infC gene encodes the translation initiation factor IF3 in bovine Pasteuella multocida,but it whether or not regulation to the virulence and cross-protection in P.multocida is still not well understood.In this study,the infC gene mutant(△infC)derived from bovine P.multocida type A strain CQ2 was constructed using by homologous recombination method.Compared with wild strain,the △infC showed significant increasing in biofilm formation,but the capsule produc-tion,virulence and bacterial loading in organs were significant decreased,and the IL-1β secretion of mouse peritoneal macrophage increased.Along with the infC gene deletion,the expression of genes related to capsule synthesis and LPS synthesis and transport were significantly down-regulated,while that of genes related to biofilm synthesis and outer membrane protein were significantly up-regulated.The inactivated vaccines of wild type and mutant were prepared and mice were immu-nized twice then challenged with wild type strains,respectively.The immuno-protection rate of△in fC inactivated vaccine against bovine P.multocida type A,B and F were 100.0％,83.3％and 0.0％,respectively,and the immuno-protection rate that against rabbit type P.multocida was 33.3％.The results indicated that infC gene could affect the virulence of P.multocida by regula-ting the production of capsule and the expressions of virulence related factors,and the deletion of infC gene conferred a certain cross-protection property of strains.This study provided a certain foundation for the development of P.multocida vaccine.

Objective:Based on the forgetting curve principle to construct a model for the popularization of cardiopulmonary resuscitation among college students and validate its effectiveness, providing reference for the promotion of cardiopulmonary resuscitation.Methods:A randomized controlled trial was conducted. In October 2021, a multistage random sampling method was used to select 262 freshmen and sophomores from the School of Mathematics and Physics at Leshan Normal University as the control group, and 262 freshmen and sophomores from the School of Electronic Information and Artificial Intelligence as the experimental group.The control group received conventional training, while the experimental group, in addition to conventional training, received cardiopulmonary resuscitation retraining based on the Ebbinghaus forgetting curve at three time intervals: within 1 week, within 1 month, and within 1 to 6 months after training. The retraining included online self-learning, on-site observation reinforcement, and social practice sublimation. The cardiopulmonary resuscitation knowledge awareness rate and cardiopulmonary resuscitation skill assessment of the two groups were compared before training, immediately after training, and 6 months after training.Results:The final number of participants was 256 in the control group (126 males and 130 females) and 224 in the experimental group (110 males and 114 females). There were no statistically significant differences in cardiopulmonary resuscitation knowledge awareness rate and cardiopulmonary resuscitation skill scores between the two groups before and immediately after training (both P>0.05). Immediately after training, the average cardiopulmonary resuscitation knowledge awareness rate in the experimental and control groups were 91% (1 626/1 792) and 88% (1 806/2 048), respectively, which were significantly higher than the pre-training rates of 55% (989/1 792) and 55% (1 129/2 048) ( χ2=8.46, 7.30, both P<0.01). The cardiopulmonary resuscitation skill scores were (42.00 ± 3.55) and (41.68 ± 3.40), respectively, higher than the pre-training scores of (15.41 ± 4.85) and (15.92 ± 4.66) ( t=84.47, 45.56, both P<0.05). Six months after training, the awareness rates of determination of consciousness and cardiac arrest judgment, cardiopulmonary resuscitation steps, chest compression location, frequency, depth, and the ratio of chest compressions to artificial ventilation in the experimental group in college students were 90% (201/224), 93% (208/224), 92% (206/224), 93% (208/224), 89% (200/224), and 94% (210/224), respectively, higher than those in the control group, which were 62% (158/256), 71% (182/256), 77% (198/256), 73% (186/256), 70% (178/256), and 69% (176/256) ( χ2 values were 4.21-7.03, all P<0.01). The cardiopulmonary resuscitation skill score in the experimental group was (49.77 ± 3.68), higher than the control group′s (34.95 ± 4.79) ( t=38.25, P<0.01). The proportion of students confident in using cardiopulmonary resuscitation to save others was 92% (206/224) in the experimental group, higher than 58% (148/256) in the control group ( χ2=7.68, P<0.01). Six months after training, the cardiopulmonary resuscitation skill score in the experimental group was (49.77 ± 3.68), higher than the immediate post-training score of (42.00 ± 3.55), whereas the control group's cardiopulmonary resuscitation skill score was (34.95 ± 4.79), lower than the immediate post-training score of (41.68 ± 3.40) ( t=22.74, 18.33, both P<0.01). Conclusions:The cardiopulmonary resuscitation popularization model for college students based on the forgetting curve is conducive to mastering cardiopulmonary resuscitation knowledge and skills among trained college students, enhancing their confidence in using cardiopulmonary resuscitation techniques to rescue others. Moreover, it helps save the human, financial, and material resources required for cardiopulmonary resuscitation training to a certain extent and is worthy of promotion.

Objective:To establish a scoring system for preoperative prediction of the malignant potential of gastric stromal tumors based on gastroscopic and endoscopic ultrasound features, along with validation.Methods:A total of 286 patients who were treated in Jiangsu Province Hospital of Chinese Medicine from January 1, 2017 to December 31, 2023 and diagnosed as having gastric stromal tumors by postoperative pathology were enrolled in the modeling group. According to National Institutes of Health classification system, 227 very-low/low-risk patients were classified into the low malignant potential (LMP) group, and the 59 intermediate/high-risk patients into the high malignant potential (HMP) group. LASSO regression analysis was performed to identify predictive factors for HMP gastric stromal tumors, and a nomogram prediction model was developed. Internal validation using the Bootstrap method was performed on the modeling group, and external validation was performed on data from 85 patients who were treated and diagnosed as having gastric stromal tumors by postoperative pathology in Taizhou People's Hospital from January 1, 2021 to December 31, 2023. The receiver operator characteristic (ROC) curves, calibration curves, and decision curve analyses were employed in both the modeling and external validation groups.Results:Tumor size (coef=0.755), tumor shape (coef=0.015), tumor location (coef=0.008), growth pattern (coef=-0.026), cystic change (coef=0.685), and surface unceration change (coef=-0.545) were the independent predictive factors for HMP gastric stromal tumors. The nomogram-based prediction model constructed using these factors achieved an area under the ROC curve of 0.959 (95% CI: 0.898-0.903) in the modeling group and 0.959 (95% CI: 0.857-1.000) in the external validation group. The model demonstrated good accuracy (0.917) and a Kappa value of 0.737 in internal validation. Calibration curve and decision curve analyses indicated strong calibration and high net benefit in both the modeling and the external validation groups. Conclusion:Tumor size, tumor shape, tumor location, growth pattern, cystic change, and surface ulceration change are independent predictive factors for HMP gastric stromal tumors. The nomogram model developed based on these factors offers effective and convenient visualization for clinicians to predict the malignant potential of gastric stromal tumors preoperatively.

ObjectiveTo clarify the feasibility of the 3D scanning volume method for distal upper limb volume measurement, and to analyze its scorer reliability and criterion-related validity. MethodsFrom January to March, 2022, a therapist (operator A) who had not been exposed to 3D scanning volume method and water displacement method was trained to use a handheld 3D laser scanner and a spilt cup to measure the volume of a PVC distal upper limb model. The operation time of 30 operations of each method was recorded. The learning curves of the two methods were plotted using cumulative sum (CUSUM) analysis. The curve was cut into the learning stage and the mastery stage by the vertex of peak. The times required to reach the mastery stage and the operation time of the mastery stage for the two methods were recorded. A total of 20 healthy subjects were recruited from Huashan Hospital of Fudan University. Two trained therapists (operator A and operator B) measured the bilateral distal upper limb volume using a handheld 3D laser scanner, and operator A measured the bilateral distal upper limb volume using a spilt cup. ResultsThe fitting learning curve of the 3D scanning volume method (R² = 0.984) reached its peak after eight times of operation; while that of the water displacement method (R² = 0.494) reached its peak after five times of operation. At mastery stage, the operator spent less time using 3D scanning volume method than using water displacement method (P < 0.05). The intraclass correlation coefficient between the two operators were both 0.979 for bilateral distal upper limb volume measure (P < 0.001). The Pearson coefficients was above 0.979 between 3D scanning volume method and water displacement method (P < 0.001). ConclusionA therapist can master the use of the 3D scanning volume method after eight times of operation, and the operation time of 3D scanning volume method is shorter than that of water displacement method at mastery stage. The 3D scanning volume method is well reliable and valid, that can be used as an alternative to the water displacement method for distal upper limb volumetric measurement.

Due to the reduction of patient cooperation and the limitation of clinical diseases, general practitioners lack effective acceptation training in the training process. To improve this situation, this study applies teacher standardized patients (TSP) combined case-based learning (CBL) to the acceptation ability training of general practitioners in medically unexplained physical symptoms (MUPS). And the acceptation ability and communication ability of 15 general practitioners were evaluated before, during and after the training. It shows that the application of "TSP + CBL" in the acceptation ability training of general practitioners in MUPS can effectively improve the acceptation ability and communication ability of general practitioners.

Objective:To identify and screen the differential methylation genes in patients with cholangiocarcinoma and to predict the prognosis of patients with CCA.Methods:Cholangiocarcinoma tissues and paracancerous tissues of 8 patients with cholangiocarcinoma in Fujian Provincial Hospital from October 2019 to May 2020 were selected for 850K methylation sequencing analysis to obtain differentially methylated genes. The 2018 genome-wide methylation data and clinical information of 36 patients with cholangiocarcinoma were download from The Cancer Genome Atlas (TCGA) database, the 2012 cholangiocarcinoma methylation data (GSE32879) were download from the Gene Expression Omnibus (GEO) database, and the 2018 TCGA database differential survival genomic data of overall survival (OS) and disease-free survival (DFS) of cholangiocarcinoma were download from the GEPIA2 database. The differentially methylated positions (DMP) and differentially methylated regions (DMR) results of 850K methylation sequencing analysis of submitted samples, methylated genes in TCGA and GEO databases, and cholangiocarcinoma survival genes of samples were jointly submitted for testing, multi-data set analysis was performed by the Sangerbox VENN tool, and common differentially methylated genes were obtained by intersection screening. The minimum P value method was used to determine the cut-off value of gene expression in Sangerbox, and the patients were divided into high and low expression groups of differentially methylated genes. The OS, DFS, disease-specific survival (DSS), disease-free interval (DFI) and progression-free interval (PFI) of cholangiocarcinoma patients were compared between the two groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. Results:A total of 121 954 DMP were identified by 850K methylation sequencing of cholangiocarcinoma tissues and paracancerous tissues of 8 patients; a total of 1 399 differentially methylated genes were identified in DMR, and the common prognosis related genes glucosaminyl (N-acetyl) transferase 1 (GCNT1) and neurotrophic receptor tyrosine kinase 3 (NTRK3) were identified by intersection identification. The expression of GCNT1 in the cholangiocarcinoma tissues was higher than that in the paracancerous tissues, and the difference was statistically significant ( P = 0.040). The expression of NTRK3 in cholangiocarcinoma tissues was higher than that in the paracancerous tissues, but the difference was not statistically significant ( P = 0.790). The minimum P value method was used to predict the prognosis of patients with cholangiocarcinoma based on the combined expression of GCNT1 and NTRK3, and the order was based on the sum of the expression levels of the two genes. When 30% of the ranking was taken as the cut-off value, the difference in DFS between the high expression group and the low expression group in cholangiocarcinoma was the most significant ( P < 0.001); there was no significant difference in OS between the two groups ( P = 0.065). The results of GO functional analysis showed that GCNT1 was involved in protein glycosylation, macromolecule glycosylation, glycosylation, glycoprotein biosynthetic process, glycoprotein metabolic process, transferase activity and transferring glycosyl groups, protein O-linked glycosylation, O-glycan processing, etc., and NTRK3 was involved in neurotrophin signaling pathway, Ras signaling pathway, EGFR tyrosine kinase inhibitor resistance, ErbB signaling pathway, phospholipase D signaling pathway, central carbon metabolism in cancer, natural killer cell mediated cytotoxicity, etc. The results of KEGG analysis showed that GCNT1 was mainly associated with system functions such as mucin-type O-glycan biosynthesis and metabolic pathways, and NTRK3 was mainly associated with cell surface receptor pathways, intracellular signal transduction, positive regulation of stimulatory responses, transmembrane receptor protein tyrosine kinase signaling pathway, enzyme-linked receptor protein signaling pathway, MAPK signaling pathway cascade and regulation, protein phosphorylation signal transduction and other system functions. Conclusions:The expressions of differentially methylated genes GCTNT1 and NTRK3 in cholangiocarcinoma have certain predictive effects on the prognosis of patients with cholangiocarcinoma.

Background: No currently available biomarkers or treatment regimens fully meet therapeutic needs of type 1 diabetes mellitus (T1DM). Circular RNA (circRNA) is a recently identified class of stable noncoding RNA that have been documented as potential biomarkers for various diseases. Our objective was to identify and analyze plasma circRNAs altered in T1DM. Methods: We used microarray to screen differentially expressed plasma circRNAs in patients with new onset T1DM (n=3) and age-/gender-matched healthy controls (n=3). Then, we selected six candidates with highest fold-change and validated them by quantitative real-time polymerase chain reaction in independent human cohort samples (n=12). Bioinformatic tools were adopted to predict putative microRNAs (miRNAs) sponged by these validated circRNAs and their downstream messenger RNAs (mRNAs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to gain further insights into T1DM pathogenesis. Results: We identified 68 differentially expressed circRNAs, with 61 and seven being up- and downregulated respectively. Four of the six selected candidates were successfully validated. Curations of their predicted interacting miRNAs revealed critical roles in inflammation and pathogenesis of autoimmune disorders. Functional relations were visualized by a circRNA-miRNA-mRNA network. GO and KEGG analyses identified multiple inflammation-related processes that could be potentially associated with T1DM pathogenesis, including cytokine-cytokine receptor interaction, inflammatory mediator regulation of transient receptor potential channels and leukocyte activation involved in immune response. Conclusion Our study report, for the first time, a profile of differentially expressed plasma circRNAs in new onset T1DM. Further in silico annotations and bioinformatics analyses supported future application of circRNAs as novel biomarkers of T1DM.

No currently available biomarkers or treatment regimens fully meet therapeutic needs of type 1 diabetes mellitus (T1DM). Circular RNA (circRNA) is a recently identified class of stable noncoding RNA that have been documented as potential biomarkers for various diseases. Our objective was to identify and analyze plasma circRNAs altered in T1DM.

Objective To identify the effect of stereotactic body radiation therapy (SBRT) on the survival of patients with recurrent pancreatic cancer after surgery.Methods The data of 104 patients with recurrent pancreatic cancer after surgery who underwent SBRT in the Department of Radiation Oncology of Changhai Hospital,Navy Medical University from February 2012 to December 2016 were retrospectively analyzed.The prescription doses ranged from 35-40 Gy/4-8 f.Survival analysis was performed using the Kaplan-Meier method,and relevant factors affecting patients' survival were screened by the Cox proportional hazards model.Results The median overall survival (OS) and progression free survival (PFS) was 12.5 (11.0-14.0) months and 7.3 (6.0-8.7) months,respectively,while the 1-year rate of OS and PFS was 55.8％ and 22.1％,respectively.Multivariate analysis indicated that tumor stage,biological effect dose (α/β =10,BED10),the decrease of CA19-9 level after treatment,and follow-up chemotherapy were all related factors affecting overall survival;tumor stage,BED10,the degree of pain relief and the decrease of CA19-9 level after treatment were related factors affecting PFS.Conclusions Patients suffering recurrent pancreatic cancer with early tumor stage,normal CA19-9 level and mild pain before treatment could be better treated by SBRT,BED10 ≥60 Gy and follow-up chemotherapy after radiotherapy can prolong the survival of patients.

Objective: To investigate the regulation of curculigoside on osteogenic differentiation of MG63 and the protective effect on osteoporosis model mice. Methods: The effects of curculigoside on the survival rate of dexamethasone or H₂O₂ treated MG63 were detected by methyl thiazolyl tetrazolium (MTT). The specimens were divided into six groups: blank control group, blank administration group, model group (dexamethasone or H₂O₂ treatment group), low dose group (dexamethasone or H₂O₂+1.0 μmol/L curculigoside), medium dose group (dexamethasone or H₂O₂+2.5 μmol/L curculigoside) and high dose group (dexamethasone or H₂O₂+5.0 μmol/L curculigoside), the sample size of each group was 10. Western blotting was used to detect the expression of osteogenic differentiation-related proteins [type Ⅰ collagen, integrin β1, osteoblast-specific transcription factor (Osterix), osteocalcin and osteopontin] in MG63 cells after 1, 7 and 14 days incubated with 0, 1.0, 2.5 and 5.0 μmol/L of curculigoside. The sample size for each group at each time point was six. The experimental mice were divided into 4 groups: blank group, model group (dexamethasone treatment group), curculigoside low-dose group (dexamethasone+5 mg/kg curculigoside) and high-dose group (dexamethasone+45 mg/kg curculigoside), twenty each. After treatment, the tibia of the mice in each group were subjected to sacral HE staining. The number of osteoclasts was counted, and the levels of oxidative related factors in serum were determined by enzyme-linked immunosorbent assay (ELISA). Results: The MTT results showed that compared with the blank control group [(100±3.7)%], the cell survival rate decreased to (44.1±5.7)% after treatment with dexamethasone, and the survival rate increased to (79.7±3.8)% after treatment with 5.0 μmol/L of curculigoside. The cell survival rate decreased to (59.1±4.7)% after H₂O₂ treatment, and the survival rate increased to (80.8±3.5)% after treatment with 2.5 μmol/L of curculigoside. The results of Western blotting showed that the expression of type Ⅰ collagen and integrin β1 in MG63 cells was significantly increased after 1.0, 2.5 and 5.0 μmol/L of curculigoside for 1, 7 and 14 days compared with 0 μmol/L of curculigo side for the same period. After increasing (P<0.05), the expression of Osterix and osteocalcin was significantly increased after 1 day of incubation (P<0.05). However, compared with 0 μmol/L curculigoside treatment, the expression of osteopontin in MG63 cells was not significantly different after incubation with 1.0, 2.5, 5.0 μmol/L of curculigoside for 7 and 14 days (P>0.05). Compared with the blank group, the number of tibia osteoclasts in the osteoporosis model group increased. In the low-dose and high-dose groups of curculigoside, the tibia cortex was more continuous and the number of osteoclasts decreased. Compared with the blank group, the activity of oxygen in the osteoporosis model group was significantly increased (P<0.05), and superoxide dimutase and catalase were significantly decreased (P<0.05). Conclusions Curculigoside promotes the differentiation of MG63 cells by increasing the expression of osteoblast differentiation-related proteins, and has a certain therapeutic effect on dexamethasone-induced osteoporosis mice.