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.

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 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 investigate the feasibility of laparoscopic simple prostatectomy (LSP) in the treatment of large volume benign prostate hyperplasia (BPH). 【Methods】 Clinical and follow-up data of 30 patients with large volume BPH treated with LSP in our hospital during Feb.2019 and Dec.2021 were retrospectively analyzed. All patients underwent extraperitoneal LSP operation. The perioperative and 1-12 month postoperative follow-up data were analyzed. 【Results】 The average prostate volume was (92.4±38.9) mL, operation time (125±45) min, and weight of resected prostate (60.25±16.90) g. The hemoglobin decreased by (12.21±7.25) g/d after operation. No blood transfusion was needed. There was no need for bladder irrigation after operation in 21 cases (70%), and 9 cases (30%) had bladder irrigation time of (0.95±0.49) d. The postoperative catheter indwelling time was (6.92±2.51) d, and hospital stay (5.36±1.63) d. During the follow-up of (9.25±5.4) months, there was 1 case of postoperative intestinal obstruction (Clavien-Dindo grade II), 1 case of transient urinary incontinence (Clavien-Dindo grade I), and 1 case of delayed hematuria (Clavien-Dindo grade I). No urethral stricture occurred. The maximum urinary flow rate (Qmax), post-void residual urine volume (PVR), International Prostate Symptom Score (IPSS) and quality of life (QoL) 3 months after operation were significantly improved compared with those before operation (P<0.05). There was no significant difference in sexual function before and after operation (P>0.05). 【Conclusion】 LSP is safe and effective in the treatment of large volume BPH. It has advantages of complete resection of glands, minor bleeding and short postoperative bladder irrigation time. However, it still needs to be confirmed by a prospective control study of large samples.

We conducted a prospective study to assess the non-inferiority of adjuvant chemotherapy alone versus adjuvant concurrent chemoradiotherapy (CCRT) as an alternative strategy for patients with early-stage (FIGO 2009 stage IB-IIA) cervical cancer having risk factors after surgery. The condition was assessed in terms of prognosis, adverse effects, and quality of life. This randomized trial involved nine centers across China. Eligible patients were randomized to receive adjuvant chemotherapy or CCRT after surgery. The primary end-point was progression-free survival (PFS). From December 2012 to December 2014, 337 patients were subjected to randomization. Final analysis included 329 patients, including 165 in the adjuvant chemotherapy group and 164 in the adjuvant CCRT group. The median follow-up was 72.1 months. The three-year PFS rates were both 91.9%, and the five-year OS was 90.6% versus 90.0% in adjuvant chemotherapy and CCRT groups, respectively. No significant differences were observed in the PFS or OS between groups. The adjusted HR for PFS was 0.854 (95% confidence interval 0.415-1.757; P = 0.667) favoring adjuvant chemotherapy, excluding the predefined non-inferiority boundary of 1.9. The chemotherapy group showed a tendency toward good quality of life. In comparison with post-operative adjuvant CCRT, adjuvant chemotherapy treatment showed non-inferior efficacy in patients with early-stage cervical cancer having pathological risk factors. Adjuvant chemotherapy alone is a favorable alternative post-operative treatment.
Female ; Humans ; Uterine Cervical Neoplasms/drug therapy* ; Prospective Studies ; Quality of Life ; Neoplasm Staging ; Chemoradiotherapy ; Chemotherapy, Adjuvant/adverse effects* ; Adjuvants, Immunologic ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use* ; Retrospective Studies

Objective:To explore the effects of cyclosporine A (CsA) on the clinical outcomes of patients with unexplained repeated implantation failure (URIF) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods:A retrospective cohort study was conducted, and the data of URIF patients at the Center for Human Reproduction and Genetics of the Affiliated Suzhou Hospital of Nanjing Medical University from April 2016 to March 2020 was analyzed. Totally 94 cycles with CsA application were enrolled, and 188 cycles of control group were selected according to age, body mass index (BMI), basal follicle-stimulating hormone (FSH), the number of high-quality embryos transferred and embryo stage matched with CsA group. The general conditions, characteristics of previous failed transfer cycles and the de novo transfer cycles, clinical outcomes, obstetric and neonatal complications were compared between the two groups, and regression analysis of CsA and clinical outcomes. Results:1) There were no statistical differences between the two groups in baseline characteristics such as age, BMI, basal FSH, duration of infertility, indications for IVF/ICSI, number of previous failed cycles, number of high-quality embryos and blastocyst transferred rate (all P>0.05). There were also no significant differences in the number of embryos transferred, the number of high-quality embryos, the rate of transferred blastocyst and the endometrial thickness on transfer day of the de novo transfer cycles (all P>0.05). 2) The embryo implantation rate, the clinical pregnancy rate and the live birth rate in CsA group [53.39% (63/118), 58.51% (55/94), 45.74% (43/94)] were distinctly higher than those of control group [38.43% (93/242), 45.74% (86/188), 33.51% (63/188)]. The differences were statistically significant ( P=0.007, P=0.043, P=0.046). While there were no differences in miscarriage rate and ectopic pregnancy rate between the two groups (all P>0.05). 3) The differences of the rates of preterm birth, multiple pregnancy, obstetric and neonatal complications were all not statistically significant between the two groups (all P>0.05). No birth defects were observed in both groups. 4) The multivariate logistic regression analysis showed that CsA was an independent promoter of clinical pregnancy (after adjusted OR=1.694, 95% CI:1.019-2.816, P=0.042) and live birth (after adjusted OR=1.700, 95% CI:1.012-2.853, P=0.045) in URIF patients after adjusting for age, BMI, basal FSH, the number of embryos transferred and the endometrial thickness on transfer day. Conclusion:CsA showed remarkably enhancement on embryo implantation rate, clinical pregnancy rate and live birth rate of de novo embryo transfer in patients with URIF, without increasing the risk of obstetric and pediatric complications. CsA application may be used as an effective treatment for URIF patients.

Objective:To explore the effects of cyclosporine A (CsA) on the clinical outcomes of patients with unexplained repeated implantation failure (URIF) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods:A retrospective cohort study was conducted, and the data of URIF patients at the Center for Human Reproduction and Genetics of the Affiliated Suzhou Hospital of Nanjing Medical University from April 2016 to March 2020 was analyzed. Totally 94 cycles with CsA application were enrolled, and 188 cycles of control group were selected according to age, body mass index (BMI), basal follicle-stimulating hormone (FSH), the number of high-quality embryos transferred and embryo stage matched with CsA group. The general conditions, characteristics of previous failed transfer cycles and the de novo transfer cycles, clinical outcomes, obstetric and neonatal complications were compared between the two groups, and regression analysis of CsA and clinical outcomes. Results:1) There were no statistical differences between the two groups in baseline characteristics such as age, BMI, basal FSH, duration of infertility, indications for IVF/ICSI, number of previous failed cycles, number of high-quality embryos and blastocyst transferred rate (all P>0.05). There were also no significant differences in the number of embryos transferred, the number of high-quality embryos, the rate of transferred blastocyst and the endometrial thickness on transfer day of the de novo transfer cycles (all P>0.05). 2) The embryo implantation rate, the clinical pregnancy rate and the live birth rate in CsA group [53.39% (63/118), 58.51% (55/94), 45.74% (43/94)] were distinctly higher than those of control group [38.43% (93/242), 45.74% (86/188), 33.51% (63/188)]. The differences were statistically significant ( P=0.007, P=0.043, P=0.046). While there were no differences in miscarriage rate and ectopic pregnancy rate between the two groups (all P>0.05). 3) The differences of the rates of preterm birth, multiple pregnancy, obstetric and neonatal complications were all not statistically significant between the two groups (all P>0.05). No birth defects were observed in both groups. 4) The multivariate logistic regression analysis showed that CsA was an independent promoter of clinical pregnancy (after adjusted OR=1.694, 95% CI:1.019-2.816, P=0.042) and live birth (after adjusted OR=1.700, 95% CI:1.012-2.853, P=0.045) in URIF patients after adjusting for age, BMI, basal FSH, the number of embryos transferred and the endometrial thickness on transfer day. Conclusion:CsA showed remarkably enhancement on embryo implantation rate, clinical pregnancy rate and live birth rate of de novo embryo transfer in patients with URIF, without increasing the risk of obstetric and pediatric complications. CsA application may be used as an effective treatment for URIF patients.

Objective:To investigate the status quo of the knowledge, attitude and practice (KAP) of standardized insulin injection at home in Shanghai community diabetic patients and analyze its influencing factors.Methods:In this cross-sectional study, totally 1 947 diabetic patients who signed to be managed by 21 community health service centers in Fengxian District, Shanghai from May to October 2020 who used insulin pen injections at home were selected by continuous enrolment. They were investigated with a general information questionnaire and a questionnaire on KAP of standardized insulin injection. Univariate analysis and multiple linear regression were used to analyze the influencing factors of patients' KAP of standardized insulin injection at home. A total of 1 947 questionnaires were distributed, and 1 697 valid questionnaires were recovered, with an effective recovery rate of 87.2%.Results:The total KAP score of insulin standard injection in the 1 697 diabetic patients was (51.01±6.81) , which was at a moderate level. Among all the items in the questionnaire, the 3 items with the lowest compliance rate were: checking the injection site every time before insulin injection (14.4%) , agreeing to put the discarded insulin needles into the capped hard-shell container (16.1%) , agreeing to and implementing insulin pen needle replacement after each use (20.6%) . Multiple linear regression analysis showed that age, education, type of medical insurance, and whether they had received insulin injection guidance were the influencing factors to the KAP of standardized insulin injection at home ( P<0.01) , which accounted for 8.6% of the total variation. Conclusions:Community medical staff should pay attention to the health education of insulin injection at home for diabetic patients, improve the content and form of education based on the characteristics of the community population, thereby improving the compliance rate of injection site inspection, replacement of insulin pen needles and standardized disposal of discarded needles, further standardizing insulin injection behavior, and improving clinical outcomes such as glycemic control in patients.

ObjectiveThe type 2C protein phosphatases （PP2C） are involved in numerous plant signal transduction pathways. They mainly participate in plant stress response and regulate second metabolites biosynthesis via negatively regulating MAPK signaling pathway. Herein，we were to identify and analyze PP2C （CsPP2C） gene family from hemp genome，in hope of providing comprehensive insights for studying CsPP2C function during the development of hemp. MethodMolecular Evolutionary Genetics Analysis （MAGA）-X was used to construct phylogenetic tree. Expert Protein Analysis System （ExPASy），WoLF PSORT，Multiple EM for Motif Elicitation （MEME），Batch Conserved Domain Search （Batch-CD-Search），PlantCare，and TBtools were used，respectively，to predict CsPP2C physicochemical properties，subcellular localization，conserved motifs，protein structure，cis-element in promoter and collinearity with Arabidopsis PP2C. Cannabis sativa transcriptome and Real-time polymerase chain reaction（Real-time PCR） were used to analyze and verify gene expressions，respectively. ResultFifty-two CsPP2C with conserved domains were identified from the entire genome of hemp，encoding proteins ranging from 244 to 1 089 aa in length and with molecular weights ranging from 26.76 to 122.53 kDa. Those genes were mainly distributed in the nucleus，cytoplasm and chloroplast. The 47 CsPP2C were divided into 10 subfamilies，and the remaining 5 were not clustered. Seven pairs of homologous genes between hemp and Arabidopsis thaliana were identified according to collinear analysis. The light-responsive elements and abscisic acid elements are most abundant in the prediction. The gene expression heat map showed varied expression pattern of CsPP2C in different tissues. Real-time PCR results of three CsPP2C were consistent with transcriptome data. Moreover，alternative splicing analysis showed that some CsPP2C had alternative-splicing genes during evolution. ConclusionWe predicted and analyzed CsPP2C gene family in genomic scale and showed that CsPP2C are involved in many biological processes，whereby provides foundation for CsPP2C functional study.