Objective To explore a novel method for early lung cancer screening based on exhaled breath analysis. Methods This study enrolled patients with suspected pulmonary malignancies and healthy individuals undergoing physical examinations at Sir Run Run Shaw Hospital, Zhejiang University School of Medicine (Qingchun and Qiantang campuses) from September 2023 to June 2024. Enrolled subjects were categorized into a lung cancer group, a benign nodule/tumor group, and a healthy control group. Exhaled breath samples were collected using a sensor array constructed from multiple graphene composite materials to capture breath fingerprints. Based on the collected data, screening and diagnostic models for lung cancer were developed and their performance was evaluated. Results A total of 4 580 subjects were included. Among them, 3 195 were pathologically diagnosed with pulmonary malignancies, including 1 394 males and 1 801 females with a mean age of (58.93±12.37) years, 599 were diagnosed with benign nodules/tumors including 339 males and 260 females with a mean age of (57.10±11.06) years, and 786 were healthy controls with no pulmonary nodules detected on chest CT including 420 males and 366 females with a mean age of (29.75±9.32) years. There were 4 031 patients in the training set and 549 patients in the external testing set. The screening model for high-risk populations (distinguishing patients with lung cancer/high-risk pulmonary nodules from healthy individuals) demonstrated excellent performance, with an area under the receiver operating characteristic curve (AUC) of 0.926. At the optimal Youden’s index (cutoff threshold of 63.5%), the external testing set achieved a specificity of 85.2%, a sensitivity of 88.4%, and an accuracy of 86.8%. The diagnostic model (distinguishing patients with lung cancer/premalignant lesions from those with benign pulmonary nodules/healthy individuals) achieved an AUC of 0.818. At its optimal Youden’s index (cutoff threshold of 47.0%), the external testing set showed a specificity of 71.7%, a sensitivity of 77.3%, and an accuracy of 74.5%. Conclusion The non-invasive breath analysis platform based on a sensor array, developed in this study, can achieve rapid and relatively accurate lung cancer screening by analyzing breath fingerprints. This confirms the feasibility of this technology for early lung cancer screening and holds promise for facilitating the early detection and intervention of lung cancer.

Objective:To explore the application effects of the blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" in Medical Immunology. Methods:Eight-year program clinical medical students who were enrolled at Peking Union Medical College in 2016, 2017, and 2018 were selected as the research subjects. An anonymous questionnaire survey was used to analyze students' multidimensional evaluations of the new teaching methodology. The percentage of frontier hotspot topics in the "Immunology Forum" was used to analyze the students' mastery of cutting-edge knowledge in immunology. The number of "Immunology Forum" related "College Students Innovative Training Plan Program" from 2020 to 2023 was used to analyze the effectiveness of this new teaching method in cultivating students' scientific research and innovation abilities. Statistical analysis was conducted using SPSS 25.0 software, and the normality of all continuous variables was assessed using the Shapiro-Wilk test.Results:The questionnaire survey showed 100.00% satisfaction with the course and 95.90% recognition of the new teaching method. More than 90% of students agreed that the new teaching method improved their learning ability, research ability, innovation ability, internal drive, and academic communication ability. The average proportion of hotspot topics in the "Immunology Forum" was 90.37%±7.12%, which was significantly higher than the proportion of non-hotspot topics (5.67%±3.12%). The average number of topics related to "Immunology Forum" in the "College Students Innovative Training Plan Program" was 17.67±1.15 per session, which was significantly higher than the number of topics not related to "Immunology Forum" (8.00±1.73).Conclusions:The blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" can help students timely grasp the cutting-edge knowledge of immunology, cultivate their learning ability, internal drive, academic communication ability, innovation ability, and research ability, and lay a foundation for students to further explore their scientific research and innovation activities.

Objective To target at the NKG2A-HLA-E inhibitory axis,a pluripotent stem cell(iPSC)-derived geneti-cally engineered natural killer cells(NK cells)with NKG2A knockout(NKG2A KO-iNK)were prepared and then their tumor-killing efficacy was evaluated in vitro.Methods NKG2A was knocked out in iPSCs using gene-editing technology.These cells were then differentiated into NKG2A KO-iNK cells.Surface markers at each differentiation stage were analyzed by flow cytometry.Western blot confirmed NKG2A knockout,and flow cytometry assessed expres-sion of activating receptors(NKG2D)and natural cytotoxicity receptors(NKp30,NKp44,NKp46)in NKG2A KO-iNK cells.Cytotoxic activity against tumor cell lines with varying human leukocyte antigen E(HLA-E)expression level was evaluated via lactate dehydrogenase(LDH)release assay.Results Co-transfection of iPSCs with Cas9 pro-tein and three small-guide RNAs(sgRNAs)targeting at exons 1 and 2 of the KLRC1 gene(encoding NKG2A)suc-cessfully generated monoclonal NKG2A-knockout iPSCs(NKG2A KO-iPSCs)with a single T-base insertion in exon 1.During iPSC differentiation into NK cells,CD34 expression reached 30％-50％at the embryoid body(EB)stage(day 8),while CD56 and CD 16 expression exceeded 80％by day 28.Western blot confirmed complete NKG2A knockout in NKG2A KO-iNK cells.Flow cytometry revealed comparable expression level of activating receptor NKG2D and cytotox-icity receptors(NKp30,NKp44,NKp46)between NKG2A KO-iNK and wild-type iNK(WT-iNK)cells.The LDH assay results indicated that the cytotoxic activity of NKG2A KO-iNK cells against the HLA-E highly-expressed B-cell precursor leukemia cell line Nalm6 cells was significantly higher than that of WT-iNK cells,while there was no signif-icant difference between them and human myeloma cell line H929 cells with low HLA-E expression and human hepa-tocellular carcinoma cell line HepG2 cells with almost no HLA-E expression.Interferon-γ(IFN-γ)pretreatment up regulated HLA-E expression in Nalm6 cells,further amplifying NKG2A KO-iNK-mediated cytotoxicity.Conclusions By disrupting the NKG2A-HLA-E inhibitory axis,NKG2A KO-iNK cells exhibit markedly enhanced in vitro cytotoxic-ity against HLA-E-high tumor cells.This result highlights their potential function as a novel adoptive cell therapy strategy for cancers reliant on HLA-E-mediated immune evasion.

Bone is a highly calcified and vascularized tissue. The vascular system plays a vital role in supporting bone growth and repair, such as the provision of nutrients, growth factors, and metabolic waste transfer. Moreover, the additional functions of the bone vasculature, such as the secretion of various factors and the regulation of bone-related signaling pathways, are essential for maintaining bone health. In the bone microenvironment, bone tissue cells play a critical role in regulating angiogenesis, including osteoblasts, bone marrow mesenchymal stem cells (BMSCs), and osteoclasts. Osteogenesis and bone angiogenesis are closely linked. The decrease in osteogenesis and bone angiogenesis caused by aging leads to osteoporosis. Long noncoding RNAs (lncRNAs) are involved in various physiological processes, including osteogenesis and angiogenesis. Recent studies have shown that lncRNAs could mediate the crosstalk between angiogenesis and osteogenesis. However, the mechanism by which lncRNAs regulate angiogenesis‒osteogenesis crosstalk remains unclear. In this review, we describe in detail the ways in which lncRNAs regulate the crosstalk between osteogenesis and angiogenesis to promote bone health, aiming to provide new directions for the study of the mechanism by which lncRNAs regulate bone metabolism.
RNA, Long Noncoding/physiology* ; Osteogenesis/physiology* ; Humans ; Neovascularization, Physiologic/genetics* ; Bone and Bones/metabolism* ; Animals ; Mesenchymal Stem Cells ; Signal Transduction ; Osteoblasts ; Osteoclasts ; Angiogenesis

Objective:To explore the application effects of the blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" in Medical Immunology. Methods:Eight-year program clinical medical students who were enrolled at Peking Union Medical College in 2016, 2017, and 2018 were selected as the research subjects. An anonymous questionnaire survey was used to analyze students' multidimensional evaluations of the new teaching methodology. The percentage of frontier hotspot topics in the "Immunology Forum" was used to analyze the students' mastery of cutting-edge knowledge in immunology. The number of "Immunology Forum" related "College Students Innovative Training Plan Program" from 2020 to 2023 was used to analyze the effectiveness of this new teaching method in cultivating students' scientific research and innovation abilities. Statistical analysis was conducted using SPSS 25.0 software, and the normality of all continuous variables was assessed using the Shapiro-Wilk test.Results:The questionnaire survey showed 100.00% satisfaction with the course and 95.90% recognition of the new teaching method. More than 90% of students agreed that the new teaching method improved their learning ability, research ability, innovation ability, internal drive, and academic communication ability. The average proportion of hotspot topics in the "Immunology Forum" was 90.37%±7.12%, which was significantly higher than the proportion of non-hotspot topics (5.67%±3.12%). The average number of topics related to "Immunology Forum" in the "College Students Innovative Training Plan Program" was 17.67±1.15 per session, which was significantly higher than the number of topics not related to "Immunology Forum" (8.00±1.73).Conclusions:The blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" can help students timely grasp the cutting-edge knowledge of immunology, cultivate their learning ability, internal drive, academic communication ability, innovation ability, and research ability, and lay a foundation for students to further explore their scientific research and innovation activities.

Objective:To develop a treatment-related quality of life scale for Chinese patients with multiple myeloma (MM) and to test its reliability and validity.Methods:The initial scale was constructed through a literature search, Delphi expert correspondence, and cognitive testing. This study conducted a preliminary survey of 379 patients with MM and a formal survey of 865 patients from the hematology departments of 155 hospitals nationwide from February 2024 to March 2024. The final scale was obtained after conducting item analysis and reliability and validity tests on the initial scale.Results:The constructed scale contains 36 items covering six domains: physiological, psychological, social, treatment side effects, general health, and others. In the preliminary survey, the Cronbach’s alpha coefficient of each item ranged from 0.597 to 0.939, and the test-retest reliability was 0.747 ( P<0.001). Exploratory factor analysis extracted eight common factors with a cumulative variance contribution of 60.058%. In the formal survey, the Cronbach’s alpha coefficient of each item ranged from 0.484 to 0.930, and the test-retest reliability was 0.835 ( P<0.001). Confirmatory factor analysis revealed a comparative fit index of 0.750, a root-mean-square error of approximation of 0.090, and a root-mean-square residual of 0.067. Conclusion:The treatment-related quality of life scale for Chinese patients with MM designed in this study exhibited good reliability and validity, reflecting the impact of treatment on the quality of life of patients. This scale can provide a reference to clinicians for assessing the disease status of patients.

Objective:To develop a treatment-related quality of life scale for Chinese patients with multiple myeloma (MM) and to test its reliability and validity.Methods:The initial scale was constructed through a literature search, Delphi expert correspondence, and cognitive testing. This study conducted a preliminary survey of 379 patients with MM and a formal survey of 865 patients from the hematology departments of 155 hospitals nationwide from February 2024 to March 2024. The final scale was obtained after conducting item analysis and reliability and validity tests on the initial scale.Results:The constructed scale contains 36 items covering six domains: physiological, psychological, social, treatment side effects, general health, and others. In the preliminary survey, the Cronbach’s alpha coefficient of each item ranged from 0.597 to 0.939, and the test-retest reliability was 0.747 ( P<0.001). Exploratory factor analysis extracted eight common factors with a cumulative variance contribution of 60.058%. In the formal survey, the Cronbach’s alpha coefficient of each item ranged from 0.484 to 0.930, and the test-retest reliability was 0.835 ( P<0.001). Confirmatory factor analysis revealed a comparative fit index of 0.750, a root-mean-square error of approximation of 0.090, and a root-mean-square residual of 0.067. Conclusion:The treatment-related quality of life scale for Chinese patients with MM designed in this study exhibited good reliability and validity, reflecting the impact of treatment on the quality of life of patients. This scale can provide a reference to clinicians for assessing the disease status of patients.

AIM:Study the effects of icariin-astragaloside IV-puerarin mixture(Yin-Huang-Ge mixture,YHG)on cognitive dysfunction and brain tissue ferroptosis in Alzheimer disease(AD)model mice,and explore its mecha-nism.METHODS:APP/PS1 mice were randomly divided into APP/PS1 group,icariin-astragaloside IV-puerarin mixture(APP/PS1+YHG)group,and idebenone(APP/PS1+IDE)group using a random number table method.C57BL/6J mice of the same age were selected as the normal group.After one month of continuous administration,Morris water maze was used to test the learning and memory abilities of mice.Nissl staining was used to observe the morphological changes in the hippocampus of mice.The ultrastructure of neurons in each group of mice was observed under electron microscopy.West-ern blot was used to detect the expression of FSP1 protein in hippocampal tissue.ELISA detection of coenzyme Q10(CoQ10),CoQ10H2,and 4-hydroxynonenal(4-HNE)content.Biochemical reagent kit is used to detect malondialde-hyde(MDA)content.RESULTS:Compared with the normal group,APP/PS1 mice showed decreased learning and mem-ory abilities,with loosely arranged and irregularly shaped hippocampal CA3 neurons.Hippocampal neurons exhibit mito-chondrial shrinkage,incomplete cristae,and increased membrane density.The expression of FSP1 protein in the brain de-creased(P＜0.05).The levels of CoQ10 and CoQ10H2 were both reduced(P＜0.01).The levels of 4-HNE and MDA in-creased(P＜0.01).After using icariin-astragaloside IV-puerarin mixture and idebenone,the learning and memory abili-ties of mice were improved.The neuronal structure in the hippocampal CA3 region is more tightly arranged and has a regu-lar shape compared to the model group.The mitochondrial structure is relatively clear,the mitochondrial membrane is rel-atively normal,and the cristae are relatively intact.The expression of FSP1 protein in the brain increased(P＜0.05).The levels of CoQ10 and CoQ10H2 both increased(P＜0.05).The levels of 4-HNE and MDA were significantly reduced(P＜0.01).CONCLUSION:The icariin-astragaloside IV-puerarin mixture can improve cognition in AD mice,and its mecha-nism may be related to inhibiting ferroptosis by activating the FSP1/CoQ10 signaling pathway.

AIM:Study the effects of icariin-astragaloside IV-puerarin mixture(Yin-Huang-Ge mixture,YHG)on cognitive dysfunction and brain tissue ferroptosis in Alzheimer disease(AD)model mice,and explore its mecha-nism.METHODS:APP/PS1 mice were randomly divided into APP/PS1 group,icariin-astragaloside IV-puerarin mixture(APP/PS1+YHG)group,and idebenone(APP/PS1+IDE)group using a random number table method.C57BL/6J mice of the same age were selected as the normal group.After one month of continuous administration,Morris water maze was used to test the learning and memory abilities of mice.Nissl staining was used to observe the morphological changes in the hippocampus of mice.The ultrastructure of neurons in each group of mice was observed under electron microscopy.West-ern blot was used to detect the expression of FSP1 protein in hippocampal tissue.ELISA detection of coenzyme Q10(CoQ10),CoQ10H2,and 4-hydroxynonenal(4-HNE)content.Biochemical reagent kit is used to detect malondialde-hyde(MDA)content.RESULTS:Compared with the normal group,APP/PS1 mice showed decreased learning and mem-ory abilities,with loosely arranged and irregularly shaped hippocampal CA3 neurons.Hippocampal neurons exhibit mito-chondrial shrinkage,incomplete cristae,and increased membrane density.The expression of FSP1 protein in the brain de-creased(P＜0.05).The levels of CoQ10 and CoQ10H2 were both reduced(P＜0.01).The levels of 4-HNE and MDA in-creased(P＜0.01).After using icariin-astragaloside IV-puerarin mixture and idebenone,the learning and memory abili-ties of mice were improved.The neuronal structure in the hippocampal CA3 region is more tightly arranged and has a regu-lar shape compared to the model group.The mitochondrial structure is relatively clear,the mitochondrial membrane is rel-atively normal,and the cristae are relatively intact.The expression of FSP1 protein in the brain increased(P＜0.05).The levels of CoQ10 and CoQ10H2 both increased(P＜0.05).The levels of 4-HNE and MDA were significantly reduced(P＜0.01).CONCLUSION:The icariin-astragaloside IV-puerarin mixture can improve cognition in AD mice,and its mecha-nism may be related to inhibiting ferroptosis by activating the FSP1/CoQ10 signaling pathway.

In order to develop a positive quality control products for the detection of porcine repro-ductive and respiratory syndrome virus(PRRSV)nucleic acid by real-time fluorescent quantitative PCR(RT-qPCR),the positive quality control products of PRRSV-1 and PRRSV-2 M genes were prepared using armored RNA technology of MS2 phage.PRRSV-1 and PRRSV-2 M genes were amplified,purified and recovered,and ligated into pET28b vector containing MS2 mature enzyme protein gene and capsid protein.After transformed into BL21(DE3),the gene products were in-duced by IPTG and purified by PEG6000 precipitation method to prepare the armored RNA virus-like particles(AR-PRRSV)containing PRRSV M gene.Following the performance evaluation,as the positive quality control products of PRRSV-1 and PRRSV-2 M genes,AR-PRRSV1M and AR-PRRSV2M were calculated using YY/T 1652-2019 standard.Results showed that it had a good u-niformity,stable storage for the armored virus-like particles at-20,4,25 ℃ for 60 d,and 37 ℃ for 30 d.The prepared armored virus-like particles AR-PRRSV1M and AR-PRRSV2M were deter-mined by digital quantitative PCR(ddPCR)after preliminary quantification by RT-qPCR.The 104 copies/μL of AR-PRRSV1M and AR-PRRSV2M ddPCR fixation was(1.33+0.50)× 104 cop-ies/μL.The above results indicates that the AR-PRRSVM can be used as the quality control of the whole detection process(nucleic acid extraction,reverse transcription and RT-qPCR).