BACKGROUND:Cationic liposome-mediated celltransfection is reliable and repeatable. However the transfection efficiency is often low.
OBJECTIVE:To study the optimized methods for gene transfection mediated by liposome into N2a cells (mouse neuroblastma cells).
METHODS:Using traditional adherent method and improved suspension method, 500 ng recombinant plasmid pcDNA3-GFP carrying green fluorescence protein was transfected into N2a cells in 24-wel culture plate, which was mediated by 1.5μL Lipofectamine?LTX Reagent. The expression of green fluorescent protein was observed by inverted fluorescence microscope, and the transfection efficiencies at different transfection ways were calculated. By using improved suspension transfection method, 500 ng plasmid DNA was transfected with different doses of Lipofectamine?LTX Reagent (1.0, 1.5, 2.0, 2.5μL). The optimal ratio of liposome and DNA was explored.
RESULTS AND CONCLUSION:The transfection efficiency of suspension transfection method was significantly higher than that of the tranditional adherent method (P<0.01) when using 1.5μL liposome/500 ng DNA. The transfection efficiency of the 1.0, 1.5, 2.0, 2.5μL Lipofectamine?LTX on 500 ng plasmid DNA was respectively (76.60±3.85)%, (80.00±4.17)%, (88.00±5.89)%, (54.96±4.23)%. It showed the 500 ng DNA and 2.0μL liposome achieve the highest transfection efficiency.

Objective To set up the quality standard of Shiwei Shujin Huoluo Powder. Methods Rhizoma Polygoni cuspidati and Cortex Cinnamomi, the main ingredients of the powder, were identified by thin layer chromatography ( TLC) , and the content of emodin in the powder was detected by high performance liquid chromatography (HPLC) . Results The spots showed by TLC were clear without interference of the negative control. HPLC results showed that linear range of emodin was 9.30 ~ 46.52 μg·mL-1, the recovery of emodin was 96.22%, and RSD was 1.76% ( N=6) . Conclusion The established method is reliable and accurate, and can be used for quality control of Shiwei Shujin Huoluo Powder.

Objective To measure the present situation of medical students' learning adaptability and find the influencing factors in order to revise Learning Adaptability Scale for medical students.Methods A stratified sampling method was used to collect 1 180 medical students from Harbin Medical University.Using the internal consistency reliability test,the reliability of the revised student learning adaptability scale was verified.Five common factors were extracted by the exploratory factor analysis of the principal analysis method,and Logistic regression was used to analyze the influencing factors of medical student learning adaptability.Results The revised Student Learning Adaptability Scale (Cronbach's Alpha=0.914) consists of 27 measurement items.It includes five dimensions,namely self-directed learning,information utilization,environment choice,stresses response and goal orientation.The results showed that the adaptability of medical students was moderate (3.28 ± 0.52),and the influencing factors included job position (OR =1.496,95％ CI=1.114-2.009),academic achievement (OR =1.638,95 ％ CI=1.386-1.936),satisfaction with learning (OR=2.160,95％CI=1.745-2.673),professional satisfaction (OR=1.369,95％CI=1.092-1.718),professional interest (OR =1.426,95％ CI=1.186-1.715) and employment prospects (OR =1.526,95％ CI=1.251-1.862).Conclusion The reliability and validity of the Student Learning Adaptability Scale is appropriate.Medical students' learning adaptability is influenced by many factors.

Objective:To construct of evaluation index system and theory-skill-humanity (TSH) model for core competence of radiation specialist nurses, to provide reference for clinical training of radiation specialist nurses.Methods:In January 2023, the draft of the evaluation index system of core competence for radiation specialist nurses was development by literature reviews and semi-structured interviews; from February to March 2023, 20 experts were consulted with 2 rounds of Delphi consultations to determine the index system and determine indicator weights based on the importance assigned by experts to each indicator.Results:Among 20 experts, 17 were female and 3 were male, aged 35-65 (46.48 ± 8.19) years old. The effective recovery rate of the questionnaires was 100.0% (20/20) and 95.0% (19/20) in the first and second round, respectively. The authority coefficient of the experts was 0.937 in the second round, the Kendall coefficients of concordance in the first-level indexes, second-level indexes and third-level indexes were 0.448, 0.407, 0.464, respectively, the difference was statistically significant ( χ2 = 53.60, 148.81, 58.18, all P<0.001). The final evaluation index system of the core competence of radiation specialist nurses included 3 first-level indicators, 11 second-level indicators and 56 third-level indicators. Conclusions:The evaluation index system of the core competence of radiation specialist nurses constructed based on TSH model in the present study has certain feasibility and practicality, which can be used as the basis for the training and evaluation of radiation specialized nurses.

Objective:To explore the differences in clinical characteristics and treatment outcomes between patients with type A and type B alcohol dependence, and to find the independent risk factors of relapse.Methods:Alcohol-dependent male patients attending the Addiction Medicine Center of Beijing Huilongguan Hospital from January 2018 to December 2020 were selected for the study and divided into type A alcohol-dependent group ( n=77) and type B alcohol-dependent group ( n=87). All patients were given acute detoxification treatment and were followed up after treatment on relapse to drinking. Differences in demographic and clinical data were compared between the two groups, and differences in treatment outcomes between the two groups at different time points over 3 months were compared. Patients were divided into relapse group and non-relapse group according to whether they drank again after 3 months. Logistic regression model was established to screen the risk factors of relapse of alcohol-dependent patients by SPSS 25.0 software. Results:There was no significant difference between the two types of patients in years of education, marital status, smoking status and working status(all P>0.05), but the proportion of co-residents( χ2=5.69, P=0.017) and the proportion of positive family history of alcoholism were significant difference between the two type of patients( χ2=13.32, P<0.001). There were statistically significant differences between the two types of patients in the onset time( t=-7.28, P<0.001), the first drinking age( t=-2.36, P=0.020), the proportion of drinking in the morning( χ2=7.83, P=0.005), psychotic symptoms( χ2=4.31, P=0.038), convulsions after withdrawal( χ2=5.30, P=0.021), and alcohol use disorder identification test(AUDIT) score( t=4.30, P<0.001). At the 4th and 8th weekend of the follow-up, there were statistically significant differences in drinking frequency(0(0, 3), 0(0, 0), Z=-4.13, P<0.001; 3(0, 3), 0(0, 3), Z=-4.42, P<0.001) and relapse rate (40(45.98%), 9(11.69%), χ2=22.92, P<0.001; 61(70.11%), 24(31.17%), χ2=24.82, P<0.001) between the two types of alcohol dependence patients after drinking again. After 12-week follow-up, there were statistically significant differences between the two types of alcohol-dependent patients in the interval of first drinking(20(7, 30)d, 88(38, 90)d, Z=-7.83, P<0.001), the cumulative duration of abstinence(4(0, 8)weeks, 12(4, 12)weeks, Z=-5.13, P<0.001), the cumulative rate of abstinence(71(81.60%), 25(32.47%), χ2=40.62, P<0.001), the frequency of drinking after abstinence(3(3, 3), 0(0, 3), Z=-5.54, P<0.001), and the reduction of daily average alcohol consumption( t=3.36, P<0.001). Logistic regression model showed that type B alcohol dependence ( OR=3.121, P=0.03, 95% CI: 1.12-8.72) and AUDIT score ( OR=1.498, P<0.01, 95% CI: 1.29-1.74) were the risk factors for relapse of alcohol-dependent patients. Conclusions:Patients with type A and type B alcohol dependence have obvious differences in clinical characteristics and treatment outcomes, and type B alcohol dependence is independent risk factor for relapse to drinking in alcohol-dependent patients, which validate the rationality and necessity of alcohol dependence subtypes.

Objective To explore the situation of sedentary behavior and how it correlated to overweight/obesity among children and adolescents in Chengdu,China.Methods A total of 2 211 children and adolescents aged 7-15 years old were recruited in Chengdu city by using the cluster random sampling method.This was a cross-sectional study which aimed to collect data on anthropometry including height and weight and information on basic characters and sedentary behavior by using a self-designed questionnaire.Results According to IOTF criteria,the reporting rate of factor as ‘watching TV ≥2 hours per day on weekend’ showed statistically significant differences among normal,overweight and obese groups (34.80％,4.21％ and 1.92％ respectively,P=0.016 0) in boys.The reporting rates of ‘doing homework ≥2 hours per day’ were 12.17％,0.89％ and 0.49％ respectively (P=0.036 0) and statistically different among the three groups,in girls.Similar results were obtained in boys based on the WGOC criteria.However,there was no statistical difference seen on sedentary behaviors among the three groups in girls according to WGOC criteria.After stratified by age groups,rates on ‘ watching TV on weekend’ were statistically different among normal,overweight and obese group in 13-15 age group in girls (P=0.035 1).Physical activity level (PAL) was positively associated with WHR among both boys and girls when grouped according to PAL (Schofield).However,it was positively associated with WHR only seen in boys according to Chinese Nutrition Society (CNS) criteria.Conclusion The reporting rates of sedentary behavior in obese children were higher than that in overweight/normal ones.However,the correlation of sedentary behaviors on overweight/obesity called for further studies.

Objective To develop Chinese Children Dietary Index,the overall diet quality of children and adolescents; the association between overall diet quality and overweight/obesity among children in Chengdu.Methods Chinese Children Dietary Index was formulated based on the current Chinese Dietary Guidelines and Food Guide Pagoda for children and adolescents,consisting of 12components.1 997 children and adolescents aged 7-15 years in Chengdu were recruited and their diet quality was evaluated according to Chinese Food Guide Pagoda.Differences in percent body fat (PBF),fat mass index (FMI),fat-free mass index (FFMI) between 3 categories as low,moderate,and high diet quality,were tested.Results The mean Chinese Children Dietary Index score for our population was 59.36 ± 11.50.The diet quality of 13-15 years old adolescents was the worst.7-9years old children with moderate diet quality had highest PBF (P=0.04),while 10-12 years old children with higher diet quality had the lowest FFMI (P=0.002).Conclusion The overall diet quality of children and adolescents in Chengdu could be reflected by Chinese Children Dietary Index.There was association between overall diet quality and overweight/obesity among children and adolescents.However,this association warrants more in depth evaluation in future studies.

Innovative self-efficacy is the degree of self-confidence in the individual's ability to per-form innovative activities. Although the theory of innovation self-efficacy is shorter, but as an important indicator of innovation ability measurement, it has been accepted by scholars in various countries, and innovative self-efficacy provides a new perspective for the cultivation of innovative talents. The research of innovation self-efficacy is still the initial stage. Chinese and foreign scholars have made some progress in measuring tools, antecedents, aftereffect and development, but there are still obvious differences and defi-ciencies,and the research results are not abundant.The future research will focus on the measurement tools, influencing factors and research direction of three aspects of systematic research to improve the theory of innovation self-efficacy.

Purpose: Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease, and Th17 cells are key factors in the pathogenesis of human inflammatory conditions, such as RA. Catalpol (CAT), a component in Rehmanniae Radix (RR), has been found to regulate human immunity. However, the effects of CAT on Th17 cell differentiation and improvement of RA are not clear. Materials and Methods: Collagen-induced arthritis (CIA) mice were constructed to detect the effects of CAT on arthritis and Th17 cells. The effect of CAT on Th17 differentiation was evaluated with let-7g-5p transfection experiments. Flow cytometry was used to detect the proportion of Th17 cells after CAT treatment. Levels of interleukin-17 and RORγt were assessed by qRT-PCR and enzyme-linked immunosorbent assay. The expression of signal transducer and activator of transcription 3 (STAT3) was determined by qRT-PCR and Western blot. Results: We found that the proportion of Th17 cells was negatively associated with let-7g-5p expression in CIA mice. In in vitro experiments, CAT suppressed traditional differentiation of Th17 cells. Simultaneously, CAT significantly decreased Tregs-to-Th17 cells transdifferentiation. Our results demonstrated that CAT inhibited Tregs-to-Th17 cells transdifferentiation by up-regulating let-7g-5p and that the suppressive effect of CAT on traditional differentiation of Th17 cells is not related with let-7-5p. Conclusion Our data indicate that CAT may be a potential modulator of Tregs-to-Th17 cells transdifferentiation by up-regulating let-7g-5p to reduce the expression of STAT3. These results provide new directions for research into RA treatment.

Objective To evaluate the efficacy and safety of high intensity focused ultrasound(HFIU)combined with endometrial ablation in the treatment of adenomyosis(AM)patients with heavy menstrual bleeding(HMB).Method A total of 199 patients with AM combined with HMB who underwent HIFU treatment at the Obstetrics and Gynecology Center of Affiliated Hospital of Chengdu University of TCM from July 2020 to September 2022 were selected and divided into two groups according to the ablation range.The combined ablation group included 80 cases with lesion and endometrial ablation(endometrial ablation rate≥30%),while the lesion ablation group 119 cases with simple lesion ablation.A 1∶1 propensity score matching was performed on the two groups,resulting in 59 patients in each group.A 6-month follow-up was conducted.The clinical manifestations(menstrual volume,degree of dysmenorrhea,uterine and lesion volume),quality of life(MS-QOL,UFS-QOL),and postoperative adverse reactions between the two groups before and 6 months after surgery was compared.Results Menstrual volume,degree of dysmenorrhea,uterine and lesion volume,MS-QOL score,and UFS-QOL score of the both groups were improved 6 months after surgery,when compared to those before surgery(P<0.05).Significantly reduced menstrual volume,degree of dysmenorrhea,and lower MS-QOL score were observed in the combined ablation group when compared with the lesion ablation group(P<0.05),but there was no statistical significance in uterine and lesion volume and UFS-QOL score(P>0.05).There was no difference in the incidence of adverse reactions between the two groups(P>0.05),but the combined ablation group had longer vaginal bleeding and fluid flow time(P<0.05).Conclusion The combined HIFU and endometrial ablation can effectively reduce menstrual flow,alleviate dysmenorrhea,and improve quality of life in patients with AM combined with HMB.Although it increases vaginal bleeding and fluid flow time,reasonable symptomatic management can effectively prevent the occurrence of adverse consequences such as infection.