) in mice, providing direct experimental evidence for further clinical applications. In vivo, continuous ISO infusion was administered to mice, and ventricular myocytes were utilized to explore LGZG?s potential mechanism of action on the ?1-adrenergic receptor (?1-AR)/Gs/G protein-coupled receptor kinases (GRKs)/?-arrestin signaling deflection system in the heart. The findings reveal that LGZG significantly reduced the messenger ribonucleic acid (mRNA) expression of hypertrophy-related biomarkers [atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP)] and improved cardiac remodeling and left ventricular diastolic function in mice with ISO-induced CHF. Furthermore, LGZG inhibited the overactivation of Gs/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling and downregulated the downstream transcriptional activity of cAMP-response element binding protein (CREB) and the expression of the coactivator CBP/P300. Notably, LGZG downregulated the expression of ?-arrestin1 and GRK 2/3/5 while upregulating the expression of ?1-AR and ?-arrestin2. These results suggest that LGZG inhibits Gs/cAMP/PKA signaling and ?-arrestin/GRK-mediated desensitization and internalization of ?1-AR, potentially exerting cardioprotective effects through the synergistic regulation of the ?1-AR/Gs/GRKs/?-arrestin signaling deflection system via multiple pathways.
Animals ; Heart Failure/genetics* ; Signal Transduction/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Mice ; Male ; G-Protein-Coupled Receptor Kinases/genetics* ; Mice, Inbred C57BL ; Humans ; Isoproterenol ; Arrestins/genetics* ; Chronic Disease

Objective To explore the potential categories of exercise perception in elderly hip arthroplasty(HA)patients,analyze the prediction factors of different categories,and provide references for clinical nursing.Methods A convenience sampling method was used to select elderly HA patients treated in 6 orthopedic wards of 2 tertiary A hospitals in Guilin from October 2024 to February 2025.The general data questionnaire,Exercise Benefits and Barriers Perception Scale for Hip/Knee Throplasty Patients,Tampa Scale of Kinesiophobia,the 5-Factor Modified Frailty Index,and Self-Efficacy for Rehabilitation Outcome Scale were used for investigation.Latent profile analysis was used to identify the potential categories of exercise perception of elderly HA patients,and decision tree model was used to explore the core predictive factors of different profile.Results 222 valid questionnaires were collected,with an effective response rate of 96.52％.A total of 222 elderly HA patients were divided into 3 profiles:high benefit-low barrier group(19.82％),low benefit-high barrier group(22.07％),and mild barrier group(58.11％).Frailty,kinesophobia,rehabilitation self-efficacy,residence,educational level and daily exercise were significant predictors of latent profiles(P＜0.05),and the frailty was located in the root node of the decision tree model.Conclusion The level of exercise perception in elderly HA patients needs to be improved,and shows population heterogeneity.Medical staff should prioritize interventions for low benefit-high barrier and mild-barrier groups,implementing targeted strategies based on profile characteristics and predictive factors to improve their exercise perception.

Objective To explore the potential categories of exercise perception in elderly hip arthroplasty(HA)patients,analyze the prediction factors of different categories,and provide references for clinical nursing.Methods A convenience sampling method was used to select elderly HA patients treated in 6 orthopedic wards of 2 tertiary A hospitals in Guilin from October 2024 to February 2025.The general data questionnaire,Exercise Benefits and Barriers Perception Scale for Hip/Knee Throplasty Patients,Tampa Scale of Kinesiophobia,the 5-Factor Modified Frailty Index,and Self-Efficacy for Rehabilitation Outcome Scale were used for investigation.Latent profile analysis was used to identify the potential categories of exercise perception of elderly HA patients,and decision tree model was used to explore the core predictive factors of different profile.Results 222 valid questionnaires were collected,with an effective response rate of 96.52％.A total of 222 elderly HA patients were divided into 3 profiles:high benefit-low barrier group(19.82％),low benefit-high barrier group(22.07％),and mild barrier group(58.11％).Frailty,kinesophobia,rehabilitation self-efficacy,residence,educational level and daily exercise were significant predictors of latent profiles(P＜0.05),and the frailty was located in the root node of the decision tree model.Conclusion The level of exercise perception in elderly HA patients needs to be improved,and shows population heterogeneity.Medical staff should prioritize interventions for low benefit-high barrier and mild-barrier groups,implementing targeted strategies based on profile characteristics and predictive factors to improve their exercise perception.

Objective:To explore the feasibility and adaptability of laparoscopy in the surgical treatment of splenic aneurysm.Methods:The data of 28 patients with splenic aneurysms who underwent laparoscopic surgery in the Department of Hepatobiliary and Pancreatic Surgery of the Third Affiliated Hospital of Soochow University from January 2010 to December 2023 were retrospectively collected and analyzed. Among them, there were 13 males and 15 females, with the age of (57.3±7.7) years. All patients underwent laparoscopic splenic aneurysm resection, and whether to perform splenic artery anastomosis or splenectomy was determined based on the intraoperative situation. Collect the long diameter and location of splenic aneurysms, intraoperative blood loss, operation time, postoperative complications (bleeding, pancreatic fistula, splenic infarction, and splenic vein and portal vein thrombosis), postoperative hospital stay and hospitalization expenses of the patients.Results:All patients successfully underwentlaparoscopic surgery for splenic aneurysms. Aneurysms were located at the origin of the splenic artery in 3 cases (10.7%), in the middle in 8 cases (28.6%), and at the tail in 17 cases (60.7%). The long diameter of the aneurysms was (3.1±1.7) cm. Among the 28 patients, 18 cases (64.3%) underwent splenic aneurysm resection alone, 5 cases (17.9%) underwent splenic aneurysm resection combined with end-to-end anastomosis of the splenic artery, and 5 cases (17.9%) underwent splenic aneurysm resection combined with splenectomy. The operation time of 28 patients was (124.3±55.1) min, the intraoperative blood loss was 100.0 (50.0, 162.5) ml, the postoperative hospital stay was (10.9±3.8) days, and the hospitalization cost was (3.7±1.2) wanyuan. Among the 28 patients, 5 cases (17.9%) developed pancreatic fistula, 1 case (3.6%) had partial splenic infarction, and 1 case (3.6%) had portal vein and splenic vein thrombosis after the operation.Conclusion:Laparoscopic surgery for splenic aneurysm is safe and feasible, with less surgical trauma and quick postoperative recovery.

Objective To reveal the pharmacodynamic material basis of Kaixinsan by using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)and the integrated analysis of"chemical component spectrum-plasma exposure component spectrum-mitochondrial function".Methods Through a review of literature,databases,and previous studies,the chemical components of ginseng,polygala,poria,and acorus were systematically cataloged.A qualitative analysis method for the chemical constituents in the aqueous extract of Kaixinsan was developed,allowing for the identification of its chemical components.A qualitative analysis for rat plasma based on HPLC-MS/MS was established,which was applied to analyze the plasma exposure component spectrum following oral administration of Kaixinsan aqueous extract in rats.Aerobic respiration was evaluated using a seahorse cell energy metabolism analyzer,and the effect of key components of Kaixinsan on mitochondrial aerobic respiration was assessed.Results Four main types of components were identified in the Kaixinsan aqueous extract,including saponins,oligosaccharide esters,xanthones,and triterpenes,comprising a total of 231 identified compounds.Analysis of rat plasma 30 minutes after gavage with Kaixinsan identified 55 compounds.The analysis revealed that ginsenoside Rg1,3,6'-disinapoylsucrose,polygalaxanthone Ⅲ and poricoic acid B could significantly enhance mitochondrial respiratory capacity using in vitro cellular assays to detect aerobic respiration of four main components entered blood.Conclusions Saponins,oligosaccharide esters,xanthones,and triterpenes may be the material basis for the pharmacological effect of Kaixinsan by improving mitochondrial function.The integrated analysis of"chemical component spectrum-plasma exposure component spectrum-mitochondrial function"provides a new approach for in-depth exploration of the material basis underlying the efficacy of traditional Chinese medicine.

OBJECTIVE To analyze the metabolites of Zhideke granules and speculate its metabolic pathway in rats in vivo. METHODS Male SD rats were randomly divided into blank group and administration group （Zhideke granules， 9.45 g/kg）； they were given ultrapure water or relevant medicine， twice a day， every 6-8 h， for 3 consecutive days. Serum， urine and feces samples of rats were collected， and their metabolites were identified by UPLC-Q-Exactive-MS technique after intragastric administration of Zhideke granules； their metabolic pathways were speculated. RESULTS After intragastric administration of Zhideke granules， 16 prototype components （i.g. irisflorentin， baicalin， chlorogenic acid） and 11 metabolites （i.g. hydration products of kaempferol or luteolin， methylation products of chlorogenic acid， and hydroxylation products of baicalin） were identified in serum， urine and feces of rats. Among them， 8 prototype components and 4 metabolites were identified in serum samples； 10 prototype components and 7 metabolites were identified in urine samples； 8 prototype components and 5 metabolites were identified in the fecal samples. CONCLUSIONS The metabolites of Zhideke granules in rats mainly include baicalin， irisflorentin，chlorogenic acid， and the main metabolic pathways included methylation， hydroxylation， glucuronidation.

OBJECTIVE To investigate the regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib. METHODS Using human liver cancer cell Huh7 as subject， the lenvatinib-resist cell model （Huh7-LR） was generated by the low-dose gradient method combined with long-term administration. The sensitivity of parental cell Huh7 and drug-resistant cell Huh7-LR to lenvatinib was detected by using CCK-8 assay and flow cytometry. Western blot assay and GFP-mCherry-LC3 plasmid transfection were performed to detect the expression levels of autophagic protein Beclin-1， autophagic adapter protein sequestosome 1 （p62）， microtubule-associated protein 1 light chain 3 （LC3） and autophagic level. Furthermore， an autophagy activation model was constructed by cell starvation， the protein expression of p62 and autophagy level were detected by using Western blot assay and GFP-mCherry-LC3 plasmid transfection， and the effect of autophagy activation on the sensitivity of Huh7-LR cells to lenvatinib was detected by flow cytometry. RESULTS Compared with parental cells， the drug resistance index of Huh7-LR cells was 6.2； protein expression of p62 was increased significantly， while apoptotic rate， protein expression of Beclin-1 and LC3Ⅱ/ LC3Ⅰ ratio were all reduced significantly （P＜0.05 or P＜0.01）； the level of autophagy was decreased to some extent. Autophagy activation could significantly increase the protein expression of p62 in Huh7-LR cells （P＜0.05） and autophagy level， and significantly increase its apoptotic rate （P＜0.05）. CONCLUSIONS Autophagy is involved in lenvatinib resistance， and activating autophagy can reverse the resistance of liver cancer cells to lenvatinib to some extent.