Nanomaterials-mediated lysosomal regulation:a robust protein-clearance approach for the treatment of Alzheimer’s disease

Alzheimer’s disease is a debilitating,progressive neurodegenerative disorder characterized by the progressive accumulation of abnormal proteins,including amyloid plaques and intracellular tau tangles,primarily within the brain.Lysosomes,crucial intracellular organelles responsible for protein degradation,play a key role in maintaining cellular homeostasis.Some studies have suggested a link between the dysregulation of the lysosomal system and pathogenesis of neurodegenerative diseases,including Alzheimer’s disease.Restoring the normal physiological function of lysosomes hold the potential to reduce the pathological burden and improve the symptoms of Alzheimer’s disease.Currently,the efficacy of drugs in treating Alzheimer’s disease is limited,with major challenges in drug delivery efficiency and targeting.Recently,nanomaterials have gained widespread use in Alzheimer’s disease drug research owing to their favorable physical and chemical properties.This review aims to provide a comprehensive overview of recent advances in using nanomaterials(polymeric nanomaterials,nanoemulsions,and carbon-based nanomaterials)to enhance lysosomal function in treating Alzheimer’s disease.This review also explores new concepts and potential therapeutic strategies for Alzheimer’s disease through the integration of nanomaterials and modulation of lysosomal function.In conclusion,this review emphasizes the potential of nanomaterials in modulating lysosomal function to improve the pathological features of Alzheimer’s disease.The application of nanotechnology to the development of Alzheimer’s disease drugs brings new ideas and approaches for future treatment of this disease.

Lupenone improves motor dysfunction in spinal cord injury mice through inhibiting the inflammasome activation and pyroptosis in microglia via the nuclear factor kappa B pathway

Spinal cord injury-induced motor dysfunction is associated with neuroinflammation.Studies have shown that the triterpenoid lupenone,a natural product found in various plants,has a remarkable anti-inflammatory effect in the context of chronic inflammation.However,the effects of lupenone on acute inflammation induced by spinal cord injury remain unknown.In this study,we established an impact-induced mouse model of spinal cord injury,and then treated the injured mice with lupenone(8 mg/kg,twice a day)by intrape ritoneal injection.We also treated BV2 cells with lipopolysaccharide and adenosine5’-triphosphate to simulate the inflammatory response after spinal cord injury.Our res ults showed that lupenone reduced IKBa activation and p65 nuclear translocation,inhibited NLRP3 inflammasome function by modulating nuclear factor kappa B,and enhanced the conve rsion of proinflammatory M1 mic roglial cells into anti-inflammatory M2 microglial cells.Furthermore,lupenone decreased NLRP3 inflammasome activation,NLRP3-induced mic roglial cell polarization,and microglia pyroptosis by inhibiting the nuclear factor kappa B pathway.These findings suggest that lupenone protects against spinal cord injury by inhibiting inflammasomes.

Biomaterials-based anti-inflammatory treatment strategies for Alzheimer's disease

The current therapeutic drugs for Alzheimer's disease only improve symptoms,they do not delay disease progression.Therefo re,there is an urgent need for new effective drugs.The underlying pathogenic factors of Alzheimer's disease are not clear,but neuroinflammation can link various hypotheses of Alzheimer's disease;hence,targeting neuroinflammation may be a new hope for Alzheimer's disease treatment.Inhibiting inflammation can restore neuronal function,promote neuro regeneration,reduce the pathological burden of Alzheimer's disease,and improve or even reverse symptoms of Alzheimer's disease.This review focuses on the relationship between inflammation and various pathological hypotheses of Alzheimer's disease;reports the mechanisms and characteristics of small-molecule drugs(e.g.,nonsteroidal anti-inflammatory drugs,neurosteroids,and plant extracts);macromolecule drugs(e.g.,peptides,proteins,and gene therapeutics);and nanocarriers(e.g.,lipid-based nanoparticles,polymeric nanoparticles,nanoemulsions,and inorganic nanoparticles)in the treatment of Alzheimer's disease.The review also makes recommendations for the prospective development of anti-inflammatory strategies based on nanocarriers for the treatment of Alzheimer's disease.

Reinfection and Risk Factors of SARS-CoV-2 during an Omicron Wave 2022 in Shanghai

Multiple waves of coronavirus disease 2019(COVID-19) outbreaks have affected numerous countries worldwide. The first case of SARS-CoV-2 reinfection was reported in Hong Kong in August 2020^([1]), leading to increased interest in the effectiveness of natural immunity acquired from primary infection. While data reports vary across countries, all findings indicate that prior SARS-CoV-2 infection provides substantial protection against reinfection^([2]). However, natural immunity from infection with previous non-Omicron or early Omicron sub-lineages offers lower levels of protection against Omicron reinfection, with rates below 60%^([3]) and approximately 75%[4], respectively.

Polydatin ameliorates hepatic ischemia-reperfusion injury by modulating macrophage polarization

Background:Polydatin,a glucoside of resveratrol,has shown protective effects against various diseases.However,little is known about its effect on hepatic ischemia-reperfusion(I/R)injury.This study aimed to elucidate whether polydatin protects liver against I/R-induced injury and to explore the underlying mechanism.Methods:After gavage feeding polydatin once daily for a week,mice underwent a partial hepatic I/R procedure.Serum alanine aminotransferase(ALT)/aspartate aminotransferase(AST),hematoxylin-eosin(H&E)and TdT-mediated dUTP nick-end labeling(TUNEL)staining were used to evaluate liver injury.The severity related to the inflammatory response and reactive oxygen species(ROS)production was also investigated.Furthermore,immunofluorescence and Western blotting were used to detect macrophage polarization and the NF-κB signaling pathway in macrophages.Results:Compared with the I/R group,polydatin pretreatment significantly attenuated I/R-induced liver damage and apoptosis.The oxidative stress marker(dihydroethidium fluorescence,malondialdehyde,superoxide dismutase and glutathione peroxidase)and I/R related inflammatory cytokines(interleukin1β,interleukin-10 and tumor necrosis factor-α)were significantly suppressed after polydatin treatment.In addition,the result of immunofluorescence indicated that polydatin reduced the polarization of macrophages toward M1 macrophages both in vivo and in vitro.Western blotting showed that polydatin inhibited the pro-inflammatory function of RAW264.7 via down-regulating the NF-κB signaling pathway.Conclusions:Polydatin protects the liver from I/R injury by remodeling macrophage polarization via NFκB signaling.

Hounsfield units in assessing bone mineral density in ankylosing spondylitis patients with cervical fracture-dislocation

BACKGROUND Cervical spine fracture-dislocations in patients with ankylosing spondylitis(AS)are mostly unstable and require surgery.However,osteoporosis,one of the comorbidities for AS,could lead to detrimental prognoses.There are few accurate assessments of bone mineral density in AS patients.AIM To analyze Hounsfield units(HUs)for assessing bone mineral density in AS patients with cervical fracture-dislocation.METHODS The HUs from C2 to C7 of 51 patients obtained from computed tomography(CT)scans and three-dimensional reconstruction of the cervical spine were independently assessed by two trained spinal surgeons and statistically analyzed.Inter-reader reliability and agreement were assessed by interclass correlation coefficient.RESULTS The HUs decreased gradually from C2 to C7.The mean values of the left and right levels were significantly higher than those in the middle.Among the 51 patients,25 patients(49.02%)may be diagnosed with osteoporosis,and 16 patients(31.37%)may be diagnosed with osteopenia.CONCLUSION The HUs obtained by cervical spine CT are feasible for assessing bone mineral density with excellent agreement in AS patients with cervical fracture-dislocation.

DAPK2 promotes autophagy to accelerate the progression of ossification of the posterior longitudinal ligament through the mTORC1 complex

Background:Ossification of the posterior longitudinal ligament(OPLL)is a prevalent condition in orthopedics.While death-associated protein kinase 2(DAPK2)is known to play roles in cellular apoptosis and autophagy,its specific contributions to the advancement of OPLL are not well understood.Methods:Ligament fibroblasts were harvested from patients diagnosed with OPLL.Techniques such as real-time reverse transcriptasepolymerase chain reaction(RT-qPCR)and Western blot analysis were employed to assess DAPK2 levels in both ligament tissues and cultured fibroblasts.The extent of osteogenic differentiation in these cells was evaluated using an alizarin red S(ARS)staining.Additionally,the expression of ossification markers and autophagy markers was quantified.The autophagic activity was further analyzed through LC3 immunofluorescence and transmission electron microscopy(TEM).An in vivo heterotopic bone formation assay was conducted in mice to assess the role of DAPK2 in ossification.Results:Elevated DAPK2 expression was confirmed in both OPLL patient tissues and derived fibroblasts,in contrast to non-OPLL controls.Silencing of DAPK2 significantly curtailed osteogenic differentiation and autophagy in these fibroblasts,evidenced by decreased levels of LC3,and Beclin1,and reduced autophagosome formation.Additionally,DAPK2 was found to inhibit the mechanistic target of the rapamycin complex 1(mTORC1)complex’s activity.In vivo studies demonstrated that DAPK2 facilitates ossification,and this effect could be counteracted by the mTORC1 inhibitor rapamycin.Conclusion:DAPK2 enhances autophagy and osteogenic processes in OPLL through modulation of the mTORC1 pathway.

Cardiac infiltration of diffuse large B-cell lymphoma manifesting as sustained ventricular tachycardia:a case report

Cardiac tumors are rare.However,cardiac metastases can occur in up to 10%of patients with cancer.Among cardiac neoplasms,metastases are much more common than primary cardiac tumors.[1]Metastatic cardiac neoplasms most frequently metastasize from the respiratory system.

Multifaceted roles of lymphatic and blood endothelial cells in the tumor microenvironment of hepatocellular carcinoma:A comprehensive review

The tumor microenvironment is a complex network of cells,extracellular matrix,and signaling molecules that plays a critical role in tumor progression and metastasis.Lymphatic and blood vessels are major routes for solid tumor metastasis and essential parts of tumor drainage conduits.However,recent studies have shown that lymphatic endothelial cells(LECs)and blood endothelial cells(BECs)also play multifaceted roles in the tumor microenvironment beyond their structural functions,particularly in hepatocellular carcinoma(HCC).This comprehensive review summarizes the diverse roles played by LECs and BECs in HCC,including their involvement in angiogenesis,immune modulation,lymphangiogenesis,and metastasis.By providing a detailed account of the complex interplay between LECs,BECs,and tumor cells,this review aims to shed light on future research directions regarding the immune regulatory function of LECs and potential therapeutic targets for HCC.

MH-STRALP:A scoring system for prognostication in patients with upper gastrointestinal bleeding

BACKGROUND Upper gastrointestinal bleeding(UGIB)is a common medical emergency and early assessment of its outcomes is vital for treatment decisions.AIM To develop a new scoring system to predict its prognosis.METHODS In this retrospective study,692 patients with UGIB were enrolled from two cen-ters and divided into a training(n=591)and a validation cohort(n=101).The clinical data were collected to develop new prognostic prediction models.The en-dpoint was compound outcome defined as(1)demand for emergency surgery or vascular intervention,(2)being transferred to the intensive care unit,or(3)death during hos-pitalization.The models’predictive ability was compared with previously esta-blished scores by receiver operating characteristic(ROC)curves.RESULTS Totally 22.2%(131/591)patients in the training cohort and 22.8%(23/101)in the validation cohort presented poor outcomes.Based on the stepwise-forward Lo-gistic regression analysis,eight predictors were integrated to determine a new post-endoscopic prognostic scoring system(MH-STRALP);a nomogram was de-termined to present the model.Compared with the previous scores(GBS,Rock-all,ABC,AIMS65,and PNED score),MH-STRALP showed the best prognostic prediction ability with area under the ROC curves(AUROCs)of 0.899 and 0.826 in the training and validation cohorts,respectively.According to the calibration cur-ve,decision curve analysis,and internal cross-validation,the nomogram showed good calibration ability and net clinical benefit in both cohorts.After removing the endoscopic indicators,the pre-endoscopic model(pre-MH-STRALP score)was conducted.Similarly,the pre-MHSTRALP score showed better predictive value(AUROCs of 0.868 and 0.767 in the training and validation cohorts,respectively)than the other pre-endoscopic scores.CONCLUSION The MH-STRALP score and pre-MH-STRALP score are simple,convenient,and accurate tools for prognosis prediction of UGIB,and may be applied for early decision on its management strategies.

Preservation of superior rectal artery in laparoscopic colectomy:The best choice for slow transit constipation?

Laparoscopic colectomy with ileorectal anastomosis may be beneficial for patients with slow transit constipation who do not respond to conservative treatment,particularly if the superior rectal artery(SRA)is preserved.Several important concerns have been addressed in this commentary.It is important to first go over the definition of surgical procedure as it is used in this text.Second,the current study lacked a control group that had SRA preservation.Thirdly,it would be best to use a prospective,randomized controlled study.Lastly,a description of the mesenteric defect’s state following a laparoscopic colectomy is necessary.

Preliminary study on the preparation of lyophilized acellular nerve scaffold complexes from rabbit sciatic nerves with human umbilical cord mesenchymal stem cells

BACKGROUND The gold standard of care for patients with severe peripheral nerve injury is autologous nerve grafting;however,autologous nerve grafts are usually limited for patients because of the limited number of autologous nerve sources and the loss of neurosensory sensation in the donor area,whereas allogeneic or xenografts are even more limited by immune rejection.Tissue-engineered peripheral nerve scaffolds,with the morphology and structure of natural nerves and complex biological signals,hold the most promise as ideal peripheral nerve“replacements”.AIM To prepare allogenic peripheral nerve scaffolds using a low-toxicity decellularization method,and use human umbilical cord mesenchymal stem cells(hUCMSCs)as seed cells to cultivate scaffold-cell complexes for the repair of injured peripheral nerves.METHODS After obtaining sciatic nerves from New Zealand rabbits,an optimal acellular scaffold preparation scheme was established by mechanical separation,varying lyophilization cycles,and trypsin and DNase digestion at different times.The scaffolds were evaluated by hematoxylin and eosin(HE)and luxol fast blue(LFB)staining.The maximum load,durability,and elastic modulus of the acellular scaffolds were assessed using a universal material testing machine.The acellular scaffolds were implanted into the dorsal erector spinae muscle of SD rats and the scaffold degradation and systemic inflammatory reactions were observed at 3 days,1 week,3 weeks,and 6 weeks following surgery to determine the histocompatibility between xenografts.The effect of acellular scaffold extracts on fibroblast proliferation was assessed using an MTT assay to measure the cytotoxicity of the scaffold residual reagents.In addition,the umbilical cord from cesarean section fetuses was collected,and the Wharton’s jelly(WJ)was separated into culture cells and confirm the osteogenic and adipogenic differentiation of mesenchymal stem cells(MSCs)and hUC-MSCs.The cultured cells were induced to differentiate into Schwann cells by the antioxidant-growth factor induction method,and the differentiated cells and the myelinogenic properties were identified.RESULTS The experiments effectively decellularized the sciatic nerve of the New Zealand rabbits.After comparing the completed acellular scaffolds among the groups,the optimal decellularization preparation steps were established as follows:Mechanical separation of the epineurium,two cycles of lyophilization-rewarming,trypsin digestion for 5 hours,and DNase digestion for 10 hours.After HE staining,no residual nuclear components were evident on the scaffold,whereas the extracellular matrix remained intact.LFB staining showed a significant decrease in myelin sheath composition of the scaffold compared with that before preparation.Biomechanical testing revealed that the maximum tensile strength,elastic modulus,and durability of the acellular scaffold were reduced compared with normal peripheral nerves.Based on the histocompatibility test,the immune response of the recipient SD rats to the scaffold New Zealand rabbits began to decline3 weeks following surgery,and there was no significant rejection after 6 weeks.The MTT assay revealed that the acellular reagent extract had no obvious effects on cell proliferation.The cells were successfully isolated,cultured,and passaged from human umbilical cord WJ by MSC medium,and their ability to differentiate into Schwann-like cells was demonstrated by morphological and immunohistochemical identification.The differentiated cells could also myelinate in vitro.CONCLUSION The acellular peripheral nerve scaffold with complete cell removal and intact matrix may be prepared by combining lyophilization and enzyme digestion.The resulting scaffold exhibited good histocompatibility and low cytotoxicity.In addition,hUC-MSCs have the potential to differentiate into Schwann-like cells with myelinogenic ability following in vitro induction.

Sorafenib inhibits ossification of the posterior longitudinal ligament by blocking LOXL2-mediated vascularization

Ossification of the Posterior Longitudinal Ligament(OPLL)is a degenerative hyperostosis disease characterized by the transformation of the soft and elastic vertebral ligament into bone,resulting in limited spinal mobility and nerve compression.Employing both bulk and single-cell RNA sequencing,we elucidate the molecular characteristics,cellular components,and their evolution during the OPLL process at a single-cell resolution,and validate these findings in clinical samples.This study also uncovers the capability of ligament stem cells to exhibit endothelial cell-like phenotypes in vitro and in vivo.Notably,our study identifies LOXL2 as a key regulator in this process.Through gain-and loss-of-function studies,we elucidate the role of LOXL2 in the endothelial-like differentiation of ligament cells.It acts via the HIF1A pathway,promoting the secretion of downstream VEGFA and PDGF-BB.This function is not related to the enzymatic activity of LOXL2.Furthermore,we identify sorafenib,a broad-spectrum tyrosine kinase inhibitor,as an effective suppressor of LOXL2-mediated vascular morphogenesis.By disrupting the coupling between vascularization and osteogenesis,sorafenib demonstrates significant inhibition of OPLL progression in both BMP-induced and enpp1 deficiency-induced animal models while having no discernible effect on normal bone mass.These findings underscore the potential of sorafenib as a therapeutic intervention for OPLL.

Ultrasound-assisted isolation:A new method to isolate stromal vascular fraction

Background:The stromal vascular fraction(SVF),a cluster of stem and progenitor cells isolated from adipose tissue,holds significant promise for application in regenerative medicine.However,the existing methods for SVF isolation are time-consuming and expensive.Thus,in this study,we explored a new method of SVF extrac-tion-ultrasound-assisted SVF isolation(USASI)-and compared the viability and characteristics of SVF isolated using different methods.Methods:SVF extraction methods using different combinations of ultrasound power,ultrasound time,collagenase dosage,and collagenase digestion time were compared with those of the control group(collagenase digestion method).The cell yield and vitality of the SVF were evaluated via cell counting and trypan blue staining.The cell components and immunophenotypes of freshly isolated SVF were analyzed using flow cytometry.The prolifer-ative capacity and differentiation potential of the SVF were also identified.Results:Ultrasonication at 95 W-20 kHz for 30 s followed by digestion with 0.15%collagenase for 30 min was identified as the most suitable parameter for the USASI method in isolating SVF,as recommended based on the evaluation of various tested conditions.The USASI method significantly reduced the collagenase dosage and shortened the digestion time.Compared to the collagenase digestion method,the USASI method had a higher cell yield and cell viability,with no adverse effects on cell components,proliferative capacity,or multipotential differentiation capacity.Conclusion:With reduced processing time,lower collagenase dosage,and increased cell yield without impairing the viability and characteristics of SVF,USASI holds the potential to emerge as a time-saving and cost-effective method for future clinical applications.

Mechanisms and Prevention of Acceleration Exposure-Induced Cardiovascular Injuries

Background: The development of high-performance flighter aircraft requires better anti-G ability of aircrew. Under the influence of continuous ±Gz or ±Gx acceleration, aircrew can suffer from cardiovascular change or injuries, which pose a huge threat to flight safety. In recent years, some achievements have been made in the physiological research of sustained ±Gz or ±Gx acceleration, especially the research on the cardiovascular physiological mechanism of sustained ±Gz or ±Gx acceleration. This paper analyzes the research progress. Methods: Original studies of any related fields will be included in the analysis. Searching of databases and manual scanning of the reference lists of the articles found during the original search will be performed. The following data items of included studies will be abstracted for analysis: publication year, first author, country, institution, journal and research priorities. Results: 28 papers were included in this study. The annual number of publications showed an ascending tendency as a whole and reached its peak in 2014 with a total of 6 documents. The country with the most publications was China. The journals of these papers focused on different fields, including military medicine, integrative medicine, preventive medicine, space medicine, environmental medicine and pharmacology. Research priorities mainly covered cardiac arrhythmia, cardiac structure, cardiovascular function, myocardial injury and myocardial enzyme. Conclusions: This paper analyzes the progress of research in the cardiovascular compensatory response. It is expected to provide reference for subsequent exploration of the mechanism of sustained ±Gz or ±Gx acceleration and the selection of ways to protect against anti-G.

A Chinese Multi-Specialty Delphi Consensus to Optimize RAASi Usage and Hyperkalaemia Management in Patients with Chronic Kidney Disease and Heart Failure

Objective Variations are present in common clinical practices regarding best practice in managing hyperkalaemia(HK),there is therefore a need to establish a multi-specialty approach to optimal renin angiotension-aldosterone system inhibitors(RAASi)usage and HK management in patients with chronic kidney disease(CKD)&heart failure(HF).This study aimed to establish a multi-speciality approach to the optimal use of RAASi and the management of HK in patients with CKD and HF.Methods A steering expert group of cardiology and nephrology experts across China were convened to discuss challenges to HK management through a nominal group technique.The group then created a list of 41 statements for a consensus questionnaire,which was distributed for a further survey in extended panel group of cardiologists and nephrologists across China.Consensus was assessed using a modified Delphi technique,with agreement defined as"strong"(≥75%and<90%)and"very strong"(≥90%).The steering group,data collection,and analysis were aided by an independent facilitator.Results A total of 150 responses from 21 provinces across China were recruited in the survey.Respondents were comprised of an even split(n=75,50%)between cardiologists and nephrologists.All 41 statements achieved the 75%consensus agreement threshold,of which 27 statements attained very strong consensus(≥90%agreement)and 14 attained strong consensus(agreement between 75%and 90%).Conclusion Based on the agreement levels from respondents,the steering group agreed a set of recommendations intended to improve patient outcomes in the use of RAASi therapy and HK management in China.

Development and Innovation of Modern Microvascular Anastomoses

High-quality microvascular anastomosis is the foundation of successful microsurgery and one of the most important basic skills for microsurgeons. The traditional manual suture is recognized as the “gold standard” for microvascular anastomosis, but it still has problems such as long operation time and easy to cause vascular damage. In order to improve the success rate of microvascular anastomosis, reduce surgical complications and make the prognosis of patients better. In order to improve the success rate of microvascular anastomosis and reduce the surgical complications, scholars at home and abroad have developed some new vascular anastomosis techniques that are simple, fast and minimally invasive while improving the traditional surgical suturing methods. In this paper, we review the microvascular anastomosis, and its application research in two methods of traditional hand suture and non-suture anastomosis, in order to promote the application development of microvascular anastomosis.

Multi-omics analysis of human tendon adhesion reveals that ACKR1-regulated macrophage migration is involved in regeneration

Tendon adhesion is a common complication after tendon injury with the development of accumulated fibrotic tissues without effective anti-fibrotic therapies,resulting in severe disability.Macrophages are widely recognized as a fibrotic trigger during peritendinous adhesion formation.However,different clusters of macrophages have various functions and receive multiple regulation,which are both still unknown.In our current study,multi-omics analysis including single-cell RNA sequencing and proteomics was performed on both human and mouse tendon adhesion tissue at different stages after tendon injury.The transcriptomes of over 74000 human single cells were profiled.As results,we found that SPP1^(+)macrophages,RGCC^(+)endothelial cells,ACKR1^(+)endothelial cells and ADAM12^(+)fibroblasts participated in tendon adhesion formation.Interestingly,despite specific fibrotic clusters in tendon adhesion,FOLR2^(+)macrophages were identified as an antifibrotic cluster by in vitro experiments using human cells.Furthermore,ACKR1 was verified to regulate FOLR2^(+)macrophages migration at the injured peritendinous site by transplantation of bone marrow from Lysm-Cre;R26R^(tdTomato) mice to lethally irradiated Ackr1^(-/-)mice(Ackr1^(-/-)chimeras;deficient in ACKR1)and control mice(WT chimeras).Compared with WT chimeras,the decline of FOLR2^(+)macrophages was also observed,indicating that ACKR1 was specifically involved in FOLR2^(+)macrophages migration.Taken together,our study not only characterized the fibrosis microenvironment landscape of tendon adhesion by multi-omics analysis,but also uncovered a novel antifibrotic cluster of macrophages and their origin.These results provide potential therapeutic targets against human tendon adhesion.

Evaluation of the Effectiveness of the “1 + 1 + 1” Method in Theoretical Teaching of Emergency Pericardiocentesis

Background: Pericardial effusion may progress to cardiac tamponade when pressure around the heart increases to a level comparable to that of the right and left atria. Patients with cardiac tamponade need timely completion of emergency pericardiocentesis to relieve the threat to the patient’s life, and to save valuable time for patients who need emergency thoracotomy and pericardial window drainage. Pericardiocentesis is a necessary clinical skill for residents in standardized training. In addition, nurses who are familiar with this technology can better assist clinicians to perform this operation. In order to make the medical staff quickly master the theoretical knowledge of emergency pericardiocentesis, we designed a “1 + 1 + 1” teaching method for the theoretical teaching of emergency pericardiocentesis. Objective: This study aims to explore the effectiveness of the “1 + 1 + 1” teaching method in the theoretical teaching of emergency pericardiocentesis. Methods: We used an English teaching video of emergency pericardiocentesis and applied the “1 + 1 + 1” teaching method for theoretical teaching. A questionnaire survey was conducted before and after the lecture among 19 medical staff of different years of service to understand their mastery of the theoretical content of emergency pericardiocentesis before and after the lecture. According to the years of service, the medical staff were divided into three groups: 1 - 3 years (Group A), 4 - 10 years (Group B), and over 10 years (Group C), and the changes in the mastery of various contents by the overall medical staff and each group were statistically analyzed. Results: Before the lecture, the number of people who mastered the indications, contraindications, most commonly used methods, and common complications of emergency pericardiocentesis were 15, 12, 16, and 17, respectively, whereas after the lecture, these numbers increased to 17, 19, 19, and 19, respectively. The overall mastery before and after the lecture was statistically significant (p Conclusion: The “1 + 1 + 1” teaching method can effectively improve the overall mastery level of medical staff’s theoretical knowledge of emergency pericardiocentesis, especially in improving the mastery of contraindications of this operation.

原发性胆汁性肝硬化患者中自身抗原丙酮酸脱氢酶特异性CD8+CTL表位预测及鉴定

目的鉴定原发性胆汁性肝硬化(PBC)患者中自身抗原丙酮酸脱氢酶(PDC-E2)上的HLA-A觹0201限制性CD8+CTL表位,为临床探索特异性免疫治疗奠定基础。方法应用数据库SYFPEITHI预测PDC-E2上两段涵盖B细胞表位和CD4+T细胞表位的氨基酸序列(163～184aa和36～49aa)附近可能存在的HLA-A觹0201限制性T细胞表位,再通过T2细胞株、细胞增殖试验和细胞毒性检测分别分析各抗原肽与HLA-A觹0201的结合力、诱导患者外周血单个核细胞(PBMCs)增殖能力及抗原肽诱导的抗原特异性T细胞杀伤毒性,逐步鉴定HLA-A觹0201限制性CD8+CTL细胞表位。结果数据库初步预测到5个可能性较大的HLA-A觹0201限制性抗原肽,其中两个抗原肽(159～167aa和165～174aa)显示与T2细胞上HLA-A觹0201分子有较高的亲和力,这两个抗原肽能刺激大部分HLA-A觹0201阳性PBC患者PBMC增殖,并且由其诱导产生的CTL具有特异杀伤活性。结论位于PDC-E2内酯酰区上的KLSEGDLLA穴159～167aa)和LLAEIETDKA穴165～174aa雪是PBC患者体内HLA-A觹0201限制性的CD8+CTL表位。