Pathogenesis of chronic enteropathy associated with the SLCO2A1 gene:Hypotheses and conundrums

Chronic enteropathy associated with the SLCO2A1 gene(CEAS)is a complex gastroenterological condition characterized by multiple ulcers in the small intestine with chronic bleeding and protein loss.This review explores the potential mechanisms underlying the pathogenesis of CEAS,focusing on the role of SLCO2A1-encoded prostaglandin transporter OATP2A1 and its impact on prostaglandin E2(PGE2)levels.Studies have suggested that elevated PGE2 levels contribute to mucosal damage,inflammation,and disruption of the intestinal barrier.The effects of PGE2 on macrophage activation and Maxi-Cl channel functionality,as well as its interaction with nonsteroidal anti-inflammatory drugs play crucial roles in the progression of CEAS.Understanding the balance between its protective and pro-inflammatory effects and the complex interactions within the gastrointestinal tract can shed light on potential therapeutic targets for CEAS and guide the development of novel,targeted therapies.

Performance Parameters:Demobilization Antibiotic Resistant Bacteria(ARB)and Carrying Genes(ARG)in Wastewater Disinfection

The UV irradiation is used for removing Antibiotic Resistant Bacteria(ARB)and Antibiotic Resistance Genes(ARG)from wastewater treatment.Bacteriophages are viruses that infect within bacteria,are recognized for bacterial control.The influence of some parameters in quantification and performance influencing of pathogen demobilization could be considered in disinfection of wastewater.The comparison of Polyvalent phage(NE1)versus Coliphage(NE4)in suppressing a bacterium Escherichia coli(NDM-1:b-lactam-resistant)with UV irradiation was observed the efficacy in reduction of cells in the disinfection and parameter process.The results with the effect of UV-C irradiation on NDM-1 infected with 1%of NE4 showed a decrease of cells from 8×10^(6)to 2×10^(5)in 60 min with UV-C dose.The NDM1(E.coli)was infected with 1%of NE4(Polyvalent Phage)under magnetic stirring for 1 h,the cells count was 8×10^(6).After 1 h in UV-C e×posure,the cells number reached 3×10^(5).The NDM1 that was e×posed in 1 h of UV-C irradiation and then was infected with 1%of NE4.Cells counting were done 24 h after this procedure.These cells were e×posed in UV-C and showed a reduction in the number of cells from 1×10^(8)to 4×10^(5)after 60 min.The results indicate that bacteriophages can mitigate bacteria species,and combined the conventional water disinfection technologies that can support the microbial safety control strategies.

Apolipoprotein E gene polymorphism and susceptibility to intracerebral hemorrhage:A meta-analysis

OBJECTIVE： To evaluate the relationship between apolipoprotein E （ApoE） gene polymorphism and susceptibility to intracerebral hemorrhage （ICH） in Chinese population by a meta-analysis. METHODS： Related literature regarding control analysis between ICH and control groups was collected. Independent case-control studies published between 1989 and 2007 that had complete data were included; and articles not closely related to the topic were excluded. The meta-analysis software, RevMan 4.2, was applied to analyze the odds ratio （OR） value in those studies included in the analysis to assess the relationship between susceptibility to ICH and ApoE polymorphism. RESULTS： Eight papers which were in accordance with the inclusion criteria were selected, and a total of 1 249 ICH cases and 1 329 controls were involved. Meta-analysis results showed that with the wildtype E3/3 as a reference, the OR values （95% confidence interval） of intracerebral hemorrhage for subjects carrying E2/2, E3/2, E4/2, E4/3, and E4/4 were 1.15 （0.60–2.21）, 1.00 （0.79–1.28）, 3.01 （1.73–5.23）, 1.78 （1.41–2.24） and 1.94 （1.03–3.65）, respectively. The combined OR values （95% confidence interval） of intracerebral hemorrhage for ε4 and ε2 carriers were 1.53 （1.16–2.01）, and 0.93 （0.69–1.25）. CONCLUSION： The results suggest that ApoE polymorphism is significantly associated with susceptibility to intracerebral hemorrhage and that ε4 carriers have a higher risk for intracerebral hemorrhage than others.

Correlation of E-selectin gene polymorphisms with risk of ischemic stroke A meta-analysis

OBJECTIVE： To investigate the association of E-selectin $128R polymorphisms with ischemic stroke. DATA SOURCES： A computer-based online search was conducted in PubMed, Elsevier, Ovid Database, the China National Knowledge Infrastructure, and Wanfang Database between January 1998 and December 2010. STUDY SELECTION： Case-controlled studies addressing the association of the E-selectin polymorphism and ischemic stroke were included in this review. The genotype distribution complied with the Hardy-Weinberg genetic equilibrium. The included reports were evaluated by two authors for strict quality screening. Meta-analysis software, REVMAN 5.1, was used to investigate heterogeneity, pooled odds ratio （OR） and 95% confidence interval （CO in individual studies. MAIN OUTCOME MEASURES： Genotype and allele distributions at the E-selectin $128R site. RESULTS： Six case-controlled studies were included after screening and application of inclusion and exclusion criteria. There was no heterogeneity in the genotype and allele frequencies, and no publication bias was found. Meta-analysis of the pooled data showed that the OR value of the （AC＋CC）/AA genotype was 1.93 （95% CI： 1.55 2.41, Z= 5.80, P 〈 0.000 01）, and the ORfor the C/A allele was 1.80 （95% CI： 1.47 2.22, Z= 5.59, P 〈 0.000 01） in the ischemic stroke group, compared with control group. Results of pooled data in Chinese subjects showed that the ORvalue of （AC＋CC）/AA was 2.36 （95% CI： 1.68 3.31, Z = 4.99, P 〈 0.000 01）, and the OR value of the C/A allele was 2.25 （95% CI： 1.63 3.12, Z= 4.89, P 〈 0.000 01）. CONCLUSION： Polymorphism of E-selectin S128R was significantly associated with susceptibility to ischemic stroke; the AC and CC genotypes as well as the C allele may be factors associated with susceptibility to ischemic stroke.

Genome-wide identification and expression analysis of Gossypium RING-H2 finger E3 ligase genes revealed their roles in fiber development,and phytohormone and abiotic stress responses

Background： RING H2 finger E3 ligase （RH2FE3） genes encode cysteine rich proteins that mediate E3 ubiquitin ligase activity and degrade target substrates. The roles of these genes in plant responses to phytohormones and abiotic stresses are well documented in various species, but their roles in cotton fiber development are poorly understood. To date, genome wide identification and expression analyses of Gossypium hirsutum RH2FE3 genes have not been reported. Methods： We performed computational identification, structural and phylogenetic analyses, chromosomal distribution analysis and estimated KJKs values of G hirsutum RH2FE3 genes. Orthologous and paralogous gene pairs were identified by all versus all BLASTP searches. We predicted cis regulatory elements and analyzed microarray data sets to generate heatmaps at different development stages. Tissue specific expression in cotton fiber, and hormonal and abiotic stress responses were determined by quantitative real time polymerase chain reaction （qRT PCR） analysis. Results： We investigated 140 G hirsutum, 80 G. orboreum, and evolutionary mechanisms and compared them with orthologs 89 G. roimondii putative RH2FB genes and their in Arobidopsis and rice. A domain based analysis of the G hirsutum RH2FE3 genes predicted conserved signature motifs and gene structures. Chromosomal localization showed the genes were distributed across all G hirsutum chromosomes, and 60 duplication events （4 tandem and 56 segmental duplications） and 98 orthologs were detected, cis elements were detected in the promoter regions of G hirsutum RH2FE3 genes. Microarray data and qRT PCR analyses showed that G hirsutum RH2FE3 genes were strongly correlated with cotton fiber development. Additionally, almost all the （brassinolide, gibberellic acid （GA）, indole 3-acetic acid drought, and salt）. dentified genes were up regulated in response to phytohormones （IAA）, and salicylic acid （SA）） and abiotic stresses （cold, heat, Conclusions： The genome wide identification, comprehensive analysis, and characterization of conserved domains and gene structures, as well as phylogenetic analysis, cis element prediction, and expression profile analysis of G hirsutum RH2FE3 genes and their roles in cotton fiber development and responses to plant hormones and abiotic stresses are reported here for the first time. Our findings will contribute to the genome wide analysis of putative RH2FE3 genes in other species and lay a foundation for future physiological and functional research on G hirsutum RH2FE3 genes.

Study on Genetic Inactivation Bacterial Ghosts of Pasteurella multocida based PhiX174 Gene E Lysis Cassette Mediated

[Objective] To study on genetic inactivation bacterial ghosts of Pasteurella multocida based PhiX174 gene E lysis cassette mediated. [ Method ] Recombinant pPBA1100-e was constructed by which the gene E of bacteriophage Phix174 and temperature sensitivity expressing control system hybridized with plasmid pPBA1100 by genetic engineering method. Recombinant was transformed to Pasteurella multocida and lysis gene E expressed by temperature induction. Recombinant was detected by restriction endonuclease. Cell morphology of bacterial ghost of Pasteurella mul- tocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysis. I Result~ The results indicated that the recombination plasmid presented three bands by restriction endonuclease and agarose electrophoresis and that molecular weight of every band ac- corded with theoretical value. The result of SEM observing showed that recombination plasmid expressed successfully in P. multocida and produced bacterial ghost. The result of CFU detecting demonstrated that inactivation ratio of P. multocida reached 99 per cent. ~Conclusion~ This study pro- vided technical bases for the preparation of antigen vaccine of natural bacterial outer membrane protein.

Sequence analysis of human papillomavirus type 16 E6E7 gene from cervical carcinoma biopsies of Chinese patients in Shandong province

Objective To study the structure specificity of HPV16E6E7 gene from cervical carcinoma biopsies of patients in Shandong province. Methods The tissue DNAs were extracted from cervical carcinoma biopsies of 14 patients of Chinese from Shandong province. By PCR method using HPV multiple primers, the HPV types were identified in cervical carcinoma tissues, Using the tissue DNA of the 2 cases with the infection of HPV16 type only as templates, HPV16E6E7 gene was amplified by PCR respectively, then inserted the HPV16E6E7 gene into pALTER I vector, and obtained the recombinant plasmid pALTER HPV16E6E7. The constructs were sequenced using Sanger method, and then compared with the sequence of HPV16E6E7 gene of the prototype. Results Sequencing results showed that HPV16E6E7 gene in the 2 cases had sequence diversity from that of the prototype, and a total of five nucleotide exchanges were detected, four of these led to amino acid exchanges. Conclusion There are structure differences between the HPV16E6E7 of Chinese and that of the prototype.

Cloning and Sequence Analysis of E Gene from Chicken Flavivirus Isolate CJD05

[ Objective] This experiment aimed to find out the origin and genetic evolution relationship of chicken flavivirus （CFV） CJD05 strain in Fujian Province. [Method] A pair of primers were designed and synthesized according to the sequences of E gene from Duck flavivirus （DFV） iso- late BYD-1. E gene of CFV isolate CJD05 was specially amplified and its sequences were analyzed. [Result] The target bar which was cloned from CFV isolate C, JD05 was I 503 bp. Homology analysis was conducted to compare E gene nucleotide sequence of CFV isolate CJD05 with DFV iso- late BYD-I and goose flavivirus （GFV） isolate JS804. Results indicated that isolate nuclectide homologies were 99.2% and 99.3%, and amino acid homologies were 99.0% and 98.6% respectively. [Conclusion] CFV isolate C, JD05, DFV isolate BYD-1 and GFV isolate JS804 were highly homologous. The homology of CFV isolate CJD05 with Tembusu virus （TMUV） was higher than with other arthropod-borne flaviviruses.

The Analysis of Human Papillomavirus Type 16 E6/E7 Genetic Variability in Jingjiang, Jiangsu Province, China

Human papillomavirus 16 (HPV-16) is a major high-risk type causing cervical cancer. The E6 and E7 genes in HPV-16 are the major virulent genes in HPV, and we wanted to investigate the polymorphism of E6 and E7 genes of HPV-16 originated from Jingjiang, Jiangsu province. In research, HPV-16 sample was collected, and the E6 and E7 genes fragments were amplified by PCR assay. The sequences of E6 and E7 genes were used for phylogenetic analysis by Mega 5.0 software. By comparison, the major mutations of E6 gene were T178G (41.67%) and G658A (17%), as well as C58G, T61A and G188C (14%), T61C and C656A (11%). The mutation of E7 gene was mainly C491A and T935A (23%);G514C, G937C and G519C (11%). The 68 nucleotide site deficiency (69.4%) of E6 and 535 nucleotide site deficiency (23%) of E7 were dominant. In our study, we did not find the relevance between the E6 and E7 genes mutations and pathologic severity. But we think the E6 and E7 genes mutation of HPV-16 might affect vaccine prevention and treatment effect.

三酰甘油与APOE基因多态性在短暂性脑缺血发作患者发生急性脑梗死中的交互作用研究

目的探讨三酰甘油(TG)与载脂蛋白E(APOE)基因多态性在短暂性脑缺血发作(TIA)患者发生急性脑梗死中的交互作用。方法对2019年1月—2021年6月349例TIA进行6个月随访,根据是否发生急性脑梗死分为发生组、未发生组,比较2组一般资料、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、TG、APOE基因多态性,并比较不同TG水平患者APOE基因多态性,使用多因素Logistic回归分析TIA发生急性脑梗死的影响因素,使用交互作用系数γ分析TG与APOE基因多态性的交互作用。结果随访6个月,共45例(12.89%)TIA患者发生急性脑梗死。发生组TG高于未发生组(P<0.01)。发生组及TG升高患者E3/3基因型、ε3等位基因频率分别低于未发生组及TG正常患者,E3/4基因型、ε4等位基因频率分别高于未发生组及TG正常患者(P<0.05)。校正TIA持续时间、TIA发作频率后,TG、APOE基因型E3/3、E3/4及等位基因ε4仍是TIA发生急性脑梗死的独立影响因素(P<0.01)。TG升高与APOE基因型E3/4在TIA发生急性脑梗死中呈正向交互作用,且交互作用符合次相乘模型。结论TG升高与APOE基因型E3/4在TIA发生急性脑梗死中呈正向交互作用,且交互作用符合次相乘模型,TG升高可增强APOE基因型E3/4对TIA发生急性脑梗死的易感性。

Association of apolipoprotein E gene polymorphism with the occurrence and therapeutic effect of ischemic stroke

At present,ischemic stroke seriously affects people's life and health,and its occurrence,development and therapeutic effect are affected by many factors.With the deep research on ischemic cerebral apoplexy disease,people have a deeper understanding of its virulence genes.The apolipoprotein E genotype is the research focus recently,its genetic type is not only involved in the occurrence and development of ischemic cerebral apoplexy,but also causes different therapeatic effects.In this paper,we reviewed the relationship between apolipoprotein E gene polymorphism and lipid metabolism and atherosclerosis in ischemic stroke,as well as the differences in the therapeutic effects of thrombolysis,thrombectomy and lipid-lowering among different genotypes.

蛋内注射硒和维生素E对1日龄肉鸡抗氧化性能和免疫性能的影响

为研究胚蛋内注射硒和维生素E对1日龄肉鸡抗氧化性能和免疫性能的影响,试验选取240枚重量相近的黄羽肉鸡种蛋,随机分为4组:对照组(生理盐水)、硒组(20 mg硒/枚)、维生素E组(5 mg维生素E/枚)和混合组(20 mg硒+5 mg维生素E/枚),孵化至10胚龄时,于胚蛋卵黄囊内注射四种不同物质,统计出雏情况并检测1日龄肉鸡相关指标.结果显示,与对照组相比,混合注射硒和维生素E显著增加了1日龄肉鸡胸肌指数和肝指数(P<0.05),三个试验组均显著增加了脾指数(P<0.05);维生素E组血清过氧化氢酶(CAT)、硒组血清谷胱甘肽过氧化物酶(GSH⁃Px)和混合组血清CAT、GSH⁃Px、超氧化物歧化酶(SOD)的活性显著升高(P<0.05);在肝中,维生素E组GSH⁃Px和硒组SOD活性显著升高(P<0.05);维生素E组肝CAT、SOD、GSH⁃Px、核因子红细胞系2相关因子2(Nrf2)、肿瘤坏死因子⁃α(TNF⁃α)和干扰素⁃γ(IFN⁃γ)等基因表达显著上调(P<0.05),混合组肝CAT、Nrf2和白细胞介素⁃1β(IL⁃1β)的基因表达显著上调(P<0.05).结果表明,蛋内注射硒和维生素E显著提高了1日龄肉鸡的抗氧化和免疫性能,并促进了雏鸡组织器官的生长发育,硒和维生素E联合注射效果优于单独注射.该研究为硒和维生素E在早期应用于鸡胚,从而提高雏鸡的抗氧化和免疫性能提供了科学理论依据.

Prediction of Tumor Microenvironment Characteristics and Treatment Response in Lung Squamous Cell Carcinoma by Pseudogene OR7E47P-related Immune Genes

Objective Pseudogenes are initially regarded as nonfunctional genomic sequences,but some pseudogenes regulate tumor initiation and progression by interacting with other genes to modulate their transcriptional activities.Olfactory receptor family 7 subfamily E member 47 pseudogene(OR7E47P)is expressed broadly in lung tissues and has been identified as a positive regulator in the tumor microenvironment(TME)of lung adenocarcinoma(LUAD).This study aimed to elucidate the correlation between OR7E47P and tumor immunity in lung squamous cell carcinoma(LUSC).Methods Clinical and molecular information from The Cancer Genome Atlas(TCGA)LUSC cohort was used to identify OR7E47P-related immune genes(ORIGs)by weighted gene correlation network analysis(WGCNA).Based on the ORIGs,2 OR7E47P clusters were identified using non-negative matrix factorization(NMF)clustering,and the stability of the clustering was tested by an extreme gradient boosting classifier(XGBoost).LASSO-Cox and stepwise regressions were applied to further select prognostic ORIGs and to construct a predictive model(ORPScore)for immunotherapy.The Botling cohorts and 8 immunotherapy cohorts(the Samstein,Braun,Jung,Gide,IMvigor210,Lauss,Van Allen,and Cho cohorts)were included as independent validation cohorts.Results OR7E47P expression was positively correlated with immune cell infiltration and enrichment of immune-related pathways in LUSC.A total of 57 ORIGs were identified to classify the patients into 2 OR7E47P clusters(Cluster 1 and Cluster 2)with distinct immune,mutation,and stromal programs.Compared to Cluster 1,Cluster 2 had more infiltration by immune and stromal cells,lower mutation rates of driver genes,and higher expression of immune-related proteins.The clustering performed well in the internal and 5 external validation cohorts.Based on the 7 ORIGs(HOPX,STX2,WFS,DUSP22,SLFN13,GGCT,and CCSER2),the ORPScore was constructed to predict the prognosis and the treatment response.In addition,the ORPScore was a better prognostic factor and correlated positively with the immunotherapeutic response in cancer patients.The area under the curve values ranged from 0.584 to 0.805 in the 6 independent immunotherapy cohorts.Conclusion Our study suggests a significant correlation between OR7E47P and TME modulation in LUSC.ORIGs can be applied to molecularly stratify patients,and the ORPScore may serve as a biomarker for clinical decision-making regarding individualized prognostication and immunotherapy.

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.

高产乙醇肺炎克雷伯adhE缺失菌株构建及对其产乙醇能力的影响

目的探究基因adhE缺失对高产乙醇肺炎克雷伯菌株生长能力,乙醇脱氢酶活性及产乙醇能力的影响。方法以高产乙醇肺炎克雷伯菌株TH1为背景,利用温敏型质粒介导的同源重组技术构建基因adhE的缺失菌株。通过PCR扩增获取基因adhE片段并构建于pGEM-Teasy表达载体获得回补质粒,导入ΔadhE缺失菌株得到回补菌株,并将pGEM-Teasy空质粒导入高产乙醇肺炎克雷伯野生菌株及ΔadhE缺失菌株作为空载对照。在液体LB培养基中测定各菌株的生长曲线,使用试剂盒检测各菌株的乙醇脱氢酶活性并通过顶空法检测各菌株的乙醇产量。结果使用PCR技术明确ΔadhE缺失菌株与相关回补菌株构建成功。基因adhE缺失后,厌氧条件下高产乙醇肺炎克雷伯菌株的生长速度显著降低,而回补菌株的生长速度显著升高(P<0.05)。同时,相比于野生型菌株,ΔadhE缺失菌株的乙醇脱氢酶活性与乙醇产量显著降低,而回补菌株的乙醇脱氢酶活性与乙醇产量恢复到了野生型菌株的水平(P<0.05)。结论基因adhE的缺失减缓了高产乙醇肺炎克雷伯菌株厌氧条件下的生长速度,同时显著降低了高产乙醇肺炎克雷伯菌株的产乙醇能力。

E型钙黏蛋白对胃癌AGS细胞增殖、侵袭和基因表达谱的影响

目的观察E型钙黏蛋白(E-cadherin)对胃癌AGS细胞增殖、侵袭、迁移及顺铂(DDP)化疗敏感性的影响,筛选E-cadherin可能调控的下游基因与机制。方法培养胃癌AGS细胞(对照组)和E-cadherin过表达的AGS细胞(E-cadherin过表达组)至对数生长期后,采用CCK-8方法检测2组细胞活力,采用集落形成实验检测细胞增殖能力,采用Transwell方法检测细胞侵袭和迁移能力,采用基因芯片比较分析E-cadherin过表达前后细胞基因表达谱变化,筛选差异表达基因,用生物信息学方法对差异表达基因进行基因本体(GO)注释和京都基因与基因组百科全书(KEGG)信号通路富集分析。结果与对照组比较,E-cadherin过表达组细胞活力下降更为显著(F组间=101.674,P<0.001;F组内=9.712,P=0.001;F交互=9.712,P=0.001),细胞集落数量减少(t=11.452,P<0.001),迁移和侵袭细胞数量减少(细胞迁移:t=3.994,P=0.016;细胞侵袭:t=2.971,P=0.041);在DDP作用下,与对照组比较,E-cadherin过表达组细胞活力下降更为显著(F组间=24.312,P<0.001;F组内=327.222,P<0.001;F交互=14.445,P<0.001),细胞迁移和侵袭细胞数量减少(细胞迁移:t=4.253,P=0.013;细胞侵袭:t=3.944,P=0.017);E-cadherin过表达后,共筛选出差异表达基因3882个,包括552个lncRNA、272个circRNA和3058个mRNA;GO注释显示,差异表达基因主要富集在GTP酶活性的正向调节、细胞外基质生成、血管生成等生物学过程;KEGG通路富集分析显示,差异表达基因主要富集在细胞外基质受体相互作用、丝裂原活化蛋白激酶信号通路、补体与凝血级联通路等。结论E-cadherin表达通过调节基因组变化抑制胃癌细胞增殖、迁移、侵袭,为胃癌个体化治疗策略研究提供了分子基础。

基于H&E图像和基因表达数据的多模态深度学习模型预测胃癌生存风险

胃癌作为高发恶性肿瘤,其致死率近年来居高不下,因此精准预测胃癌患者的生存风险对于治疗至关重要。文章提出了一种基于多模态深度学习的预测模型,旨在评估胃癌患者的生存风险。该模型整合了H&E(Hematoxylin-Eosin staining)染色图像和基因表达数据,首先,采用ResNet18卷积神经网络模型提取深层H&E图像信息,将其编码为一维特征向量。其次,采用多模态紧凑型双线性池化方法,将图像特征与基因表达数据进行融合,用于预测胃癌患者的风险分数。在TCGA的胃癌样本中,该模型的一致性指数(c-index)为0.70。在测试集上进行的Kaplan-Meier分析结果显示,模型成功地区分出高风险群和低风险群。结果表明,该模型在区分胃癌患者风险层次方面表现出色,具有显著优势。

Microbial and enzyme technology: An efficient and convenient method for MiniPrep analysis of recombinant plasmids

Minipreparation (MiniPrep) analysis is an essential step for obtaining a recombinant plasmid that carries a DNA insert containing a gene of interest. The most commonly used method for this involves cultivation of transformed Escherichia coli (E. coli) in liquid medium, brief centrifugation for precipitation of bacterial pellets, and subsequent lysis of the pellets. This process is time-consuming and laborious, especially when the sample number is high. Here, we describe a more convenient method for MiniPrep analysis that utilizes solid medium-based cultivation of bacteria.

Exercise-induced modulation of miR-149-5p and MMP9 in LPS-triggered diabetic myoblast ER stress: licorice glycoside E as a potential therapeutic target

Background:This study explores the relationship between endoplasmic reticulum(ER)stress and diabetes,particularly focusing on the impact of physical exercise on ER stress mechanisms and identifying potential therapeutic drugs and targets for diabetes-related sepsis.The research also incorporates traditional physical therapy perspectives,emphasizing the genomic insights gained from exercise therapy in disease management and prevention.Methods:Gene analysis was conducted on the GSE168796 and GSE94717 datasets to identify ER stress-related genes.Gene interactions and immune cell correlations were mapped using GeneCard and STRING databases.A screening of 2,456 compounds from the TCMSP database was performed to identify potential therapeutic agents,with a focus on their docking potential.Techniques such as luciferase reporter gene assay and RNA interference were used to examine the interactions between microRNA-149-5p and MMP9.Results:The study identified 2,006 differentially expressed genes and 616 miRNAs.Key genes like MMP9,TNF-α,and IL1B were linked to an immunosuppressive state.Licorice glycoside E demonstrated high affinity for MMP9,suggesting its potential effectiveness in treating diabetes.The constructed miRNA network highlighted the regulatory roles of MMP9,IL1B,IFNG,and TNF-α.Experimental evidence confirmed the binding of microRNA-149-5p to MMP9,impacting apoptosis in diabetic cells.Conclusion:The findings highlight the regulatory role of microRNA-149-5p in managing MMP9,a crucial gene in diabetes pathophysiology.Licorice glycoside E emerges as a promising treatment option for diabetes,especially targeting MMP9 affected by ER stress.The study also underscores the significance of physical exercise in modulating ER stress pathways in diabetes management,bridging traditional physical therapy and modern scientific understanding.Our study has limitations.It focuses on the microRNA-149-5p-MMP9 network in sepsis,using cell-based methods without animal or clinical trials.Despite strong in vitro findings,in vivo studies are needed to confirm licorice glycoside E’s therapeutic potential and understand the microRNA-149-5p-MMP9 dynamics in real conditions.

APOE多态性在炎症因子诱导的神经毒性反应性星形胶质细胞中的差异作用

目的:探讨载脂蛋白E(APOE)基因多态性在神经毒性反应性星形胶质细胞中的差异作用,为阿尔茨海默病(AD)发病机制的研究提供理论依据。方法:体外分离培养APOE基因敲除小鼠(APOE^(-/-))原代皮层星形胶质细胞,免疫荧光染色法鉴定细胞纯度。构建人APOE3和APOE4重组过表达质粒,分别转染至原代APOE^(-/-)星形胶质细胞中,以APOE^(-/-)原代细胞为对照,采用Western blotting法检测细胞中APOE和神经胶质酸性蛋白(GFAP)蛋白表达水平,采用酶联免疫吸附试验(ELISA)法检测细胞培养上清中APOE水平。采用白细胞介素1α(IL-1α)、肿瘤坏死因子(TNF)和补体C1q联合刺激转染APOE3和转染APOE4的原代星形胶质细胞制备炎症模型,分为APOE3+PBS组、APOE4+PBS组、APOE3+IL-1α+TNF+CC1q组和APOE4+IL-1α+TNF+C1q组,采用细胞免疫荧光染色法观察各组细胞形态表现,采用实时荧光定量PCR(RT-qPCR)法检测各组细胞中磷脂酰肌醇蛋白聚糖4(Gpc4)、磷脂酰肌醇蛋白聚糖6(Gpc6)、血小板反应蛋白1(Thbs1)、血小板反应蛋白2(Thbs2)、酸性分泌蛋白类似蛋白1(Sparcl1)、胶质细胞源性神经营养因子(GDNF)、C3和S100钙结合蛋白B(S100B)mRNA表达水平,微球吞噬实验检测各组细胞吞噬能力,Western blotting法检测各组细胞中B细胞淋巴瘤2(Bcl-2)和含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)蛋白表达水平。结果:与APOE^(-/-)组比较,转染APOE3和APOE4组细胞中APOE和GFAP蛋白表达水平及细胞培养上清中APOE水平明显升高(P<0.01)。荧光显微镜下观察,分别与APOE3+PBS组和APOE4+PBS组比较,APOE3+IL-1α+TNF+Cq1组和APOE4+IL-1α+TNF+Cq1组星形胶质细胞突起变短,胞体变大;与APOE3+IL-1α+TNF+Cq1组比较,APOE4+IL-1α+TNF+Cq1组星形胶质细胞突起更短。分别与APOE3+PBS组和APOE4+PBS组比较,APOE3+IL-1α+TNF+Cq1组和APOE4+IL-1α+TNF+Cq1组细胞中Gpc4、Gpc6、Thbs1、Thbs2和Sparcl1 mRNA表达水平明显降低(P<0.01);与APOE3+IL-1α+TNF+Cq1组比较,APOE4+IL-1α+TNF+Cq1组细胞中Gpc4、Gpc6、Thbs1、Thbs2和Sparcl1 mRNA表达水平明显降低(P<0.05或P<0.01)。分别与APOE3+PBS组和APOE4+PBS组比较,APOE3+IL-1α+TNF+Cq1组和APOE4+IL-1α+TNF+Cq1组细胞中GDNF mRNA表达水平明显降低(P<0.01),C3和S100B mRNA表达水平明显升高(P<0.01);与APOE3+IL-1α+TNF+Cq1组比较,APOE4+IL-1α+TNF+Cq1组细胞中GDNFmRNA表达水平明显降低(P<0.05),C3和S100B mRNA表达水平明显升高(P<0.05)。分别与APOE3+PBS组和APOE4+PBS组比较,APOE3+IL-1α+TNF+Cq1组和APOE4+IL-1α+TNF+Cq1组细胞吞噬微珠数量明显减少;与APOE3+IL-1α+TNF+Cq1组比较,APOE4+IL-1α+TNF+Cq1组细胞吞噬微珠数量明显减少。分别与APOE3+PBS组和APOE4+PBS组比较,APOE3+IL-1α+TNF+Cq1组和APOE4+IL-1α+TNF+Cq1组细胞中Bcl-2蛋白表达水平明显降低(P<0.05或P<0.01),Caspase-3蛋白表达水平明显升高(P<0.01);与APOE3+IL-1α+TNF+Cq1组比较,APOE4+IL-1α+TNF+Cq1组细胞中Bcl-2蛋白表达水平明显降低(P<0.01),Caspase-3蛋白表达水平明显升高(P<0.05)。结论:APOE4基因型比APOE3诱导炎症因子能力更强,可诱导神经毒性反应性星形胶质细胞表型,增加神经毒性,影响星形胶质细胞凋亡,从而加剧神经元损伤。