Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Objective:To explore the impacts of smoking on the need for respiratory support and mortality in patients hospitalized with pneumonia.Methods:A total of 24 367 patients hospitalized with pneumonia from 2009 to 2017 in the China Kadoorie Biobank, were included. Smoking status was self-reported, and data regarding respiratory support during hospitalization and mortality during follow-up were obtained from medical claims and death registries, respectively. OR, HR, and 95% CI were calculated and adjusted for potential confounders using logistic regression models and Cox proportional hazards regression models, respectively. Results:Among males, current smokers or those who quit smoking due to illness had higher risks of requiring respiratory support ( OR=1.15, 95% CI: 1.03-1.29), 1-year mortality ( HR=1.66, 95% CI: 1.32-2.08), and 5-year mortality ( HR=1.32, 95% CI: 1.13-1.54) following pneumonia hospitalization compared to nonsmokers. Male smokers who started smoking at a younger age or with longer smoking duration had the highest mortality risks (trend test both P<0.05). Female current smokers or those who quit smoking due to illness had higher risks of 1-year mortality ( HR=1.62, 95% CI: 1.17-2.23) and 5-year mortality ( HR=1.33, 95% CI: 1.06-1.67). We found no statistically significant difference in 90-day mortality between current smokers/those who quit smoking due to illness and nonsmokers. Conclusions:Smoking was associated with higher risks of requiring respiratory support and mortality in patients hospitalized with pneumonia, especially among males and heavy smokers. These findings highlight the need for targeted strategies to promote smoking cessation in patients hospitalized with pneumonia.

Abdominal aortic aneurysm (AAA) is an extremely dangerous aortic dilating disease, with a high mortality rate once ruptured. With the advancement of imaging techniques such as computed tomography angiography (CTA), the detection rate of this disease has increased correspondingly. Early detection and timely treatment are crucial for reducing mortality and improving patient prognosis. Compared to traditional open surgery, endovascular aortic repair (EVAR) for AAA is minimally invasive, using a stent graft to isolate the aneurysm sac as a whole, preventing arterial blood from entering the sac and further preventing the aneurysm from progressing or rupturing. Due to the characteristics of EVAR, precise preoperative assessment of AAA anatomy is required to develop a surgical plan for selecting the appropriate stent diameter and length. However, current attention on preoperative assessment of EVAR is more focused on the neck landing zone of the aneurysm, its shape, calcification, etc., while there is relatively less attention and lack of uniform standards for the selection and positioning judgment of the distal landing zone of the iliac artery. And, a growing number of studies have shown that improper treatment of the distal landing zone may lead to complications such as type Ⅰb endoleak and restenosis after stent implantation, affecting the prognosis of patients. This article provides a review of the current domestic and international methods and progress in the preoperative assessment of the distal landing zone for EVAR.

Objective To investigate the characteristics of epilepsy onset based on the theory of five circuits and six qi in traditional Chinese medicine(TCM).Methods A total of 8 389 epilepsy patients from a community-based natural population cohort study from UK Biobank were included.Frequency and constituent ratio analyses were performed to describe the distribution of epilepsy onset in relation to the five circuits and six qi elements,and statistical inferences were made using chi-square tests.Results The analysis revealed statistically significant differences existed in epilepsy onset across different heavenly stems,earthly branches,recombinant yearly circuit,celestial control and terrestrial effect,and dominant qi(all P＜0.05).From the perspective of circuits,the highest incidence occurred in years corresponding to the"Ji"and"Hai"years,while the lowest incidence was observed in"Bing"and"Mao"years.Years dominated by wood circuit and wind manifestations exhibited the highest incidence,whereas years with cold manifestations showed the lowest incidence,suggesting the liver as the primary pathological site in epilepsy.From the perspective of qi,the third dominant qi was most frequently associated with onset.Regarding presiding and subordinate qi,years with taiyin damp earth presiding and taiyang cold water subordinate had the highest incidence,while years with yangming dry metal presiding and shaoyang monarch fire subordinate had the lowest incidence,indicating that cold-damp years and midsummer periods are potential triggers for epilepsy.Conclusion There is a certain regularity in the epilepsy onset correlated with the features of five circuits and six qi,which can guide prevention and clinical diagnosis and treatment of epilepsy.

Objective:To explore the impacts of smoking on the need for respiratory support and mortality in patients hospitalized with pneumonia.Methods:A total of 24 367 patients hospitalized with pneumonia from 2009 to 2017 in the China Kadoorie Biobank, were included. Smoking status was self-reported, and data regarding respiratory support during hospitalization and mortality during follow-up were obtained from medical claims and death registries, respectively. OR, HR, and 95% CI were calculated and adjusted for potential confounders using logistic regression models and Cox proportional hazards regression models, respectively. Results:Among males, current smokers or those who quit smoking due to illness had higher risks of requiring respiratory support ( OR=1.15, 95% CI: 1.03-1.29), 1-year mortality ( HR=1.66, 95% CI: 1.32-2.08), and 5-year mortality ( HR=1.32, 95% CI: 1.13-1.54) following pneumonia hospitalization compared to nonsmokers. Male smokers who started smoking at a younger age or with longer smoking duration had the highest mortality risks (trend test both P<0.05). Female current smokers or those who quit smoking due to illness had higher risks of 1-year mortality ( HR=1.62, 95% CI: 1.17-2.23) and 5-year mortality ( HR=1.33, 95% CI: 1.06-1.67). We found no statistically significant difference in 90-day mortality between current smokers/those who quit smoking due to illness and nonsmokers. Conclusions:Smoking was associated with higher risks of requiring respiratory support and mortality in patients hospitalized with pneumonia, especially among males and heavy smokers. These findings highlight the need for targeted strategies to promote smoking cessation in patients hospitalized with pneumonia.

Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Abdominal aortic aneurysm (AAA) is an extremely dangerous aortic dilating disease, with a high mortality rate once ruptured. With the advancement of imaging techniques such as computed tomography angiography (CTA), the detection rate of this disease has increased correspondingly. Early detection and timely treatment are crucial for reducing mortality and improving patient prognosis. Compared to traditional open surgery, endovascular aortic repair (EVAR) for AAA is minimally invasive, using a stent graft to isolate the aneurysm sac as a whole, preventing arterial blood from entering the sac and further preventing the aneurysm from progressing or rupturing. Due to the characteristics of EVAR, precise preoperative assessment of AAA anatomy is required to develop a surgical plan for selecting the appropriate stent diameter and length. However, current attention on preoperative assessment of EVAR is more focused on the neck landing zone of the aneurysm, its shape, calcification, etc., while there is relatively less attention and lack of uniform standards for the selection and positioning judgment of the distal landing zone of the iliac artery. And, a growing number of studies have shown that improper treatment of the distal landing zone may lead to complications such as type Ⅰb endoleak and restenosis after stent implantation, affecting the prognosis of patients. This article provides a review of the current domestic and international methods and progress in the preoperative assessment of the distal landing zone for EVAR.

Objective To explore the mechanism of Yangxue Qingnao Granules(Siwu Decoction modified)in the treatment of hypertension based on network pharmacology and molecular docking.Methods The chemical composition analysis results of Yangxue Qingnao Granules in the early stage of the research group were used as the basis for the screening of active compounds.The oral bioavailability≥30%and drug-likeness≥0.18 were used as the screening conditions,and the blood components were supplemented in combination with the literature.TCMSP,chemical professional database and SWISS database were used to predict the targets of potential active compounds of Yangxue Qingnao Granules.Hypertension-related targets were obtained through GeneCards and DiGSeE databases.The intersection of the targets related to hypertension disease and the targets of the potential active compounds of Yangxue Qingnao Granules(common targets)is the potential target of Yangxue Qingnao Granules for the treatment of hypertension.The potential targets were matched with the potential active compounds of Yangxue Qingnao Granules to obtain the antihypertensive active compounds of Yangxue Qingnao Granules.PPI analysis was performed on the potential targets of serum brain granules in the treatment of hypertension through the STRING database,and the core targets were screened according to the degree value.The David database was used to analyze the GO function and KEGG pathway enrichment of the core targets.The core targets with the top six degrees were selected as the docking target proteins,and molecular docking verification was performed with the antihypertensive active compounds.Results A total of 32 potential active compounds,161 active compound targets and 1 539 hypertension-related targets were obtained.After intersection,88 potential targets(common targets)of Yangxue Qingnao Granules in the treatment of hypertension were obtained,involving 29 antihypertensive active compounds.PPI analysis screened 14 core targets:PPARG,ACHE,IL4,CCL2,JUN,NOS3,APP,IL1B,CAT,PTGS2,CASP3,TP53,TNF,IL6,involving 158 GO entries and 13 signaling pathways.Five key active ingredients,chlorogenic acid,rosmarinic acid,paeoniflorin catechinic acid and aloe emodin,were obtained by molecular docking,which were combined with PTGS2,CASP3,TNF,CAT,TP53 and IL6,respectively.Conclusion Yangxue Qingnao Granules may act on core targets such as PTGS2 and CASP3 through key active components such as chlorogenic acid and rosmarinic acid,regulate key pathways such as TNF signaling pathway,MAPK signaling pathway and Toll-like receptor signaling pathway,and play a role in the treatment of hypertension through anti-inflammatory effects.

Diabetic peripheral neuropathy(DPN)is a common and devastating complication of diabetes,for which effective therapies are currently lacking.Disturbed energy status plays a crucial role in DPN pathogenesis.However,the integrated profile of energy metabolism,especially the central carbohy-drate metabolism,remains unclear in DPN.Here,we developed a metabolomics approach by targeting 56 metabolites using high-performance ion chromatography-tandem mass spectrometry(HPIC-MS/MS)to illustrate the integrative characteristics of central carbohydrate metabolism in patients with DPN and streptozotocin-induced DPN rats.Furthermore,JinMaiTong(JMT),a traditional Chinese medicine(TCM)formula,was found to be effective for DPN,improving the peripheral neurological function and alleviating the neuropathology of DPN rats even after demyelination and axonal degeneration.JMT ameliorated DPN by regulating the aberrant energy balance and mitochondrial functions,including excessive glycolysis restoration,tricarboxylic acid cycle improvement,and increased adenosine triphosphate(ATP)generation.Bioenergetic profile was aberrant in cultured rat Schwann cells under high-glucose conditions,which was remarkably corrected by JMT treatment.In-vivo and in-vitro studies revealed that these effects of JMT were mainly attributed to the activation of adenosine monophosphate(AMP)-activated protein kinase(AMPK)and downstream peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PGC-1α).Our results expand the thera-peutic framework for DPN and suggest the integrative modulation of energy metabolism using TCMs,such as JMT,as an effective strategy for its treatment.

African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10％.The total co-incidence rate with real-time fluorescence quantitative PCR was 92％(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.