ALKBH5 suppresses autophagic flux via N6-methyladenosine demethylation of ZKSCAN3 mRNA in acute pancreatitis

BACKGROUND Increasing evidence has demonstrated that N6-methyladenosine(m6A)RNA modification plays an essential role in a wide range of pathological conditions.Impaired autophagy is a critical hallmark of acute pancreatitis(AP).AIM To explore the role of the m6A modification of ZKSCAN3 in the regulation of autophagy in AP.METHODS The AP mouse cell model was established by cerulein-treated mouse pancreatic acinar cells(MPC-83),and the results were confirmed by the levels of amylase and inflammatory factors.Autophagy activity was evaluated by specific identification of the autophagy-related microstructure and the expression of autophagy-related genes.ZKSCAN3 and ALKBH5 were knocked down to study the function in AP.A m6A RNA binding protein immunoprecipitation assay was used to study how the m6A modification of ZKSCAN3 mRNA is regulated by ALKBH.RESULTS The increased expression of amylase and inflammatory factors in the supernatant and the accumulation of autophagic vacuoles verified that the AP mouse cell model was established.The downregulation of LAMP2 and upregulation of LC3-II/I and SQSTM1 demonstrated that autophagy was impaired in AP.The expression of ZKSCAN3 was upregulated in AP.Inhibition of ZKSCAN3 increased the expression of LAMP2 and decreased the expression of the inflammatory factors,LC3-II/I and SQSTM1.Furthermore,ALKBH5 was upregulated in AP.Knockdown of ALKBH5 downregulated ZKSCAN3 expression and restored decreased autophagic flux in AP.Notably,the bioinformatic analysis revealed 23 potential m6A modification sites on ZKSCAN3 mRNA.The m6A modification of ZKSCAN3 mRNA was significantly decreased in AP.Knockdown of ALKBH5 increased the modification of ZKSCAN3 mRNA,which confirmed that ALKBH5 upregulated ZKSCAN3 expression in a m6A-dependent manner.CONCLUSION ALKBH5 inhibits autophagic flux through m6A demethylation of ZKSCAN3 mRNA in AP,thereby aggravating the severity of the disease.

Effects of drought on non-structural carbohydrates and C,N,and P stoichiometric characteristics of Pinus yunnanensis seedlings

To study non-structural carbohydrate character-istics and nutrient utilization strategies of Pinus yunnanen-sis under continuous drought conditions,2-year-old seed-lings were planted in pots with appropriate water,light and moderate and severe drought treatments[(80±5),(65±5),(50±5),and(35±5)%of field water-holding capacity].Non-structural carbohydrates,carbon(C),nitrogen(N),and phosphorus(P)concentrations were measured in each plant component.The results show that:(1)With increasing drought,non-structural carbohydrates gradually increased in leaves,stems,and coarse roots,while gradually decreased in fine roots;(2)C concentrations of all were relatively stable under different stress levels.Phosphorous utilization of each component increased under light and moderate drought conditions,while N and P utilization efficiency of each plant component decreased under severe drought.Growth was mainly restricted by N,first decreasing and then increasing with increased drought;(3)There was a correlation between the levels of non-structural carbohydrates and C,N,and P in each component.Changes in N concentration affected the interconversion between soluble sugar and starch,which play a regulatory role in the fluctuation of the concentration of non-structural carbohydrates;and,(4)Plasticity analysis showed that P.yunnanensis seedlings responded to drought mainly by altering starch concentration,the ratio of soluble sugar to starch in leaves and stems,and further by alter-ing N and P utilization efficiencies.Overall,these results suggest that the physiological activities of all organs of P.yunnanensis seedlings are restricted under drought and that trade-offs exist between different physiological indicators and organs.Our findings are helpful in understanding non-structural carbohydrate and nutrient adaptation mechanisms under drought in P.yunnanensis seedlings.

Constructing long-cycling crystalline C_(3)N_(4)-based carbonaceous anodes for sodium-ion battery via N configuration control

Carbon nitrides with two-dimensional layered structures and high theoretical capacities are attractive as anode materials for sodium-ion batteries while their low crystallinity and insufficient structural stability strongly restrict their practical applications.Coupling carbon nitrides with conductive carbon may relieve these issues.However,little is known about the influence of nitrogen(N)configurations on the interactions between carbon and C_(3)N_(4),which is fundamentally critical for guiding the precise design of advanced C_(3)N_(4)-related electrodes.Herein,highly crystalline C_(3)N_(4)(poly(triazine imide),PTI)based all-carbon composites were developed by molten salt strategy.More importantly,the vital role of pyrrolic-N for enhancing charge transfer and boosting Na+storage of C_(3)N_(4)-based composites,which was confirmed by both theoretical and experimental evidence,was spot-highlighted for the first time.By elaborately controlling the salt composition,the composite with high pyrrolic-N and minimized graphitic-N content was obtained.Profiting from the formation of highly crystalline PTI and electrochemically favorable pyrrolic-N configurations,the composite delivered an unusual reverse growth and record-level cycling stability even after 5000 cycles along with high reversible capacity and outstanding full-cell capacity retention.This work broadens the energy storage applications of C_(3)N_(4) and provides new prospects for the design of advanced all-carbon electrodes.

A Novel Foreign Object Detection Method in Transmission Lines Based on Improved YOLOv8n

The rapid pace of urban development has resulted in the widespread presence of construction equipment andincreasingly complex conditions in transmission corridors. These conditions pose a serious threat to the safeoperation of the power grid.Machine vision technology, particularly object recognition technology, has beenwidelyemployed to identify foreign objects in transmission line images. Despite its wide application, the technique faceslimitations due to the complex environmental background and other auxiliary factors. To address these challenges,this study introduces an improved YOLOv8n. The traditional stepwise convolution and pooling layers are replacedwith a spatial-depth convolution (SPD-Conv) module, aiming to improve the algorithm’s efficacy in recognizinglow-resolution and small-size objects. The algorithm’s feature extraction network is improved by using a LargeSelective Kernel (LSK) attention mechanism, which enhances the ability to extract relevant features. Additionally,the SIoU Loss function is used instead of the Complete Intersection over Union (CIoU) Loss to facilitate fasterconvergence of the algorithm. Through experimental verification, the improved YOLOv8n model achieves adetection accuracy of 88.8% on the test set. The recognition accuracy of cranes is improved by 2.9%, which isa significant enhancement compared to the unimproved algorithm. This improvement effectively enhances theaccuracy of recognizing foreign objects on transmission lines and proves the effectiveness of the new algorithm.

Clinical outcome and prognostic factors of T4N0M0 colon cancer after R0 resection:A retrospective study

BACKGROUND Paradoxically,patients with T4N0M0(stage II,no lymph node metastasis)colon cancer have a worse prognosis than those with T2N1-2M0(stage III).However,no previous report has addressed this issue.AIM To screen prognostic risk factors for T4N0M0 colon cancer and construct a prognostic nomogram model for these patients.METHODS Two hundred patients with T4N0M0 colon cancer were treated at Tianjin Medical University General Hospital between January 2017 and December 2021,of which 112 patients were assigned to the training cohort,and the remaining 88 patients were assigned to the validation cohort.Differences between the training and validation groups were analyzed.The training cohort was subjected to multi-variate analysis to select prognostic risk factors for T4N0M0 colon cancer,followed by the construction of a nomogram model.RESULTS The 3-year overall survival(OS)rates were 86.2%and 74.4%for the training and validation cohorts,respectively.Enterostomy(P=0.000),T stage(P=0.001),right hemicolon(P=0.025),irregular review(P=0.040),and carbohydrate antigen 199(CA199)(P=0.011)were independent risk factors of OS in patients with T4N0M0 colon cancer.A nomogram model with good concordance and accuracy was constructed.CONCLUSION Enterostomy,T stage,right hemicolon,irregular review,and CA199 were independent risk factors for OS in patients with T4N0M0 colon cancer.The nomogram model exhibited good agreement and accuracy.

Advances of N6-methyladenosine modification on circular RNA in hepatocellular carcinoma

N6-methyladenosine(m6A)is a reversible epigenetic modification, which is one of the most abundant modifiers in eukaryotic cells and has been commonly reported in messenger RNAs and non-coding RNAs. The processing modification of m6A regulates RNA transcription, processing, splicing, degradation, and translation, and plays an important role in the biological process of tumors. Circular RNA, which lacks the 5' cap structure, has been mistakenly regarded as a "junk sequence" generated by accidental shearing during the transcription process. However, it has been found that circRNAs can be involved in tumor invasion and metastasis through microRNAs, binding proteins, translated peptides, and m6A modifications. In this paper, we reviewed the role of m6A modifications in circRNA regulation and their functions in hepatocellular carcinoma and discussed their potential clinical applications and future development in this field.

N6-methyladenosine methylation regulates the tumor microenvironment of Epstein-Barr virus-associated gastric cancer

BACKGROUND N6-methyladenosine(m6A)methylation modification exists in Epstein-Barr virus(EBV)primary infection,latency,and lytic reactivation.It also modifies EBV latent genes and lytic genes.EBV-associated gastric cancer(EBVaGC)is a distinctive molecular subtype of GC.We hypothesized EBV and m6A methylation regulators interact with each other in EBVaGC to differentiate it from other types of GC.AIM To investigate the mechanisms of m6A methylation regulators in EBVaGC to determine the differentiating factors from other types of GC.METHODS First,The Cancer Gene Atlas and Gene Expression Omnibus databases were used to analyze the expression pattern of m6A methylation regulators between EBVaGC and EBV-negative GC(EBVnGC).Second,we identified Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment of m6A-related differentially expressed genes.We quantified the relative abundance of immune cells and inflammatory factors in the tumor microenvironment(TME).Finally,cell counting kit-8 cell proliferation test,transwell test,and flow cytometry were used to verify the effect of insulin-like growth factor binding protein 1(IGFBP1)in EBVaGC cell lines.RESULTS m6A methylation regulators were involved in the occurrence and development of EBVaGC.Compared with EBVnGC,the expression levels of m6A methylation regulators Wilms tumor 1-associated protein,RNA binding motif protein 15B,CBL proto-oncogene like 1,leucine rich pentatricopeptide repeat containing,heterogeneous nuclear ribonucleoprotein A2B1,IGFBP1,and insulin-like growth factor 2 binding protein 1 were significantly downregulated in EBVaGC(P<0.05).The overall survival rate of EBVaGC patients with a lower expression level of IGFBP1 was significantly higher(P=0.046).GO and KEGG functional enrichment analyses showed that the immunity pathways were significantly activated and rich in immune cell infiltration in EBVaGC.Compared with EBVnGC,the infiltration of activated CD4+T cells,activated CD8+T cells,monocytes,activated dendritic cells,and plasmacytoid dendritic cells were significantly upregulated in EBVaGC(P<0.001).In EBVaGC,the expression level of proinflammatory factors interleukin(IL)-17,IL-21,and interferon-γ and immunosuppressive factor IL-10 were significantly increased(P<0.05).In vitro experiments demonstrated that the expression level of IGFBP1 was significantly lower in an EBVaGC cell line(SNU719)than in an EBVnGC cell line(AGS)(P<0.05).IGFBP1 overexpression significantly attenuated proliferation and migration and promoted the apoptosis levels in SNU719.Interfering IGFBP1 significantly promoted proliferation and migration and attenuated the apoptosis levels in AGS.CONCLUSION m6A regulators could remodel the TME of EBVaGC,which is classified as an immune-inflamed phenotype and referred to as a“hot”tumor.Among these regulators,we demonstrated that IGFBP1 affected proliferation,migration,and apoptosis.

Toxoplasmic Chorioretinitis: About a Case in an Immunocompetent Adult at the Renaissance University Hospital Centre (UHC) in N’Djamena (Chad) and Review of the Literature

Toxoplasmic chorioretinitis also known as ocular toxoplasmosis is a parasitic infection caused by the obligately intracellular multiplying protozoan called Toxoplasma gondii. Active toxoplasmic chorioretinitis is a rare condition in immunocompetent people. We report a case of a 47-year-old patient who received reduced right visual acuity, left hemiparesis and eye pain. Examination of the right anterior segment of the eye is unremarkable. In the fundus of the right eye, there was a focus of active toxoplasmic chorioretinitis located in the macula at 1.5 mm papillary diameter next to old scar lesions. The ophthalmological examination of the left eye was unremarkable. The paraclinical assessment carried out on the patient shows a positive Remington test on two occasions three weeks apart. The clinical outcome after initiation of treatment was favorable with a healing of the active site within four weeks.

Long non-coding RNA GATA6-AS1 is mediated by N6-methyladenosine methylation and inhibits the proliferation and metastasis of gastric cancer

BACKGROUND Through experimental research on the biological function of GATA6-AS1,it was confirmed that GATA6-AS1 can inhibit the proliferation,invasion,and migration of gastric cancer cells,suggesting that GATA6-AS1 plays a role as an anti-oncogene in the occurrence and development of gastric cancer.Further experi-ments confirmed that the overexpression of fat mass and obesity-associated protein(FTO)inhibited the expression of GATA6-AS1,thereby promoting the occurrence and development of gastric cancer.AIM To investigate the effects of GATA6-AS1 on the proliferation,invasion and migration of gastric cancer cells and its mechanism of action.METHODS We used bioinformatics methods to analyze the Cancer Genome Atlas(https://portal.gdc.cancer.gov/.The Cancer Genome Atlas)and download expression data for GATA6-AS1 in gastric cancer tissue and normal tissue.We also constructed a GATA6-AS1 lentivirus overexpression vector which was transfected into gastric cancer cells to investigate its effects on proliferation,migration and invasion,and thereby clarify the expression of GATA6-AS1 in gastric cancer and its biological role in the genesis and development of gastric cancer.Next,we used a database(http://starbase.sysu.edu.cn/starbase2/)to analysis GATA6-AS1 whether by m6A methylation modify regulation and predict the methyltransferases that may methylate GATA6-AS1.Furthermore,RNA immunoprecipitation experiments confirmed that GATA6-AS1 was able to bind to the m6A methylation modification enzyme.These data allowed us to clarify the ability of m6A methylase to influence the action of GATA6-AS1 and its role in the occurrence and development of gastric cancer.RESULTS Low expression levels of GATA6-AS1 were detected in gastric cancer.We also determined the effects of GATA6-AS1 overexpression on the biological function of gastric cancer cells.GATA6-AS1 had strong binding ability with the m6A demethylase FTO,which was expressed at high levels in gastric cancer and negatively correlated with the expression of GATA6-AS1.Following transfection with siRNA to knock down the expression of FTO,the expression levels of GATA6-AS1 were up-regulated.Finally,the proliferation,migration and invasion of gastric cancer cells were all inhibited following the knockdown of FTO expression.CONCLUSION During the occurrence and development of gastric cancer,the overexpression of FTO may inhibit the expression of GATA6-AS1,thus promoting the proliferation and metastasis of gastric cancer.

Lateral root elongation in maize is related to auxin synthesis and transportation mediated by N metabolism under a mixed NO_(3)^(–) and NH_(4)^(+) supply

A mixed nitrate (NO_(3)^(–)) and ammonium (NH_(4)^(+)) supply can promote root growth in maize (Zea mays),however,the changes in root morphology and the related physiological mechanism under different N forms are still unclear.Here,maize seedlings were grown hydroponically with three N supplied in three different forms (NO_(3)^(–)only,75/25 NO_(3)^(–)/NH_(4)^(+)and NH_(4)^(+)only).Compared with sole NO_(3)^(–)or NH_(4)^(+),the mixed N supply increased the total root length of maize but did not affect the number of axial roots.The main reason was the increased total lateral root length,while the average lateral root (LR) length in each axle was only slightly increased.In addition,the average LR density of 2nd whorl crown root under mixed N was also increased.Compared with sole nitrate,mixed N could improve the N metabolism of roots (such as the N influx rate,nitrate reductase (NR) and glutamine synthase (GS)enzyme activities and total amino content of the roots).Experiments with exogenously added NR and GS inhibitors suggested that the increase in the average LR length under mixed N was related to the process of N assimilation,and whether the NR mediated NO synthesis participates in this process needs further exploration.Meanwhile,an investigation of the changes in root-shoot ratio and carbon (C) concentration showed that C transportation from shoots to roots may not be the key factor in mediating lateral root elongation,and the changes in the sugar concentration in roots further proved this conclusion.Furthermore,the synthesis and transportation of auxin in axial roots may play a key role in lateral root elongation,in which the expression of ZmPIN1B and ZmPIN9 may be involved in this pathway.This study preliminarily clarified the changes in root morphology and explored the possible physiological mechanism under a mixed N supply in maize,which may provide some theoretical basis for the cultivation of crop varieties with high N efficiency.

S/N对硫化物型自养反硝化性能及NO-2-N积累的影响

以Na 2S为硫源,采用批次实验探究不同初始S/N(摩尔比)对硫化物自养反硝化性能及NO_(2)^(-)-N积累的影响。结果表明,当S/N≤2.80时,NO_(3)^(-)-N及NO-X-N(NO_(2)^(-)-N+NO_(3)^(-)-N)去除效果随S/N的升高而显著提升,S/N由0.35升高到2.80时,前6 h内NO_(3)^(-)-N及NO-X-N去除率分别提高了3.7倍及3.6倍,当S/N继续增至4.20时,NO_(3)^(-)-N及NO-X-N去除效果较S/N为2.80时变化不大;当S/N≤0.70时系统内可以获得一定的NO_(2)^(-)-N积累,反应结束时NO_(2)^(-)-N积累率(0.35,0.7)分别可达47.9%及24.5%,当S/N≥1.40时难以稳定积累NO_(2)^(-)-N,NO_(2)^(-)-N最高积累率(1.40,2.80,4.20)分别为55.0%,33.3%,39.7%。S/N≤0.70时可获得一定的NO_(2)^(-)-N积累,而S/N在1.40~2.80范围内时,升高S/N可以提供更多电子用于反硝化,缓解各种反硝化酶对于电子的竞争,降低中间产物积累,提升脱氮效果。

Nanopore测序揭示m6A修饰铁死亡相关基因在中晚期头颈部鳞状细胞癌中的表达及临床意义

目的研究N6-甲基腺苷(m6A)修饰在中晚期头颈部鳞状细胞癌(HNSCC)的作用和基因相关通路,为头颈部HNSCC患者的个体化精准治疗提出新的观点及研究参考。方法选取3对临床中晚期HNSCC患者的癌及其癌旁组织(6个样本)进行测序,从癌和癌旁组织的转录组、m6A相关差异基因中筛选出两者共同的差异基因,通过基因本体(GO)分析、pathway分析及蛋白相互作用(PPI)分析,找出其中的枢纽(hub)基因,随后对网络中的基因进行基因集变异分析(GSVA),从而得到m6A修饰影响的基因及通路。结果从癌与癌旁组织的差异基因中筛选出96个转录组m6A相关的上调基因、159个m6A相关下调基因;信号通路分析发现PPAR通路、免疫相关通路下调,代谢通路上调;构建PPI后发现,CDK1呈现上调趋势,而DCN及ARG2均呈现下调趋势;GSVA分析发现PI3K/AKT信号通路在癌组织中上调;PPI中的hub基因共表达分析发现AGR2和CDK1在m6A调控下,与铁死亡相关基因联系很密切。结论m6A修饰与FOXA1、RPL8、DNC、CDK1、ApoE、POSTN、YWHAZ、MMP13等铁死亡相关基因紧密相关,m6A修饰可能通过铁死亡相关基因影响免疫、代谢相关通路,从而在中晚期HNSCC中起重要作用。

射干止咳胶囊体内外抗甲型H1N1流感病毒作用初探

目的初步探讨射干止咳胶囊对甲型H1N1流感病毒的体内外抗病毒作用。方法通过细胞病变效应(cytopathic effect,CPE)结合MTT法考察射干止咳胶囊体外抑制甲型H1N1流感病毒活性的作用;采用小鼠滴鼻法感染甲型H1N1流感病毒为模型,观察射干止咳胶囊对甲型H1N1流感病毒的体内抗病毒作用。结果体外抗病毒试验结果显示,射干止咳胶囊对甲型H1N1流感病毒的最高抑制率为11.86%,提示在攻毒剂量为100TCID_(50)的条件下,射干止咳胶囊在体外对甲型H1N1流感病毒的抑制作用不明显。小鼠滴鼻感染5LD_(50)的甲型H1N1流感病毒,射干止咳胶囊低、中、高剂量组小鼠的死亡率分别为30%、10%、30%,死亡保护率达50%、70%、50%,平均生存天数分别为13 d、14 d、13 d,说明各给药组对攻毒小鼠具有较好的死亡保护作用。结论射干止咳胶囊体外抗甲型H1N1流感病毒的作用不显著,但射干止咳胶囊各剂量组对甲型H1N1流感病毒攻毒致小鼠死亡却具有明显的保护作用。

两种咔唑基-吡啶-N-氧化物内盐荧光极性探针研究

合成了4-(9H-咔唑-9-基)吡啶1-氧化物(CPNO)和4-(4-(9H-咔唑-9-基)苯基)吡啶1-氧化物(CPPNO)两种咔唑基-吡啶-N-氧化物内盐,测定了它们在不同溶剂中的紫外-可见吸收和荧光光谱,均表现出对溶剂极性较好的敏感性。计算表明,两个化合物都具有较大的激发态偶极矩,是化合物溶剂极性敏感性的原因。研究为开发新型的荧光极性探针提供了一种新思路。

甲型流感病毒H1N1型灭活病毒标准物质研究

以人工培养真实病毒为原料,研制了一种甲型流感病毒H1N1型灭活病毒标准物质。该标准物质经化学灭活及灭活效果验证后,联合多家实验室采用数字PCR定量检测方法对拷贝数浓度进行定值,对标准物质的均匀性、稳定性及不确定度进行了评估。结果显示该标准物质均匀性良好,在-80~-70℃保存条件下稳定保存5个月,标准物质最终定值结果为(1384.5±249.3)copies/μL(k=2)。该标准物质可为H1N1型甲型流感病毒检测的全过程提供计量溯源和质量控制支撑,有助于提升相关核酸检测结果的准确性和可靠性。

广西H9N2亚型禽流感病毒疫苗候选株的筛选

【目的】筛选出免疫原性更优的H9N2亚型禽流感病毒(AIV)疫苗候选株用于新型疫苗研发,为加强对H9N2亚型AIV的防控提供技术支持。【方法】以2000—2020年广西地区分离获得的36株H9N2亚型AIV分离株为试验材料,依据HA基因遗传进化分析选择12株不同年份、不同地域及不同宿主来源的H9N2亚型AIV代表株,与目前使用的商品化H9亚型AIV疫苗株(SS株)进行交叉血凝抑制试验(HI)及抗原性分析,根据抗原分析结果选择其中具有不同抗原差异的代表株制备油乳剂灭活疫苗,并分别与商品化H9亚型AIV疫苗(SS株)进行交叉免疫保护试验。【结果】在36株H9N2亚型AIV广西分离株中有31株属于Y280-Like(9.4.1),其中26株属于分支I、5株属于分支II,另外5株属于G1-Like(h9.4.1),与我国常用疫苗株分属于不同进化分支。12株H9N2亚型AIV广西代表性分离株灭活后的HA效价≥7 log2,病毒灭活效果良好,可用于制备油乳剂灭活疫苗,且制备的油乳剂灭活疫苗稳定性和安全性良好。根据交互HI抗体滴度计算出各毒株间的抗原相关值(R),结果发现A/chicken/Guangxi/C227/2015(H9N2)株(简称C227株)与其他毒株的抗原性无明显差异,对应的R在0.72~0.93间波动;疫苗交叉保护试验结果也表明,C227株制备的油乳剂灭活疫苗对不同毒株的免疫保护率均高于80%,且免疫保护效果优于A/chicken/Guangxi/CX/2013(H9N2)株(简称CX13株),说明C227株更适合作为H9N2亚型AIV灭活疫苗的候选株。【结论】基于抗原性分析和免疫原性测定筛选出的A/chicken/Guangxi/C227/2015(H9N2)株对广西地区绝大部分H9N2亚型AIV流行株的免疫保护效果良好,可作为疫苗候选株用于研制新型H9N2亚型AIV疫苗。

减氮对华北地区麦玉轮作农田土壤N2O排放调控机理

【目的】探究减氮对华北地区麦玉轮作农田土壤N2O排放调控机制。【方法】冬小麦季和夏玉米季均以不施氮为对照(CK),设置2个施氮量,分别为常规施氮量(纯氮300 kg/hm2,N2)、减氮20%(纯氮240 kg/hm2,N1),研究施氮量对麦玉轮作农田土壤性质及N2O排放通量影响,基于逐步回归分析,研究减氮对麦玉轮作农田土壤N2O排放调控机制。【结果】(1)减氮有效降低了N2O排放通量,且夏玉米季N2O排放通量远高于冬小麦季,夏玉米季由施肥引起的N2O排放量较高。(2)冬小麦季N2O排放通量与环境因子逐步回归公式为:N2O排放通量=181.952+1.450×硝态氮+8.401×铵态氮-0.514×电导率;硝态氮、铵态氮会对冬小麦季N2O排放通量产生显著的正向影响,而电导率会对冬小麦季N2O排放通量产生显著的负向影响。(3)夏玉米季N2O排放通量与环境因子逐步回归公式为:N2O排放通量=-354.606+4.592×硝态氮+157.848×铵态氮;硝态氮、铵态氮会对夏玉米季N2O排放通量产生显著的正向影响。【结论】综上可知,适量减氮显著降低夏玉米季N2O累积排放量和增温潜势,应加强夏玉米季农田水肥管理。

H9N2 AIV灭活疫苗免疫SPF鸡攻毒后肺组织免疫相关基因表达变化研究

[目的]本试验旨在探究H9N2亚型禽流感病毒攻击对H9灭活疫苗免疫SPF鸡呼吸系统的影响。[方法]选用36只3周龄SPF鸡,随机分为对照组(Con)、攻毒组(Flu)、免疫后攻毒组(Vac+Flu)。以每只0.4 mL剂量免疫接种H9灭活疫苗,3周后以10~6 EID_(50)的病毒量进行攻毒,采集拭子、血清、气管、肺组织等样品,通过血凝抑制(HI)试验、RT-PCR、荧光定量PCR(qPCR)等方法检测血清中HI抗体滴度、肺脏流感病毒M基因拷贝数以及免疫相关基因CD4、CD8、GATA3、T-bet、IL-4、IL-13、TNF-α、IFN-γ、Perforin、Granzyme、FasL、Fas等的表达量;并利用HE染色、免疫组织化学的方法观察气管、肺脏的病理变化及病毒特异性抗原NP蛋白的分布特征。[结果]HI试验结果显示,SPF鸡疫苗免疫后可产生较高的H9N2 AIV抗体效价。免疫保护试验和RT-PCR结果显示,SPF鸡疫苗免疫后攻毒仍能检测到机体排毒和流感病毒M基因在肺脏组织中的表达。HE染色和免疫组化结果显示,接种H9N2 AIV灭活疫苗后能够明显减轻SPF鸡气管和肺脏的病理损伤,降低气管、肺脏中的病毒载量。荧光定量PCR结果显示,接种H9N2 AIV灭活疫苗后能够提高SPF鸡肺脏中Th1、Th2细胞因子分泌水平,促进穿孔素/颗粒酶途径相关基因表达进而增强肺脏中细胞毒性反应。[结论]H9N2疫苗免疫鸡感染H9病毒后虽然仍存在排毒现象,但其抗病毒免疫系统活跃、其主要器官病毒载量降低,建议按照流感疫苗免疫程序进行免疫接种,提高对流感的防控水平。

蛋白酶N1对小鼠心肌细胞增殖的作用及机制

目的探讨蛋白酶N1(PKN1)对小鼠心肌细胞增殖的作用及机制。方法使用异氟烷麻醉1日龄小鼠2只,分离小鼠心肌细胞,并分为对照组和干扰组,干扰组心肌细胞转染PKN1干扰片段,对照组细胞转染对照片段。按照随机数字表法将10只小鼠分为正常组和观察组,每组5只。观察组小鼠于心脏原位注射PKN1干扰腺病毒,正常组小鼠于心脏原位注射空载腺病毒。采用免疫荧光法检测4组小鼠心肌细胞或心肌组织中Ki-67阳性表达率,以Ki-67阳性表达率表示心肌细胞增殖能力;采用实时荧光定量聚合酶链式反应法检测对照组和干扰组小鼠心肌细胞中PKN1 mRNA表达;采用Western blot法检测对照组和干扰组小鼠心肌细胞中PKN1和cyclin D1蛋白表达。结果干扰组心肌细胞中Ki-67阳性表达率均显著低于对照组(t=11.201,P<0.01),观察组小鼠心肌组织中Ki-67阳性表达率显著低于正常(t=11.851,P<0.01)。干扰组心肌细胞中PKN1 mRNA相对表达量显著低于对照组(t=7.022,P<0.01)。干扰组心肌细胞中PKN1和cyclin D1蛋白相对表达量均显著低于对照组(t=5.762、6.884,P<0.01)。结论小鼠心肌细胞中PKN1表达降低可抑制cyclin D1蛋白表达,从而抑制心肌细胞增殖。

N6-甲基腺苷相关调节因子与骨关节炎:生物信息学和实验验证分析

背景:越来越多证据表明N6-甲基腺苷(N6-methyladenosine,m6A)调节因子与骨关节炎密切相关,被认为是防治骨关节炎新方向,但具体作用机制不明。目的:通过对骨关节炎基因芯片数据集进行生物信息学分析,探讨m6A对骨关节炎的作用,解析骨关节炎发病机制。方法:首先利用R软件提取GEO数据库中GSE1919数据集中骨关节炎相关m6A调节因子及其表达量,进而对提取结果行基因差异分析及GO、KEGG富集分析;接着对PPI网络拓扑学分析结果和机器学习结果取交集得到m6A关键调节因子,并通过体外细胞实验验证。结果与结论:①提取得到16个骨关节炎相关m6A调节因子表达量,通过差异分析获得ZC3H13、YTHDC1、YTHDF3、HNRNPC等11个m6A差异调节因子;②GO富集分析显示,骨关节炎相关m6A差异调节因子在生物过程中主要于mRNA转运、RNA分解代谢、胰岛素样生长因子受体信号通路调控等发挥作用;③KEGG富集分析显示,差异调节因子主要参与p53、白细胞介素17和AMPK信号通路;④综合PPI网络拓扑学分析和机器学习结果获得m6A关键调节因子——YTHDC1;⑤体外细胞实验结果表明,m6A关键调节因子——YTHDC1在对照组与骨关节炎组中表达存在显著差异(P<0.05);⑥结果显示,YTHDC1与骨关节炎发生发展密切相关,有望成为m6A治疗骨关节炎的分子靶点。