BACKGROUND: Molecular subtyping is an essential complementarity after pathological analyses for targeted therapy. This study aimed to investigate the consistency of next-generation sequencing (NGS) results between circulating tumor DNA (ctDNA)-based and tissue-based in non-small cell lung cancer (NSCLC) and identify the patient characteristics that favor ctDNA testing. METHODS: Patients who diagnosed with NSCLC and received both ctDNA- and cancer tissue-based NGS before surgery or systemic treatment in Lung Cancer Center, Sichuan University West China Hospital between December 2017 and August 2022 were enrolled. A 425-cancer panel with a HiSeq 4000 NGS platform was used for NGS. The unweighted Cohen's kappa coefficient was employed to discriminate the high-concordance group from the low-concordance group with a cutoff value of 0.6. Six machine learning models were used to identify patient characteristics that relate to high concordance between ctDNA-based and tissue-based NGS. RESULTS: A total of 85 patients were enrolled, of which 22.4% (19/85) had stage III disease and 56.5% (48/85) had stage IV disease. Forty-four patients (51.8%) showed consistent gene mutation types between ctDNA-based and tissue-based NGS, while one patient (1.2%) tested negative in both approaches. Patients with advanced diseases and metastases to other organs would be suitable for the ctDNA-based NGS, and the generalized linear model showed that T stage, M stage, and tumor mutation burden were the critical discriminators to predict the consistency of results between ctDNA-based and tissue-based NGS. CONCLUSION ctDNA-based NGS showed comparable detection performance in the targeted gene mutations compared with tissue-based NGS, and it could be considered in advanced or metastatic NSCLC.
Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; Circulating Tumor DNA/blood* ; High-Throughput Nucleotide Sequencing/methods* ; Female ; Male ; Lung Neoplasms/pathology* ; Middle Aged ; Mutation/genetics* ; Aged ; Adult ; Aged, 80 and over

Objective Through integrative bioinformatics analysis of multi-source transcriptomic data, potential biomarkers to asthma epithelial cells were identified. The expression of these candidate target was subsequently validated in lung tissues and epithelial cells from asthma models. Methods The gene expression profile data of epithelial cells from three asthma patient cohorts and corresponding healthy controls were integrated from the Gene Expression Omnibus (GEO) database. Differential expression analysis and gene co-expression network analysis were performed to identify key genes and biological pathways associated with asthma. The key genes were validated in lung tissues and epithelial cells in asthma animal models. Results Differential gene expression analysis revealed 1121 upregulated and 1484 downregulated genes in epithelial cells from asthma patients compared with healthy controls. The biological pathway enrichment analysis revealed that the upregulated genes were mainly involved in glycosylation processes, whereas the downregulated genes were mainly associated with immune cell differentiation process. The gene co-expression network analysis revealed that module 9, enriched in glycosylation-related pathways, was significantly positively correlated with asthma, whereas module 17, associated with insulin and other signaling pathways, showed a significant negative correlation with asthma. We identified the genes of polypeptide N-acetylgalactosaminyltransferase 5 (GALNT5), pyrroline-5-carboxylate reductase 1 (PYCR1), and carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) as key genes within module 9, all of which were significantly upregulated in asthma. Finally, we validated that the expression levels of GALNT5, PYCR1, and CEACAM5 were significantly upregulated in epithelial cells from asthmatic lung tissue. Additionally, using a rat asthma model, we further confirmed that the protein levels of these three genes were significantly upregulated in lung tissues of the model group. Conclusion Through data integration and experimental validation, this study identified key genes and biological pathways closely associated with asthma pathogenesis. These findings provide a novel theoretical basis and potential targets for the diagnosis and treatment of asthma.
Asthma/metabolism* ; Humans ; Epithelial Cells/metabolism* ; Animals ; Biomarkers/metabolism* ; Gene Expression Profiling ; Transcriptome ; Gene Regulatory Networks ; Rats ; Computational Biology

Objective This study aims to achieve high-yield functional expression of the human voltage-gated potassium channel K_V3.1 using an Escherichia coli cell-free protein synthesis system, thereby providing a novel synthetic approach for drug screening, structural analysis and functional characterization of K_V3.1. Methods K_V3.1 was expressed in an Escherichia coli cell-free protein synthesis system for 10 hours in the presence of peptide surfactant A₆K. The secondary structure of K_V3.1 was analyzed by circular dichroism spectroscopy. The potassium channel activity of the recombinant protein liposome K_V3.1-A₆K was investigated using fluorescent dyes Oxonol VI as indicators, which are capable of reflecting alterations in membrane potential. Results Soluble K_V3.1 protein was successfully synthesized, achieving a purified yield of up to 1.2 mg/mL via an Escherichia coli cell-free protein synthesis system. Circular dichroism spectroscopy revealed that K_V3.1 exhibited characteristic α-helical secondary structures. Membrane potential fluorescence assays demonstrated that the K_V3.1-A₆K proteoliposomes, which were reconstructed with surfactant peptide A₆K, exhibited remarkable potassium ion permeability. Conclusion This study successfully achieved high-yield expression of human K_V3.1 with activity using an Escherichia coli-based cell-free protein synthesis system. This innovative method not only significantly enhances the expression yield of K_V3.1, but also maintains its functional activity, thereby establishing a novel and efficient synthetic platform for drug screening and advancing our understanding of structure-function relationships in K_V3.1 research.
Humans ; Escherichia coli/metabolism* ; Shaw Potassium Channels/biosynthesis* ; Cell-Free System ; Circular Dichroism ; Protein Biosynthesis ; Recombinant Proteins/metabolism* ; Membrane Potentials ; Shab Potassium Channels

Objective To compare the efficacy of 2 L and 3 L polyethylene glycol(PEG)electrolyte solution for bowel preparation in a real-world setting.Methods A real-world,single-center cohort study was conducted on the individuals undergoing colonoscopy in Department of Gastroenterology of Chengdu Third People's Hospital between May and October 2023.Based on our inclusion and exclusion criteria,they were given 2 L(n=4 684)and 3 L(n=3 700)PEG electrolyte solution for bowel preparation.The primary outcome indicator was the adequacy of bowel preparation by Boston bowel preparation score(BBPS).Secondary outcome indicators included the BBPS score,polyp detection rate(PDR),tolerability,compliance,and incidence of adverse events.Results The adequacy rate of bowel preparation was 94.35％in the 3 L group,significantly higher than that of the 2 L group(91.29％,P＜0.001).The 3 L group obtained a higher BBPS score then the 2 L group(6.92±1.06 vs 6.81±1.14,P＜0.001).But there was no statistical difference in the PDR between the 2 groups(P=0.073).And the rate of PEG completion(P=0.810),administration of low residue diet as required(P=0.094)or use of dimethicone(P=0.072)were comparable between the 2 groups.However,the incidences of vomiting(4.5％vs 3.2％,P=0.002),abdominal discomfort(5.0％vs 3.9％,P=0.011)and sleep disturbance(18.0％vs 14.6％,P＜0.001)were obviously higher in the 3 L group than the 2 L group.Conclusion In a real-world setting,2 L PEG is a considerably safe and effective regimen for bowel preparation.

Objective To investigate the expression of circular RNA ubiquitin-associated protein 2(circ-UBAP2)and RAS-related C3 botulinum toxin substrate 1(Rac1)in cervical cancer tissues and their relation-ship with radiotherapy efficacy and prognosis.Methods A total of 96 patients with cervical cancer admitted to the hospital from January 2019 to May 2021 were selected as the research subjects.Detect the expression of circ-UBAP2 and Rac1 in the cancer tissues and adjacent tissues of the patients.The clinical data of the patients were recorded.According to the radiotherapy efficacy,they were divided into patients with effective radiother-apy(n=62)and patients with ineffective radiotherapy(n=34),and the relationship between the expressions of circ-UBAP2 and Rac1 and the radiotherapy efficacy was analyzed.The survival conditions of the patients were followed up for 3 years and they were divided into the survival group(n=81)and the death group(n=15),and the clinical data of the two groups were compared.The relationship between the expressions of circ-UBAP2 and Rac1 and the survival prognosis of patients was analyzed by Kaplan-Meier.The risk factors influ-encing the prognosis of patients were analyzed by Cox regression.Results The circ-UBAP2 expression and the positive rates of Rac1 expression in cancer tissues were both higher than those in adjacent tissues,and the difference was statistically significant(P＜0.05).The effective rate of radiotherapy in patients with high ex-pression of circ-UBAP2 and Rac1 in cancer tissues was lower than that in patients with low expression of circ-UBAP2 and Rac1,and the difference was statistically significant(P＜0.05).The 3-year overall survival rate and average survival duration of patients with high expression of circ-UBAP2 and Rac1 in cancer tissues were lower than those of patients with low expression of circ-UBAP2 and Rac1,and the difference was statistically significant(P＜0.05).There were statistically significant differences in clinical stage,tumor differentiation degree,expressions of circ-UBAP2 and Rac1,and the proportion of lymph node metastasis between the two groups(P＜0.05).The results of Cox regression analysis showed that clinical stage Ⅱ B-Ⅲ,poorly differen-tiated tumor degree,high expression of circ-UBAP2 and Rac1,and lymph node metastasis were risk factors af-fecting the prognosis of patients(P＜0.05).Conclusion circ-UBAP2 and Rac1 are abnormally highly ex-pressed in cervical cancer tissues and are related to the radiotherapy efficacy and prognosis.

ObjectiveTo investigate the levels of serum antibodies against novel coronavirus （SARS-CoV-2） in healthcare workers after one month of natural infection， to explore the influencing factors and their correlations with the levels of antibodies， and to provide reference for strengthening the protection of healthcare workers and preventive intervention in Pudong New Area in Shanghai. MethodsVenous blood samples were collected from 1 102 medical staff in Pudong hospitals one month after infection. The serum levels of new coronavirus specific antibodies IgM， IgG and neutralizing antibodies were detected by chemiluminescent immunoassay. The information of gender， age， position， infection severity， vaccination， basic diseases and use of immunosuppressants were obtained by questionnaire to explore the influencing factors and their correlation with the antibody level. ResultsOne month after natural infection， 99.00% （1 091/1 102） of the subjects were found to be positive for IgG antibody against the new coronavirus， 17.79% （196/1 102） of the subjects were IgM antibody positive， and 99.00% （1 091/1 102） of the samples were positive for the neutralizing antibody. The level of antibody might be influenced by the severity of infection， the time of the last dose of vaccination， and the long-term use of immunosuppressants. The more severe the disease， the stronger the neutralizing antibody response. The antibody level in the people who received the final dose of vaccine within 6 months was higher than that of the people who received the vaccine 6 months ago， and the difference was statistically significant. The antibody levels were low in the subjects who received long-term immunosuppressants. ConclusionThe specific IgM， IgG and neutralizing antibody were found， one month after infection， in the medical workers in Pudong New Area， Shanghai， and the antibody titers were high， which had a good protective effect. The antibody level of the people who were vaccinated within 6 months was higher， it is recommended that people who receive the last vacination more than 6 months should be re-vaccinated with the booster vaccine， to improve the autoimmunity against the novel coronavirus.

Objective:To analyze the risk factors for central lymph node metastasis (CLNM) in patients with papillary thyroid cancer (PTC) aged 55 years and above, and to construct a predictive model with columnar graph.Methods:This retrospective study included 406 PTC patients aged 55 and above, treated at the First Affiliated Hospital of Zhengzhou University from Nov. 2019 to Feb. 2022. Data on demographic characteristics, disease features, and laboratory test results were collected. Univariate and multivariate logistic regression analyses were used to identify risk factors for CLNM and develop a clinical prediction model and nomogram.Results:The study involved 406 patients, divided into a modeling group (285 patients) and a validation group (121 patients). The predictive model identified independent risk factors for CLNM. In the modeling group, the model demonstrated a ROC AUC of 0.769, with 82.6% sensitivity, 63.0% specificity, and 67.7% accuracy. The validation group showed 66.7% sensitivity, 74.5% specificity, and 72.7% accuracy, with an AUC of 0.760. Hosmer-Lemeshow tests indicated good fit in both groups. Decision curve analysis confirmed the model's clinical decision-making value, showing better performance than traditional strategies and good generalizability and reliability.Conclusions:Sex, maximum tumor diameter, bilateral involvement of thyroid lobes, clinically evident cervical lymph nodes, and local invasion are independent predictive factors for CLNM in patients over 55 with papillary thyroid carcinoma (PTC). A clinical risk stratification nomogram model based on these risk factors demonstrates good predictive performance.

Objective To analyze the clinical characteristics of C-TI-RADS 3 thyroid nodules with a diameter greater than 2 cm and explore their correlation with gender,nodule ingredient,contralateral cancer presence,diffuse echo changes,TPOAB and TGAB.Methods A retrospective analysis was made on the clinical and pathological information of 94 patients with thyroid nodules who were admitted to our department from September 2022 to March 2023.All the patients underwent cytological and/or histopathological examinations.The proportions of TBS I category,benign tumors,low-risk tumors,and malignant tumors were calculated.The proportion of TBS type Ⅰ,benign tumors,low-risk tumors,and malignant tumors was quantified.Subsequently,a comparative analysis was conducted among the benign,low-risk,and malignant groups in terms of clinical characteristics including gender distribution,nodule composition,contralateral cancer occurrence,diffuse echo changes presence,as well as TPOAB and TGAB levels.Results Seven cases in TBS I category were excluded.Among the remaining 87 cases with confirmed pathology results for nodules,there were 72 benign cases(38 cytology cases and 34 histology cases),5 low-risk thyroid tumors(2 cytology cases and 3 histology cases),10 malignant cases(8 PTC cases,1 FTC case,and 1 MTC case).There was a significant difference in nodule ingredient(cystic/solid)between different pathological types(x2=10.369,P=0.006).However,no statistical significance was found in terms of gender,diffuse echo changes,contralateral cancer presence,TPOAB or TGAB(P＞0.05).Further analysis showed that the proportion of solid component was higher in low-risk tumors than in benign nodules(x2=9.571,P=0.002).No statistical significance was found between malignant nodules and low-risk nodules(x2=2.143,P=0.143),or between malignant nodules and benign nodules(x2=2.165,P=0.141).Conclusion Although TI-RADS 3 nodules are generally considered as potentially benign according to various versions of thyroid imaging reporting and data system,malignant nodules still account for a certain proportion.Attention should be paid to thyroid nodules with a typical ultrasonic signs,such as cystic nodules,thyroid follicular tumors and medullary thyroid carcinoma.Ultrasound guided fine needle aspiration cytopathology is necessary for evaluating benign and malignant nodules.It is necessary to pay attention to unsatisfactory or undiagnosable specimens to improve the accuracy of diagnosis.

Objective:To investigate the clinical effect of proper management of inferior pancreaticoduodenal artery (IPDA) in laparoscopic pancreaticoduodenectomy (LPD).Methods:This is a retrospective case series study. The clinical and pathological data of 70 patients who received LPD due to pancreatic head tumors, periampullary tumors, or distal common bile duct tumors in the Pancreatic Center of the Second Clinical College of Guangzhou University of Chinese Medicine from January to December 2022 were retrospectively collected. There were 47 males(67.1%) and 23 females(32.9%),aged (59.9±12.8)years(range:13 to 87 years).The procedure of IPDA exposure was as follows:a middle approach was utilized to expose the right half of superior mesenteric artery(SMA) and its right branches between the SMA and superior mesenteric vein(SMV) in superior colonic region. In the subcolonic region,SMA trunk exposure via dissection along the jejunal artery from feet to head and identification the association between IPDA and jejunal artery were prior to IPDA root ligation and dissection. The safety and efficacy of intraoperative IPDA handling were assessed based on surgical videos. Follow-up was carried out in outpatient clinic or by telephone, and outpatient follow-up was conducted once every 1 to 3 months after surgery.Results:The percentage of total LPD was 98.6%(69/70),with all patients achieving R0 resection. Nine cases(12.9%) were involved in combined vascular resection and reconstruction,with 1 case (1.4%) requiring additional upper abdominal incision for vascular and gastrointestinal reconstruction,while the remaining eight cases (11.4%) were completed laparoscopically. The operative time was (432.7±115.4)minutes(range:282 to 727 minutes), and the blood loss was (140.0±125.7)ml(range:20 to 800 ml). Only two patients(2.9%) received fresh frozen plasma transfusion,with an average volume of 650 ml. Reliable ligation and safe handling of the IPDA were achieved in 91.4%(64/70) of cases, with 8.6%(6/70) suffering from IPDA injury-related bleeding. No one was converted to opened surgery. Pathologically,the mean tumor size was (3.3±1.6)cm (range:1 to 7 cm),and the mean number of harvested lymph nodes was 17.0±7.3(range:0 to 46). Lymph node metastasis was observed in 13 cases (18.6%). Five cases (13.2%) developed grade B pancreatic fistula,while no grade C pancreatic fistula occurred. Other complications included bile leakage in one case(1.4%),delayed gastric emptying in two cases(2.9%), lymphatic leakage in 2 cases(2.9%),intra-abdominal infection in 9 cases(12.9%),and fat liquefaction of surgical incision in 1 case(1.4%). Two cases(2.9%) experienced postoperative intra-abdominal bleeding,one due to mesangial bleeding of lesser curvature of the stomach and the other due to oozing from the hepatic arterial sheath. These bleeding events were not concerned with IPDA. The average length of postoperative hospital stay was (15.2±4.6)days(range:9 to 28 days).Conclusion:Proper intraoperative management of IPDA in LPD might reduce IPDA-related bleeding during and after surgery and improve the safety of LPD.

Objective To establish a scientific training program that takes into account both anaerobic and aerobic training for pilots,and to explore the appropriate ratio of aerobic and anaerobic training.Methods According to the physical examination standards for pilots,a total of 16 healthy subjects aged 18-24 were selected from two batches.The two batches of subjects were trained with different aerobic and anaerobic ratios.Training period was 3 months.The changes in cardiopulmonary function of the subjects before and after training were evaluated using the cardiopulmonary function exercise testing system(CPET),and the changes in anaerobic capacity were evaluated using changes in strength as an indicator.Results After training,the weight load of the subjects in the two training programs,including barbell squats,leg flexion and hard pull,and barbell under 10RM and 3RM,was significantly increased(P<0.001),and there was no statistically significant difference in anaerobic strength growth between the two groups.The results of CPET showed that the maximum load,maximum heart rate,and respiratory quotient in the two groups were significantly increased after than before the training(P<0.01).The maximum load(Experiment group 1:29.12±19.69,Experiment group 2:72.00±46.24)and respiratory quotient(Experiment grouop 1:0.11±0.09,Experiment group 2:0.28±0.16)of the subjects in experiment group 2 before and after training were greater than those in experiment group 1.The difference was statistically significant(P<0.05).Conclusion The anaerobic and aerobic capacities of the subjects in the experiment group 2 are effectively improved,indicating that ratio of aerobic and anaerobic of the training scheme is better.