Objective To investigate the relationship between the expression of nuclear factor ⅠB(NFⅠB)in myeloid cells and intestinal inflammation by constructing NFⅠB conditional gene knockout(cKO)mice.Methods Human Protein Atlas database,Genotype-Tissue Expression database,and FANTOM5 database were used to investigate the expression of NFⅠB in inflammatory cells.NFⅠB-floxed mice were constructed using CRISPR/Cas9 technology and hybridized with LyZ2-Cre transgenic mice.Myeloid specific NFⅠB cKO mice(NFⅠBfl/flLyz2-Cre)were obtained by self-crossing the progeny.After the genotype identification of mice by agarose gel electrophoresis,4 NFⅠB cKO mice of C57BL/6N strain were selected as experimental group,and 4 non-cKO mice were selected as control group.Both groups were induced with 2.5％dextran sulfate sodium salt(DSS)under the same condition to establish a chronic colitis model,and the severity of colitis was evaluated by clinical manifestations and histopathology.Results Analysis showed that NFⅠB was expressed in both myeloid granulocytes and monocytes,and the highest expression was found in neutrophils.NFⅠB cKO mice were successfully constructed using CRISPR/Cas9 technology and Cre-loxP system.DSS-induced enteritis NFⅠB cKO mice developed diarrhea,gross blood stools,reduced activity,and weight loss in a short time.The gross examination of the intestines showed that the colon of the NFⅠB cKO mice was significantly shorter than that of the non-cKO mice([8.23±0.35]cm vs[10.30±0.36]cm,P＜0.01).Intestinal H-E staining showed changes in mucosal glandular structure and connective tissue hyperplasia with extensive inflammatory cell infiltration in NFⅠB cKO mice.The histological score of NFⅠB cKO mice was significantly higher than that of non-cKO mice(4.25±0.50 vs 0.50±0.58,P＜0.01).Intestinal immunohistochemical staining showed that more CD11b positive cells were recruited in NFⅠB cKO mice than non-cKO mice.Conclusion Myeloid specific NFⅠB cKO mice have been successfully constructed,and NFⅠB in myeloid cells can reduce infiltration of immune cells(granulocytes or/and monocytes)to inhibit intestinal inflammation.

In recent decades, the global community has encountered several significant viral outbreaks, including the Ebola epidemic in West Africa, the Zika virus epidemic in South America, and the recent worldwide COVID-19 pandemic. In these instances, the deployment of effective vaccines has been instrumental in protecting public health. Nevertheless, as new challenges emerge in the prevention and management of infectious diseases, the traditional model of global vaccine development confronts both unprecedented opportunities and challenges. These circumstances underscore the limitations inherent in conventional vaccine development, particularly the protracted timelines and substantial costs involved. This article examines innovative approaches in contemporary vaccine clinical trials, investigates randomization techniques specific to vaccine studies, and delineates essential statistical considerations pertinent to vaccine trial design. The objective is to provide scientific support for vaccine development and to foster ongoing innovation and optimization within the realm of vaccine research and development.

In recent decades, the global community has encountered several significant viral outbreaks, including the Ebola epidemic in West Africa, the Zika virus epidemic in South America, and the recent worldwide COVID-19 pandemic. In these instances, the deployment of effective vaccines has been instrumental in protecting public health. Nevertheless, as new challenges emerge in the prevention and management of infectious diseases, the traditional model of global vaccine development confronts both unprecedented opportunities and challenges. These circumstances underscore the limitations inherent in conventional vaccine development, particularly the protracted timelines and substantial costs involved. This article examines innovative approaches in contemporary vaccine clinical trials, investigates randomization techniques specific to vaccine studies, and delineates essential statistical considerations pertinent to vaccine trial design. The objective is to provide scientific support for vaccine development and to foster ongoing innovation and optimization within the realm of vaccine research and development.

Long non-coding RNA(lncRNA)is a type of non-protein-coding RNA with a length of at least 200 nucleotides.Ex-isting evidence shows that lncRNA affects the growth and development of humans and the occurrence of diseases through various mech-anisms,and plays an irreplaceable role in the differentiation and activation of immune cells and the development process of autoim-mune diseases.In recent years,lncRNA H19 has been found to play a unique regulatory function in pathological processes related to autoimmune diseases,such as inflammation and fibrosis.Therefore,the molecular mechanism involved in H19 is expected to become a potential target for the treatment of autoimmune diseases.In this review,we discussed the related mechanisms of H19 involved in the pathogenesis of autoimmune diseases based on the known functions of H19 and summarized the research results of H19 in several com-mon autoimmune diseases.

Objective To screen long non-coding RNA(lncRNA)associated with disulfidptosis and investigate the immune landscape between lncRNA and pancreatic cancer,for effective guidance in clinical practice.Methods The normal and pancreatic cancer tissue samples were obtained from The Cancer Genome Atlas database,and the lncRNA associated with disulfidptosis was identified based on the Cox and LASSO regression analyses.A risk prognosis model was constructed,and its predictive performance was verified using comprehensive methods.An accurate nomogram was construted to predict the prognosis of patients with pancreatic cancer.The biological differences were analyzed via Gene Ontology,Gene Set Enrichment Analysis,and an immunoassay.The immunotherapy response was estimated using the tumor mutational burden(TMB)score.Results A total of 251 disulfidptosis-related lncRNAs were successfully identified,and three groups of lncRNAs were selected as the reference for the risk model.Pathway analysis showed that immune-related pathways were associated with disulfidptosis-related lncRNA risk models.The risk score was significantly correlated with immune cell infiltration and the ESTIMATE score.Patients with higher risk scores had elevated TMB,indicating that high-risk patients exhibited a better immune checkpoint blockade response.Conclusion The findings of this study contribute to a deeper understanding of disulfidpto-sis-related lncRNA and provide a potential therapeutic strategy for pancreatic cancer.

Humans ; COVID-19 ; HIV Infections/drug therapy* ; Risk Factors

【Objective】 To establish RH gene mRNA sequencing method based on nanopores sequencing and to explore the RHD and RHCE mRNA transcripts in D positive and D^el individuals. 【Methods】 From March 2021 to May 2022, 5 RhD positive samples and 5 D^el samples screened out by hospitals in Chengdu were sent to our laboratory for futher examination. The erythrocytes and buff coat were isolated, then DNA and RNA were extracted.All 10 samples were genotyped by PCR-SSP. After the mRNA was reversely transcribed into cDNA, the full-length mRNA of RHD and RHCE genes were simultaneously amplified by a pair of primers. Sanger sequencing and third-generation sequencing technology based on Nanopore were used to sequence the amplified products, and the types and expressions of different splices of RHD and RHCE gene mRNA transcripts were analyzed. 【Results】 The method established in this study can simultaneously amplify the full length transcripts of RHD and RHCE. Ten different RHD gene mRNA transcripts and nine RHCE gene mRNA transcripts were detected in 10 samples. RHD full-length transcript (RHD-201) can be detected in RhD D^el type, but the expression amount was significantly lower than that in RhD positive samples. The expression amount of transcript RHD-207 (Del789) in D^el samples was significantly higher than that in RhD positive samples. The transcript RHD-208 (Del8910+ 213) was only detected in RhD D^el type individuals, and no significant difference was found between other RHD transcripts and all RHCE transcripts in the two phenotypes. 【Conclusion】 In this study, an analytical method for sequencing full-length transcript isomers of RHD and RHCE mRNA via the third generation was successfully established, and complex alternative splicing patterns were found in RHD and RHCE genes, providing a new method for the study of alternative splicing of blood group gene variants mRNA.

Pancreaticoduodenectomy (PD) is a major operation of abdominal surgery, with high risk, complex operation, more complications and high mortality. Pancreatic fistula is the main cause of death after PD. Due to its complexity and obstinacy, pancreatic fistula has become the top challenge of pancreatic surgery. The authors review the relevant literature and summarize the prevention and management of pancreatic fistula after PD, such as the optimization strategies for pancreatic anastomosis, the use of external stents, prophylactic drains in surgical field, the applica-tion of somatostatin and its analogues, and whole-course nutrition management, based on the precise and comprehensive opinion, in order to provide reference for improving the prevention and treatment of pancreatic fistula after PD.

Primary liver cancer is one of the most severe cancer burdens around the world. Metabolic reprogramming is one phenotype of cancer， and blood metabolic markers are closely associated with metabolic reprogramming and can predict the risk of recurrence and survival or assess the treatment response of liver cancer， with important significance in the stratified management of patients， the development of rational treatment strategies， and the improvement of patient prognosis. By reviewing the recent studies on blood metabolomics in assessing the treatment response or predicting the prognosis of liver cancer， this article summarizes the blood metabolites with predictive significance and their mechanism of action and analyzes the current research status， existing problems， and prospects of this field. It is believed that the metabolites， such as aromatic amino acids， lipids， and bile acids， have an important clinical value in predicting the prognosis of liver cancer， and metabolomics technology has great potential in finding useful metabolites， but there are still many issues to be solved， such as technical limitations， insufficient studies， and multiple influencing factors.

【Objective】 To understand the memory phenotype and function of SARS-CoV-2 reactive CD4+ T cells in healthy individuals. 【Methods】 In this study, SARS-CoV-2-derived peptides were used to stimulate PBMC from participants.SARS-CoV-2 reactive memory T cells were detected by intracellular staining and flow cytometry, and memory phenotype analysis was performed.CBA was used to detect cytokine secretion after SARS-CoV-2-derived peptides stimulation to evaluate the function of SARS-CoV-2 reactive memory T cells. 【Results】 We found that SARS-CoV-2 reactive CD4+ memory T cells could be detected in 40% (6/15) healthy donors.Phenotypic analysis of memory showed that these T cells were mainly composed of central memory T cells(82.2%), and other memory cells accounted for 17.8%.Compared with negative control, IL-10 was significantly decreased after stimulation of SARS-CoV-2-derived peptides (P<0.05), while the secretion of IFNγ, TNFα, IL-2 and IL-4 showed no significant difference. 【Conclusions】 SARS-CoV-2 reactive CD4+ memory T cells are present in healthy individuals from China.