Lung cancer remains one of the most prevalent and lethal malignancies worldwide. The advancement of its precise diagnosis and therapeutic development urgently requires in vitro models that can highly recapitulate the pathophysiological characteristics of human tissues. Organ-on-a-chip has emerged as a novel technological platform that integrates microfluidic engineering, biomaterials, and other engineering strategies with organoid culture. This platform enables precise control over the cellular microenvironment, thereby closely mimicking the three-dimensional structure and physiological functions of human organs in vitro. Organ-on-a-chip systems demonstrate significant advantages in cancer research, developmental biology, and disease modeling, as they not only preserve the heterogeneity and pathological features of patient samples but also support co-culture of various cell types to reconstruct the tumor microenvironment (TME). However, standardized construction methods and integrated analytical strategies for this technology in lung cancer research remain to be further refined. This review systematically elaborates on the key technical principles of organ-on-a-chip and its recent advances in lung cancer modeling, drug screening, and immunotherapy research. It aims to provide a theoretical foundation and technical perspective for promoting the deeper application of organ-on-a-chip in precision medicine and translational research for lung cancer.
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Humans ; Lung Neoplasms/drug therapy* ; Organoids/drug effects* ; Lab-On-A-Chip Devices ; Animals ; Tumor Microenvironment

Objective:To study the clinical manifestations and genetic characteristics of neonatal-onset primary mitochondrial disease (PMD) caused by nuclear gene mutations.Methods:From May 2020 to March 2022, the clinical data, genetic results and follow-up information of neonates with PMD admitted to the Department of Neonatology of our two hospitals were retrospectively analyzed.Results:A total of 4 patients were enrolled, all with hyperlactatemia and metabolic acidosis. In case 1, the fetal cranial MRI showed agenesis of corpus callosum. In case 2, echocardiography after birth indicated hypertrophic cardiomyopathy. Whole exome sequencing found the following mutations: EARS2 nuclear gene c.1294C>T and c.971G>T variants, COA6 nuclear gene c.411_412insAAAG variant, ACAD9 nuclear gene c.1278+1G>A and c.895A>T variants, FOXRED1 nuclear gene c.1054C>T and c.3dup variants. Mitochondrial second-generation sequencing and multiplex ligation-dependent probe amplification showed no abnormalities. Cases 1 and 3 died during the neonatal period. Case 2 died at 2-year-and-2-month of age. Case 4 was followed up to 1 year of age with developmental delay.Conclusions:The main phenotypes of neonatal-onset PMD caused by nuclear gene mutations are hyperlactatemia, refractory metabolic acidosis and cardiomyopathy, which have a poor prognosis. Proactive genetic tests are helpful for early diagnosis.

Objective:To discuss the effect of Aspergillus fumigatus(Af)on DNA damage and interleukin(IL)-33 expression in the human bronchial epithelial cells,and to clarify its related mechanism.Methods:Different concentrations(1,5,and 10 mg·L-1)of Af were used to stimulate the bronchial epithelial BEAS-2B cells to select the appropriate stimulation concentration.When the BEAS-2B cells were treated with N-acetylcysteine(NAC)and Af,the cells were divided into control group,Af group,NAC group,and Af+NAC group.When the BEAS-2B cells were treated with DNA double-strand break repair inhibitor NU7441 and Af,the cells were divided into control group,Af group,NU7441 group,and Af+NU7441 group.The comet assay was used to detect the percentages of comet tail DNA of cells in various groups;immunofluorescence method was used to detect the expression levels of DNA damage-related protein phosphorylated H2AX(yH2AX)in the cells in various groups;2,7-dichlorofluorescein diacetate(DCFH-DA)fluorescence probe was used to detect the levels of reactive oxygen species(ROS)in the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of interleukih-33(IL-33),thymic stromal lymphopoietin(TSLP),and interleukih-25(IL-25)mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of phosphorylated nuclear factor κB(p-NF-κB),phosphorylated ataxia telangiectasia mutated(p-ATM),and γH2AX proteins in the cells in various groups.Results:Compared with control group,the percentage of comet tail DNA and the expression level of γH2AX in the cells in 1 mg·L-1 Af group showed no significant difference(P＞0.05),while the percentage of comet tail DNA and the expression level of γH2AX in the cells in 5 mg·L-1 Af group were significantly increased(P＜0.01);compared with 5 mg·L-1 Af group,the percentage of comet tail DNA and the expression level of γH2AX in the cells in 10 mg·L-1 Af group were significantly increased(P＜0.01).Compared with control group,the ROS levels in the bronchial epithelial cells in 1 mg·L-1 Af group was significantly increased(P＜0.05);compared with 1 mg·L-1 Af group,the ROS level in the cells in 5 mg·L-1 Af group was significantly increased(P＜0.01);compared with 5 mg·L-1 Af group,the ROS level in the cells in 10 mg·L-1 Af group was significantly increased(P＜0.05).After treatment of NAC,compared with Af group,the percentage of comet tail DNA(P＜0.01),the expression level of γH2AX(P＜0.05),and the ROS level(P＜0.01)in the cells in Af+NAC group were significantly decreased;after treatment of NU7441,compared with Af group,the percentage of comet tail DNA and the expression level of yH2AX in the cells in Af+NU7441 group were significantly increased(P＜0.01).The RT-qPCR results showed that after treatment of NAC,compared with control group,the expression level of IL-33 mRNA in the cells in Af group was significantly increased(P＜0.05);compared with Af group,the expression level of IL-33 mRNA in the cells in Af+NAC group was significantly decreased(P＜0.05);after treatment of NU7441,compared with Af group,the expression level of IL-33 mRNA in the cells in Af+NU7441 group was significantly increased(P＜0.05).The Western blotting results showed that after treatment of NAC,compared with control group,the expression levels of p-NF-κB,p-ATM,and γH2AX proteins in the cells in Af group were significantly increased(P＜0.05);after treatment of NU7441,compared with Af group,the expression levels of p-NF-κB,p-ATM,and γH2AX proteins in the cells in Af+NAC group were significantly decreased(P＜0.05);After treat ment of NU7441,compared with Af group,the expression levels of p-NF-κB,p-ATM,and γH2AX proteins in the cells in Af+NU7441 group were significantly increased(P＜0.05).Conclusion:Af promotes the IL-33 expression in the human bronchial epithelial cells by causing DNA damage,and its mechanism may be related to the activation of ATM/NF-κB signaling pathway.

Objective To explore the pathological features of lung tissue in mild chronic obstructive pulmonary disease(COPD)and their association with inflammatory factors.Methods A total of 70 patients who underwent surgery for small lung nodule were prospectively included,and were divided into the normal group(n=10),the mild COPD group(n=50)and the moderate and severe COPD group(n=10).The pathological changes of lung tissue were evaluated after HE,Masson and EVG staining.The expression levels of SMA,Actin and CD31 proteins were detected by immunohistochemistry staining.Tumor necrosis factor α(TNF-α),interleukin-10(IL-10)protein and mRNA levels were detected by immunohistochemistry and qPCR.Results Pulmonary tissue in mild COPD showed widening of alveolar septum,dilation of small airways,mild thickening of blood vessel wall and inflammatory reaction dominated by lymphocyte infiltration.Immunohistochemistry staining showed that contents of SMA and Actin proteins in mild COPD lung tissue were higher than those in the normal group(P＜0.05).In addition,the TNF-α mRNA and the positive rate of TNF-α in lung tissue of mild COPD were significantly higher than those in the normal group,while the IL-10 mRNA was significantly lower than that of the normal group(all P＜0.05).SMA and Actin were positively correlated with the positive expression of inflammatory cytokine TNF-α,but negatively correlated with the positive expression of IL-10(all P＜0.05).Conclusion The main pathological changes of lung tissue in mild COPD include small lung blood vessel remodeling ocharacterized by thickening of small blood vessel smooth muscle layer and lymphocyte-dominated inflammatory response,while the increase of pro-inflammatory factor TNF-α and decrease of anti-inflammatory factor IL-10 are associated with pathological changes of COPD.

AIM: To analyze the chemical ingredients of Qingxin-zishen prescription decoction (QZPD) and predict its main pharmacodynamic substances and mechanism in the prevention and treatment of menopause syndrome (MPS) with the help of high performance liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-Q-TOF/MS) combined with network pharmacology. METHODS: The chemical ingredients of QZPD were identified after analyzing the retention time, exact mass, secondary mass spectrometry fragmentation and other information obtained from HPLC-Q-TOF/MS and comparing them with the established chemical ingredients database and the literatures. The targets of ingredients in QZPD were predicted by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and SwissTargetPrediction database. The disease targets of MPS were obtained through Online Mendelian Inheritance in Man (OMIM) and GeneCards Database. Gene ontology (GO) function enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of potential targets were analyzed with the Metascape database. Cytoscape 3.7.2 software was used to construct the network of active components-key targets-pathways. AutoDockTools 4.2.5 software was applied in the molecular docking verification between the key active components and key targets. RESULTS: A total of 83 components were identified in QZPD and 847 drug targets were predicted. After intersection them with 3 050 disease targets, 395 common targets were obtained. After network topology analysis, 74 key targets were obtained, involving mitogen-activated protein kinase (MAPK), phosphatidylinositol-3 kinase/protein kinase B (PI3K/Akt), transforming growth factor-β (TGF-β) and other signaling pathways. Molecular docking analysis results indicated that 23 key active components, such as berberine, epiberberine, coptisine, geissoschizine methyl ether, liensinine, norcoclaurine, palmatine, quercetin, and luteolin, had good binding activity with several of the key targets. CONCLUSION: This study preliminarily identifies the potential effective chemical ingredients of QZPD, predicts its targets in the prevention and treatment of MPS, which provides supporting information for the further study of the pharmacodynamic substances and mechanisms of QZPD.

The main components of the medicinal plant Ilex pubescens Hook. et Arn. are the pentacyclic triterpenoids with various chemical structures. The modification of the triterpenoid skeleton is closely related to a class of key enzymes downstream of the biosynthetic pathway, cytochrome P450 monooxygenase(CYP450). In this study, unigenes with a length of more than 1100 bp were retrieved from the previously obtained Ilex pubescens transcriptome data, and a cytochrome P450 gene(Unigene 0036170)proposed to have the oxidative function of pentacyclic triterpene C-28 was screened through phylogenetic tree analysis. The gene was named as IpAO2. The open reading frame of IpAO2 is 1443 bp, encoding 480 amino acids. RNA was extracted from the tender leaves of Ilex pubescens, and cDNA was obtained by reverse transcription using the RNA as the template. The specific primers AO2-F and AO2-R were designed according to the DNA sequence of IpAO2, and the IpAO2 gene was amplified by PCR using high fidelity enzyme. The IpAO2 gene was ligated into the pEASY cloning vector to obtain pEASY-IpAO2 plasmid. The ligation product was transformed into E. coli Trans1-T1 competent cells and positive clones were screened. The pESC-TRP plasmid was digested with restriction endonucleases to obtain a linearized vector. Primers V-AO2-F and V-AO2-R were designed and the DNA fragment was amplified by PCR using the pEASY-IpAO2 plasmid as template. The DNA fragment was ligated to the vector pESC-TRP to obtain pESC-TRP-IpAO2 recombinant plasmid. The pESC-TRP-IpAO2 plasmid was transformed into a strain of S. cerevisiae capable of efficiently synthesizing amyrin, and the expression of IpAO2 protein was induced with galactose. After extracting the total protein of recombinant S. cerevisiae, Western blot analysis was carried out through the 6×His antigen tag linked to the C-terminus of the target protein. The results showed that the recombinant protein was synthesized in accordance with the expected size. After 7 days of induction and cultivation, the cells of recombinant yeast were collected. The metabolites of cells were extracted and detected by GC-MS after silanization. Through GC-MS analysis of recombinant yeast metabolites, it was found that the protein encoded by the IpAO2 gene can oxidize α-amyrin and β-amyrin to ursolic acid and oleanolic acid, respectively, indicating that IpAO2 is a pentacyclic triterpene C-28 oxidase gene. Meanwhile, the transmembrane domain and protein tertiary structure of the IpAO2 were predicted using the Phyre2 online tool, the results showing that IpAO2 is a transmembrane protein. This study is of great significance to elucidate the biosynthetic mechanism of triterpenoids of Ilex pubescens, and lays a foundation for the further utilization of the metabolic engineering to produce triterpenoids of Ilex pubescens in S. cerevisiae.

Objective To establish the high myopic choroidal neovascularization (CNV) in guinea pigs and to investigate the role of (matrix metalloproteinase-2,MMP-2) and tissue inhibitor-2 of metalloproteinase (TIMP-2) in high myopic CNV.Methods Seventy-two 2-week-old guinea pigs were randomized into control group (n =36) and high myopia group (n =36).Right eyes were indued form deprivation high myopia for 6 weeks.Thirty guinea pigs were randomly selected in each group,and CNV were induced in the right eyes br the 532 nm laser.MMP-2 and TIMP-2 expressions were investigated by immunohistochemistry pre-laser and 7,14,21,28,35 days after laser induction,respectively,while MMP-2 and TIMP-2 relative expression levels in retinal pigment epithelium (RPE)-choroid-sclera complex were detected by real-time PCR.Integral opitical density (IODs) of positive expression and mRNA relative expression levels of these factors were performed by statistical analyses.Results The expressions of MMP-2 and TIMP-2 were up-regulated in the two groups after laser photocoagulation through immunohistochemistry and real-time PCR examinations.Expression of MMP-2 peaked at 21 d and TIMP-2 at 28 d,respectively.IODs of positive expression and mRNA relative expression levels of MMP-2 were higher in high myopia group than those in control group at each inspective time point,and the differences were statistically significant (P ＜ 0.05).TIMP-2 expression was significantly reduced in high myopia group compared with control group before laser photocoagulation (P＜0,05),while there was no significant difference between the two groups at each time point after laser photocoagulation.Conclusions MMP-2 and TIMP-2 were closely related to the formation of high-myopic CNV.Balance disorders of MMP-2 and TIMP-2 might participate in the occurrence and development of high-myopic CNV.

Objective To probe into the effect of diversified teaching methods in the teaching of di-agnostics and clinical probation.Methods One hundred clinical undergraduates were randomly divided in-to experimental class (fifty students) and traditional class (fifty students). The experimental class was based on the textbook of diagnostics.It used PBL to share teaching resources with teachers and students,and used electronic simulation and standardized patients to carry out clinical probation in combination with clinical practice. Through the objective structured clinical examination and questionnaire survey, students' learning situation of two classes was analyzed. R 3.3.3 was used to perform t test or χ2test for the data of two classes. Result The evaluation results show that the average scores of the students in the experimental class OSCE are excellent, and the average scores of the tests are higher than those of the traditional class, and the difference is statistically significant (P<0.05). The result of questionnaire showed that the students of experimental class were more satisfied than the traditional class students (P<0.05) in all aspects. Conclusions Diversified teaching methods are beneficial to cultivating students' theoretical practice trans-formation ability,clinical thinking ability and independent problem solving ability.

Multiple myeloma is a monoclonal plasma cell malignant proliferative disease,accounting for about 10％ of malignaut tumors of blood system.In recent years,due to the application of hematopoietic stem cell transplantation and the application of MDT by combining new drug such as thalidomide,lenalidomide and bortezomib,the average survival of MM patients was significantly longer than before.The treatment strategy should be individualized before treatment according to the patient's age,physical condition and complications.In this paper,the treatment strategy of multiple myeloma patients were reviewed.