OBJECTIVE To optimize ethanol extraction process of Yihuang powder. METHODS An orthogonal experiment was designed by reflux extraction with ethanol volume fraction， liquid-to-material ratio， and extraction time as investigation factors. The parameters used were the contents of hesperidin， nobiletin， tangeretin， gallic acid， chebulagic acid， chebulinic acid， liquiritin， glycyrrhizin， eugenol， and the paste-forming rate. The analytic hierarchy process （AHP） was used to calculate the comprehensive score. The optimal ethanol extraction process parameters of Yihuang powder were determined by verifying the results predicted by orthogonal experiment and genetic algorithm （GA）-back propagation neural network （BP neural network）. RESULTS The optimal ethanol extraction process parameters， as optimized by orthogonal experiment， were as follows: ethanol volume fraction of 60%， liquid-solid ratio of 14∶1 （mL/g)， extraction time of 90 min， and extraction for 2 times. The comprehensive score obtained by verification was 79.19. Meanwhile， the optimal ethanol extraction process parameters， optimized by GA-BP neural network， were ethanol volume fraction of 65%， liquid-solid ratio of 14∶1 （mL/g )， extraction time of 60 min， and extraction for 2 times. The comprehensive score obtained by verification was 85.30， higher than the results obtained from orthogonal experiment. CONCLUSIONS The optimization method of orthogonal experiment combined with GA-BP neural network is superior to the traditional orthogonal experiment optimization method. The optimized ethanol extraction process of Yihuang powder is stable and reliable.

Objective:To explore the effect of communication competence and psychological resilience on job burnout among Operating Room nurses.Methods:From March to June 2023, randomized clustering sampling was used to select 138 registered Operating Room nurses from four ClassⅢ Grade A hospitals in Taizhou for investigation. The survey was conducted using the general information questionnaire, Operating Room Nurses' Job Stressor Scale, Chinese version of the Connor-Davidson Resilience Scale, Nurses' Clinic Communication Competence Scale, and Maslach Burnout Inventory-General Survey. Hierarchical linear regression analysis was used to explore the effects of communication competence and psychological resilience on job burnout among Operating Room nurses.Results:A total of 138 questionnaires were sent out, and 133 valid questionnaires were collected, with a valid response rate of 96.38% (133/138). Among 133 Operating Room nurses, the job burnout score was (56.35±9.28), and the communication competence, psychological resilience, and work stress scale scores were (196.71±18.92), (78.09±18.31), and (96.37±22.47), respectively. Pearson correlation showed that job burnout among Operating Room nurses was negatively correlated with psychological resilience ( r=-0.475, P<0.01) and communication competence ( r=-0.241, P<0.01), and positively correlated with work stress ( r=0.360, P<0.01). Hierarchical linear regression analysis showed that, after controlling for other variables, psychological resilience and communication competence were the influencing factors of job burnout among Operating Room nurses ( P<0.01), which could explain 17.70% of the variation. Conclusions:The level of job burnout among Operating Room nurses is relatively high, and psychological resilience and communication competence are independent influencing factors. Managers can provide psychological counseling and support services for Operating Room nurses, offer communication competence training programs, and prevent and reduce job burnout among Operating Room nurses.

Objective To explore and analyze the prescription patterns of Professor Zhou Min in treating primary liver cancer at different stages according to the China Liver Cancer Staging (CNLC) system. Methods The clinical records of outpatients with primary liver cancer treated by Professor Zhou Min were collected and entered into the Traditional Chinese Medicine Inheritance Support System (Version 2.50) to establish a database. Data mining methods such as frequency analysis, drug association analysis, and cluster analysis were employed, the pathogenesis of primary liver cancer the prescription patterns at different stages was explored and medication rules were analyzed according to Professor Zhou Min's experience in treating liver cancer at various CNLC stages. Results A total of 202 prescriptions from 113 patients with primary liver cancer were collected, involving 230 traditional Chinese medicines. The high-frequency drugs and drug combinations at each stage were identified. The drugs with higher frequencies at each stage included Fuling, Chenpi, Yiyiren, fried Baishu, and Fabanxia. For stage Ⅰ, high-frequency drugs also included Zhongjiefeng, Xiangfu, Jiangcan, and Jianghuang. For stages Ⅱ and Ⅲ, high-frequency drugs further encompassed Zhongjiefeng, Xianhecao, Banzhilian, Baihua Sheshecao, Jiangcan, Zeqi, Xiangfu, and Maidong. For stage Ⅳ, high-frequency drugs also include Maydis stigma, Huoxiang, fried Maiya, Jineijin, and fried Guya. The majority of the drugs were cold in nature, with sweet and bitter tastes being the most common, and their meridian tropism were mostly distributed in the spleen and stomach meridians. The drug combinations with higher frequencies at each stage were mostly derived from Sijunzi Decoction and Erchen Decoction. The drug efficacies were mainly heat-clearing and dampness-resolving. Cluster analysis screened out new prescriptions with unique characteristics at each stage. Conclusion By performing data mining on the prescriptions used by Professor Zhou Min in treating primary liver cancer at various CNLC stages through the Traditional Chinese Medicine Inheritance Platform, combined with his understanding of the pathogenesis and clinical experience of the disease, the pathogenesis characteristics of primary liver cancer are summarized as dampness-heat, phlegm, and toxin accumulation, as well as qi and yin deficiency. The basic treatment methods established are heat-clearing and dampness-resolving, spleen-invigorating and yin-nourishing, with an emphasis on strengthening the body resistance to eliminate pathogenic factors and stage-based treatment. Flexible prescriptions and medications are used for different complications.

Flavanone 3-hydroxylase (F3H) is a key enzyme in the synthesis of phycocyanidins. In this experiment, the petals of red Rhododendron hybridum Hort. at different developmental stages were used as experimental materials. The R. hybridum flavanone 3-hydroxylase (RhF3H) gene was cloned using reverse transcription PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) techniques, and bioinformatics analyses were performed. Petal RhF3H gene expression at different developmental stages were analyzed by using quantitative real-time polymerase chain reaction (qRT-PCR). A pET-28a-RhF3H prokaryotic expression vector was constructed for the preparation and purification of RhF3H protein. A pCAMBIA1302-RhF3H overexpression vector was constructed for genetic transformation in Arabidopsis thaliana by Agrobacterium-mediated method. The results showed that the R. hybridum Hort. RhF3H gene is 1 245 bp long, with an open reading frame of 1 092 bp, encoding 363 amino acids. It contains a Fe²⁺ binding motif and a 2-ketoglutarate binding motif of the dioxygenase superfamily. Phylogenetic analysis showed that the R. hybridum RhF3H protein is most closely related to the Vaccinium corymbosum F3H protein. qRT-PCR analysis showed that the expression level of the red R. hybridum RhF3H gene tended to increase and then decrease in the petals at different developmental stages, with the highest expression at middle opening stage. The results of the prokaryotic expression showed that the size of the induced protein of the constructed prokaryotic expression vector pET-28a-RhF3H was about 40 kDa, which was similar to the theoretical value. Transgenic RhF3H Arabidopsis thaliana plants were successfully obtained, and PCR identification and β-glucuronidase (GUS) staining demonstrated that the RhF3H gene was integrated into the genome of A. thaliana plants. qRT-PCR, total flavonoid and anthocyanin contentanalysis showed that RhF3H was significantly higher expressed in the transgenic A. thaliana relative to that of the wild type, and its total flavonoid and anthocyanin content were significantly increased. This study provides a theoretical basis for investigating the function of RhF3H gene, as well as for studying the molecular mechanism of flower color in R. simsiib Planch.
Arabidopsis/metabolism* ; Rhododendron/metabolism* ; Amino Acid Sequence ; Anthocyanins/metabolism* ; Phylogeny ; Flavonoids/metabolism* ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism*

OBJECTIVE To compare the similarities and differences between raw and different preparations of Terminalia chebula based on fingerprint， antioxidant spectrum-effect correlation and multi-component contents， and to provide a reference for searching for modern processing methods of T. chebula that are similar to classical ancient methods. METHODS Ten batches of raw and different preparations of T. chebula （single stir-fried products， bran-roasted products， sand-scorched products， ash-roasted products， stir-fried charcoal products， and wine-steamed products） were used as test samples. The high-performance liquid chromatography （HPLC） fingerprints of different samples were established by using the Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）， the chromatographic peaks were identified， and chemometrics analysis was carried out. At the same time， HPLC method was used to determine the contents of 8 identified components. The antioxidant capacity of raw and different preparations of T. chebula was determined by DPPH free radical scavenging method， and the spectrum- effect relationship was analyzed. RESULTS A total of 20 common peaks were identified in the fingerprints of the raw and different preparations of T. chebula， and the similarity of each sample was ＞0.9. Nine common peaks were identified from the raw and different preparations of T. chebula， including chromatographic peak 2 （chebulic acid）， 3 （gallic acid）， 6 （punicalagin A）， 8 （punicalagin B）， 12 （corilagin）， 15 （chebulagic acid）， 18 （ellagic acid）， 19 （1，2，3，4，6-O-pentagalloyl glucose）， 20 （chebulinic acid）， etc. Compared with crude drug， the contents of the above 8 components （punicalagin A and B are recorded as punicalagin） in different preparations of T. chebula were changed， and the changes of the contents of the stir-fried charcoal and wine-steamed products were more obvious than those of other processed products. Chemometric analysis showed that the fingerprints of stir-fried charcoal and wine-steamed products of T. chebula were obviously distinguished from other processed products， and the fingerprint information of raw products and other processed products of T. chebula was partially overlapped. Four main differential components （chebulinic acid， chebulagic acid， gallic acid， ellagic acid） were obtained between raw and processed products of T. chebula； and four main effective components （chebulinic acid， chebulagic acid， gallic acid， corilagin） were obtained by analyzing the spectrum-effect relationship of antioxidant activity. The single stir-fried product of T. chebula showed the strongest antioxidant activity. CONCLUSIONS The single stir-frying method is a modern processing method of T. chebula which is similar to the classical ancient method and is more excellent.

Phenylalaninammo-nialyase (PAL) is a key enzyme in the synthesis of methyl benzoate - a plant aroma compound. In order to understand the function of this enzyme in the formation of fragrance in the scented Rhododendron species-Rhododendron fortunei, we cloned a gene encoding this enzyme and subsequently examined the gene expression patterns and the profile of enzyme activity during development in various tissues. The full length of RhPAL gene was cloned by reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The expression levels of RhPAL gene were measured by real-time quantitative reverse transcription PCR (qRT-PCR) and the amount of phenylalanine and cinnamic acid were assayed with LC-MS. The results showed that the ORF sequence of RhPAL gene amplified from the cDNA templates of flower buds had 2 145 bp, encoding 715 amino acids, and shared 90% homology to the PAL amino acid sequences from other species. qRT-PCR analysis showed that the expression of RhPAL in petals during flowering kept in rising even until the flowers wilted. The expression of RhPAL in pistil was much higher than that in stamen, while the expression in the younger leaves was higher than in old leaves. However, the expression level was relatively lower in petal and stamen compared to that in leaves. We also measured the PAL activity by Enzyme-linked immuno sorbent assay in the petals of flowers at different flowering stages. The results showed that PAL activity reached the highest at the bud stage and then decreased gradually to the lowest when the flowers wilted, which followed a similar trend in the emission of the flower fragrance. The phenylalanine and cinnamic acid contents measured by LC-MS were highly correlated to the expression level of RhPAL in various tissues and at different flowering stages, implying that RhPAL plays an important role in the formation of the flower fragrance. This work may facilitate the breeding and improvement of new fragrant Rhododendron cultivars.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; Flowers/genetics* ; Rhododendron/genetics*

Objective: To create a model to predict nosocomial infections in emergency intensive care units (EICU), so as to provide insights into early identification and interventions among patients with nosocomial infections. Methods: All nosocomial infections were collected from patients hospitalized in the EICU of a large tertiary hospital from 2017 to 2020. The 2017-2019 data were selected as the training set to create a logistic regression model, and the fitting effectiveness of the predictive model was evaluated using Hosmer-Lemeshow test. The 2020 data were selected as the test set to evaluate the external validation of the predictive model. In addition, the value of the model for prediction of nosocomial infections was examined using the receiver operating characteristic (ROC) curve analysis. Results : Totally 1 546 inpatients in EICU were enrolled, and the prevalence of nosocomial infections was 7.18%. Multivariable logistic regression analysis identified hospital stay duration of >7 days (OR=21.845, 95%CI: 7.901-60.398), use of ventilators (OR=3.405, 95%CI: 1.335-8.682), and surgery (OR=1.854, 95%CI: 1.121-3.064) as risk factors of nosocomial infections. The predictive model was p=ey/(1+ey), y=-6.105+(3.084×duration of hospital stay)+(1.225×use of ventilators)+(0.617×surgery). The area under ROC curve was 0.806 (95%CI: 0.774-0.838) for the training set and 0.723 (95%CI: 0.623-0.823) for the test set, and if the 0.065 cut-off of the predictive model created by the training set was included in the test set, the predictive value yield a 0.739 sensitivity and 0.642 specificity for prediction of nosocomial infections among patients hospitalized in EICU. Conclusion The created predictive model for nosocomial infections among patients hospitalized in EICU presents a high accuracy, which shows a satisfactory predictive value for high-risk nosocomial infections.

Terpene synthase (TPS) plays important roles in the synthesis of terpenoids which are the main fragrances in Rhododendron flowers. To understand the function of TPS genes in terpenoid metabolism in relation to flower aroma formation, we identified all TPS gene family members in Rhododendron by analyzing its genome database. We then used a transcriptomic approach to analyze the differential gene expression patterns of TPS gene family members in the scented flower Rhododendron fortunei compared to the non-scented flower Rhododendron 'Nova Zembla'. The contents of terpenoid compounds in petals of the above two Rhododendron species at different developmental stages were also measured by using qRT-PCR and head space-solid phase micro-extraction combined with gas chromatography-mass spectrometry. Our results showed that a total of 47 RsTPS members, with individual lengths ranged from 591 to 2 634 bp, were identified in the Rhododendron genome. The number of exons in RsTPS gene ranged from 3 to 12, while the length of each protein encoded ranged from 196 to 877 amino acids. Members of the RsTPS family are mainly distributed in the chloroplast and cytoplasm. Phylogenetic analysis showed that RsTPS genes can be clustered into 5 subgroups. Seven gene family members can be functionally annotated as TPS gene family since they were temporally and spatially expressed as shown in the transcriptome data. Notably, TPS1, TPS10, TPS12 and TPS13 in Rhododendron fortunei were expressed highly in flower buds reached the peak in the full blossoming. Correlation analysis between gene expression levels and terpenoid content indicates that the expression levels of TPS1, TPS4, TPS9, TPS10, TPS12 and TPS13 were positively correlated with the content of terpenoids in the petals of R. fortunei at all flower developmental stages, suggesting that these six genes might be involved in the aroma formation in R. fortunei.
Rhododendron/metabolism* ; Phylogeny ; Terpenes/metabolism* ; Family ; Gene Expression Regulation, Plant

Objective: To assess the use of statins and low-density lipoprotein cholesterol (LDL-C) levels at admission in hospitalized patients aged 75 years and older with acute coronary syndrome (ACS) in China. Methods: Data used in this study derived from the Improving Care for Cardiovascular Disease in China (CCC)-ACS project, a nationwide registry with 150 tertiary hospitals reporting details of clinical information of ACS patients. This study enrolled patients 75 years and older with ACS in CCC-ACS project from November 2014 to June 2017. Patients were divided into two groups according to the history of atherosclerotic cardiovascular disease (ASCVD). Pre-hospital statin use, LDL-C levels at admission and prescription of statins at discharge were reported. Results: A total of 10 899 patients 75 years and older with ACS were enrolled. The median age was 79 years and 58.7% (6 397 cases) were male. Among patients with history of ASCVD, 33.9% (1 028 cases) of them received statins before hospitalization. Among patients without history of ASCVD, 12.7% (996/7 871) received statins before hospitalization. The mean level of LDL-C was (2.4±0.9) mmol/L and LDL-C was <1.8 mmol/L in 24.7% (747 cases) of patients with history of ASCVD. The mean level of LDL-C was (2.6±0.9) mmol/L and LDL-C was <2.6 mmol/L in 51.7% (4 072 cases) of patients without history of ASCVD. At discharge, 91.2% (9 524/10 488) of patients were prescribed with statins in patients without contraindications for statin. Conclusion In elderly patients with recurrent ASCVD, there was an inadequate statin use before hospitalization and most patients did not reach the LDL-C target level when they had the recurrent events. In the elderly ACS patients without history of ASCVD, more than half of the patients had an ideal LDL-C level. It seems that ideal LDL-C level for primary prevention of ACS in elderly people needs to be reevaluated with further studies.

Objective: To evaluate the current status and related factors of influenza vaccination among health care workers (HCWs) in tertiary hospitals of Xining city after the implementation of the free influenza vaccination policy. Methods: In August 2018, the cluster sampling method was used to select four medical institutions in Xining that had previously conducted investigations and interventions. All HCWs(excluding logistic staff) in each medical institution were included in the study. A total of 3 260 valid respondents were included. Questionnaires were used to collect the demographic characteristics, influenza and influenza vaccination awareness, implementation of free policy in the influenza epidemic season from 2017 to 2018, influenza vaccination status, awareness of influenza vaccination schedule and free policy. The multivariate logistic regression model was used to analyze related factors of influenza vaccination. Results: The age of respondents was (31.41±5.00) years. The influenza vaccination rate was 6.80% (226/3 260) in 2017-2018 influenza epidemic season. After controlling for related factors, the awareness of the influenza vaccination schedule (OR=17.05, 95%CI: 5.86-49.59), vaccination frequency (OR=8.22, 95%CI: 2.98-22.61) and the free policy (OR=3.15, 95%CI: 1.49-6.67) had higher vaccination rate. Conclusion The influenza vaccination rate of HCWs in tertiary hospitals of Xining city was low. Increasing the awareness of the vaccination schedule, frequency and free policy may promote the influenza vaccination rate of HCWs.