Objective:To compare the effectiveness between three methods for purifying the immunoglobulin of egg yolk(IgY) which are polyethylene glycol (PEG) method, chloroform extraction method and chloroform / PEG method, and to provide basis for obtaining the batch of IgY.Methods:The inactivated vaccine of Vibrio parahemolyticus (V. parahemolyticus) was prepared and the hens were immunized by multi-point intramuscular injection.The eggs were collected and the IgY was purified by PEG method, chloroform extraction method and chloroform/PEG method.The protein extraction rate, the IgY titer and the purity of the antibody which purified by different methods were detected.Furthermore, the operation process, cost and safety of the three methods were analyzed.Results:The protein contents of the extraction belonging three methods from high to low in turn were chloroform extraction method, chloroform/PEG method, and PEG method.There was no significant difference in the antibody titer between three methods, and the tiler of chloroform extraction method was slightly high.The purities of purified antibody from high to low in turn were PEG method, chloroform/PEG method and chloroform method.The PEG method had better security but relatively lower extraction efficiency and higher cost.The chloroform/PEG method had high extraction efficiency and good antibody purity.Conclusion:The PEG method is suitable for a small amount of extraction in the laboratory.The chloroform/PEG method is appropriate for extracting the high quality IgY in a batch as it has high extraction efficiency and good antibody purity.

bjective　To analyze the drug resistance screening status and drug resistance influencing factors of high-risk groups of drug-resistant pulmonary tuberculosis in Qingdao, and to understand the inclusion of rifampicin patients in treatment, so as to provide a reference for the prevention and treatment of drug-resistant pulmonary tuberculosis. Methods　The medical records of 726 cases of drug-resistant pulmonary tuberculosis among high-risk populations registered in Qingdao from 2018 to 2022 were obtained from the National Health Insurance Information System of the China Center for Disease Control and Prevention. The drug resistance to five anti-tuberculosis drugs, namely isoniazid (INH), rifampicin (RFP), ethambutol (EMB), levofloxacin (Lfx), and amikacin (Am), in the high-risk populations of drug-resistant pulmonary tuberculosis was analyzed. Univariate and multivariate logistic regression were used toidentify factors influencing rifampicin resistance, and the detection and inclusion of treatment for rifampicin-resistant patients were evaluated. Results　Of the 726 subjects, 278 were drug-resistant, with a total drug resistance rate of 38.29%. The drug resistance for the five anti-tuberculosis drugs in descending order was: INH 25.90%(188/726), RFP 22.87%(166/726), Lfx 14.19%(103/726), EMB 11.29%(82/726), Am 2.48%(18/726). Analysis of the drug resistance spectrum showed that among those resistant to one drug, RFP was most common, accounting for 13.67% (38/278); among those resistant to two drugs, INH+RFP was predominant, accounting for 15.83% (44/278); among those resistant to three drugs, INH+RFP+Lfx was most frequent, at 7.19% (22/278); and among those resistant to four drugs, INH+RFP+EMB+Lfx was highest, at 6.12% (17/278). Multivariate logistic regression analysis of rifampicin resistance showed that compared with patients under 25 years of age, the risk of developing rifampicin resistance was lower in the groups aged 45 to under 65 and those aged 65 and above (OR=0.356, 95%CI: 0.181-0.700; OR=0.352, 95%CI: 0.170-0.729). Compared with migrant patients in other provinces, local patients from within the same county or district had a lower risk of developing rifampicin resistance (OR=0.599, 95%CI:0.383-0.962). Compared with patients who were smear-positive at the end of the second month of initial treatment, the risk of developing rifampicin resistance was higher in patients with relapse/return, failure of retreatment/chronic, and other categories of patients (OR=9.380, 95%CI:3.717-23.671;OR=25.749, 95%CI:8.037-82.490; OR=36.651, 95%CI:8.438-159.201). Conclusions　The situation of drug-resistant pulmonary tuberculosis in Qingdao cannot be ignored. Individuals under 25 years old, migrants from other provinces, and patients with relapse/return, failure of retreatment/chronic, and other categories are significant risk factors for developing rifampicin resistance in the high-risk groups of drug-resistant pulmonary tuberculosis.

Objective:To observe any effect of electroacupuncture applied to the Jialianquan points in treating post-stroke oropharyngeal dysphagia.Methods:Forty-five stroke survivors with oropharyngeal dysphagia were randomly divided into an electroacupuncture group ( n=15), a neuromuscular electrical stimulation group ( n=15) and a control group ( n=15). All groups received 30 minutes of routine swallowing function training 5 times a week for 3 weeks. The electroacupuncture group was additionally provided with 30 minutes of electroacupuncture applied to the Jialianquan (CV23) points, and the neuromuscular electrical stimulation group instead received 30 minutes of neuromuscular electrical stimulation over the bilateral submental muscles. Videofluoroscopic swallowing studies were performed before and after the 3 weeks of treatment. Standardized swallowing assessment was conducted producing functional oral intake scale ratings, modified barium swallow impairment profiles and the penetration-aspiration scale scores. Surface electromyography was also employed to evaluate submental muscle functioning through measuring the swallowing time, average EMG (AEMG) value and peak amplitude. Results:After the treatment, significant improvement was observed in all of the evaluations with both groups, but the average scores were significantly better in the electroacupuncture and neuromuscular electrical stimulation groups compared with the control group and significantly better in the electroacupuncture group than in the neuromuscular electrical stimulation group.Conclusion:Electroacupuncture at the Jialianquan point can significantly improve the swallowing of stroke survivors with oropharyngeal dysphagia. It is more effective than neuromuscular electric stimulation.

Objective:To observe any effect of transcranial direct current stimulation (tDCS) on the cognition of stroke survivors and the integrity of their white matter fibers.Methods:Thirty persons with post-stroke cognitive impairment (PSCI) were randomly divided into an experimental group ( n=15) and a control group ( n=15). In addition to basic drug therapy and routine cognition training, the experimental group received 20 minutes of tDCS daily, 5 days per week for 3 weeks, while the control group received sham tDCS stimulation. Before and after the treatment, both groups′ cognitive functioning was evaluated using the mini-mental state examination (MMSE) and the Montreal cognitive assessment scale (MoCA). Their ability in the activities of daily living (ADL) was quantified using the modified Barthel index (MBI). Diffusion tensor imaging (DTI) was employed to observe any changes in the integrity of their white matter fibers. Results:The average MMSE, MOCA and MBI scores of both groups had improved significantly after the treatment, but the improvement in the experimental group was significantly greater than among the controls. The average fractional anisotroposy value of the affected inferior fronto-occipital fasciculus in both groups was positively correlated with the group′s average MMSE score and MoCA score.Conclusion:tDCS can effectively improve the cognition and functioning in the activities of daily living of stroke survivors. Its mechanism may be related to improving the integrity of the white matter fibers involved.

Objective: To establish a method for the determination of 1-methoxy-2-propanol in urine using headspace solid phase micro-extraction coupled with gas chromatography. Methods: The 1-methoxy-2-propanol was enriched by headspace solid phase micro-extraction fiber coated with carbene/polydimethylsiloxane (CAR/PDMS) . Single factor rotation method was used to optimize the conditions of extraction temperature, salt amount, and extraction time. The separation was performed on DB-5 (30 m×0.32 mm×0.25 μm) capillary column and detected with flame ionization detector. The quantification was based on the standard curve. Results: The concentration of 1-methoxy-2-propanol in urine was linear in the range of 0.50-10.0 mg/L, and the linear correlation coefficient was 0.9993. The detection limit of the method was 0.14 mg/L, and the limit of quantification was 0.45 mg/L. The recovery was 85.8% to 104.7%, and the RSD of intra- and inter-batch precision were 3.25%-6.65% and 0.81%-3.96%, respectively. Conclusion The method is high sensitivity and simple operation, and is suitable for the determination of 1-methoxy-2-propanol in urine of occupational exposure population.

Objective To establish a lncRNA prognostic risk model for gastrointestinal tumors based on the TCGA database and evaluate the prognosis of patients. Methods We collected the data of patients with esophageal cancer, gastric cancer, colon cancer and rectal cancer in the TCGA database. Univariate Cox analysis, Lasso and multivariate Cox analysis were performed to construct the prognostic risk scoring model. The model was validated and tested for independence. Time-dependent ROC curve analysis was performed to evaluate the clinical application value of the model. Results We established a prognostic risk model based on 13 lncRNAs. The three-year AUC of the training set and the validation set were 0.746 and 0.704, respectively. The pan-cancer data set was divided into high- and low-risk groups for survival analysis. The 5-year survival rate of the low-risk group was significantly higher than that of the high-risk group; among all cancer types, the five-year survival rates of the low-risk group were higher than those of the high-risk group. Multivariate Cox analysis showed that the risk score could be an independent indicator of prognosis. Conclusion The 13-gene prognostic risk score model is constructed successfully. The risk score obtained by this model can be used as an independent prognostic predictor of the patients with gastrointestinal cancer.

MicroRNAs(miRNAs)are important regulators of gene expression.The large-scale detection and profiling of miRNAs have been accelerated with the development of high-throughput small RNA sequencing(sRNA-Seq)techniques and bioinformatics tools.However,generating high-quality comprehensive miRNA annotations remains challenging due to the intrinsic complexity of sRNA-Seq data and inherent limitations of existing miRNA prediction tools.Here,we present iwa-miRNA,a Galaxy-based framework that can facilitate miRNA annotation in plant species by combining computational analysis and manual curation.iwa-miRNA is specifically designed to generate a comprehensive list of miRNA candidates,bridging the gap between already annotated miRNAs provided by public miRNA databases and new predictions from sRNA-Seq datasets.It can also assist users in selecting promising miRNA candidates in an interactive mode,contributing to the accessibility and reproducibility of genome-wide miRNA annotation.iwa-miRNA is user-friendly and can be easily deployed as a web application for researchers without programming experience.With flexible,interactive,and easy-to-use features,iwa-miRNA is a valu-able tool for the annotation of miRNAs in plant species with reference genomes.We also illustrate the application of iwa-miRNA for miRNA annotation using data from plant species with varying genomic complexity.

ObjectiveTo clarify the relationship between childhood maltreatment and insomnia in middle school students， and to explore the mediating role of post-traumatic stress disorder （PTSD） symptoms and the moderating role of mental resilience involvement. MethodsFrom April to May 2021， a total of 3 412 students in 3 middle schools in a city in western China were selected using convenient sampling method， and all students were assessed by Childhood Trauma Questionnaire （CTQ）， Post Traumatic Stress Disorder Scale （PCL）， Insomnia Severity Index （ISI） and mental resilience scale. The moderated mediating effect analysis was conducted using Process v3.4. ResultsAmong the middle school students， CTQ scores were positively correlated with PCL and ISI scores （r=0.540， 0.320， P<0.05）， and the scores of CTQ， PCL and ISI were negatively correlated with the score of mental resilience scale （r=-0.049， -0.193， -0.132， P<0.05）. PTSD symptoms exhibited a partial mediating effect on the relationship between general childhood maltreatment and insomnia （β=0.161， P<0.05）， accounting for 80.5% of the total effect. ConclusionAmong the middle school students， the relationship between childhood maltreatment and insomnia is partially mediated via PTSD symptoms， and mental resilience exerts a moderating role between childhood maltreatment and PTSD symptoms.

ObjectiveTo support intelligent clinical decision-making in traditional Chinese medicine（TCM）， this study utilized ontology and knowledge graph construction techniques to achieve the IT application of clinical practice guidelines. MethodBased on the principles of findability， accessibility， interoperability， and reusability （FAIR principles）， this study employed ontology techniques to construct an ontology for TCM clinical practice guidelines and built a knowledge graph using coronary heart disease as an example. Based on the Checklist for Reporting Practice Guidelines in Traditional Chinese Medicine and Recommendation Grading in TCM Clinical Guidelines/Consensus （T/CAS 530—2021），the ontology of TCM clinical practice guidelines was constructed using the seven-step ontology construction method. On this basis，the TCM diagnosis and treatment data from the Guidelines for the Diagnosis and Treatment of Stable Angina Pectoris in Coronary Heart Disease were stored in Neo4j in the form of triples through knowledge extraction，integration，and storage. ResultThe information in the clinical practice guidelines was divided into three categories： onset and prevention information， diagnosis information， and treatment information， and the TCM clinical practice guideline ontology was constructed. A total of 27 concepts related to TCM clinical diagnosis and treatment and 14 data attributes were obtained， and 12 conceptual relationships including hierarchical relationships and object attributes were established. By taking coronary heart disease as an example and the TCM clinical practice guideline ontology as the model layer， the knowledge map of TCM diagnosis and treatment guidelines for stable angina pectoris in coronary heart disease with 276 nodes and 336 relationships was constructed， realizing the visual display and query of the guideline content. ConclusionThe ontology of TCM clinical practice guidelines and the knowledge graph of stable angina pectoris in coronary heart disease constructed by combining the seven-step ontology construction method and Neo4j graph database technology are efficient and flexible，providing an intelligent TCM diagnosis and treatment scheme and promoting the standardization and objectification of TCM diagnosis and treatment.