Hypoxia is an important feature of solid tumors microenvironment, which is closely related to tumor highly aggressive, metastatic characteristics and poor prognosis. Therefore, it is of great significance to establish an efficient, rapid, sensitive and non-invasive method for detection of cell endogenous hypoxia and exogenous hypoxia. In this study, a ratio fluorescence probe was designed to detect endogenous and exogenous hypoxia levels, and the ratio fluorescence imaging was used to detect intracellular hypoxia. The oxygen sensitive fluorescent dye tris(4,7-biphenyl- 1,10)-phenanthroline) ruthenium dichloride ([(Ru(dpp)(3))]Cl-2) was loaded on mesoporous yolk-shell organosilicon nanoparticle (MYSN) doped with fluorescein isothiocyanate (FITC). The linear response range of the probe was 5-290 mu mol/L, the response time was 1 min, and the detection limit was as low as 4.68 mu mol/L. Cell experiments showed that the as-prepared probe could be significantly taken up, exhibiting high specificity and high stability in fluorescence imaging of endogenous and exogenous hypoxia.

An outbreak of a novel coronavirus was reported in Wuhan, China, in late 2019. It has spread rapidly through China and many other countries, causing a global pandemic. Since February 2020, over 28 countries/regions have reported confirmed cases. Individuals with the infection known as coronavirus disease-19 (COVID-19) have similar clinical features as severe acute respiratory syndrome first encountered 17 years ago, with fever, cough, and upper airway congestion, along with high production of proinflammatory cytokines (PICs), which form a cytokine storm. PICs induced by COVID-19 include interleukin (IL)-6, IL-17, and monocyte chemoattractant protein-1. The production of cytokines is regulated by activated nuclear factor-kB and involves downstream pathways such as Janus kinase/signal transducers and activators transcription. Protein expression is also regulated by post-translational modification of chromosomal markers. Lysine residues in the peptide tails stretching out from the core of histones bind the sequence upstream of the coding portion of genomic DNA. Covalent modification, particularly methylation, activates or represses gene transcription. PICs have been reported to be induced by histone modification and stimulate exudation of hyaluronic acid, which is implicated in the occurrence of COVID-19. These findings indicate the impact of the expression of PICs on the pathogenesis and therapeutic targeting of COVID-19.

目的 本文通过细胞功能学实验探讨循环miR-3182和其靶基因磷酯磷酸化酶4(LPPR4)在成骨细胞分化成熟中的调控作用.方法 本文通过对GEO数据GSE63446及GSE62402分析循环MicroRNA(miRNA)和组织表达谱信使RNA(mRNA)分别在人体低水平骨密度(BMD)与高水平BMD的表达谱的差异.对差异化表达的miRNA及其潜在调控靶点在表达谱芯片数据进行调控比对.对可能调控BMD水平的LPPR4基因进行过表达和微小RNA(siRNA)干扰方法研究其对人成骨细胞hFOB1.19分化成熟的研究.结果 外周循环miR-3182在低BMD人群中显著高表达,其在区分高BMD和低BMD人群有非常好的敏感性和特异性.慢病毒质粒载体介导的miR-3182高表达显著降低细胞LPPR4基因mRNA水平和蛋白水平,并抑制成骨细胞hFOB1.19矿化结节形成.LPPR4蛋白高表达显著增加人成骨细胞hFOB1.19矿化结节的形成率和分化标志物OPG表达水平.结论 受miR-3182调控的LPPR4基因促进成骨细胞分化.

The National Medical Licensing Examination has become one of the most important indicator s to measure the teaching quality of medical colleges and universities. In this paper, by analyzing the status of pathophysiology in National Medical Licensing Examination and the current problems existing in pathophysiology teaching, the author proposed a scheme of reform in the pathophysiology teaching based on Medical Licensing Examination, including changing teaching idea, optimizing teaching content, reforming teaching means and adjusting the assessment methods . This reform aims to make the pathophysiology teaching really serve the needs of clinical application.

病理生理学是联系基础医学和临床医学的桥梁学科,提高其教学质量是重中之重.在病理生理学教学中尝试采用PBL与TBL相结合教学方法,将其与传统教学模式进行对比,探索新型教学方法的作用效果.结果表明,采用PBL与TBL相结合教学方法有利于提高教学质量,一定程度上提高学生的学习成绩,并提高学习主观能动性、自学能力、综合分析能力和创新意识等素质,加强了团队合作能力和交际能力.

探讨微课结合PBL教学法在病理生理学病例讨论教学中的应用效果,寻求提高教学质量的有效方法.以2个病例为例,采用微课结合PBL教学方案实施,采取问卷调查和对照实验的研究方法,对教学效果进行分析.实验班平均成绩比对照班高出2.51分;得分为75-80分数段,实验班比对照班高11.75％;而75分以下,对照班比实验班高出18.3％.在病理生理学病例讨论教学中,采用微课结合PBL教学法的效果优于PBL教学法.

病理生理学被称为临床执业医师资格考试的"影子学科".文章回顾性分析了近年来临床执业医师资格考试中涉及的病理生理学知识点,并就病理生理学在基础综合中的"强化作用"、临床综合中的"桥梁作用"及临床思维培养中的"促进作用"进行了阐述,以期为深化病理生理学教学改革提供导向作用.

本文在"互联网+"背景下,对微课和移动学习进行了分析,针对病理生理学课程的特点与需求,研究设计基于移动互联网的微课教学资源平台-移动微课,并提出了相应的学习模式及具体实施方案.该平台的建设及学习模式的应用,是经典教学模式的有益补充,可为病理生理学课程教学改革提供的参考.

BACKGROUND:Kruppel family member zinc binding protein 89 (ZBP-89), also known as ZNF148, regulates Bak expression via binding to GC-rich promoter domain. It is not clear if other GC-rich binding factors, such as Sp family members, can interact with ZBPp-89 on Bak expression. This study aims to elucidate the mechanism of Bak expression regulation by ZBP-89 and Sp proteins, based on in vitro experiment and The Cancer Genome Atlas (TCGA) hepatocellular carcinoma (HCC) data cohort.METHODS:We downloaded TCGA hepatocellular carcinoma (HCC) cohort data to analysis the association of Bak transcription level with ZBP-89 and Sp proteins transcription level. HCC cell lines and liver immortal non-tumour cell lines were used for mechanism study, including western blotting analysis, expression vector mediated gene expression and siRNA interference.RESULTS:Results showed that cancer tissues have higher Bak transcription level compared with adjacent non-cancer tissues. Bak transcription level was correlated with Sp1 and Sp3 expression level, while no correlation was found in ZBP-89 and Bak, neither Sp2 nor Sp4. Mithramycin A (MMA) induced Bak expression in a dose-dependent manner. Western blotting results showed Sp1 overexpression increased Bak expression both in liver immortal non-tumour cells and HCC cells. Interference Sp1 expression could inhibit Bak expression alone. ZBP-89 siRNA suppressed Bak expression even in the presence of MMA treatment and S1 overexpression. Additionally, Bak and Sp1 level were associated with HCC patient survival.CONCLUSIONS:Bak expression required ZBP-89 and Sp1 cooperative regulation simultaneously.

Objective To screen the extraction method to obtain the total alkaloids of Squilla oratoria,and investigate the effects of the total alkaloids on nasopharyngeal carcinoma cell line CNE-2Z.Methods The acid extraction and alkaline deposition,and ethyl acetate extraction were used in the extraction and separation of the ethanol extract of Squilla oratoria and fresh Squilla oratoria,and then the supernatant extract (L for short) and deposition extract (S for short) were collected.Thinlayer chromatography (TLC) and chemical reactions were used to explore the deploying condition and determine the nature preliminarily.Following,the CCK-8 method was used to observe each extract on the growth and proliferation of human nasopharyngeal carcinoma CNE-2Z cells in vitro.Results The maximum alkaloids were extracted with the acid extraction and alkaline deposition,and ethyl acetate extraction was directly extracted from fresh Squilla oratoria.The chloroformcyclohexane system was the best deploying system for the total alkaloids of Squilla oratoria.The results of color identification by TLC showed that the precipitation extract (S for short) belong to alkaloids.Through the CCK-8 method screening,it was discovered that S1 and S2 had the strong inhibitory action on the human nasopharyngeal carcinoma CNE-2Z cell growth,but the function of L1 and L2 relatives was not obvious.The IC50(s) of cell viability on nasopharyngeal carcinoma CNE-2Z cells after treated with the total alkaloids ofSquilla oratoria S1 and S2 for 48 h were (71.18±0.56) μg/mL and (49.31± 0.65)μg/mL,respectively.Conclusions The method of extraction of the total alkaloids from fresh Squilla oratoria by the acid extraction and alkaline deposition,and ethyl acetate extraction is stable,convenient and efficient.The total alkaloids of Squilla oratoria have an inhibitory effect on the cell growth of CNE-2Z cells,which has a good development and application prospect.

Objective To extract and analyze the anti-tumor function of the total alkaloids of Squilla oratoria against the human hep-atoma HepG2 cell. Methods The total alkaloids of Squilla oratoria were extracted by acid extraction and alkaline deposition,then isolated by ethyl acetate;CCK-8 assay,plate clone formation assay,flow cytometry,were performed to measure the effect of the total alkaloids of Squilla oratoria on the human hepatoma HepG2 cell. Results The IC50 of cell viability on HepG2 cells after treated with S for 24 h,48 h,and 72 h was 746. 72 μg/mL,47. 58 μg/mL or 26. 32 μg/mL,respectively. After treated with S (0 μg/mL, 10 μg/mL,50 μg/mL and 100 μg/mL)for two weeks,the cloning efficiency of HepG2 cells was 56. 1%,35. 7%,14. 1% or 0,re-spectively;After treated with S for 48 h ,the human hepatoma HepG 2 cells in S stage was increased from 26. 14% to 40. 80%. Conclusion The total alkaloids of Squilla oratoria has an inhibitory effect on the cell growth in HepG2 cells through the Pathways of cell cycle arrest.

Guanidinium-functionalized molecules are commonly studied for their use as pharmaceutically active compounds and drugs carriers. Herein, four cyclometalated iridium(III) complexes containing guanidinium ligands have been synthesized and characterized as potential anticancer agents. These complexes exhibit moderate antitumor activity in HeLa, MCF-7, HepG2, CNE-2, and A549 human tumor cells. Interestingly, all complexes showed higher cytotoxicity than cisplatin against a cisplatin-resistant cell line A549R, and less cytotoxicity on the nontumorigenic LO2 cells. Intracellular distribution studies suggest that these complexes are selectively localized in the mitochondria. Mechanism studies indicate that these complexes arrested the cell cycle in the G0/G1 phase and can influence mitochondrial integrity, inducing cancer cell death through reactive oxygen species (ROS)-dependent pathways.

基础医学概论对基础医学各学科知识进行了重组融合,是针对医学院校非医学专业学生开设的一门综合性课程.文章阐述了基础医学概论教学实践中存在的主要问题,如内容多而学时有限、专业针对性不强,学生学习积极性不高、教学和考核模式单一等.并针对这些问题,就教学过程中如何加强课程整合、优化教学内容、改进教学手段和考核方式等方面进行了有益的探索和实践.

Background: Epigallocatechin-3-gallate (EGCG) has exhibited antitumor properties in several types of cancers, including nasopharyngeal carcinoma (NPC), but the molecular mechanisms underlying this function remain incompletely understood. The aim of the present study was to characterize the global impact of EGCG on the expression of microRNAs (miRNAs) in NPC cells. Methods: Using microarray analysis, the alterations of miRNA expression profiles were investigated in EGCG-treated CNE2 cells. Furthermore, the target genes and signaling pathways regulated by EGCG-specific miRNAs were identified using target prediction program and gene ontology analysis. Results: A total of 14 miRNAs exhibited >2-fold expression changes in a dose-dependent manner after treatment with 20 μmol/L and 40 μmol/L EGCG. Totally 43, 49, and 52 target genes from these differentially expressed miRNAs were associated with the apoptosis, cell cycle regulation, and cell proliferation, respectively. A total of 66 signaling pathways, primarily involved in cancer development and lipid and glucose metabolism, were shown to be regulated by EGCG-specific miRNAs. Conclusion: EGCG induces considerable alterations of miRNA expression profiles in CNE2 cells, which provides mechanistic insights into cellular responses and antitumor activity mediated by EGCG.

This study aims to identify prognostic microRNAs (miRNAs) biomarkers for diagnosis and survival of hepatocellular carcinoma (HCC) based on large patients cohort analysis. HCC patient cohort data were downloaded from The Cancer Genome Atlas, including paired HCC and adjacent non-cancer tissues. Receiver operating characteristic curve method was used to classify cancer and non-cancer tissues according to microRNAs expression levels. The aberrant microRNAs expression level were ranked and risked for building a prognostic miRNAs signature model. Kaplan-Meier survival was used to analyze the differences among various risk factors in accordance with miRNAs ranking scores. The study showed 33-miRNA signature, 11 were down-regulated and 22 were up-regulated through comparison between cancer samples and non-cancer samples. The maximum correct classification rate is up to 98.7%. Five microRNAs, hsa-mir-3677, hsa-mir-421, hsa-mir-326, hsa-mir-424 and hsa-mir-511-2, significantly correlated with patient survival. The survival rate and time negatively associated with lowering miRNAs index. In the low risk group, over 70% patients showed 5 years survival, while none patients survived longer than 5 years in the high risk group. MiR-424, miR-326 and miR-511 could be applied for HCC diagnostic biomarkers. These five miRNAs were significantly associated with lysosome pathway and D-Glutamine and D-glutamate metabolism pathway via Kyoto Encyclopedia of Genes and Genomes pathway analysis and Gene Ontology annotation. Conclusively, the five miRNAs expression signature could be used as HCC prognostic and diagnostic biomarkers.

目的:探讨思维导图教学模式在病理生理学“教”与“学”中的应用.方法:对照组采用传统的教学模式,实验组采用思维导图教学模式.通过问卷调查、考试成绩和学生座谈综合分析“教”与“学”的效果.结果:思维导图教学模式能够激发学生的学习兴趣、提高学习效率,培养发散性思维.但是,与对照组相比,考试成绩没有统计学差异.结论:思维导图教学模式可以在病理生理学教学中推广,但是需要更合理的评价标准.

AIM:To study the effect of epigallocatechin-3-gallate (EGCG) on the proliferation of human naso-pharyngeal carcinoma ( NPC) cells, and to explore its mechanism by targeting miR-34a.METHODS: Nasopharyngeal carcinoma CNE-2Z cells were treated with various concentrations of EGCG .The ability of cell proliferation was detected by CCK-8 assay, 5-ethynyl-2-deoxyuridine (EdU) incorporation assay and colony-forming assay.The cell cycle distributions were analyzed by flow cytometry .The protein levels of P53 and Notch1 were detected by Western blot .The expression of miR-34a and Notch1 mRNA was measured by real-time PCR.RESULTS:EGCG effectively inhibited the proliferation and colony formation of CNE-2Z cells in a dose-dependent manner , which was related to its induction of cell cycle arrest at G 0/G1 phase.The expression of P53 and miR-34a in CNE-2Z cells was significantly increased after treated with EGCG , while the expression of Notch1 at mRNA and protein levels was markedly suppressed .CONCLUSION:EGCG induces cell cycle arrest and suppresses cell proliferation by regulating the P 53/miR-34a/Notch1 pathway in NPC cells.

Objective To study the inhibitory effects of 20(R)- ginsenoside Rg3 on vasculogenic mimicry and migration of human nasopharyngeal carcinoma CNE - 2 cell line in vitro. Methods CNE - 2 cells treated with different concentrations of Rg3(0 μmol·L - 1 ,38 μmol·L - 1 ,76 μmol·L - 1 and 114 μmol·L - 1 )were assayed with transwell assay and anti - angiogenic test for testing the potential of migration and tube - like structure(TLSs)formation,respec-tively. Meanwhile,the expression of COX - 2,HIF - 1α,VEGF and Fascin1 proteins were detected by Western blotting. Results The formation of TLSs in CNE - 2 cells were significantly inhibited by Rg3 in concentration dependent manners (P < 0. 01);so was the migration ability of CNE - 2 cells(P < 0. 05). At the same time,the expression of COX - 2, HIF - 1α,VEGF and Fascin1 in CNE - 2 cells were down - regulated by Rg3. A significantly positive correlation between the expression of Fascin1 and the migration ability was observed(P < 0. 05),so was the significantly positive correlation between the expression of COX - 2,HIF - 1α and VEGF and the inhibition of the TLS formation(P < 0. 05). Conclusion Rg3 can inhibit the vasculogenic mimicry and migration of CNE - 2 cells in vitro,via the down - regulation of COX - 2, HIF - 1α,VEGF and Fascin1.

目的:尝试多元互动式教学法在生理科学实验课教学中的应用,以提高实验教学质量.方法:随机抽取广东医学院2008级临床医学本科两个班学生为研究对象,通过对学生实验技能考核成绩以及调查问卷结果进行分析,比较传统实验教学方法与多元互动式教学方法的教学效果.结果:86.7％(26/30)的学生对多元互动式教学模式的评价较好,实验组平均总评成绩明显高于对照组(P＜0.01),实验组动手操作能力、实验观察能力及综合分析能力得到提高(P＜0.01).结论:多元互动式教学模式有利于调动学生的学习积极性,提高实验教学质量.