Splicing factors are proteins that mediate the RNA splicing process, playing a significant role in tumor metastasis by regulating molecular mechanisms and selective splicing patterns of RNA. They are closely associated with the occurrence and development of hepatocellular carcinoma, promoting tumor formation through various mechanisms such as cell proliferation, apoptosis, migration, enhanced metastatic potential, treatment resistance, and immune evasion. Splicing factors also play a crucial role in the diagnosis, treatment, and prognosis of hepatocellular carcinoma. Despite considerable advancements in splicing factor research in hepatocellular carcinoma, the precise mechanisms by which they are utilized remain unknown. Further investigation is necessary to elucidate their precise contribution to the pathogenesis of hepatocellular carcinoma.

医学教育 医学生物化学是一门医学专业基础课程,研究内容包括生物体的分子结构与功能、物质代谢与调节及其在生命活动中的作用. 在全面推进中国特色社会主义现代化过程中,各行各业都离不开更多又"红"又"专"的高素质人才.

Hepatocellular carcinoma (HCC) is one of the most malignant tumors with the highest incidence and mortality in the world, causing a serious burden on society. Pyruvate kinase M2 (PKM2) is one of the principal metabolic enzymes involved in glycolysis. Studies have shown that PKM2 is highly expressed in HCC and can be translocated to the nucleus, where it interacts with various transcription factors and proteins such as hypoxia-inducible factor-1α, sterol regulatory element-binding protein 1a, signal transducer and activator of transcription 3, nuclear factor erythroid 2-like 2 and histone H3, exerting non-metabolic enzyme functions to regulate the cell cycle, proliferation, apoptosis, immune escape, migration, and invasion, as well as HCC angiogenesis and tumor microenvironment. This review is focused on the recent progress of PKM2 interacting with various transcription factors and proteins affecting the onset and development of HCC, as well as natural drugs and noncoding RNA impacting diverse biological functions of liver cancer cells by regulating PKM2 non-metabolic enzyme functions, thereby providing valuable directions for the prognosis improvement and molecular targeted therapy of HCC in the future.

Cervical cancer (CC) ranks the fourth in gynecologic cancers. The incidence and mortality of CC has been decreased due to the cancer screening and early treatments in recent years, but the prognosis of CC patients at advanced stage is still sorrowful. Whether PSME3 exerted a role in the radioresistance of CC cells remains to be investigated. In this study, the expression of PSME3 in mRNA and protein levels was measured by RT-qPCR and western blot analysis, and increased expression of PSME3 in CC tissues and cells was observed. CCK-8 and colony formation assay revealed that the cell viability and proliferation of Hela and CaSki cells treated with different doses of X-ray was reduced due to the depletion of PSME3, indicating that silencing of PSME3 enhanced the radiosensitivity of CC cells. In addition, repair on DNA damage in CC cells was enhanced by PSME3 and the damage was attenuated by PSME3. Besides, the expression of glycolysis-related proteins (GLUT1, PGC-1α, LDHA and HK2) were enhanced by PSME3 but reduced by silencing PSME3 in CC cells. PSME3 restraint attenuated the levels of glucose consumption and lactate production, suggesting PSME3 depletion suppressed abnormal glycolysis of CC cells. Mechanically, PSME3 increased the PARP1 expression via elevating c-myc. Finally, we observed PSME3 attenuation inhibited CC growth in vivo. In conclusion, PSME3 enhanced radioresistance and aerobic glycolysis in CC by regulating PARP1, which might shed a light into the function of PSME3 in CC treatment.

2018年,国务院印发《关于推动创新创业高质量发展打造"双创"升级版的意见》中明确表示要强化大学生的创新创业教育培训,把创新创业教育和实践课程纳入高校必修课体系[1-3].2021年10月,国务院办公厅印发《关于进一步支持大学生创新创业的指导意见》中强调要深化高校创新创业教育改革,将创新创业教育贯穿人才培养全过程,建立以创新创业为导向的新型人才培养模式[4].

目的 探讨研究长链非编码RNA LINC00675对肝癌细胞生物学行为的影响及其分子机制.方法 通过qRT-PCR检测LINC00675在肝癌组织以及细胞中表达量,并检测LINC00675在肝癌细胞HepG3与Huh7细胞质及细胞核中的表达分布.通过在HepG3与Huh7中稳定转染慢病毒过表达LINC00675建立细胞模型.通过CCK-8与流式细胞学检测LINC00675对肝癌细胞增殖、凋亡行为的影响,并通过检测细胞模型中葡萄糖摄取量以及乳酸产量变化监测细胞Warburg效应的变化,运用带有AGO2抗体的RNA免疫沉淀技术评估LINC00675与mRNA的潜在结合能力.使用LncBase Predicted v.2 DIANA工具预测LINC00675下游mRNA靶基因.运用RNA pull-down技术检测LINC00675与miR-665结合的可能性.运用荧光素酶实验验证LINC00675与miR-665的靶向结合,共转染实验验证miR-665介导了LINC00675对肝癌生物学行为的影响.通过使用JASPAR数据库预测LINC00675上游调控基因.通过构建NR2C2过表达以及敲低细胞模型,并使用qRT-PCR实验检测细胞模型中的LINC00675表达量的变化.运用荧光素酶实验验证LINC00675启动子与NR2C2的靶向结合位点.结果 LINC00675在肝癌细胞系中低表达,且主要表达于细胞质中.过表达LINC00675抑制肝癌细胞的增殖能力与Warburg效应,并促进肝癌细胞凋亡.NR2C2介导的LINC00675在肝癌细胞中靶向结合miR-665,负向调控miR-665的表达,并通过结合miR-665来调控肝癌细胞的生物学行为.结论 NR2C2介导的LINC00675通过靶向结合miR-665调控肝癌细胞的增殖和凋亡的生物学行为.

课程质量直接决定人才培养质量,一流课程建设在人才培养中发挥着重要作用.教学团队从教学理念、课程教学团队、课程目标、教学设计、课程内容、教学组织、课程管理与评价方面着手推动课程建设,提升人才培养质量.

科研型教学模式是培养学生科研素养的有效途径,但参与科研项目是否有利于促进学生的学习成绩,这有待进一步研究.我们在生物化学和分子生物学教学过程中实施了一种改进型的科研型教学模式,即科研项目趋动型课堂教学模式.在这一教学模式中,我们将教学内容分成8大模块、9个讲座、6个实验,课程中充分结合学生的兴趣和需要,让学生学会科研数据的分析方法,让学生有充分的获得感.通过比较参与与不参与科研项目设计和实验的学生的期末考试成绩,分析参与科研项目对学生成绩的可能影响,同时比较学习态度对学习成绩的影响.结果发现,参与科研项目和学习态度好的学生的成绩均明显优于不参与科研项目和学习态度不好的学生(P＜0.05).研究结果表明,科研型课堂教学模式能很好地调动学生学习积极性,值得推广应用.

课程整合和教学模式革新成为了当代大学生学生能力提高和学业负担减低的主要手段.生物化学、分子生物学、细胞生物学、遗传学、表观遗传学是生命科学和医学学科的主要基础课程.这些人为分割的各门独立课程不仅内容上有交叉重叠,而且增加学生学业负担,不利于学生科研素养和科学创新能力提高.我们通过转变教育理念和开展教学实践,将以上课程及其理论和实验内容整合成为了一门《细胞、分子和基因》课程,并配套以线上线下科研型模块化教学模式.现将课程整合的设计理念、具体实施流程、整合的实施方案(包括教学内容模块)以及整合的效果评价总结如下,以期为想要进行课程整合的其他学科提供参考.

目的 关于异常寡核苷酸结合折叠域蛋白基因(OBGs)通过微小染色体维持(MCM)复合物影响肝细胞癌DNA复制起始的报道较少.文中旨在探讨逆转录相关基因(RTGs)在肝细胞癌中的作用及可变剪接和单核苷酸位点变异(SNV)与基因表达异常的相关性. 方法 选取150只洁净级昆明小鼠,采用随机数字表法取100只,双前肢腋下注射对数生长期肝细胞癌细胞株H22等渗盐水悬液作为H22组,余50只注射0.2 mL不含H22的等渗盐水作为对照组.从H22组小鼠中选出成瘤小鼠,解剖取出瘤体,从对照组小鼠中取健康肝组织.提取H22组和对照组总RNA,分析差异表达基因.筛选差异表达的逆转录相关DEGs(RDEGs),对RDEGs进行GO和KEGG分析.对RDEGs编码蛋白进行互作分析,对RDEGs多态性与基因表达进行相关性分析. 结果 肝细胞癌中有193个差异表达的RTGs,共参与2个生物学程序、3个细胞组分、1个分子功能、3个信号通路和3个功能部位;其功能主要集中在DNA复制,尤其复制起始中OBGs参与的MCM复合体及端粒复合体构建.可变剪接分析结果显示,4个RDEGs在基因的3个位点发生了差异表达的可变剪接,且可变剪接与相应基因的表达呈正相关.SNV和INDEL分析显示肝细胞癌组织中共有157个RDEGs,发生1541个SNV和78个INDEL位点的改变;上述基因位点的改变主要发生在基因的6个部位,即外显子区、内含子区、基因间区、基因下游区、基因上游区和拼接区;共有28个基因的SNV改变差异有统计学意义(P<0.05).基因位点多态性分析结果显示,5个OBGs在肝细胞癌中出现了SNV现象,而健康肝组织中则没有SNV. 结论 RTGs中的OBGs可能在肝细胞癌中通过自身的基因多态性突变引起MCM复合体及端粒复合体的改变,从而调控DNA复制起始及保护端粒的完整性,进而在癌细胞的增殖过程中发挥重要作用.可变剪接及SNV则可能是一些基因表达的重要调控因素.

目的:探讨模型小鼠肝细胞癌组织(TM)内差异表达的外泌体来源RNA剪接基因(exosome-derived RNA-splicing-related DEGs,ERDs)的变化图谱及AS、SNV和INDEL与基因表达的关系.方法:以健康小鼠肝组织(生理微环境,PM)、模型小鼠无瘤体肝组织(细胞癌前微环境,PTM)和模型小鼠肝细胞癌组织(肿瘤微环境,TM)为材料,分别提取各组样本中的总RNA进行RNA-seq和生物信息学分析,并通过RT-qPCR验证实验结果.结果:与PTM比较,TM中有158个高表达的ERDs;与PM比较,TM中有141个高表达的ERDs;两类对比样本中有21个相同的ERDs在TM中高表达.这21个ERDs主要参与了4个生物学过程(BP)、5个细胞组分(CC)、2个分子功能(MF)和1个信号通路(KEGG pathway).其功能主要富集在剪接体的功能上,尤其是在剪接体执行剪接过程中ploy(A)RNA结合、核来源的第二步剪接小体的催化作用以及剪接体snRNP组装.结论:TM中高表达的ERDs主要通过poly(A)RNA结合异常,介导错误的RNA剪接和剪接体snRNP组装,可能导致mRNA错乱剪接进而影响模型小鼠肝细胞癌的发生发展.同时,引起基因多态性的可变剪切(AS)、单核苷酸位点变异(SNV)及插入缺失标记(INDEL)与TM中高表达的RNA剪接具有关联性.

目的:探索肝细胞癌(HCC)来源血液相关外泌体组分参与天然免疫的可能机制.方法:以肝细胞癌模型小鼠的瘤体组织和癌旁肝组织为实验材料,分别提取各组织中的总RNA,通过RNA测序(RNA-seq)和生物信息学手段分析两样本中HCC源血液相关外泌体差异表达基因,并用RT-qPCR验证实验结果.结果:发现在瘤体组织和癌旁肝组织中存在81个HCC源血液相关外泌体差异表达基因.对这些差异表达基因功能富集,共富集到10个生物学过程、4个细胞组分、3个分子功能和2个KEGG通路.进一步分析发现它们主要参与天然免疫反应、血液凝固、炎症反应等生物学过程,以及受体结合、丝氨酸型肽酶活性等分子功能.其中33个HCC源血液相关外泌体差异表达基因参与了补体和凝血级联信号通路.结论:HCC源血液相关外泌体组分可能主要以受体结合、丝氨酸型肽酶催化活性等,通过补体和凝血级联信号途径,参与HCC组织中的天然免疫反应.

目的 探讨线粒体源外泌体基因参与肝细胞癌(HCC)病理过程的可能分子机制.方法 通过RNA-seq和生物信息学手段,分析HCC中差异表达的线粒体源外泌体基因(DEMEs)及其生物学功能.结果 HCC组织中共鉴定出93个DEMEs,其中74个表达下调,19个表达上调.表达下调的DEMEs经功能富集,共富集到4个生物学过程(biological process,BP)、4个细胞组分(cellular component,CC)、2个分子活性(molecular function,MF)、1个信号通路(KEGG pathway,KG)和4个蛋白位点序列特征(UP_SEQ_FEATURE,US).它们参与了氧化还原、脂质代谢等过程,且大量DEMEs还具有线粒体转运肽功能.而上调表达的DEMEs,除了富集到1个CC,即线粒体组分,还发现其中一些参与了跨膜转运、对过氧化氢和对机械刺激的反应过程,有些还具有转移酶活性和ATP结合能力,并参与癌症的发生和转移等病理过程.结论 线粒体源外泌体基因在HCC肿瘤组织与非肿瘤组织存在差异性表达,生物信息学分析显示这种差异性表达的基因涉及多个生物学过程,提示它们可能通过多种途径影响HCC的发生.

Background The tumor microenvironment (TM) in close contact with cancer cells is highly related to tumor growth and cancer metastasis. This study is to explore the biogenesis mechanism of a secondary hepatocellular carcinoma (HCC) based on the function of RNA binding proteins (RBPs)-encoding genes in the physiological microenvironment (PM). Methods The healthy and HCC mice were used to isolate the PM, pre-tumor microenvironment (PTM), and TM. The samples were analyzed using the technology of RNA-seq and bioinformatics. The differentially expressed RBPs-encoding genes (DERs) and differentially expressed DERs-associated genes (DEDs) were screened to undergo GO and KEGG analysis. Results 18 DERs and DEDs were identified in the PTM vs. PM, 87 in the TM vs. PTM, and 87 in the TM vs. PM. Those DERs and DEDs participated in the regulation of gene expression at the levels of chromatin conformation, gene activation and silencing, splicing and degradation of mRNA, biogenesis of piRNA and miRNA, ribosome assemble, and translation of proteins. Conclusion The genes encoding RBPs and the relevant genes are involved in the transformation from PM to PTM, then constructing the TM by regulating protein synthesis. This regulation included whole process of biological genetic information transmission from chromatin conformation to gene activation and silencing to mRNA splicing to ribosome assemble to translation of proteins and degradation of mRNA. The abnormality of those functions in the organic microenvironments promoted the metastasis of HCC and initiated the biogenesis of a secondary HCC in a PM when the PM encountered the invasion of cancer cells.

In order to find a teaching mode that can meet the needs of the teaching in large class, we integrate various theories and teaching methods, construct grouping micro-video competition teaching mode and implement it in the biochemistry course in 6 undergraduate classes of 2016 nursing major of Youjiang Medical University for Nationalities to test its application effect in large class. After surveying to one experimental class (n = 103) and 2 control classes (n = 210) in the form of questionnaire and analyzing the final examine scores of experimental and control contents, it is shown that students' average scores in experimental class in which the micro-video competition teaching mode is adopted and the scores of objective and subjective questions are significantly higher than those in controls (P < 0.05), while there is no significant difference in the scores of contents between experimental class and controls (P > 0.05). In particular, there is a statistical difference in the scoring rate of objective questions between experimental content and control content in the experimental class (P < 0.05), while there is no significant difference in the scoring rate of subjective questions (P > 0.05). Students prefer to affirm the micro-video competition teaching mode ( P < 0.05), while they are generally uncertain towards traditional lecturing teaching mode in questionnaire (P < 0.05). The research results show that the micro-video competition teaching mode has promoted students' memory and mastery on the experiment content. Especially, in the aspect of training students' comprehensive analysis and the ability to apply knowledge, the teaching effect of the mode is better that of traditional mode. It not only promotes the ability of students' comprehensive analysis and its application to the experiment content, but also the ability of their comprehensive analysis to other teaching content and relevant skills.

目的 探讨肿瘤微环境(TM)中趋化因子参与肝细胞癌(HCC)引起的炎症反应的可能机制.方法 提取肿瘤组织(TM组)和癌旁组织(对照组)总RNA并通过RNA-seq和生物信息学手段分析趋化因子相关差异表达基因的功能及其编码蛋白的互作情况,并以RT-qPCR验证实验结果 .结果TM中共有43个趋化因子相关差异表达基因,它们被富集到2个KEGG信号通路、5个生物学过程和2个细胞组分,其中大量趋化因子相关基因参与炎症反应、免疫反应、趋化性、细胞对IL-1的反应和细胞对肿瘤坏死因子的反应过程.尤其值得一提的是,涉及趋化性的7个基因(ACKR3、CCL20、CCL3、CCL5、CCR4、CCR6、PTGDR2)中,有6个参与细胞因子-细胞因子受体信号通路、5个参与免疫反应、4个参与炎症反应;涉及细胞因子-细胞因子受体信号通路的134个基因中有7个参与炎症反应.结论 TM中趋化因子通过与细胞因子受体互作的方式参与HCC诱导的炎症反应.

Objective To investigate whether single nucleotide polymorphisms of catechol-O-methyl transferase (COMT) of Val158Met are associated with the risk of breast cancer.Methods Gene polymorphisms of COMT were detected using di-allele-specific-amplification with artificially modified primers combined with SYBR Green I real-time polymerase chain reaction in a case-control study,which included 96 breast cancer patients (treatment group) and 116 healthy women(control group).Results The frequency of allele G in COMT gene Val158Met was 65.10％ and 71.98％ in treatment group and control group,and the frequency of allele A were 34.90％ and 28.02％ respectively.There were no differences between the two groups in allele frequencies of COMT Val158Met among Guangxi Baise population (all P ＞ 0.05).COMT Val158Met G/G genotype frequency distribution of the treatment group (38.54％) was lower than that of control group (54.31％),A/G genotype frequency distribution of the treatment group was higher(53.13％)than that of control group (35.34％).The distribution frequency differences of the two groups homozygous wild-type and heterozygous were statistically significant (all P ＜ 0.05).A/A genotype frequency distribution was relatively similar in the treatment group and control group,and they were 8.33％ and 10.35％ respectively.The breast cancer risks of Guangxi Baise women with A/G heterozygous genotype increased by 2.118 times compared with that of G/G homozygous genotype.Conclusion Gene polymorphism of COMT Val158Met may be associated with the risk of breast cancer.

目的 探索在生物化学实验教学中动物组织核酸的提取方法.方法 经过反复实践后,随机挑选两组学生(酚提取组27例,三氯醋酸提取组28例),分别以酚提取法和三氯醋酸提取法提取猪肝中的核酸,分别进行水解和鉴定核酸的各种成分(磷酸、嘌呤碱、核糖和脱氧核糖),对比分析各种成分鉴定的阳性率.结果 核酸四种成分定性鉴定,两组间磷酸、核糖和脱氧核糖对比差异无统计学意义(P＞0.05),三氯醋酸提取组的嘌呤碱明显优于酚提取组(P＜0.001).结论 三氯醋酸提取法提取动物组织中核酸,实验试剂更加安全,步骤简单,对实验设备要求不高,实验效果明显.

目的:探索翻转课堂理念的趣味教学在医学生物化学课程中的效果.方法:在课程中进行传统教学法及趣味教学法对比研究,通过考核评价教学效果,并通过网络问卷调查学生满意度.结果:趣味教学组的学生总体满意度高达99％,显著高于传统教学.趣味教学并没有显著影响学生的考试成绩(P＞0.05).结论:翻转课堂理念的趣味教学法虽然没有提高学生成绩,但极大的提高学生上课的热情及自学能力,有效地改善学生对生物化学的厌学情绪,值得进一步研究.