随着互联网技术高速发展,线上教学已成为学生学习的主流方式之一.对"互联网+"与微课混合教学模式在病原生物学教学中的应用进行探索,发现学生对"互联网+"与微课混合教学模式认同度较高,自主学习效率有所提升,但是学生教学参与度、教师信息化技术水平有待提高.

Metastasis is the major reason for poor prognosis and high fatality in hepatocellular carcinoma (HCC).Due to the "Warburg effect", an acidic tumor microenvironment (TME) exists in solid tumors and plays a critical role in cancer metastasis.Thus, clarifying the mechanism underlying the acidic TME in tumor metastasis could facilitate the development of new therapeutic strategies for HCC.Anoikis resistance is one of the most important events in the early stage of cancer metastasis.Here, we report that acidic extracellular pH (pHe) promotes autophagy of HCC cells via the AMPK/mTOR pathway.We found that autophagy induced by acidity enhances anoikis resistance of HCC cells, which could be reversed by autophagy inhibitors.Furthermore, miR-3663-3p was downregulated by acidity, and overexpression of miR-3663-3p abolished acidic pHe-induced autophagy and anoikis resistance.In summary, acidic pHe enhances anoikis resistance of HCC cells by inducing autophagy, which is regulated by miR-3663-3p.Our findings provide new insight into how the acidic TME is involved in HCC progression.

Alterations in cellular metabolism are one of the characteristics in cancer. They are not only the result of tumor progression but also the cause of cancer initiation. Pyruvate dehydrogenase kinase 4 (PDK4) is a key metabolic enzyme, which regulates cell metabolism by inhibiting pyruvate dehydrogenase (PDH). However, the function and regulating mechanism of PDK4 in HCC remain unclear. Here, we found that the expression of PDK4 was significantly decreased in HCC tissues, and its downregulation could predict poor prognosis of HCC patients. Silencing PDK4 significantly facilitated proliferation and migration of HCC cells. Knockdown of PDK4 didn't influence the oxidative phosphorylation and glycolysis capacity of HCC cells in vitro. However, knockdown of PDK4 increased expression of key lipogenic enzymes, fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD), which finally induced lipogenesis. These data suggest that PDK4 inhibits proliferation and migration of HCC cells probably via suppressing lipogenesis.

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.

Liver cancer is a leading cause of cancer morbidity and mortality worldwide, especially in China. Sorafenib (SRF) is currently the most commonly used systemic agent against advanced hepatocellular carcinoma (HCC), which is the most common type of liver cancer. However, HCC patients have only limited benefit and suffer a serious side effect from SRF. Therefore, new approaches are urgently needed to improve the therapeutic effectiveness of SRF and reduce its side effect. In our current study, we developed a self-imaging and self-delivered nanodrug with SRF and indocyanine (ICG) to improve the therapeutic effect of sorafenib against HCC. With the π-π stacking effect between SRF and ICG, a one-step nanoprecipitation method was designed to obtain the SRF/ICG nanoparticles (SINP) via self-assembly. Pluronic F127 was used to shield the SINP to further improve the stability in an aqueous environment. The stability, photothermal effect, cell uptake, ROS production, cytotoxicity, tumor imaging, and tumor-targeting and tumor-killing efficacy of the SINP were evaluated in vitro and in vivo by using an HCC cell line Huh7 and its xenograft tumor model. We found that our designed SINP showed monodisperse stability and efficient photothermal effect both in vitro and in vivo. SINP could rapidly enter Huh7 cells and achieve potent cytotoxicity under near-infrared (NIR) laser irradiation partly by producing a great amount of reactive oxygen species (ROS). SINP had significantly improved stability and blood half-life, and could specifically target tumor via the enhanced permeability and retention (EPR) effect in vivo. In addition, SINP showed improved cytotoxicity in both subcutaneous and orthotopic HCC implantation models in vivo. Overall, this rationally designed sorafenib delivery system with a very high loading capacity (33%) has considerably improved antitumor efficiency in vitro and could completely eliminate subcutaneous tumors without any regrowth in vivo. In conclusion, our self-imaging and self-delivered nanodrug could improve the efficacy of SRF and might be a potential therapy for HCC patients.

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 & AIMS Individuals with Down syndrome have a low risk for many solid tumors, prompting the search for tumor suppressor genes on human chromosome 21 (HSA21). We aimed to identify and explore potential mechanisms of tumor suppressors on HSA21 in hepatocellular carcinoma (HCC). METHODS We compared expression of HSA21 genes in 14 pairs of primary HCC and adjacent noncancer liver tissues using the Affymetrix HG-U133 Plus 2.0 array (Affymetrix, Santa Clara, CA). HCC tissues and adjacent normal liver tissues were collected from 108 patients at a hospital in China for real-time polymerase chain reaction and immunohistochemical analyses; expression levels of regulator of calcineurin 1 (RCAN1) isoform 4 (RCAN1.4) were associated with clinical features. We overexpressed RCAN1.4 from lentiviral vectors in MHCC97H and HCCLM3 cells and knocked expression down using small interfering RNAs in SMMC7721 and Huh7 cells. Cells were analyzed in proliferation, migration, and invasion assays. HCC cells that overexpressed RCAN1.4 or with RCAN1.4 knockdown were injected into livers or tail veins of nude mice; tumor growth and numbers of lung metastases were quantified. We performed bisulfite pyrosequencing and methylation-specific polymerase chain reaction analyses to analyze CpG island methylation. We measured phosphatase activity of calcineurin in HCC cells. RESULTS RCAN1.4 mRNA and protein levels were significantly decreased in primary HCC compared with adjacent noncancer liver tissues. Reduced levels of RCAN1.4 mRNA were significantly associated with advanced tumor stages, poor differentiation, larger tumor size, and vascular invasion. Kaplan-Meier survival analysis showed that patients with HCCs with lower levels of RCAN1.4 mRNA had shorter time of overall survival and time to recurrence than patients whose tumors had high levels of RCAN1.4 mRNA. In HCC cell lines, expression of RCAN1.4 significantly reduced proliferation, migration, and invasive activity. HCC cells that overexpressed RCAN1.4 formed smaller xenograft tumors, with fewer metastases and blood vessels, than control HCC cells. In HCC cells, RCAN1.4 inhibited expression of insulin-like growth factor 1 and vascular endothelial growth factor A by reducing calcineurin activity and blocking nuclear translocation of nuclear factor of activated T cells (NFAT1). HCC cells incubated with the calcineurin inhibitor cyclosporin A had decreased nuclear level of NFAT1. HCC cells had hypermethylation of a CpG island in the 5' regulatory region of RCAN1.4, which reduced its expression. CONCLUSIONS RCAN1.4 is down-regulated in HCC tissues, compared with non-tumor liver tissues. RCAN1.4 prevents cell proliferation, migration, and invasion in vitro; overexpressed RCAN1.4 in HCC cells prevents growth, angiogenesis, and metastases of xenograft tumors by inhibiting calcineurin activity and nuclear translocation of NFAT1.

Emerging evidence has indicated that deregulation of long non‐coding RNAs (lncRNAs) can contribute to the progression and metastasis of human cancer, including hepatocellular carcinoma (HCC). However, the roles of most lncRNAs in HCC remain largely unknown. Here we found a long noncoding RNA termed SchLAH (seven chromosome locus associated with HCC; also called BC035072) was generally downregulated in HCC. Low expression of SchLAH was significantly correlated with shorter overall survival of HCC patients. In vitro and in vivo assays indicated that overexpression of SchLAH inhibited the migration and lung metastasis of HCC cells. Knockdown of SchLAH by siRNA pool promoted the migration of HCC cells. RNA pull‐down and RNA immunoprecipitation assays demonstrated SchLAH physically interacted with fused in sarcoma (FUS). PCR array analysis showed that RhoA and Rac1 were the downstream effector molecules of SchLAH during HCC metastasis. Knockdown of FUS rescued the mRNA levels of RhoA and Rac1 that were repressed by SchLAH. These results suggest that SchLAH may suppress the metastasis of HCC cells by interacting with FUS, which indicates potential of SchLAH for the prognosis and treatment of HCC.

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.

The capacity of liver regeneration is critical for patients with liver diseases. However, cellular and molecular mechanisms of liver regeneration are still incompletely defined. Here, we assessed roles of LASS2 in liver regeneration following partial hepatectomy (PHx) in mice. Our results showed that protein level of LASS2 remarkably increased during liver regeneration after PHx in wildtype (WT) mice. Comparing to WT mice, liver regeneration index after PHx was significantly decreased from day 1 to day 5 in liver-specific LASS2 knockout (LASS2-LKO) mice. Interestingly, liver mass of LASS2-LKO mice could sufficiently recover at day 14 after PHx. Immunohistochemistry (IHC) and western blot analyses revealed that proliferation markers, such as PCNA and Ki67, were potently reduced during liver regeneration in LASS2-LKO mice. In addition, several cell cycle related molecules, such as cyclin A, CDK2 and p-Rb, were decreased in LASS2-LKO mice after PHx. Co-immunoprecipitation assay further revealed a decreased formation of CDK4/cyclin D1 complex after PHx in LASS2-LKO mice. However, phosphorylation of Akt was significantly activated from day 2 after PHx in LASS2-LKO mice when compared with that in WT mice, which may explain the recovery of liver mass at the late stage of liver regeneration in LASS2-LKO mice. Taken together, we conclude that LASS2 plays an important role in efficient liver regeneration in response to PHx.

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.

Chemoresistance in breast cancer has been of great interest in past studies.However, the development of rational therapeutic strategies targeting chemoresistant cells is still a challenge in clinical oncology.By integrating data from global differences of gene expression and phospho-receptor tyrosine kinases between sensitive parental cells (MCF-7) and doxorubicin-resistant cells (MCF-7/ADR), we identified Axl as a potential target for chemoresistance and metastasis in multidrug resistant breast cancer cells.We analyzed Axl expression in 57 breast cancer cell lines and detected a dramatic increase in its expression level in mesenchymal breast cancer cell lines.Axl silencing suppressed invasive and metastatic potentials of chemoresistant breast cancer cells as well as increased elimination of cancer cells when combined with doxorubicin.Furthermore, in preclinical assays, an Axl inhibitor R428 showed increased cell death upon doxorubicin treatment.Additionally, using phospho-kinase array based proteomic analysis, we identified that Akt/GSK-3β/β-catenin cascade was responsible for Axl-induced cell invasion.Nuclear translocation of β-catenin then induced transcriptional upregulation of ZEB1, which in turn regulated DNA damage repair and doxorubicin-resistance in breast cancer cells.Most importantly, Axl was correlated with its downstream targets in tumor samples and was associated with poor prognosis in breast cancer patients.These results demonstrate that Gas6/Axl axis confers aggressiveness in breast cancer and may represent a therapeutic target for chemoresistance and metastasis.

The asialoglycoprotein receptor (ASGR), which is expressed mainly in hepatocytes, is downregulated in poorly differentiated hepatocellular carcinoma (HCC). Here we investigated the role of ASGR1 in HCC metastasis as well as the possible underlying molecular mechanisms. We found that ASGR1 was downregulated in HCC tissue compared with adjacent non-tumorous liver tissue and that lower ASGR1 expression was associated with higher TNM stage and poorer prognosis in HCC patients. ASGR1 overexpression inhibited hepatoma cell migration and invasion in vitro and in vivo, while ASGR1 knockdown had the opposite effects. Furthermore, ASGR1 interacted directly with human longevity assurance homolog 2 of yeast LAG1 (LASS2). Knockdown of LASS2 attenuated the inhibitory effects of ASGR1 on hepatoma cell migration and invasion in vitro. ASGR1 decreased V-ATPase activity in hepatoma cells, and this was reversed by LASS2 knockdown. Finally, HCC patients with low LASS2 levels had poor prognosis, while those with high ASGR1 and LASS2 levels had better prognosis. Thus, ASGR1 may act as a potential metastasis suppressor in HCC, and the combination of ASGR1 and LASS2 may help predict the prognosis of HCC patients.

Longevity assurance homolog 2 of yeast LAG1 (LASS2) has been reported to act as an important tumor suppressor in the development of human cancers. However, little is known about the prognostic value of LASS2 in hepatocellular carcinoma (HCC) . In the present study, we analyzed correlation between LASS2 and TGF-β1 levels, and evaluated their prognostic values in HCC patients. We first analyzed the expression of LASS2 and TGF-β1 in two independent cohorts (test cohort: 184 HCC patients; validation cohort: 118 HCC patients) using immunohistochemistry (IHC). Kaplan-Meier survival and Cox regression analyses were executed to evaluate the prognosis of HCC. The results of IHC analysis revealed a positive correlation between the expression of LASS2 and TGF-β1. HCC Patients with low expression of LASS2 and TGF-β1 had shorter overall survival (OS) and time to recurrence (TTR) than patients with high expression of LASS2 and TGF-β1. Furthermore, combination of LASS2 and TGF-β1 was an independent and significant risk factor for OS and TTR. In conclusion, low expression of LASS2 and TGF-β1 contributes to the aggressiveness and poor prognosis of HCC, and may represent a novel prognostic biomarker for HCC patients.

Background/Aims: Non-small cell lung carcinoma (NSCLC) is the most common type of lung cancer and the cause of most cancer-related deaths. The molecular mechanisms that are involved in NSCLC development are currently not well understood. Accumulating evidence shows that histone demethylases play important roles in the regulation of pathological developmental processes in many diseases, including various types of cancers. Methods: Mitochondrial membrane potential assays, migration and invasion assays, caspase-3 and caspase-9 activity assays and western blot analysis were used in this research. Results: We found that overexpression of KDM6B, a demethylase that acts on histone H3 at lysine 27 (H3K27), inhibited cell growth by initiating mitochondria-dependent apoptosis and by attenuating the invasion-metastasis cascade in NSCLC cells. Moreover, our results showed that KDM6B directly interacted with FOXO1 and that overexpression of KDM6B promoted nuclear accumulation of FOXO1. The effects of KDM6B on cell apoptosis and metastasis were weakened by knockdown of FOXO1 expression. On the contrary, knocking down expression of KDM6B inhibited cell apoptosis and promoted cell growth by mitigating the nuclear translocation of FOXO1 in NSCLC cells. Conclusions: These findings suggest that KDM6B may act in a pro-apoptotic role in NSCLC by causing the nuclear translocation of FOXO1.

Kynurenine 3-monooxygenase (KMO) is a pivotal enzyme in the kynurenine pathway of tryptophan degradation and plays a critical role in Huntington’s and Alzheimer’s diseases. This study aimed to examine the expression of KMO in human hepatocellular carcinoma (HCC) and investigate the relationship between its expression and prognosis of HCC patients. We first analyzed KMO expression in 120 paired HCC samples (HCC tissues vs matched adjacent non-cancerous liver tissues) and 205 clinical HCC specimens using immunohistochemistry (IHC). Kaplan-Meier survival and Cox regression analyses were executed to evaluate the prognosis of HCC. The results of IHC analysis showed that KMO expression was significantly higher in HCC tissues than that in normal liver tissues (all p < 0.05). Survival and recurrence analyses showed that KMO was an independent prognostic factor for overall survival (OS) and time to recurrence (TTR) (both p<0.01). And in vitro studies revealed that KMO positively regulated proliferation, migration and invasion of HCC cells. These results suggest that KMO exhibits tumor-promoting effects towards HCC and it may serve as a novel prognostic marker in HCC.