Hippocampal gene expression in a rat model of depression after electroacupuncture at the Baihui and Yintang acupoints

Preliminary basic research and clinical findings have demonstrated that electroacupuncture ther- apy exhibits positive effects in ameliorating depression. However, most studies of the underlying mechanism are at the single gene level; there are few reports regarding the mechanism at the whole-genome level. Using a rat genomic gene-chip, we profiled hippocampal gene expression changes in rats after electroacupuncture therapy. Electroacupuncture therapy alleviated depres- sion-related manifestations in the model rats. Using gene-chip analysis, we demonstrated that electroacupuncture at Baihui （DU20） and Yintang （EX-HN3） regulates the expression of 21 genes. Real-time PCR showed that the genes Vgf, lgf2, Trnp32, Loc500373, Hifla, Folrl, Nrnb, and Rtn were upregulated or downregulated in depression and that their expression tended to nor- malize after electroacupuncture therapy. These results indicate that electroacupuncture at Baihui and Yintang modulates depression by regulating the expression of particular genes.

RNAi Plasmid Construction of PttGA 20-oxidase Gene

[Objective] Genetic Engineering technology was used to regulate the expression of PttGA20-oxidase gene thus restrained plant height growth and internode elongation for cultivating dwarfed plant.[Method] Based on the RNAi principle,the gene specific sequences of PttGA20-oxidase in the antisense and sense orientations interrupted by a gene sequence from GUS were cloned into a binary vector pBI121.The selection marker gene npt Ⅱ was replaced with bar gene to RNAi plasmid.[Result] After undergone different endonuclease restrictions,the constructed constraint vector released different segments whose sizes were similar to that of target segment,which demonstrated that the RNAi plasmid of PttGA20-oxidase gene was successfully constructed.[Conclusion] The experiment provided a new way for culturing dwarfed plant.

三氧化二砷对急性髓系白血病细胞HL-60 Cdc20及Mad2的影响

目的:探讨三氧化二砷(As_2O_3)对急性髓系白血病细胞HL-60增殖过程中Cdc20及Mad2的影响。方法:不同浓度As_2O_3处理HL-60细胞24、48、72 h后CCK-8法检测细胞增殖情况;Real-Time PCR及Western blot技术检测不同浓度As_2O_3作用于HL-60细胞48 h后Mad2和Cdc20 mRNA及蛋白表达情况。结果:As_2O_3能显著抑制HL-60细胞增殖,并具有良好的时间剂量相关性。RT-PCR和Western blot检测结果表明,As_2O_34、8、10μmol/L作用于HL-60细胞后,Mad2表达上调,Cdc20表达下降(P<0.05)。结论:As_2O_3对人急性髓系白血病HL-60细胞增殖具有较明显的抑制作用,其机制可能与上调Mad2表达和下调Cdc20表达有关。

Detections of mefA, ermB, and mphA Macrolides Resistant Genes in Bacteria Isolated from Covid-19 Patients from Selected Health Facilities in Ibadan, Nigeria

Background: COVID-19 is a disease caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Epidemiological data indicated that bacterial complications in COVID-19 would decrease clearance rate of the infecting agent and increase mortality rate. Macrolides such as Azithromycin are usually administered to COVID-19 patients as palliative treatments. Currently, a considerable number of bacterial strains have developed resistance to various antibiotics, especially macrolides. Resistance is reported to be due to possession of mefA, ermB, and mphA genes by Gram positive and Gram negative bacteria. Therefore, this study determined antibiotic resistance patterns and identify mefA, ermB and mphA macrolide-resistant genes in bacterial pathogens isolated from COVID-19 cases in Ibadan, Nigeria. Methods: 400 Nasopharyngeal samples were collected from symptomatic cases before antibiotic medication;structured questionnaires were administered to collect socio-demographic data of participants. Samples were cultured on Blood, Chocolate, MacConkey and Mannitol salt agar at 37°C for 48 hrs. Bacterial identification was performed using VITEK 2.0 ID cards and API 20E for Gram positive and negative bacteria respectively. Antibiotic Susceptibility Testing was performed using Kirby Bauer disc diffusion methods and VITEK 2.0 AST card kits. DNA of multidrug resistant bacterial isolates was extracted;resistant genes were determined using a polymerase chain reaction with specific primers. Amplified genes were detected using agarose gel electrophoresis. Results: 240 (60%) had bacterial growth and 97 (22.2%) yielded no growth. From the 240 bacterial isolates, 38 (15.83%) were multi-drug resistant including resistance to macrolides (Azithromycin) 20 (52.63%) of which were positive for either mefA or ermB, and none (0.0%) possess mphA gene;14 (36.8%) isolates had mefA gene, 10 (26.3%) isolates carried ermB gene. Conclusion: Multi-drug bacterial resistance including macrolides and quinolones was detected. Only mefA and ermB genes were detected in the bacterial isolates, especially in Gram positive organisms. The detection of mefA and ermB genes in the MDR bacterial isolates raised concern on the use of azithromycin as palliative treatment for COVID-19 symptomatic patients.

The role of FZR1 in tumorigenesis: Focus on cell-cycle control

Fizzy-related protein homolog 1 (FZR1) mainly functions as a specific activator of the anaphase-promotingcomplex/cyclosome (APC/C) in the cell cycle and controls the G0 and G1 phases of the cell cycle. We highlightrecent work that has studied the role of FZR1 in tumorigenesis, growth, differentiation, and genome stability throughcell-cycle control. We summarize the current state of knowledge regarding FZR1 structure, function, and the distinctways of APC/C dysregulation in solid tumors and hematologic malignancies. We also discuss novel approaches fortargeting the FZR1 as a cancer therapy and research area for future work.

Expression of CDC20, TOP2A in Esophageal Phosphorus Carcinoma and Its Relationship with Survival of Esophageal Carcinoma

Objective:To study the expression of CDC20,TOP2A in esophageal squamous cell carcinoma and its relationship with survival of esophageal carcinoma.Methods:65 patients with esophageal squamous cell carcinoma from January 2016 to June 2018 were selected by computer random selection.All patients were treated with radical operation.The CDC20,TOP2A expression of the patients was examined.At the same time,the relationship between 3-year survival rate and CDC20,TOP2A was analyzed by follow-up investigation.Results:the CDC20,TOP2A expression level of cancer tissue group was higher than that of adjacent tissue group and normal tissue group(P<0.05),and the CDC20,TOP2A expression level of adjacent tissue group was higher than that of normal tissue group(P<0.05).There was no significant difference in sex,age,tumor size,tumor location and CDC20,TOP2A expression level in patients with esophageal squamous cell carcinoma,P>0.05;there were differences between groups(P P>0.05)and positive proportion in TNM stage,lymphatic metastasis,invasion factor and CDC20,TOP2A expression level;there were differences in tumor differentiation,5-year survival rate and CDC20,TOP2A expression level(P P>0.05),and showed inverse proportion relationship.Conclusion:metastasis,recurrence and prognosis of esophageal squamous cell carcinoma are related to the level of CDC20,TOP2A expression.These two indexes can effectively evaluate the pathological situation of esophageal cancer and provide an important reference for the prognosis of esophageal squamous cell carcinoma patients.

马钱苷元对白血病细胞增殖及CDC20/MAD2表达的影响

目的探讨马钱苷元对人急性髓系白血病MOLM-13细胞增殖过程中Cdc20及Mad2的影响。方法不同浓度马钱苷元处理MOLM-13细胞24、48、72 h后CCK-8法检测细胞增殖情况;流式细胞术检测不同浓度马钱苷元作用于MOLM-13细胞24h后细胞周期及凋亡的变化;Western blot检测不同浓度马钱苷元作用于MOLM-13细胞48h后Mad2和Cdc20蛋白表达情况。结果马钱苷元能显著抑制MOLM-13细胞增殖,促进MOLM-13细胞凋亡,细胞周期阻滞于G2/M期,Mad2和Cdc20蛋白表达水平均降低。结论马钱苷元对人急性髓系白血Molm细胞增殖具有较明显的抑制作用,其机制可能与调节Mad2和Cdc20的蛋白表达水平,影响细胞有丝分裂有关。

CDC20通过稳定NLRP3的表达促进食管癌细胞增殖的研究

背景与目的:食管癌(esophageal carcinoma,ESCA)是死亡率较高的恶性肿瘤之一,其发生、发展的机制不明。CDC20被认为具有癌基因功能,其表达失调与肿瘤的发生、发展密切相关。CDC20在多种肿瘤中表达升高,敲低CDC20能够抑制肿瘤细胞增殖。NLRP3是炎症小体的主要成分之一,炎症小体失调与肿瘤的发生、发展也密切相关。本研究旨在探究CDC20是否通过NLRP3促进ESCA细胞增殖,同时分析其调控机制。方法:通过癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库和GTEx公共数据库分析ESCA患者中CDC20和NLRP3基因的表达水平。收集新乡医学院第一附属医院收治的80例ESCA患者的临床病理学资料和组织,通过免疫组织化学染色检测NLRP3在ESCA患者中的蛋白表达水平。本研究通过新乡医学院第一附属医院伦理委员会的审批(编号:EC-021-137)。通过短发夹RNA(short hairpin RNA,shRNA)技术检测敲低CDC20和NLRP3基因后对食管鳞状细胞癌细胞系EC9706和KYSE150增殖能力的影响。通过免疫共沉淀(Co-immunoprecipitation,Co-IP)、蛋白酶体抑制剂和泛素化实验检测CDC20是否与NLRP3相互作用,以及阐明CDC20是否通过泛素化途径调控NLRP3表达。结果:TCGA数据库分析结果显示,ESCA组织中CDC20和NLRP3 mRNA表达水平明显高于癌旁组织。免疫组织化学结果也进一步显示,与癌旁组织相比,CDC20、NLRP3在ESCA组织中蛋白表达水平升高。敲低CDC20和NLRP3基因可抑制ESCA细胞增殖。Co-IP、蛋白酶体抑制剂和泛素化实验证实CDC20通过NLRP3的LRR区与NLRP3相互作用,且CDC20通过促进NLRP3泛素化稳定其表达。结论:CDC20和NLRP3在ESCA癌组织中表达上调,且CDC20通过泛素化NLRP3稳定其表达,从而促进ESCA细胞增殖,提示CDC20和NLRP3可能是ESCA潜在的诊断靶向标志物。

CDC20在胃腺癌组织中的表达及其临床意义

目的:检测细胞分裂周期蛋白20(cell division cycle 20,CDC20)在胃腺癌组织中的表达及临床病理学意义。方法:使用GEPIA数据库分析CDC20在非肿瘤性胃黏膜组织及胃癌组织中的mRNA水平。免疫组化(SP)法检测CDC20蛋白表达,分析其与胃腺癌临床病理特征及预后的关系。结果:GEPIA分析显示,CDC20在胃腺癌组织中显著高表达(P<0.05)。免疫组化结果显示:CDC20蛋白在胃腺癌组织中的阳性表达率明显高于癌旁正常胃黏膜组织(P<0.05)。且CDC20表达与胃腺癌的淋巴结转移、TNM分期以及分化程度显著相关(P<0.05)。Cox比例风险回归模型分析显示:CDC20高表达是影响胃腺癌患者预后的独立危险因素(P<0.05)。结论:胃腺癌组织中CDC20呈高表达,与肿瘤分期和淋巴结转移显著相关,可能是胃腺癌预后的标志物。

肺腺癌关键基因筛选及CDC20在肺腺癌中的表达

目的:筛选肺腺癌的关键基因,揭示肺腺癌潜在的分子机制。方法:利用3个肺腺癌相关基因芯片数据集,共包括156例肺腺癌和106例正常肺组织,进行肺腺癌潜在关键基因的筛选。结果:通过芯片数据分析得到341个重叠差异表达基因(differentially expressed gene,DEG),DEG主要富集的生物学过程是细胞外基质组织、调控化学激酶介导的信号通路、细胞对转化生长因子刺激的反应、细胞外基质组装、调控新生血管生成等。通过蛋白互作网络(protein protein interaction,PPI)筛选出10个关键基因,其中8个基因AURKA(aurora kinase A)、CDC20(cell division cycle 20)、CDH5(cadherin 5)、COL1α1(collagenⅠ,α1)、EDN1(endothelin 1)、MMP9(matrix metalloprotein 9)、PECAM1(platelet endothelial cell adhesion molecule 1)、SPP1(secreted phosphoprotein 1)与肺腺癌预后相关。其中CDC20与肺腺癌预后相关性最高,亚组分析发现,CDC20在肺腺癌各年龄段患者中的表达均明显高于正常组,CDC20在一期肺腺癌中表达低于二期、三期、四期,CDC20在腺泡型、实性为主型、混合型、透明细胞型表达高于其他亚型,CDC20在吸烟的肺腺癌患者中的表达低于不吸烟者。免疫组化检测显示CDC20在肺腺癌组织中表达明显高于正常肺组织,且腺泡型、实体型表达高于其他亚型。结论:CDC20在肺腺癌中高表达,且其高表达与肺腺癌的不良预后显著相关。CDC20有可能成为肺腺癌治疗的新靶点。

Gene-activated matrix harboring a miR20a-expressing plasmid promotes rat cranial bone augmentation

Gene-activated matrix(GAM)has a potential usefulness in bone engineering as an alternate strategy for the lasting release of osteogenic proteins but efficient methods to generate non-viral GAM remain to be established.In this study,we investigated whether an atelocollagen-based GAM containing naked-plasmid(p)DNAs encoding microRNA(miR)20a,which may promote osteogenesis in vivo via multiple pathways associated with the osteogenic differentiation of mesenchymal stem/progenitor cells(MSCs),facilitates rat cranial bone augmentation.First,we confirmed the osteoblastic differentiation functions of generated pDNA encoding miR20a(pmiR20a)in vitro,and its transfection regulated the expression of several of target genes,such as Bambi1 and PPARc,in rat bone marrow MSCs and induced the increased expression of BMP4.Then,when GAMs fabricated by mixing 100 ll of 2%bovine atelocollagen,20mg b-TCP granules and 0.5mg(3.3 lg/ll)AcGFP plasmid-vectors encoding miR20a were transplanted to rat cranial bone surface,the promoted vertical bone augmentation was clearly recognized up to 8 weeks after transplantation,as were upregulation of VEGFs and BMP4 expressions at the early stages of transplantation.Thus,GAM-based miR delivery may provide an alternative non-viral approach by improving transgene efficacy via a small sequence that can regulate the multiple pathways.

超表达Cdc20基因不影响牛卵母细胞第一极体排出

CDC20(cell division cycle 20)是后期促进复合物(anaphase-promoting complex,APC)的共激活剂之一,也是纺锤体组装检查点(spindle assembly checkpoint,SAC)的靶点,在细胞周期调控中扮演重要角色.为探讨Cdc20在第一极体排出(first polar body extrusion,PBE I)中的作用,Cdc20基因被成功克隆并构建了真核表达载体pCdc20-Venus,随后用T7 Mmessage Mmachine Kit(Ambion)以线性化pCdc20-Venus为模板体外转录(in vitro transcription)获得带帽的Cdc20-Venus mRNA,将Ccdc20-Venus mRNA显微注射到体外培养的牛卵母细胞胞质中进行超量表达.结果表明,真核表达载体pCdc20-Venus转染HeLa细胞后能够正常表达,绿色荧光在细胞核周围呈弥散状分布;将Cdc20-Venus mRNA注射到牛卵母细胞胞质后,胞质内有绿色荧光出现.Cdc20-Venus mRNA注射组卵母细胞的PBE I率(48.9%)与Venus mRNA注射组卵母细胞的PBE I率(50.9%)和未注射组卵母细胞的PBE I率相比均没有显著差异(P>0.05).通过Cdc20在牛卵母细胞内的超量表达,结果显示,超量表达Cdc20基因对牛卵母细胞PBE I没有显著影响(P>0.05).

RNA联合干扰对肝癌细胞CDC20和HPSE表达的影响

目的探讨RNA联合干扰(RNAico)对肝癌细胞株Hepa1-6中丝裂原调控子20(CDC20)和乙酰肝素酶(HPSE)表达的抑制作用。方法设计针对小鼠肝癌细胞Hepa1-6 CDC20和HPSE mRNA的小干扰RNA,以阳离子脂质体包埋后转染鼠肝癌细胞Hepa1-6,即RNAico;转染48 h后,采用RT-PCR和Re-al-Time PCR测定CDC20、HPSE mRNA表达的变化。结果联合干扰组CDC20和HPSE mRNA表达明显抑制,与其他各组相比,在统计学上有显著性差异(P<0.05)。结论 RNAico可同时显著抑制小鼠肝癌细胞Hepa1-6中CDC20和HPSE mRNA的表达。

CDC20在细胞分裂周期中的作用

CDC20是细胞周期相关蛋白之一。在细胞分裂周期中,CDC20是纺锤体组装检查点的靶向物和有丝分裂后期促进复合体的正调控因子,在引导细胞周期中某些蛋白质的泛素化降解和确保染色体正常分离的过程中起着重要的作用。

过表达cdc20基因对绒山羊卵母细胞体外成熟的影响

为研究过表达cdc20基因对绒山羊卵母细胞体外成熟的影响,构建了Myc-cdc20载体,在体外转录成cRNA后显微注射到绒山羊卵母细胞中,观察其对绒山羊卵母细胞体外成熟过程的影响。结果表明,在对照组卵母细胞体外培养的过程中,卵母细胞培养到8 h后发生GVBD,18 h后进入MⅠ期,直到24 h后,约75%的绒山羊卵母细胞排出第一极体。卵母细胞在GV期时显微注射g Myc-cdc20 cRNA,培养10 h后,通过Western Blotting的方法检测到了MYC-CDC20融合蛋白的表达。但统计结果显示,试验组卵母细胞各阶段发育时间和成熟率与对照并无显著差异。因此,单独过表达cdc20基因不影响绒山羊卵母细胞体外成熟过程。

Host Interferon-Stimulated Gene 20 Inhibits Pseudorabies Virus Proliferation

Host interferon-stimulated gene 20(ISG20)exerts antiviral effects on viruses by degrading viral RNA or by enhancing IFN signaling.Here,we examined the role of ISG20 during pseudorabies virus(PRV)proliferation.We found that ISG20 modulates PRV replication by enhancing IFN signaling.Further,ISG20 expression was upregulated following PRV infection and poly(I:C)treatment.Ectopic expression of ISG20 inhibited PRV proliferation in PK15 cells,whereas knockdown of ISG20 promoted PRV proliferation.In addition,ISG20 expression upregulated IFN-βexpression and enhanced IFN downstream signaling during PRV infection.Notably,PRV UL24 suppressed the transcription of ISG20,thus antagonizing its antiviral effect.Further domain mapping analysis showed that the N terminus(amino acids 1-90)of UL24 was responsible for the inhibition of ISG20 transcription.Collectively,these findings characterize the role of ISG20 in suppressing PRV replication and increase the understanding of host-PRV interplay.

Expression of pathogenesis-related genes in cotton roots in response to Verticillium dahliae PAMP molecules

Verticillium wilt disease becomes a major threat to many economically important crops. It is unclear whether and how plant immunity takes place during cotton-Verticillium interaction due to the lack of marker genes. Taking advantage of cotton （Gossypium hirsutum） genome, we discovered pathogenesis-related （PR） gene families, which have been widely used as markers of immune responses in plants. To profile the expression of G. hirsutum PR genes in the process of plant immunity, we treated cotton roots with two immunogenic peptides, fig22 and nlp20 known as pathogen-associated molecular patterns, as well as three Verticillium dahliae-derived peptides, nlp20vd2, nlp23vd3, and nlp23vd4 which are highly identical to nlp20. Quantitative real-time PCR results revealed that 14 G. hirsutum PR gene （GhPR） families were induced or suppressed independently in response to fig22, nip20, nlp20va2, nlp23vd3, and nlp23vd4. Most GhPR genes are expressed highest at 3 h post incubation of immunogenic peptides. Compared to fig22 and nlp20, nlp20vd2 is more effective to trigger up-regulated expression of GhPR genes. Notably, both nlp23vd3 and nlp23vd4 are able to induce GhPR gene up-regulation, although they do not induce necrosis on cotton leaves. Thus, our results provide marker genes and new immunogenic peptides for further investigation of cotton-V, dahliae interaction.

The RNF20/40 complex regulates p53-dependent gene transcription and mRNA splicing

p53 is a key transcription factor to regulate gene transcription.However,the molecular mechanism of chromatin-associated p53 on gene transcription remains elusive.Here,using unbiased protein affinity purification,we found that the RNF20/40 complex associated with p53 on the chromatin.Further analyses indicated that p53 mediated the recruitment of the RNF20/40 complex to p53 target gene loci including p21 and PUMA loci and regulated the transcription of p21 and PUMA via the RNF20/40 complex-dependent histone H2B ubiquitination(ubH2B).Lacking the RNF20/40 complex suppressed not only ubH2B but also the generation of the mature mRNA of p21 and PUMA.Moreover,ubH2B was recognized by the ubiquitin-binding motif of pre-mRNA processing splicing factor 8(PRPF8),a subunit in the spliceosome,and PRPF8 was required for the maturation of the mRNA of p21 and PUMA.Our study unveils a novel p53-dependent pathway that regulates mRNA splicing for tumor suppression.

CDC20、TOP2A、NEK2在食管鳞癌中的表达及其与临床病理特征及预后的相关性

目的探讨CDC20、TOP2A、NEK2在食管鳞癌中表达及其与食管鳞癌临床病理特征及预后的相关性。方法选取实施根治性手术的食管鳞癌患者70例,均术中做病理取材标本,A组(癌组织)、B组(癌旁组织)、C组(正常组织),采用免疫组化法、半定量逆转录聚合酶链反应(RT-PCR)检测CDC20、TOP2A、NEK2表达情况。结果 A组CDC20、TOP2A、NEK2表达水平最高,与B、C组差异明显(P <0. 05); B组CDC20、TOP2A、NEK2表达水平高于C组,差异明显(P<0. 05)。食管鳞癌患者性别、年龄、瘤体大小、位置等因素与CDC20、TOP2A、NEK2表达无相关性(P> 0. 05); TNM分期、淋巴转移及浸润因素与CDC20、TOP2A、NEK2呈正相关性(P <0. 05);肿瘤分化与CDC20、TOP2A、NEK2阳性表达率呈负相关。TNM分期、肿瘤分化程度、淋巴转移、静脉浸润及CDC20、TOP2A、NEK2阳性表达均是影响食管鳞癌患者预后存活的危险因素。结论 CDC20、TOP2A、NEK2高表达与食管鳞癌转移、复发、预后均有密切相关,三项指标联检可准确评估食管癌病理状态,为患者预后判定提供参考。