The utilization of knowledge graphs has emerged as a prevailing trend in current medical education reform due to its advantages in visualization and personalized learning. The blended teaching still faces challenges such as knowledge fragmentation, lack of personalized learning pathways and inadequate analysis of learning situations. To evalute the learning experience, learning effect and usability of knowledge graph in the clinical microbiology examination course. The knowledge graph of the clinical microbiology examination course was constructed using natural language processing, deep learning, and other advanced technologies on the Superstar learning platform. 100 third-year students enrolled in 2021 and 2020 majoring in medical examination technology served as the experimental and control group, respectively. The theoretical test scores of the experimental group using knowledge graph for online learning were higher compared to those of the control group using MOOC. The utilization of knowledge graph assisted students in constructing an overall knowledge framework of course and achieving personalized learning and self-evaluation. And it also helped teachers accurately analyze learning situations to provide targeted offline teaching methods and optimize online learning resources. The average scores of four dimensions including the quality of work and life, perceived usefulness, perceived ease of use and user control obtained by the "Health Information Technology Usability Assessment Scale" were 4.67, 4.51, 4.32 and 4.26, respectively. The knowledge graph of clinical microbiology examination course was available, which was helpful to realize personalized learning and precise teaching and effectively solve the problems existing in blended teaching model.

IntroductionExtraction techniques that influence cell wall polysaccharides (EPS) is crucial for maximizing their bioactivity. This study evaluates ultrasound technology for extracting antioxidant polysaccharides from Geotrichum candidum LG-8, assessing its impacton antioxidant activity.MethodsUltrasound extraction of EPS from G. candidum LG-8 was optimized (18 min, pH 7.0, 40 W/cm2, 0.75 M NaCl). ABTS scavenging efficiency and monosaccharide composition of LG-EPS1 and LG-EPS3 were analyzed using Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM).ResultsThe Results showed that ultrasonic treatment markedly increased the ABTS radical scavenging efficiency of LG-8 cells by 47%. At a concentration of 1 mg/mL, the ultrasonically extracted LG-EPS1 and LG-EPS3 polysaccharides exhibited significant ABTS radical scavenging efficiencies of 26% and 51%, respectively. Monosaccharide composition analysis identified mannose and glucose in LG-EPS1, while LG-EPS3 was primarily composed of mannose. FTIR spectra verified the polysaccharides' presence, and SEM provided visual confirmation of the nanoparticle structures characteristic of LG-EPS1 and LG-EPS3.DiscussionThis research not only underscores the technological merits of ultrasound in polysaccharide extraction but also highlights the potential of G. candidum LG-8 derived polysaccharides as valuable bioactive compounds for antioxidant utilization.

The application of LG-8 and its dry fragments as zearalenone (ZEN) adsorbents was investigated. The study showed that Geotrichum candidum LG-8 and its fragments dried at 55°C or through lyophilization are able to adsorb around 80% of ZEN. However, besides in water and 55°C-drying conditions, SEM indicated that higher 90% of ZEN binding tended to occur when cell walls of fragments were intact with less adhesion among themselves. Notably, ZEN/LG-8 fragments complexes were quite stable, as only 1.262% and 1.969% of ZEN were released after successive pH treatments for 4 h and 5 min. The kinetic data signified that adsorption of ZEN onto LG-8 fragments followed well the pseudo-first-order kinetic model. Isotherm calculations showed Langmuir model was favourable and monolayer adsorption of ZEN occurred at functional binding sites on fragments surface. Therefore, we conclude that it can be an alternative biosorbent to treat water contained with ZEN, since LG-8 is low-cost biomass and its fragments have a considerable high biosorption capacity avoiding impacting final product quality and immunodeficient patients.

BackgroundAs a convenient teaching tool, virtual simulation experiment technology had been widely utilized in the field of medical education. However, virtual learning could not fully replace the benefits of in-person instruction. Therefore, finding ways to integrate both methods was crucial for achieving optimal educational outcomes. The objective of this study was to compare the effectiveness of the self-built virtual simulation and design experiment combining teaching mode and the traditional experimental teaching mode in the clinical microbiology examination experiment teaching.MethodsThis study was conducted at Shandong First Medical University in China. The experimental group consisted of 100 third-year students from the grade 2020 majoring in medical examination technology, who underwent an innovative teaching model combining virtual and real experiments. The control group comprised of 100 third-year students from the grade 2019 in the same major, who received traditional experimental teaching model. In this study, we referred to grade 2020 as cohort 2020 and grade 2019 cohort 2019. The performance of both groups was assessed via experimental and theoretical testing. Meanwhile, survey questionnaires were administered to evaluate the efficacy of the innovative experimental teaching model and students’ level of satisfaction with it. Cohort 2020 conducted a survey for modules 1 to 4, while cohort 2019 only conducted a survey for module 4, as detailed in the Appendix.ResultsThe majority of students in the experimental group expressed satisfaction with the teaching model that combined virtual and real experiments, as evidenced by their superior performance on both experimental operational skills (87.54 ± 8.93 vs. 82.39 ± 10.55) and theoretical knowledge tests (83.65 ± 9.02 vs. 80.18 ± 8.24) compared to those in the control group.ConclusionThe combination of virtual simulation experiment and design experiment in the microbiological examination of clinical specimens represented an effective pedagogical approach. The instructional approach had the potential to incite a passion for learning, enhance proficiency in standardized experimental techniques, foster the ability to integrate theory with practice, and cultivate clinical reasoning skills.

The effects of the heating conditions and water content on the structure and digestibility of wheat starch (WS)-glycerol monopalmitin (GMP) complexes were investigated. The results showed that the higher water content and the heating conditions of 90°C for 60 min after 100°C for 10 min favor the formation of more WS-GMP complexes with the greater short-range order, although the thermal transition temperatures of GWS-GMP-100 complexes are not significantly affected by the water content. Only the type I complexes were formed under the heating conditions of 90°C for 60 min. The heating conditions of 90°C for 60 min after 100°C for 10 min facilitates the formation of type II complexes, and the amounts of type II complexes increased with increasing water content. The digestion rates of WS-GMP complexes decreased slightly with increasing water content, and the extent of starch amylolysis of WS-GMP complexes significantly decreased after heating further at 90°C compared with that only heating at 100°C. The digestibility of complexes is mainly related to structural order rather than the number of complexes. This study is helpful to further understand starch-lipid complexes by showing that heating conditions and water content influence the formation of WS-GMP complexes.

虚拟仿真实验是一种现代信息化和智慧教学的重要方法,对高等医学院校教学质量的发展起到重要的推进作用.我们自主开发建设的"霍乱弧菌检测与防控虚拟仿真实验",既弥补了因生物安全问题不能开展的实验教学,也解决了微生物学检验实验教学中存在的操作标准化问题.采用"三步进阶"混合教学模式,实现以"学生为中心"的师生互动模式,创建"设计性实验报告",与育人元素有机结合,培育医学生的职业使命感.细化考核标准,实现过程性评价.探索课前启发铺垫、课中内化升华、课后巩固拓展混合式实验教学模式,有效提高学生的实验技能,实现知行合一、素能共育的教学目标.

Acute lung injury (ALI) is a kind of lung disease with acute dyspnea, pulmonary inflammation, respiratory distress, and non-cardiogenic pulmonary edema, accompanied by the mid- and end-stage characteristics of COVID-19, clinically. It is imperative to find non-toxic natural substances on preventing ALI and its complications. The animal experiments demonstrated that Lentinus edodes polysaccharides (PLE) had a potential role in alleviating ALI by inhibiting oxidative stress and inflammation, which was manifested by reducing the levels of serum lung injury indicators (C3, hs-CRP, and GGT), reducing the levels of inflammatory factors (TNF-α, IL-1β, and IL-6), and increasing the activities of antioxidant enzymes (SOD and CAT) in the lung. Furthermore, PLE had the typical characteristics of pyran-type linked by β-type glycosidic linkages. The conclusions indicated that PLE could be used as functional foods and natural drugs in preventing ALI.

目的 探索真姬菇菌糠多糖是否通过Wnt/β-catenin信号通路促进HepG2细胞凋亡.方法 采用MTT试验检测真姬菇菌糠多糖抑制HepG2细胞增殖,显微镜下观察HepG2细胞发生凋亡的形态改变;采用实时荧光定量PCR和Western blot方法检测真姬菇菌糠多糖对Wnt/β-catenin信号通路中β-catenin、Caspase-3及其下游分子Bax在基因和蛋白水平表达的影响.结果 50μg/mL的真姬菇菌糠多糖可以抑制HepG2细胞增殖(F=30.956,P<0.001),并使HepG2细胞发生明显的细胞凋亡的形态改变,能够抑制 β-catenin信号通路相关分子 β-catenin(t=3.940,P<0.05)、Caspase-3(t=3.847,P<0.05)在基因和蛋白水平的表达量,上调Bax分子(t=3.975,P<0.05)的基因和蛋白表达水平.结论 真姬菇菌糠多糖通过抑制Wnt/β-catenin信号通路及上调促凋亡分子Bax促进HepG2细胞凋亡,发挥抗肝癌HepG2细胞增殖的作用.

目的 探索香菇菌糠多糖对HepG2细胞增殖的影响,及是否通过wnt/β-catenin信号通路促进HepG2细胞的凋亡.方法 通过MTT试验检测香菇菌糠多糖对HepG2细胞增殖的影响,并观察HepG2细胞形态的变化,采用western blot检测香菇菌糠多糖对wnt/β-catenin信号通路中β-catenin、Caspase-3和Bcl-2表达的影响,以判定香菇菌糠多糖是否可以通过wnt/β-catenin信号通路促进HepG2细胞的凋亡.结果 50μg/mL的香菇菌糠多糖能够抑制HepG2细胞增殖(F=572.991,P<0.001),可使HepG2细胞发生明显皱缩,并可抑制β-catenin信号通路相关蛋白β-catenin、Caspase-3和Bcl-2的表达量.结论 香菇菌糠多糖可能通过wnt/β-catenin信号通路及其下游的Bcl-2促进HepG2细胞凋亡,从而起到抗肿瘤的作用.

目的 探索肝癌(hepatocellular carcinoma,HCC)组织HBV cccDNA T3098C突变情况并检测T3098C突变对HBsAg合成和分泌的影响.方法 通过滚环扩增(rolling cycle amplification,RCA)和PCR直接测序,检测23例HCC患者癌组织中T3098C突变情况,并对患者的术后生存时间进行分析,将野生型质粒与T3098C突变质粒分别转染Huh7和HepG2肝癌细胞,检测细胞和上清中HBsAg含量.结果 在23例癌组织中,发生T3098C突变的有9例,T3098C突变的生存分析结果显示,C3098突变型组的肝癌患者的生存时间明显长于T3098野生型组,Cox比例风险模型多因素分析发现,T3098C突变是HCC患者术后独立的保护预后预测因素;T3098C突变型组细胞内的HBsAg含量低于HBV 1.2×野生型组(F=42.91,P<0.001),差异具有统计学意义;T3098C突变型组上清和细胞内HBsAg总量低于HBV 1.2×野生型组(F=34.25,P<0.001),差异具有统计学意义.结论 T3098C突变是HCC患者独立的保护预后预测因素,这可能与其降低HBsAg的合成和促进HBsAg的外泌有关.

SummaryIt was repeatedly reported that the hepatitis B virus (HBV) T1719G mutation was very common and related to progression and malignancy of liver disease. However, its effect on viral replication efficiency remains unclear. In this study, we aimed to evaluate the function and mechanisms of the T1719G mutation on viral replication capacity. Wild‐type and T1719G mutation‐bearing HBV1.2× plasmids were transfected into Huh7 and HepG2 cells, respectively, and HBV total RNA, 3.5 kb RNA and supernatant HBV DNA were assessed using real‐time PCR, hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels were measured by time‐resolved fluoroimmunoassay. In order to assess Enh II activity and the binding capacity of HNF3β to Enh II sequence, dual‐luciferase assay and Chromatin immunoprecipitation (ChIP)‐PCR were employed, respectively. Simultaneously, the HBx or HBx‐mut (T1719G) plasmid was co‐transfected to evaluate the effect of HBx on viral replication. Our results showed that the T1719G mutation impaired viral replication efficacy compared with the wild type both by reducing Enh II activity and binding capacity of HNF3β with Enh II. And such reduction caused by T1719G mutation could be rescued by HBx protein. Our results show that the T1719G mutation decreases HBV viral replication capacity possibly by mutant HBx protein and altered Enh II activity.

目的 对一个跖骨短小症家系的成员进行全基因组测序,探究其可能的致病因素.方法 应用Illumina HiSeq X ten测序平台对一个跖骨短小症家系中2例病例和3例正常人进行测序,测序数据经过1 000 Genomes Project和dbSNP数据库过滤,筛选出突变基因.结果 跖骨短小症呈X染色体隐性遗传.在X染色体上得到1 527个特异性单核苷酸多态性位点(single nucleotide polymorphism,SNP).没有筛选到符合条件的特异性小片段的插入/缺失(insertion/deletion,In/del).经过1 000 Genomes Project和dbSNP数据库比对后,得到位于11个基因上的39个特异性SNP位点,包括CCNB3、CYSLTR1、GK、NROB1、PCYT1B、PRPS2、SHROOM4、SMS、TBL1X、TLR7、TLR8基因.结论 这些基因SNP位点在我国汉族人群中也是存在的,提示本家系疾病并非单纯由遗传因素导致.

•PLWM inhibited the secretion of HBsAg and HBeAg.•PLWM had no effect on the inhibition of HBV DNA replication, but 50 µg/ml PLWM strengthened the inhibition effect on HBV DNA replications of Lamivudine (10 µmol/l).•PLWM could inhibit cell proliferation and affect the expression of some anti-apoptosis proteins.

目的 探索乙型肝炎病毒(hepatitis B virus,HBV) G588C突变在原发性肝细胞癌(HCC)组织HBV cccDNA中的存在情况及其对HBV复制的影响. 方法 通过滚环扩增(rolling cycle amplification,RCA)和PCR扩增测序,寻找G588C突变在18对HCC患者癌与癌旁组织中的存在情况.在HBV 1.2×质粒(HBV C基因型)的基础上构建G588C突变位点,将G588C突变质粒与野生型质粒分别转染HepG2和Huh7肝癌细胞系细胞,检测上清和细胞中HBsAg含量及上清中HBV DNA水平. 结果 在18对癌与癌旁组织中G588C、突变只存在于3例癌组织中;G588C突变组上清中HBsAg 3、4d含量分别为HepG2细胞5.605±1.182,8.270±2.241,Huh细胞2.714±0.371,10.148±2.793,细胞内HBsAg 3、4d含量分别为HepG2细胞4.852±1.024,7.736±1.762,Huh细胞18.349±3.040,34.110±2.129;野生型组上清中HBsAg 3、4d含量分别为HepG2细胞8.083±1.428,13.170±0.938,Huh细胞6.231±0.373,23.971±1.573,细胞内HBsAg 3、4d含量分别为HepG2细胞2.937±0.876,4.270±1.659,Huh细胞13.498±3.06,21.010±3.488.G588C突变组上清中HBsAg含量显著低于野生型组(F=44.88,P＜0.01),G588C突变组细胞内HBsAg含量显著高于野生型组(F=34.65,P＜0.01);G588C突变组上清和细胞中HBs Ag总量与HBV 1.2×野生型组比较差异无统计学意义(F=7.69,P＞0.05);G588C突变组上清中HBsAg与细胞内HBsAg的比值显著低于HBV 1.2×野生型组(F=23.59,P＜0.05),HBV DNA水平与两组间差异无统计学意义(F=6.23,P＞0.05). 结论 G588C突变不影响HBV的复制,但影响HBsAg由细胞内向细胞外的分泌,这可能与HCC的发生发展有关.

Objectives To explore the possible pathways by which hepatitis B virus (HBV) enters peripheral blood monocytes (PBMCs) and to understand the possible mechanisms by which HBV infects PBMCs by detecting the expression of HBV receptor in PBMCs and hepatocytes. Methods In order to detect the expression of HBV receptor in PBMCs and hepatocytes, PBMCs were isolated with Ficoll-Hypaque.Total protein from PBMCs and hepatocytes from liver tumor-adjacent tissues was extracted with RIPA and then assayed using a Western blot assay. Results A Western blot assay indicated that HBV receptor protein expression was positive in hepatocytes, and the reaction band with the hNTCP antibody was located between (35～40) ×103, as expected.However, the same reaction band with the hNTCP antibody was not detected in PBMCs, and HBV receptor protein expression was negative. Conclusion PBMCs do not express HBV receptor protein, though it was found on the surface of hepatocytes, suggesting that HBV may enter PBMCs via non-receptor pathways or that HBV receptor on the surface of PBMCs differs from HBV receptor on the surface of hepatocytes.More research is needed to interpret the exact mechanism by which HBV infects PBMCs.

In present study, a quadratic regression model based on the response surface methodology (RSM) coupled with Box–Behnken design (BBD) was employed to optimize the enzyme-assisted extraction (EAE) process of enzymatic-extractable Pleurotus djamor polysaccharides (EnPPs). By solving the regression equations and analyzing the model graphs, the optimum conditions were at pH 7.36, water to material ratio 56.78, extraction time 44.77 min and extraction temperature 35.36 °C, respectively. Under these conditions, the experimental yields of EnPPs reached 3.61%, which were in good agreement with the validated values (3.57 ± 0.51%). Basic physicochemical properties including molecular weights, structural identification by FT-IR and monosaccharide compositions were processed. In addition, the in vivo inhibiting activities against oxidative stress were investigated. The results provided an alternative bioresource for the exploitation of EnPPs under EAE from P. djamor, and the EnPPs had potential effects in prevention of oxidative stress.

Objective To ascertain the molecular mechanism by which Helicobacter pylori CagA upregulates CIP2A.Methods After AGS and GES-1 cells were separately cultured to exponential phase,AGS or GES-1 cells were separately transfected with either the recombinant plasmid WT-cagA or the control vector plasmid pcDNA3.1.The cells were then treated with a signal inhibitor of B-Raf or JNK2 and transfected with the plasmid recombinant WT-cagA.The expression of CIP2A was detected with Western blotting,and the cell clonogenic potential was determined using a cell cloning assay.Cell migration was detected with a cell migration assay.Results AGS and GES-1 cells transfected with the plasmid WT-cagA had a higher level of CIP2A expression than that in the control.After treatment with a signal inhibitor of B-Raf or JNK2 and subsequent transfection with the plasmid WT-cagA,the level of CIP2A expression in the cells was decreased.The cell clonogenic potential and cell migration decreased below the cell clonogenic potential and cell migration noted in cells transfected with the plasmid WT-cagA alone.Conclusion H.pylori CagA upregulates the expression of the oncogene CIP2A through B-Raf and JNK2,affecting the cell clonogenic potential and cell migration.

Background: More than half of hepatocellular carcinomas (HCCs) are etiologically attributed to hepatitis B virus (HBV) infection, but it remains unclear whether HBV mutations are virological factors that contribute to formation of HCC or instead reflect accumulation during the progression of HBV-related disease.Methods: Rolling-cycle amplification and PCR sequencing were used to characterize covalently closed circular DNA (cccDNA) mutations in tumor tissues. Paired non-tumor tissues were used as controls.Results: High frequencies of C1653T, T1753V, and A1762T/G1764A cccDNA mutations were observed in both tumor and non-tumor tissues. T1719G, C1329A, and T3098C mutations were related to the overall survival of HCC patients. Patients with G1719 tended to be in the high Barcelona Clinic Liver Cancer stage and had lower levels of total DNA and cccDNA per cell than patients with T1719. Additionally, in vitro analysis revealed that T1719G mutation reduced viral replication efficacy. Finally, significantly higher levels of preoperative alpha-fetoprotein were observed in patients harboring the G1078T, C1653T, G1727A, C1913A, T1978C, or C3116T mutations at the cccDNA level.Conclusions: We speculated that HBV cccDNA mutations accumulated over the course of HBV-related disease development, and that some key mutations had prognostic value for patients with HBVrelated HCC. (C) 2017 Elsevier Ltd. All rights reserved.

Objective To investigate preliminarily the tissue distribution and safety of HSV-2 DNA vaccine pc-P6-gBCTL-TBK-1 in mice.Methods The recombinant plasmid pc-P6-gBCTL-TBK-1 was injected into BALB/c mice with 100 μg per mouse for three times at every other week.Heart,liver,spleen,kidney,lung,brain,blood and muscle tissues of immune mice were collected at 24 h,7 d,21 d,35 d and 49 d after inoculation,then total DNAs of tissues were extracted,and the tissue distribution of plasmid DNA and the possibility of integration with host genome were detected by PCR assay.Results At 24 h,7 d,21 d and 35 d after the last immunization,the plasmid DNA was detected in heart,liver,spleen,kidney,lung,brain and muscle tissues.At 49 d after the last immunization,the plasmid DNA was detected only in muscle tissue.The gel-purified genomic DNA was detected negative by PCR assay.Conclusion The plasmid pc-P6-gBCTL-TBK-1 can spread quickly to the heart,liver,spleen,kidney,lung,brain,blood and muscle tissues of the mice.With the extention of time,the plasmid DNA only distributes in muscle tissue of vaccination,and the DNA does not integrate with the host genome,which suggests that the plasmid is safe.

Objective To investigate the immune effects of herpes simplex virus type 2 (HSV-2) DNA vaccine of pc(pcDNA3.1-)-P6-gBCTL-TBK-1 in BALB/c mice.Methods The eukaryotic expression plasmid of pc-P6-gBCTL-TBK-1 was constructed and transfected into HeLa cells,and the expressed protein was verified by Western blotting.The BALB/c mice were immunized by the plasmids through intramuscular injection for three times,in week 0,2 and 4.In week 5,the levels of interferon (IFN-γ),interleukin-1 8 (IL-18),interleukin-4 (IL-4),interleukin-6 (IL-6) and specific HSV-2gD-P6 IgG antibody were measured by ELISA.In week 8,spleen cells of vaccinated mice were separated and the proliferation of spleen lymphocytes were detected by MTT.The activity of cytotoxic T lymphocyte (CTL) was detected by lactic dehydrogenase (LDH) method.Results After being inoculated by the plasmid of pc-P6-gBCTL-TBK-1,the mice were induced to generate higher level of specific antibody,including IFN-γ,IL-18 and IL-4;the CTL kill cells' activity and proliferation capability of splenic lymphocytes were improved.Conclusion The recombinant plasmid of pc-P6-gBCTL-TBK-1 has immunogenicity,which can induce the humoral and cellular immune responses,and TBK-1 may have good function of molecular chaperone.