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: To investigate chronic hepatitis C virus (HCV) infection's effect on gestational liver function, pregnancy and delivery complications, and neonatal development. METHODS: A total of 157 HCV antibody-positive (anti-HCV[+]) and HCV RNA(+) patients (Group C) and 121 anti-HCV(+) and HCV RNA(-) patients (Group B) were included as study participants, while 142 anti-HCV(-) and HCV RNA(-) patients (Group A) were the control group. Data on biochemical indices during pregnancy, pregnancy complications, delivery-related information, and neonatal complications were also collected. RESULTS: Elevated alanine aminotransferase (ALT) rates in Group C during early, middle, and late pregnancy were 59.87%, 43.95%, and 42.04%, respectively-significantly higher than Groups B (26.45%, 15.70%, 10.74%) and A (23.94%, 19.01%, 6.34%) ( P < 0.05). Median ALT levels in Group C were significantly higher than in Groups A and B at all pregnancy stages ( P < 0.05). No significant differences were found in neonatal malformation rates across groups ( P > 0.05). However, neonatal jaundice incidence was significantly greater in Group C (75.16%) compared to Groups A (42.25%) and B (57.02%) ( χ ² = 33.552, P < 0.001). HCV RNA positivity during pregnancy was an independent risk factor for neonatal jaundice ( OR = 2.111, 95% CI 1.242-3.588, P = 0.006). CONCLUSIONS Chronic HCV infection can affect the liver function of pregnant women, but does not increase the pregnancy or delivery complication risks. HCV RNA(+) is an independent risk factor for neonatal jaundice.
Humans ; Female ; Pregnancy ; Adult ; Pregnancy Complications, Infectious/epidemiology* ; Retrospective Studies ; Pregnancy Outcome ; Infant, Newborn ; Viremia/virology* ; Hepatitis C ; Hepacivirus/physiology* ; Hepatitis C, Chronic/virology* ; Young Adult ; Alanine Transaminase/blood*

ObjectiveTo construct the core outcome set （COS） for premature ovarian insufficiency （POI） treated with traditional chinese medicine （TCM）， thereby standardizing the effect evaluation system of POI clinical researches. MethodsDatabases including PubMed， EMbase， Cochrane Library， Web of Science， CNKI， Wanfang， VIP， SinoMed from January 2017 to December 31， 2021， and Chinese Clinical Trials Registry （ChiCTR）， ClinicalTrials.gov were searched to collect the outcomes used in clinical studies of POI. Supplemented with the patients preferred outcomes through semi-structured interviews， the outcome pool was then constructed. Finally through the Delphi survey and consensus conference， the COS for POI was obtained. ResultsA total of 554 clinical researches and 14 registered clinical research protocols were included. A semi-structured interview was conducted within 20 patients. A total of 77 outcomes were included in the pool after consolidation. After two rounds of Delphi surveys， 20 outcomes were initially included. The COS， which was finalized through expert consensus conference， consists of 8 outcomes in 3 domains， which were ovarian reserve function （follicle stimulating hormone， estradiol， anti-Mullerian hormone， sinus follicle count）， menstruation-related indicators （menstrual symptoms condition） and fertility indicators （pregnancy rate， live birth rate and the numbers of retrieved oocytes）. Through expert discussion， the corresponding measurement methods as well as the measurement time points of the COS were determined. ConclusionThis study formed COS for clinical studies of TCM in the treatment of POI， by practicing the general method of developing COS， and exploring the key points of developing COS with TCM features.

Objective This study aimed to investigate the potential relationship between urinary metals copper (Cu), arsenic (As), strontium (Sr), barium (Ba), iron (Fe), lead (Pb) and manganese (Mn) and grip strength. Methods We used linear regression models, quantile g-computation and Bayesian kernel machine regression (BKMR) to assess the relationship between metals and grip strength.Results In the multimetal linear regression, Cu (β=-2.119), As (β=-1.318), Sr (β=-2.480), Ba (β=0.781), Fe (β= 1.130) and Mn (β=-0.404) were significantly correlated with grip strength (P < 0.05). The results of the quantile g-computation showed that the risk of occurrence of grip strength reduction was -1.007 (95％ confidence interval:-1.362, -0.652; P < 0.001) when each quartile of the mixture of the seven metals was increased. Bayesian kernel function regression model analysis showed that mixtures of the seven metals had a negative overall effect on grip strength, with Cu, As and Sr being negatively associated with grip strength levels. In the total population, potential interactions were observed between As and Mn and between Cu and Mn (Pinteractions of 0.003 and 0.018, respectively).Conclusion In summary, this study suggests that combined exposure to metal mixtures is negatively associated with grip strength. Cu, Sr and As were negatively correlated with grip strength levels, and there were potential interactions between As and Mn and between Cu and Mn.

Objective:To investigate the effects of CP-31398 on proliferation and metastasis of gallbladder carcinoma cell lines expressing mutant P53GBC-SD and wild-type p53NOZ.Meth-ods:The effects of CP-31398 on the proliferation of wild-type and mutant p53 gallbladder carci-noma cells were detected by CCK8 cell proliferation assay and clonal formation assay.The effect of CP-31398 on cell cycle and apoptosis of gallbladder carcinoma was detected by flow cytometry.The effects of CP-31398 on the migration and invasion of gallbladder carcinoma cells were detected by scratch,cytoskeleton and Transwell assay.The effect of CP-31398 on the expression of p53 protein was detected by Western blot and the related mechanism was discussed.Results:In mutant P53GBC-SD cells,the proliferation of CCK8 cells showed that the proliferation rate of gallbladder cancer cells in CP-31398 treatment group was slower than that in control group(P＜0.01).The cycle experiment showed that compared with the control group,the proportion of cells in S phase in-creased in CP-31398 treatment group,and the proportion of cells in G0 and G1 phase decreased,so the cells were blocked in S phase(P＜0.001).Apoptosis experiment showed that the apoptosis rate of control group was lower than CP-31398 treatment group(P＜0.05).Transwell experiment showed that the number of cells passing through the cell compartment in the control group was higher than that in the CP-31398 treatment group.Western blot analysis showed that the expression of p53 pro-tein in CP-31398 treatment group was increased(P＜0.05).In wild-type p53NOZ cells,the prolifera-tion of CCK8 cells showed that the proliferation rate of gallbladder cancer cells in CP-31398 treat-ment group was slower than that in control group.The number of cell clones in the control group was higher than that in the CP-31398 treatment group(P＜0.001).The cycle test showed that the propor-tion of cells in G2 and M phases increased significantly,while the proportion of cells in G0 and G1 phases decreased,and the cells were blocked in G2 and M phases(P＜0.001).Apoptosis experiment showed that the apoptosis rate of control group was(14.04±3.08)％,and that of CP-31398 treat-ment group was(34.55±3.30)％(P＜0.01).Transwell experiment showed that the number of cells passing through the cell compartment in the control group was higher than that in the CP-31398 treatment group.Western blot analysis showed that the expression of p53 protein in CP-31398 treatment group was increased(P＜0.01).Conclusion:CP-31398 can effectively inhibit the prolif-eration and metastasis of gallbladder cancer cells,and is independent of p53 mutation,and CP-31398 has a good effect on NOZ gallbladder cancer cell lines expressing wild-type p53,which can be used as a new drug treatment for gallbladder cancer.

Objective To understand the disease acceptance status and related factors in human immunodeficiency virus(HIV)-infected/acquired immunodeficiency syndrom(AIDS)patients,so as to guide the clinical development of intervention measures,and to provide empirical evidence for improving clinical outcomes.Methods Convenience sampling method was used to select 555 HIV-infected/AIDS patients who received treatment in the designated AIDS treatment clinic of a hospital.General data,disease acceptance,disease self-management efficacy and clinical out-comes(such as quality of life,CD4+T lymphocyte count and HIV viral load)of the studied subjects were collected.Results The average disease acceptance of HIV-infected/AIDS patients was(26.08±5.34)points.Multiple linear regression analysis showed that religious belief and self-management efficacy were related factors affecting the di-sease acceptance of patients(both P＜0.05),which could explain the 30.4％variation in disease acceptance of HIV-infected/AIDS patients,and the disease acceptance of patients was closely related to their quality of life(P＜0.001).Conclusion HIV-infected/AIDS patients have a moderate level of disease acceptance.Medical staff should fully consider patients'religious beliefs and self-management efficacy,so as to formulate targeted intervention mea-sures to improve patients'acceptance of disease,and further promote patients'quality of life.

Objective To establish a nomogram model for efficiently predicting overall survival(OS)and cancer-specific survival(CSS)in patients with CRCHBM.Method 2239 patients from 2010 to 2019 were retrospectively analyzed from the Surveillance,Epidemiology,and End Results Program(SEER)databases and Wuhan Union Hospital Cancer Center.SEER is randomly assigned to the training and internal validation cohorts,and the Wuhan database serves as the external validation.Cox regression analyses were used to determine the independent clinicopathological prognosis factors affecting OS and CSS,and a nomogram was constructed to predict OS and CSS.The clinical utility of columnar plots was assessed using calibration curves,area under the curve(AUC),and decision curve analysis(DCA).Result OS column line graphs were constructed based on nine independent predictors:age,tumor location,degree of differentiation,tumor size,TNM stage,chemotherapy,primary focus surgery,number of lymph nodes sampled,and serum carcinoembryonic antigen(CEA)level.The C-index of the nomogram to predict the 1-,3-,and 5-year OS were 0.764,0.790,and 0.805 in the training group,0.754,0.760,and 0.801 in the internal validation group,and 0.822,0.874,and 0.906 in the external validation group.CSS column line graphs were constructed based on 3 independent predictors of TNM staging,radiotherapy and chemotherapy.The 1-,3-,and 5-year CSS AUROC values of the training group were 0.791,0.757,and 0.782,respectively.0.682,0.709,0.625 in the internal validation group and 0.759,0.702,0.755 in the external validation group,respectively.The results of receiver operating characteristic curve(ROC),ROC and DCA showed that the use of our model was more effective in predicting OS and CSS than other single clinicopathological features.Conclusion In summary,the nomogram based on significant clinicopathological features can be conveniently used to predict OS and CSS individually in patients with CRCHBM.

Objective To design an adjustable and removable nursing support for dressing patients with lower extremity injuries.Methods The adjustable nursing support was composed of a supporting plate,a cylinder,an upper adjustment mechanism and a lower fixation mechanism.The supporting plate was used to hold the leg of the patient,which had a curved st ruc t u re with a length from 40 to 80 cm;the cylinder was internally snap-fitted with a second slip sleeve to facilitate the adjustment of the support plate;the upper adjustment mechanism mainly consisted of a second sliding bar,a second cross bar and an adjustment plate;the lower fixation mechanism was mainly composed of a first clamping plate and a second clamping plate.Results The adjustable nursing support could be firmly fixed on the sickbed,and its height and angle could be adjusted according to the patient's wound position and subjective comfort.Conclusion The adjustable nursing support gains advantages in safety and patient comfort,and can be used for the dressing of patients with lower extremity injuries.[Chinese Medical Equipment Journal,2024,45(8):110-112]

Aesculus chinensis is an important medicinal and horticultural plant. Its dried mature seeds, known as "Suoluozi", are a well-known traditional Chinese medicine. Aescins are its main active components, possessing multiple pharmacological activities such as anti-inflammatory and anti-exudative effects. They are widely used in the treatment of diseases such as lumbar disc herniation, postoperative edema, and sports injuries, leading to a continuous increase in market demand in recent years. The DNA binding with one finger(Dof) family is a unique transcription factor family found in the plant kingdom. It plays a crucial role in plant growth, development, stress responses, and regulation of secondary metabolism. However, research on the Dof gene family in A. chinensis is relatively scarce. In this study, we identified 36 AcDof genes from the genome of A. chinensis and analyzed their physicochemical properties, chromosomal localization, phylogenetic relationships, gene structures, cis-acting elements, and expression patterns in different tissues. The results showed that AcDof proteins ranged from 81 to 493 amino acids in length, with molecular weights ranging from 9 270.38 to 55 015.68 and isoelectric points ranging from 4.84 to 10.2. The subcellular localization analysis revealed that 34 AcDof proteins were located in the nucleus, while the remaining two AcDof proteins were located in the chloroplasts. Phylogenetic analysis divided AcDofs into nine subgroups, and gene structure analysis indicated that all AcDof genes possessed a C2-C2 type single zinc finger domain. Gene expression analysis using transcriptome data revealed tissue-specific expression patterns among AcDof family members. Specifically, AcDof04, AcDof02, and AcDof03 exhibited specific expression in seeds, suggesting their potential involvement in the regulation of aescin biosynthesis. This study not only enhances our understanding of the Dof gene family in A. chinensis but also provides important genetic resources for further investigation of the functions and regulatory mechanisms of Dof genes in this species.
Plant Proteins/metabolism* ; Phylogeny ; Gene Expression Regulation, Plant ; Transcription Factors/metabolism* ; Gene Expression Profiling ; Multigene Family

OBJECTIVE: To explore influence of external factors of wind, cold and dampness on clinical symptoms in knee osteoarthritis (KOA) patients with different constitutions of traditional Chinese medicine. METHODS: A cross-sectional stratified study was performed to select 108 patients with GradeⅡKOA in Kellgren & Lawrence (K-L) classification, including 22 males and 86 females, aged from 47 to 75 years old with an average of (60.7±6.0) years old;body mass index(BMI) ranged from 17.87 to 31.22 kg·m^-2 with an average of (23.80±2.86) kg·m^-2. According to Classification and Judgment of TCM Physique (ZYYXH/T157-2009), the types of TCM physique were determined and divided into 4 layers according to the deficiency and actual physique. Among them, there were 24 patients without biased physique, 12 males and 12 females, aged from 51 to 73 years old with an average of(62.8±6.0) years old, BMI ranged from 17.87 to 31.14 kg·m^-2 with an average of (24.32±3.25) kg·m-2;there were 46 patients with virtual bias constitution, including 7 males and 39 females, aged from 47 to 70 years old with an average of (60.0±5.8) years old, BMI ranged from 19.38 to 31.22 kg·m^-2 with an average of(23.42±2.97) kg·m^-2;There were 26 patients with solid bias constitution, including 2 males and 24 females, aged from 48 to 75 years old with an average of (60.4±5.8) years old, BMI ranged from 21.16 to 30.76 kg·m^-2 with an average of (24.15±2.33) kg·m^-2;there were 9 patients with special constitution, 1 male and 8 female, aged from 53 to 75 years old with an average of (59.8±7.5) years old, BMI ranged from 19.26 to 26.67 kg·m^-2 with an average of (23.79±2.49) kg·m^-2. Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) was used to evaluate severity of clinical symptoms. The wind-cold-dampness external factor score was calculated through the questionnaire of wind-cold-dampness syndrome scale to evaluate degree of influence of wind-cold-dampness external factor. Pearson correlation analysis and partial correlation analysis were used to calculate the correlation coefficient between severity of external factors affecting wind, cold and dampness and severity of clinical symptoms in patients with different TCM constitution stratification. RESULTS: There was no statistical significance between total score of wind-cold-dampness and WOMAC score in patients with no biased constitution and special condition. Total wind-cold-dampness score of patients with virtual biased constitution was positively correlated with WOMAC stiffness score (r=0.327, P=0.032), and total wind-cold-dampness score of patients with solid biased constitution was positively correlated with WOMAC pain score (r=0.561, P=0.005) and WOMAC overall score (r=0.446, P=0.033). After further adjusting for the interaction of external factors of wind-cold-dampness, there was no statistical significance between wind-cold-dampness scores and WOMAC scores in patients with solid biased constitution. The score of dampness and pathogenic factors was positively correlated with WOMAC stiffness score (r=0.414, P=0.007). CONCLUSION The external factors of wind-cold dampness have different effects on the clinical symptoms of KOA patients with different TCM constitutions. Compared with other constitutions, the rigid symptoms of patients with asthenic biased constitutions are more susceptible to dampness pathogenic factors.
Aged ; Female ; Humans ; Male ; Middle Aged ; Cross-Sectional Studies ; Medicine, Chinese Traditional ; Osteoarthritis, Knee ; Syndrome ; Wind ; Cold Temperature