Abstract Schizophrenia (SCZ) is a serious psychiatric condition. While PM2.5 exposure has been linked to SCZ, the specific effects of its components remain poorly understood. This study aimed to explore the relationships between PM2.5 constituents (including BC: black carbon, OM: organic matter, SO42−: sulfate, NH4+: ammonium, and NO3−: nitrate) and SCZ. It incorporated the hospitalization records of 16,082 SCZ patients from Nanning Fifth People's Hospital, spanning from 1 January 2014, to 31 December 2023. The daily concentration data of PM2.5 and its five chemical components were sourced from Tracking Air Pollution in China (TAP). A distributed lag nonlinear model (DLNM) was employed to measure the dynamic correlation between PM2.5 components and the risk of hospitalization for SCZ. Further analysis was conducted by stratifying based on gender, age, and cold/warm seasons to identify susceptible populations. Our study revealed that OM and BC demonstrated lagged effects on SCZ hospitalization, with significant associations observed at lag 3 day(lag3)and lag4. The strongest effect was identified at lag4, with OM showing an relative risks (RR) of 1.010 (95%CI: 1.001, 1.019) and BC exhibiting a higher RR of 1.010 (1.001, 1.019). And the lag effect of the OM relative percentage was identified at lag 3 (RR = 1.013, 95% CI: 1.005–1.022). PM2.5, SO42−, and NH4+ showed lagged response trends but no statistically significant effects (p > 0.05). Subgroup analysis indicated that males, 45 years and younger, and those exposed during the warm season had higher risks associated with SO42−, OM, BC, and PM2.5. Short‐term exposure to OM is significantly related to SCZ hospitalization.

Liver cancer is a common tumor of digestive system. Hepatocellular carcinoma (HCC) is a common type of liver cancer, which has a high degree of malignancy and ranks among the top causes of cancer-related death in the world. Metabolic reprogramming is considered to be an important marker of carcinogenesis. Glucose metabolism is one of the main ways for cells to produce energy. Glycolysis, as the basic reaction of glucose metabolism, plays an important role in cell metabolism. Therefore, the regulation of glycolysis is of great significance to the proliferation and evolution of tumors. More and more non-coding RNAs (ncRNA) have been proved to play an important role in the regulation of tumor glycolysis. This article reviews the role of ncRNA in the regulation of HCC glycolysis and its related mechanisms. At the same time, the prospect of targeted therapy for HCC based on the related mechanisms of glycolysis regulation is put forward.

生物化学与分子生物学是医学专业的核心基础课,知识点相对微观和复杂.小规模限制性在线课程(small private on-line course,SPOC)是近十年来发展起来一种教学模式.SPOC结合了慕课的丰富资源和小规模特色教学的两大优势.为了提升生物化学与分子生物学课堂的教学效果和激发学生的学习主动性,文章以医科大学医科类专业为研究对象,采用异步SPOC在线课堂的方式进行教学实践,并通过统计分析三个学期异步SPOC课程实践的教学数据,发现异步SPOC在线课堂能够提高学生学习的主导性和主动性.

利用CRISPR-Cas9 基因编辑技术敲除人Huh7 肝癌细胞的血管紧张素转化酶 2(ACE2)基因,构建ACE2 基因敲除细胞株,为研究ACE2 在肝细胞癌的作用提供细胞模型.首先,对ACE2 结构域进行鉴定,利用在线网站设计两条破坏所有结构域、靶向作用于ACE2 外显子的sgRNA.其次,构建重组载体并转染肝癌细胞Huh7,嘌呤霉素筛选出单克隆细胞株.最后,免疫印迹鉴定敲除效果.结构域鉴定结果显示,在 340-520 号氨基酸位置存在Zn结合位点和激活位点,根据sgRNA靶点设计原则,采用片段敲除的方式,针对ACE2 的第 9、第 10 外显子设计两对sgRNA,并成功构建PX459-ACE2-sgRNA重组质粒.嘌呤霉素成功筛选出单克隆细胞株,并测序证实了发生片段敲除,ACE2 蛋白在敲除细胞株中不表达;成功构建ACE2 敲除的Huh7 细胞株,为日后研究ACE2 在肝细胞癌的发生机制奠定基础.

目的:基于网络药理学和生物信息学探讨补骨脂定(PSO)治疗鼻咽癌的作用及分子机制.方法:将鼻咽癌5-8F细胞分为5-8F组(0μmol/LPSO),PSO低剂量组(10μmol/L组)、中剂量组(20μmol/L组)、高剂量组(30μmol/L组).通过CCK-8、平板克隆探究PSO对鼻咽癌5-8F细胞增殖、克隆形成的影响;Pubchem、Pharmmapper、GeneCards数据库得到PSO治疗鼻咽癌的潜在靶点;DAVID数据库进行GO、KEGG分析;STRING、GEPIA数据库和Cytoscape软件构建网络图并得到核心靶点;分子对接、western blotting对核心靶点及主要通路进行验证.结果:PSO呈浓度依赖性抑制5-8F细胞的增殖;与5-8F组比较,PSO各剂量组细胞克隆形成能力降低(P＜0.05);PSO治疗鼻咽癌的潜在靶点共有66个,KEGG分析显示,与PI3K-Akt信号通路密切相关;核心靶点为 ALB、HSP90AA1、SRC、EGFR、CASP3、ANXA5、MAPK1,其中 ALB、HSP90AA 1、ANXA5 为生存相关靶点;PSO与核心靶点ALB、HSP90AA1、ANXA5具有良好的结合能力;与5-8F组比较,PSO高剂量组细胞中PI3K、p-PI3K、Akt、p-Akt蛋白水平的表达降低(P＜0.05).结论:PSO可有效抑制鼻咽癌5-8F细胞的增殖、克隆形成,其抗鼻咽癌机制主要与抑制PI3K-Akt信号通路活性有关.

肾素-血管紧张素系统(renin-angiotensin system,RAS)是人类重要的体液调节系统,对维持心血管内稳态和水电解质平衡至关重要,近年来的研究显示RAS与肿瘤发生发展、血管的形成和转移密切相关.RAS主要通过经典的ACE-AngⅡ-AT1轴以及ACE2-Ang(1-7)-Mas替代轴发挥作用,ACE-AngⅡ-AT1轴与导致肿瘤的生物学过程相关,ACE2-Ang(1-7)-Mas轴是ACE-AngⅡ-AT1轴的反向调节轴,ACE2-Ang(1-7)-Mas轴的激活对大多数肿瘤增殖和转移起抑制作用,本文现就这两条轴与肿瘤的关系进行阐述.

Nasopharyngeal carcinoma (NPC) is a malignant tumor highly prevalent in Southeast Asia. The distant metastasis and disease recurrence are still unsolved clinical problems. In recent years, traditional Chinese medicine (TCM) monomers have become significantly attractive due to their advantages. Using high throughput drug sensitivity screening, we identified gambogic acid (GA) as a common TCM monomer displaying multiple anti-NPC effects. GA could effectively inhibit the proliferation of low differentiated cells and highly metastatic cells in NPC via inducing apoptosis and G2/M cell cycle arrest. In addition, GA obviously repressed the abilities of cell clone, migration, invasion, angiogenesis and represented satisfied synergistic effects combined with chemotherapy. Importantly, we found the elevated immune checkpoint CD47 stimulated after chemotherapy was dramatically impaired by GA treatment. Mechanically, the network pharmacology analyses unraveled that the oncogenic signaling pathways including STATs were rewired by GA treatment. Taken together, our study reveals a molecular basis and provides a rationale for GA application as the treatment regime in NPC therapy in future.

人类猴痘是由猴痘病毒引起的人畜共患病.猴痘的临床表现类似于天花,但传染性不如天花.猴痘病毒通过密切接触损伤的皮肤、体液、呼吸道飞沫和被褥等受污染物而在人与人之间传播.目前有3类天花疫苗(MVA-BN、LC16、ACAM2000)可用于预防猴痘.可使用抗病毒药物(特考韦瑞、布林西多福韦、西多福韦)作为治疗手段,在存在严重疾病、免疫功能低下患者、孕妇和哺乳妇女、儿科以及当病变出现在口腔、眼睛和生殖器附近等情况时,可考虑使用抗病毒药物.本综述旨在描述当前用疫苗预防猴痘和使用抗病毒药物治疗猴痘的进展.

目的:通过网络药理学分析蝙蝠葛碱对鼻咽癌(NPC)的作用靶点及通路,并探讨其作用机制.方法:运用Pubchem、Pharmmapper、GeneCards数据库检索蝙蝠葛碱和NPC的作用靶点后并取得它们的交集靶点;使用Cytoscape软件构建"蝙蝠葛碱—NPC疾病靶点"网络图,并利用David数据库进行GO功能注释及KEGG通路富集分析.CCK-8法检测蝙蝠葛碱作用的CNE-2细胞在24h、48h、72h的增殖抑制情况.Western blotting法检测预测出的通路靶点蛋白的表达.结果:共筛选出潜在作用靶点69个,GO功能注释获得分子功能条目16个、生物过程条目24个和细胞组成条目10个,KEGG通路富集得到20条相关通路,涉及PI3K-AKT信号、癌症信号、催乳激素等信号通路.蝙蝠葛碱呈浓度和时间依赖性抑制CNE-2细胞增殖,PI3K和AKT蛋白表达明显下调(P＜0.05).结论:蝙蝠葛碱能有效抑制NPC细胞增殖,并且验证了PI3K-AKT通路为蝙蝠葛碱作用于NPC的主要通路.

目的:研究低氧条件对Hep3B肝癌细胞增殖、迁移的影响,探讨低氧对上皮—间质转化(EMT)、糖异生及脂肪酸合成相关基因的表达变化及意义.方法:克隆形成实验研究Hep3B的增殖能力,细胞划痕愈合和Transwell迁移实验研究Hep3B的迁移能力.肿瘤基因组图谱数据库(TCGA)和基因表达综合数据库(GEO)分析EMT相关基因SNAl1、SNAl2、CDH1、CDH2、ZEB1和Vimentin,糖异生相关基因PCK2、G6PC及脂肪酸合成相关基因ACACA的mRNA在肝细胞癌(HCC)与癌旁组织中的表达情况,并使用GEO数据库分析上述基因在低氧条件下的mRNA表达.实时荧光定量聚合酶链式反应(RT-qPCR)检测上述基因在常氧和低氧条件下培养的Hep3B细胞中的mRNA表达情况.结果:低氧环境可以促进Hep3B细胞的增殖和迁移能力.与癌旁组织比较,TCGA和GEO数据库均显示,ZEB1和ACACA在肝癌组织中表达上调,G6PC和PCK2则下调,SNAl2和Vimentin仅在TCGA中显示上调,SNAl1和CDH1仅在GEO显示下调.RT-qPCR结果显示,与常氧组相比,低氧环境下的Hep3B细胞中EMT相关基因SNAl1、CDH1、CDH2的表达量降低(P＜0.05),而SNAl2、Vimentin的表达量则升高(P＜0.05),糖异生相关基因PCK2、G6PC和脂肪酸合成相关基因ACACA在低氧环境中的表达量均降低(P＜0.05),而ZEB1的表达量差异无统计学意义(P＞0.05).结论:低氧微环境可以调节肝癌Hep3B细胞中EMT相关基因SNAl1、SNAl2、CDH1、CDH2和Vimentin以及代谢相关基因ACA CA、G6PC和PCK2的表达,进而促进Hep3B细胞的增殖和迁移.

Background. The importance of prostaglandin F2 receptor inhibitors (PTGFRN) in the progression of a variety of malignant tumors has been recognized in recent years. So far, no role of PTGFRN in hepatocellular carcinoma (HCC) has been reported. In this study, we focused on the possible mechanisms of PTGFRN in HCC based on the Cancer Genome Atlas (TCGA) data. Methods. The mRNA gene expression data of PTGFRN were downloaded from TCGA database to analyze the expression level of PTGFRN in HCC. According to the human protein atlas database, the expression difference of PTGFRN protein between HCC and adjacent tissues was verified. Wilcoxon signed-rank test and logistic regression were used to analyze the relationship between PTGFRN and clinicopathological characteristics. Kaplan Meier method and Cox proportional hazards model were used to explore the prognostic role of PTGFRN in HCC. The ROC curve was used to evaluate the diagnostic value of PTGFRN in HCC. Gene Set Enrichment Analysis (GSEA) was used to investigate the function of PTGFRN related Gene sets. Finally, obtain the co-expressed genes of PTGFRN through the cBioPortal database, and use the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) function enrichment analysis to further explore the role of PTGFRN in HCC regulated related pathways. Results. Analysis of mRNA expression data of 377 HCC patients showed that the expression of PTGFRN was up-regulated in HCC, which was confirmed by immunohistochemistry. The overexpression of PTGFRN was significantly correlated with clinical stage (P = 0.028) and histological grade (P = 0.027). High expression of PTGFRN was associated with poorer overall survival.. Meanwhile, multivariate Cox analysis showed that PTGFRN may be a potential independent risk factor for HCC. GSEA enrichment results showed that the up-regulated PTGFRN phenotype was concentrated in "endocytosis", "oocyte meiosis" and "ERBB signaling pathway". In addition, through the analysis of KEGG and GO pathways, we found that PTGFRN co-expressed genes are mainly involved in extracellular matrix tissue, epithelial-mesenchymal transition, cell adhesion and cell cycle, and PI3K-Akt/NF-kB signaling pathways. Conclusions. PTGFRN is highly expressed in HCC and can be used as an independent predictor of the clinical prognosis of HCC.

Objective To analyze the physicochemical properties, structure and function of melanoma-associated antigen D4 (MAGE-D4) protein, and then construct the eukaryotic expression vector of MAGE-D4. Methods The physicochemical properties, structure and function of MAGE-D4 protein were analyzed by bioinformatics. Using MAGE-D4/pMAL-C2 prokaryotic recombinant plasmid as the template, PCR product digested by restriction enzyme was connected with pEGFP-C1 eukaryotic expression plasmid and transformed into E. coli. Ligation products were identified by antibiotic screening, enzyme digestion and sequencing. Then the recombinant plasmid was transfected into A549 lung cancer cells by liposome. Results MAGE-D4 protein was an unstable hydrophilic protein without transmembrane structure and signal peptide. Its secondary structure was mainly α-helix. MAGE-D4 contained multiple functional modification sites and was mainly located in the nucleus. SLLLVILGV might be a restricted T cell epitope of HLA-A*0201 derived from MAGE-D4. The first three proteins to potentially interact with MAGE-D4 were NSMCE4A, MLANA/MART-1 and BAGE5. DNA sequencing showed that the recombinant plasmid contained full-length coding sequence (CDS) of MAGE-D4 and it could be successfully transfected into A549 lung cancer cells. Conclusion MAGE-D4 protein is an unstable nuclear protein, which may play functions by interacting with a variety of melanoma-related proteins. The peptide derived from MAGE-D4 may have strong immunogenicity. The eukaryotic expression vector of MAGE-D4 has been successfully constructed.

通过探索建设具有我校生物化学与分子生物学学科特色的"金课",全面梳理课程教学内容,拓展课程深度,将线上和线下教学的优势结合,弥补传统线下教学的缺点,解决医学相关专业学生在生化学习过程中难度过大问题,培养学生综合基础医学思维能力,有助于提高医学相关专业学生在实际工作中独立地发现问题、解决问题的能力.

目的:分析非小细胞肺癌(NSCLC)中黑素瘤相关抗原D4(MAGE-D4)基因启动子甲基化程度与MAGE-D4 mRNA表达水平及患者临床病理特征的关系.方法:采用甲基化特异性PCR(MSP)法检测38例NSCLC患者癌组织及其癌旁组织中MAGE-D4启动子甲基化程度,实时荧光定量PCR(qPCR)法检测组织中MAGE-D4 mRNA相对表达量,分析MAGE-D4启动子甲基化频率与患者临床病理特征的关系.结果:肺癌组织中MAGE-D4基因甲基化频率为23.68％,明显低于癌旁组织的86.84％(P<0.05);而癌组织中MAGE-D4 mRNA表达水平则显著高于癌旁组织(5.49±5.65 vs.1.44±1.08,P<0.05),MAGE-D4表达量与启动子甲期化程度呈负相关关系(r=-0.663,P<0.05).肿瘤直径>3 cm的肺癌患者MAGE-D4甲基化发生率明显低于肿瘤直径≤3 cm患者(P<0.05).结论:NSCLC中MAGE-D4基因转录活性的增强与其启动子低甲基化相关;肿瘤直径较大的NSCLC患者MAGE-D4甲基化发生率较低.

CRISPR-CAS9技术是新一代基因编辑技术,可简便快捷地对哺乳动物细胞进行指定基因的敲除,实现沉默外源基因表达的目的.但传统的CRISPR-CAS9技术提供的酶切方案没有特异性,致使酶切效率不稳定,影响胶回收率以及后续实验.本实验通过改变酶切体系大小,以及酶切反应时间,继而通过胶回收率及测序检测的方法验证最佳酶切条件,最后通过荧光定量PCR和Western blotting实验检测目的基因的表达量,确认是否成功敲除目的基因.研究结果说明:Bbs Ⅰ酶与px459质粒比值为1μL:500 ng,酶切体系为50 μL,酶切时间为45 min时,酶切效果最佳、胶回收率最高,且此时目的基因在细胞内的表达量为0,确定该实验条件下目的基因被成功敲除.

To investigate the gene polymorphism characteristic of HPA of the Han nationality in China and compare the HPA polymorphism between the Han population in south China and north China,we enrolled data which were about the genotypes of HPA-1,HPA-2,HPA-3,HPA-4,HPA-5,HPA-6 and HPA-15 from 28 gene polymorphism research articles in 24 districts,which were merged and managed by using statistic methods and software and then to be analyzed.We found that there was great difference between the gene frequency of a and b in HPA-1,HPA-2,HPA-3,HPA-4,HPA-5,HPA-6 gene locus,and the genotype frequency of bb was close to 0,while the frequency of a and b in HPA-3,HPA-15 gene locus were similar.In the comparison of the HPA gene polymorphism between the south and the north Han population,there was some differences in the genotypes of HPA-1,HPA-5,and the difference of the genotype frequency distribution of HPA-2 between south and north was statistically significant (9＜0.05).The objective of this study was to obtain the polymorphism features of HPA gene and set up a database for HPA to guide the clinical blood transfusion.

肾素-血管紧张素系统(rennin angiotensin system,RAS)是机体内重要的具有调节血压、水钠平衡等功能的内分泌调节系统.近年在各肝病的研究中发现,RAS失衡与各种肝病的发生与发展均有密切关系.肝组织内存在局部的完整RAS系统,当肝细胞或肝组织受到某种致病因素的刺激时,局部RAS被激活,各成分活性增强并重新分布,继而影响肝病的发展.早期研究已证实ACE-A ngⅡ-AT1R轴表达上调可加重肝脏损伤,促进各肝病的进展.新近研究发现的ACE2-Ang (1-7)-Mas轴被不断证明是ACE-A ngⅡ-AT1R轴的反向调节轴,被称为“替代RAS经典轴”,发挥拮抗ACE-A ngⅡ-AT1R轴的作用,对肝病的进展起一定的缓解或逆转作用.本研究就近年来关于RAS对各肝病的影响的相关研究做一综述,为找到更有效的缓解和治愈肝病的新方法提供新思路.

摸索用Lipofectamine 2000 (Lipo)转染质粒pEGFP-C1到肝癌细胞HepG2较为合适的转染条件.以HepG2细胞为研究对象,采用脂质体Lipofectamine 2000转染pEGFP-C1质粒,在24孔板先按每孔0.5 μg固定pEGFP-C1质粒用量,摸索Lipo在1μL、1.5 μL、2μL、2.5 μL量上较为合适的用量,确定Lipo用量后,然后固定Lipo为1μL,摸索质粒用量0.5 μg、1 μg,确定脂质体与质粒的最佳比例.此外,对转染中培养基是否含血清,Lipofectamine 2000与pEGFP-C1质粒比例为1∶0.5的基础上,扩大用量,即Lipofectamine 20002μL和pEGFP-C1质粒1μg,以及脂质体复合物孵育细胞时间进行优化.最后,采用荧光倒置显微镜观察细胞转染效率和方差分析及秩和检验的统计学方法进行统计分析.结果显示,pEGFP-C1质粒固定为0.5 μg时,Lipo用量在1μL及1.5 μL用量组转染效率最好,之后随着Lipo用量加大,转染效率下降;固定Lipo用量1μL,pEGFP-C1质粒0.5 μg时转染效率最好(p＜0.05),比例不变,扩大Lipo和质粒用量,并不增加转染效率.转染后于3h、6h、8h、12h、24 h换成正常培养基培养,转染6h后换液较好.此外,研究发现培养基是否含血清不影响转染效率.本研究表明在24孔板板中用Lipofectamine 2000转染HepG2细胞时较为合适的转染条件为每孔1 μL Lipofectamine 2000和0.5 μg质粒,脂质体与质粒的最佳比例为2∶1,血清不影响转染效率,用含血清培养基转染后6h换液培养.

Objective To assess the relevance of four autoantigens to lupus nephritis (LN), and find out some more sen-sitive and specific characteristic autoantigens. Methods Sera were acquired from Laboratory Department of the First Affiliated Hospital of Guangxi Medical University in July 2013 to July 2014, containing 30 normal sera, 30 systemic lu-pus erythematosus (SLE) patients’ sera, 30 LN patients’ sera and 30 nephrotic syndrome (NS) patients’ sera. The nor-mal group came from healthy examinees. The SLE group was collected according to SLICC Revision of the ACR Clas-sification Criteria for SLE. The LN group came from those SLE patients with renal lesions. The NS group came from NS patients without SLE. Crude lysate enzyme-linked immunosorbent assay (CrELISA) was used to detect potential anti-gens’ expression levels in each group. Bioinformatics method was used to analyze the results. Results Expression levels of A1 and A2 in SLE group (0.67±0.36), (0.31±0.18) are statistically higher than those in normal group (0.47±0.24), (0.22±0.10) and NS group (0.33±0.17), (0.19±0.10) (P<0.05), but similar to those in LN group (0.53±0.27), (0.27±0.14) (P>0.05). Conclusion Among these four autoantigens, A1 and A2 suggest higher sensibility, which make them the more possible characteristic autoantigens of SLE/LN and hopeful diagnostic basis.

为了研究Ang-(1-7)通过Mas对人肝癌细胞增殖的影响及在Mas高、低表达模型中分析其表达差异、凋亡等细胞生理活性变化,我们采用药物(Mas激活剂Ang-(1-7),Mas抑制剂A779)干预HepG2细胞,分别对Ang-(1-7)和A799进行剂量、时间效应实验,实时定量RT-PCR检测药物干预后细胞Mas的表达水平,最后流式细胞仪检测药物干预后细胞的凋亡变化.结果表明,Ang-(1-7)与A779的有效工作浓度为10-7 mol/L,Ang-(1-7)促进Mas的表达,Mas表达水平高于对照组.相反,A779抑制Mas的表达,但作用不明显,主要为拮抗Ang-(1-7)的功能.Ang-(1-7)处理组细胞凋亡明显升高,揭示Mas表达提升诱导人肝癌细胞凋亡.