Objective To investigate the feasibility of using the cell-free DNA(cfDNA)carrying tissue-specific mutations as biomarkers for assessing the severity of exercise-induced tissue injury.Methods Based on a public gene expression database,ten tissue-specific and highly-expressed genes were selected for each of ten different human tissues.A total of 34 young healthy volunteers were recruited,and their peripheral blood samples were collected after running 5 km per day for one week.Genomic DNA from leukocytes,plasma exosomal RNA,and plasma cfDNA were extracted and subjected to high-throughput sequencing respectively.Tissue-specific somatic mutations in plasma exosomal RNA were identified,and the relationship between cfDNA carrying these mutations and traditional protein biomarkers was analyzed.Results Tissue-specific cfDNA mutations related to five tissues(myocardium,skeletal muscle,intestine,stomach,and kidney)were detected in more than five volunteers.The correlation coefficients between total plasma cfDNA levels and the levels of tissue-specific protein biomarkers associated with these five tissues were less than 0.3(n=34,r=-0.51-0.28,P=0.0022-0.65).Notably,for each specific tissue,the levels of tissue-specific cfDNA mutations were positively correlated with the corresponding protein biomarker levels,and correlation coefficients were over 0.8(n=7-13,r=0.81-0.92,P=9.0×10-4-0.020).Conclusion Tissue-specific cfDNA mutations may serve as novel potential biomarkers for non-invasive evaluation of exercise-related tissue injury.

Objective:To summarize the best evidence on palliative care for end-stage heart failure patients, so as to provide ideas and references for clinical healthcare professionals and decision-makers.Methods:According to the "6S" evidence pyramid model, clinical decisions, guidelines, expert consensus, and systematic reviews on palliative care for patients with end-stage heart failure were searched from top to bottom in BMJ Best Practice, UpToDate, Joanna Briggs Institute (JBI) Evidence-Based Health Care Center Database, Guidelines International Network, National Guideline Clearinghouse, PubMed, Web of Science, China National Knowledge Infrastructure, WanFang Data, SinoMed, VIP, and other websites and databases. Two researchers evaluated the quality of the literature and extracted information.Results:A total of 15 papers were included, including 4 guidelines, 2 clinical decisions, 7 expert consensus, and 2 randomized controlled trials. Twenty-four pieces of best evidence were developed from 11 aspects of goals, place, team building and training, applicability, key components, assessment, communication and shared decision-making, quality of life improvement, symptom management, psychological and spiritual care, and hospice care.Conclusions:The summarized best evidence on palliative care for patients with end-stage heart failure is scientific and standardized, and can guide healthcare professionals to use the evidence in a targeted way that takes into account the patient's condition and wishes to improve the patient's quality of life.

Objective:To summarize the best evidence on palliative care for end-stage heart failure patients, so as to provide ideas and references for clinical healthcare professionals and decision-makers.Methods:According to the "6S" evidence pyramid model, clinical decisions, guidelines, expert consensus, and systematic reviews on palliative care for patients with end-stage heart failure were searched from top to bottom in BMJ Best Practice, UpToDate, Joanna Briggs Institute (JBI) Evidence-Based Health Care Center Database, Guidelines International Network, National Guideline Clearinghouse, PubMed, Web of Science, China National Knowledge Infrastructure, WanFang Data, SinoMed, VIP, and other websites and databases. Two researchers evaluated the quality of the literature and extracted information.Results:A total of 15 papers were included, including 4 guidelines, 2 clinical decisions, 7 expert consensus, and 2 randomized controlled trials. Twenty-four pieces of best evidence were developed from 11 aspects of goals, place, team building and training, applicability, key components, assessment, communication and shared decision-making, quality of life improvement, symptom management, psychological and spiritual care, and hospice care.Conclusions:The summarized best evidence on palliative care for patients with end-stage heart failure is scientific and standardized, and can guide healthcare professionals to use the evidence in a targeted way that takes into account the patient's condition and wishes to improve the patient's quality of life.

Objective:To retrieve, evaluate, and summarize relevant evidence on remote pulmonary rehabilitation programs for patients with Chronic Obstructive Pulmonary Disease (COPD) .Methods:Following the 6S Pyramid Evidence Model, domestic and international guideline websites, evidence-based databases, association websites, and original databases were systematically searched for literature related to remote pulmonary rehabilitation for patients with COPD, including guidelines, expert consensus, evidence summaries, clinical decisions, practice recommendations, systematic reviews, and randomized controlled trials. The search was conducted from the databases' inception to July 31, 2023. Quality assessment and data extraction were independently conducted by two researchers.Results:A total of 27 articles were included, including five guidelines, one clinical decision, four evidence summaries, one expert consensus, 14 systematic reviews, and two randomized controlled trials. A total of 27 best evidence recommendations were synthesized across seven aspects, including setting, target population, pre-intervention preparation, communication devices, remote pulmonary rehabilitation programs, maintenance exercise plans, and remote health monitoring.Conclusions:The process of forming the best evidence for remote pulmonary rehabilitation programs for patients with COPD is comprehensive and scientific. Healthcare professionals should integrate evidence-based practice with patient preferences to develop personalized remote pulmonary rehabilitation programs according to individual circumstances.

Aflatoxin B1(AFB1)is a secondary metabolite produced by Aspergillus aflatoxin and Aspergillus parasiticus.It has strong immunotoxicity and carcinogenicity and seriously harms the health of humans and livestock.Therefore,this study aims to explore the effect of selenomethionine(SeMet)in alleviating AFB1-induced thymic damage.Fifty 35-day-old rabbits were randomly divid-ed into 5 groups,with 10 rabbits in each group.They are the control group,AFB1 group,AFB1+0.2Se group(0.2 mg/kg SeMet),AFB1+0.4Se group(0.4 mg/kg SeMet)and AFB1+0.6Se group(0.6 mg/kg SeMet).The test period was 21 d.On the 17th day,each rabbit in the AFB1 group and each SeMet group was gavaged with AFB1(0.3 mg/kg)every day for 5 consecutive days.Rabbit thymus tissue was taken for HE,TUNEL and PCNA detection.ELISA was used to detect the ex-pression of inflammatory factors in thymus tissue,and GSH-Px,T-AOC and MDA kits were used to detect oxidative stress indicators in thymus tissue.The results showed that AFB1 exposure caused a very significant increase in MDA levels in rabbit thymus tissue by 116.04％,and a signifi-cant decrease in the activities of antioxidant enzymes GSH-Px and T-AOC by 29.20％and 52.17％respectively.In addition,AFB1 induced an inflammatory response in the thymus,promoting TNF-a secretion to increase by 124.71％,IL-6 by 174.72％,and IL-1β by 62.38％compared with the con-trol group.SeMet pretreatment significantly improved the pathological changes of the thymus in rabbits and reduced its oxidative stress and inflammatory response.Therefore,we confirmed that SeMet alleviates AFB1-induced thymic damage and improves the immune performance of rabbits.

Liquid biopsy is gradually being applied in the clinical diagnosis and treatment of lung cancer.At pres-ent,with the development of metabolomics,more and more metabolic biomarkers are considered as potential sensitive mark-ers reflecting the occurrence and development of tumors.This article summarizes the changes in the main metabolic pathways of lung cancer,including glucose metabolism,amino acid metabolism,lipid metabolism,sphingolipid metabolism,glycero-phospholipid metabolism,and purine metabolism.Meanwhile,this article reviews the role of metabolic biomarkers in the early diagnosis of lung cancer,predicting disease progression,and evaluating the efficacy of chemotherapy and immunotherapy,aim-ing to provide effective biomarkers for tumor diagnosis and treatment.

In the field of implementation science, the use of appropriate theories is an important basis for promoting the promotion and application of evidence-based practice. However, the specific methods of selecting theories in the research design are rarely mentioned. This paper reviews the main contents, usage methods, characteristics and applications of "Theory Comparison and Selection Tool" (T-CaST) and A Pragmatic Guide for Selecting Theoretical Approaches in the field of implementation science, aiming at providing a reference for researchers to select appropriate theories.

Objective:To construct a recombinant herpes simplex virus 2 (HSV-2) expressing enhanced green fluorescent protein (EGFP) using clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology.Methods:Four strategies for inserting exogenous EGFP gene into HSV-2 genome using CRISPR/Cas9 technology were designed: (1) conventional homology-directed repair: circular two homology arm donor-mediated gene knock-in; (2) linearized single homology arm donor-mediated gene knock-in; (3) homology-independent targeted integration; (4) conventional homology-directed repair-mediated by cell lines stably expressing Cas9 and sgRNA.Results:The recombinant virus HSV-2-EGFP was successfully constructed based on the second, the third and the fourth strategies. The second strategy was the most efficient, followed by the third and the fourth strategies. The purified recombinant virus could stably express green fluorescent protein in seven passages and shared similar growth characteristics in Vero cells to the parental virus.Conclusions:Linearized single homology arm donor could increase the efficiency of gene knock-in, and cell lines stably expressing Cas9 and sgRNA could increase the efficiency of gene knock-in mediated by homology-directed repair.

Objective:To investigate the effects of genomic location of a foreign gene in Shanghai-191 strain measles virus (MV) vector on gene expression and virus replication.Methods:The nucleotide sequence encoding S1 protein of SARS-CoV-2 was inserted at different positions in MV antigenome (the upstream of the N gene, between P and M genes, between H and L genes), and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and N, P, and L proteins, respectively. The transfected cells were lysed and the supernatants were used to infected Vero cells to harvest recombinant viruses. S1 proteins expressed by the recombinant viruses were identified by RT-PCR, indirect immunofluorescence assay, Western blot and ELISA. Growth kinetics of the recombinant viruses were analyzed.Results:Recombinant viruses were failed to be rescued when the S1 protein-coding sequence was cloned into the upstream of N gene. Two recombinant viruses, MV-M-S1 and MV-L-S1, were successfully rescued when cloning the S1 protein-coding sequence into the intergenic region between P and M genes, or H and L genes, and could express S1 protein. MV-M-S1 expressed more S1 protein than MV-L-S1, but the titer of MV-M-S1 was lower.Conclusions:Inserting a foreign gene at different positions in the MV genome might have different effects on gene expression and virus replication. This study provided reference for the subsequent construction of MV vector.

Objective:To identify the clinical significance of CDK5 in colon cancer tissues.Methods:Two hundred colon cancer tissues were tested for CDK5 expression by immunohistochemistry on tissue microarrays. The correlation between CDK5 expression and clinicopathological features, prognosis and peripheral inflammation-related cells was analyzed.Results:CDK5 was low expressed in 100 cases (50.0%), and high in another 100 cases (50.0%). Longer time to tumor progression ( P=0.026) and overall survival ( P=0.035) were observed in patients with high CDK5 expression. By multivariate analysis , the expression of CDK5 was an independent risk factor for poor prognosis ( HR=0.45,95% CI: 0.21-0.99, P=0.049). The expression of CDK5 was not related to the counts of white blood cells and neutrophils ( P>0.05). Prognosis of patients with a positive lymph node ratio less than 0.15 was significantly better than that of patients with a higher lymph node ratio ( P<0.001). Conclusions:Patients with low CDK5 expression have poor prognosis, and CDK5 expression is not related to the counts of peripheral white blood cells and neutrophils.