The roles of macrophage migration inhibitory factor in retinal diseases

Macrophage migration inhibitory factor(MIF),a multifunctional cytokine,is secreted by various cells and participates in inflammatory reactions,including innate and adaptive immunity.There are some evidences that MIF is involved in many vitreoretinal diseases.For example,MIF can exacerbate many types of uveitis;measurements of MIF levels can be used to monitor the effectiveness of uveitis treatment.MIF also alleviates trauma-induced and glaucoma-induced optic nerve damage.Furthermore,MIF is critical for retinal/choroidal neovascularization,especially complex neovascularization.MIF exacerbates retinal degeneration;thus,anti-MIF therapy may help to mitigate retinal degeneration.MIF protects uveal melanoma from attacks by natural killer cells.The mechanism underlying the effects of MIF in these diseases has been demonstrated:it binds to cluster of differentiation 74,inhibits the c-Jun N-terminal kinase pathway,and triggers mitogen-activated protein kinases,extracellular signal-regulated kinase-1/2,and the phosphoinositide-3-kinase/Akt pathway.MIF also upregulates Toll-like receptor 4 and activates the nuclear factor kappa-B signaling pathway.This review focuses on the structure and function of MIF and its receptors,including the effects of MIF on uveal inflammation,retinal degeneration,optic neuropathy,retinal/choroidal neovascularization,and uveal melanoma.

Exploration of cyclooxygenase-2 inhibitory peptides from walnut dreg proteins based on in silico and in vitro analysis

Walnut dreg protein hydrolysates(WDPHs)exhibit a variety of biological activities,however,the cyclooxygenase-2(COX-2)inhibitory peptide of WDPHs remain unclear.The aim of this study was to rapidly screen for such peptides in WDPHs through a combination of in silico and in vitro analysis.In total,1262 peptide sequences were observed by nano liquid chromatography/tandem mass spectrometry(nano LC-MS/MS)and 4 novel COX-2 inhibitory peptides(AGFP,FPGA,LFPD,and VGFP)were identified.Enzyme kinetic data indicated that AGFP,FPGA,and LFPD displayed mixed-type COX-2 inhibition,whereas VGFP was a non-competitive inhibitor.This is mainly because the peptides form hydrogen bonds and hydrophobic interactions with residues in the COX-2 active site.These results demonstrate that computer analysis combined with in vitro evaluation allows for rapid screening of COX-2 inhibitory peptides in walnut protein dregs.

Application of Structured Health Education and Combined 5A Nursing Model in Self-control of Vulnerable Chronic Diseases: Take Coronary Heart Disease as an Example

Objective:To explore the intervention effect of the Structured Health Education course and 5A nursing model for self-control of elderly patients with coronary heart disease.Methods:Using the random sampling method,124 elderly CAD patients admitted to the First Affiliated Hospital of Bengbu Medical University were randomly divided into an experimental group and a control group.The control group line routine health education,experimental group take structured health education combined with 5A nursing before and after the intervention using a coronary heart disease assessment questionnaire,coronary heart disease self-control scale evaluation of two groups of intervention,compare two groups before and after intervention blood pressure,blood sugar,body mass index,lipid index level and complications within 8 months after discharge.Results:After the course intervention,the disease cognition and self-behavior of the experimental group were higher than that of the control group,and the differences were statistically significant(all P<0.1).Conclusion:This course is suitable for elderly patients with coronary heart disease.The 5A model improves the cognitive and management ability of elderly patients to a certain extent,which is worthy of clinical application.

A Pair of New Spirocyclic Alkaloid Enantiomers with TrxR Inhibitory Activities Were Isolated from Marine-Derived Aspergillus ruber TX-M4-1

One new spirocyclic alkaloid,5-isopentenyl-cryptoechinuline D(1),along with 11 known compounds(2–12),were iso-lated from a marine fungus Aspergillus ruber TX-M4-1.The structures of compounds 1–12 were elucidated by spectroscopic evi-dences.Compound 1 was initially isolated as an enantiomer,and further separation of 1 by chiral HPLC afforded a pair of enantio-mers,including(-)-5-isopentenyl-cryptoechinuline D(1a)and(+)-5-isopentenyl-cryptoechinuline D(1b).Their absolute configura-tions were elucidated by ECD spectroscopic data.Compounds 1a,5 and 10 could inhibit thioredoxin reductase(TrxR)activity with IC50 values of 6.2,36.3 and 18.6μmol L^(-1),respectively.Surface plasmon resonance(SPR)study also demonsrated the interactions between compounds 6,8 and Niemann-Pick C1 Like 1(NPC1L1)respectively,which indicate that compounds 6 and 8 are potential NPC1L1 inhibitors.

Macrophage migration inhibitory factor facilitates astrocytic production of the CCL2 chemokine following spinal cord injury

Spinal cord injury causes accumulation of a large number of leukocytes at the lesion site where they contribute to excessive inflammation.Overproduced chemokines are responsible for the migratory process of the leukocytes,but the regulatory mechanism underlying the production of chemokines from resident cells of the spinal cord has not been fully elucidated.We examined the protein levels of macrophage migration inhibitory factor and chemokine C-C motif chemokine ligand 2 in a spinal cord contusion model at different time points following spinal cord injury.The elevation of macrophage migration inhibitory factor at the lesion site coincided with the increase of chemokine C-C motif chemokine ligand 2 abundance in astrocytes.Stimulation of primary cultured astrocytes with different concentrations of macrophage migration inhibitory factor recombinant protein induced chemokine C-C motif chemokine ligand 2 production from the cells,and the macrophage migration inhibitory factor inhibitor 4-iodo-6-phenylpyrimidine attenuated the stimulatory effect.Further investigation into the underlying mechanism on macrophage migration inhibitory factor-mediated astrocytic production of chemokine C-C motif chemokine ligand 2 revealed that macrophage migration inhibitory factor activated intracellular JNK signaling through binding with CD74 receptor.Administration of the macrophage migration inhibitory factor inhibitor 4-iodo-6-phenylpyrimidine following spinal cord injury resulted in the reduction of chemokine C-C motif chemokine ligand 2-recruited microglia/macrophages at the lesion site and remarkably improved the hindlimb locomotor function of rats.Our results have provided insights into the functions of astrocyte-activated chemokines in the recruitment of leukocytes and may be beneficial to develop interventions targeting chemokine C-C motif chemokine ligand 2 for neuroinflammation after spinal cord injury.

Self-Control Training Decreased Intensity of Penalty Toward Previous Offender

Previous studies have found that self-control training was effective in improving an individual’s self-control,which plays an important role in inhibiting negative emotions.However,it is unclear whether self-control training can facilitate refraining from retaliation.This study randomly assigned participants(N=55)to a training condi-tion(building self-control by avoiding sweets)or a control condition.Before and after training,participants com-pleted the Transgression-Related Interpersonal Motivations Inventory-18(TRIM-18)and a modified Taylor aggression task once each.Participants in the training condition inflicted more low-intensity penalties on the pre-vious offender compared to control participants.Participants in the training condition reported lower revenge scores after training than before training.These results provide preliminary support that providing people who wish to forgive self-control training might promote forgiveness.

Paradoxical roles of inhibitory autapse and excitatory synapse in formation of counterintuitive anticipated synchronization

Different from the common delayed synchronization(DS)in which response appears after stimulation,anticipated synchronization(AS)in unidirectionally coupled neurons denotes a counterintuitive phenomenon in which response of the receiver neuron appears before stimulation of the sender neuron,showing an interesting function of brain to anticipate the future.The dynamical mechanism for the AS remains unclear due to complex dynamics of inhibitory and excitatory modulations.In this article,the paradoxical roles of excitatory synapse and inhibitory autapse in the formation of AS are acquired.Firstly,in addition to the common roles such that inhibitory modulation delays and excitatory modulation advances spike,paradoxical roles of excitatory stimulation to delay spike via type-II phase response and of inhibitory autapse to advance spike are obtained in suitable parameter regions,extending the dynamics and functions of the excitatory and inhibitory modulations.Secondly,AS is related to the paradoxical roles of the excitatory and inhibitory modulations,presenting deep understandings to the AS.Inhibitory autapse induces spike of the receiver neuron advanced to appear before that of the sender neuron at first,and then excitatory synapse plays a delay role to prevent the spike further advanced,resulting in the AS as the advance and delay effects realize a dynamic balance.Lastly,inhibitory autapse with strong advance,middle advance,and weak advance and delay effects induce phase drift(spike of the receiver neuron advances continuously),AS,and DS,respectively,presenting comprehensive relationships between AS and other behaviors.The results present potential measures to modulate AS related to brain function.

Novel insight into the formation and inhibition mechanism of dipeptidyl peptidase-Ⅳ inhibitory peptides from fermented mandarin fish(Chouguiyu)

Fermented foods are a potential source to produce novel dipeptidyl peptidase-IV inhibitory peptides(D4IPs).In this study,the fermented mandarin fish(Chouguiyu)was used to screen D4IPs and their formation mechanism was studied by metagenomics and peptidomics.A total of 400 D4IPs with DPP-IV inhibition structure and high hydrophobicity were identified.The correlation network map showed that Lactococcus,Bacillus,Lysobacter,Pelagivirga,Kocuria,Escherichia,Streptococcus,and Peptostreptococcus were significantly correlated with the most D4IPs.Four stable D4IPs,including KAGARALTDAETAT,GEKVDFDDIQK,VVDADEMYLKGK,and GQKDSYVGDEAQ were respectively from the precursor proteins parvalbumin,troponin,myosin,and actin,and were mainly formed by the hydrolysis of subtilisin(EC 3.4.21.62),aspartic proteinase(EC 3.4.23.1),thermolysin(EC 3.4.24.27),oligopeptidase B(EC 3.4.21.83),and proteinase P1(EC 3.4.21.96)from Bacillus,Kocuria,Lysobacter,Lactococcus,and Peptostreptococcus.The inhibition mainly resulted from the hydrogen bond and salt bridge between D4IPs and DPP-IV enzyme.This study provides important information on the proteases and related microbial strains to directionally prepare D4IPs in Chouguiyu.

Inhibitory effect induced by fractional Gaussian noise in neuronal system

We discover a phenomenon of inhibition effect induced by fractional Gaussian noise in a neuronal system. Firstly,essential properties of fractional Brownian motion(fBm) and generation of fractional Gaussian noise(fGn) are presented,and representative sample paths of fBm and corresponding spectral density of fGn are discussed at different Hurst indexes.Next, we consider the effect of fGn on neuronal firing, and observe that neuronal firing decreases first and then increases with increasing noise intensity and Hurst index of fGn by studying the time series evolution. To further quantify the inhibitory effect of fGn, by introducing the average discharge rate, we investigate the effects of noise and external current on neuronal firing, and find the occurrence of inhibitory effect about noise intensity and Hurst index of f Gn at a certain level of current. Moreover, the inhibition effect is not easy to occur when the noise intensity and Hurst index are too large or too small. In view of opposite action mechanism compared with stochastic resonance, this suppression phenomenon is called inverse stochastic resonance(ISR). Finally, the inhibitory effect induced by fGn is further verified based on the inter-spike intervals(ISIs) in the neuronal system. Our work lays a solid foundation for future study of non-Gaussian-type noise on neuronal systems.

Characterization of SARS-COV-2 main protease inhibitory peptides from Ulva prolifera proteins

The main protease(M^(pro))is essential for the replication of SARS-COV-2 and therefore represents a promising anti-viral target.In this study,we screened M^(pro)inhibitory peptides from Ulva prolifera protein on in-silico proteolysis.Cytotoxicity analysis using the online toxic prediction tool ToxinPred revealed that all the peptides were non-cytotoxic.The hexapeptide(SSGFID)exhibited high M^(pro)inhibitory activity in molecular docking and its IC_(50)value was 139.40±0.82μmol/L in vitro according to fluorescence resonance energy transfer assay(FRET).Quantitative real-time(qRT-)PCR results show that SSGFID could stimulate the expression of mitosis-related factors,including nuclear factor-κB,cyclin D1,and cyclin-dependent kinase 4,to promote the proliferation of mice splenocytes.Stability study revealed that SSGFID showed resistance against pepsin and trypsin but lost D(Asp)after pretreatment at121℃ for 15 min.Besides,SSGFID was mainly transported through the Caco-2 cell monolayer by the peptide transporter PepT1 and passive-mediated transport during the transport study.Unfortunately,the peptide was also degraded by Caco-2 intracellular enzymes,and the transfer rate of intact peptide was4.2%.Furthermore,Lineweaver–Burk plots demonstrated that SSGFID possessed a mixed inhibitory characteristic with M^(pro).Our study indicated the potential of Ulva prolifera as antiviral and immuneenhancing functional food ingredients and nutraceuticals.

Effects of Continuous Non-Invasive Blood Pressure Monitoring on Intraoperative Hemodynamics and Postoperative Myocardial Injury in Craniotomy:Comparison Between Groups Based on Self-Control and Propensity Score Matching

Objective:To explore the effect of continuous non-invasive blood pressure monitoring on intraoperative hemodynamics and postoperative myocardial injury in craniotomy.Methods:120 cases of elective craniotomy were divided into the self-control group(continuous non-invasive blood pressure monitoring and intermittent cuff non-invasive blood pressure monitoring,CNAP group)and propensity score matching group(only intermittent cuff non-invasive blood pressure measurement in previous craniotomy,PSM group);Goal-directed hemodynamic management in CNAP group included heart rate(HR),blood pressure(BP),stroke volume(SV),stroke variability(SVV),and systemic vascular resistance index(SVRI).The main index is to compare the troponin level within 72 hours after operation between the CNAP group and the PSM group;The secondary indicators are the comparison of the hemodynamic conditions between the CNAP group and the PSM at 10 specific time points.Results:The incidence of postoperative myocardial injury in the CNAP group was significantly lower than that in the PSM group(12%vs.30%,P=0.01);in the CNAP group hypotensive episodes(6 vs.3,P=0.01),positive balance of fluid therapy(700 vs.500 mL,P<0.001),more use of vasoactive drugs(29 vs.18,P=0.04),more stable hemodynamics medical status(P=0.03)were recorded.Conclusion:The hemodynamic management strategy based on continuous non-invasive blood pressure monitoring can reduce the incidence of myocardial injury after elective craniotomy and maintain a more stable hemodynamic state.

基于转录组学分析揭示蜡样芽胞杆菌(Bacillus cereus)AR1002阻控黄曲霉生长的生理机制研究

黄曲霉(Aspergillus flavus)极易侵染油料等农产品,其次级代谢产物黄曲霉毒素严重危害人体健康,阻控黄曲霉污染已成为亟待解决的国际难题。为探明蜡样芽胞杆菌(Bacillus cereus)AR1002抑制黄曲霉生长、阻控黄曲霉污染的生理及分子机制。该研究采用AR1002代谢产物对黄曲霉进行处理,并对黄曲霉生长及产孢表型进行鉴定,通过显微结构及转录组学分析,对AR1002的抑菌机制进行了初探。蜡样芽胞杆菌AR1002代谢物可抑制黄曲霉菌丝生长、干物质积累分别达24.73%,65.00%,减少孢子数量达98.80%;AR1002通过抑制glpA和AYR1等关键基因表达调控黄曲霉甘油磷酸代谢,通过抑制SEC61A、RAD23、ATP13A1、UBE2G2等关键基因的表达调控内质网中蛋白质加工;此外,AR1002代谢产物通过下调RAD51和RAD54B等DNA修复机制相关基因,抑制黄曲霉同源重组及非同源性末端连接过程,最终达到抑制黄曲霉生长及产孢的目的。上述结果表明,蜡样芽胞杆菌(Bacillus cereus)AR1002有望应用于实际生产中以应对黄曲霉污染问题。

IFN-α对禽网状内皮组织增殖病病毒体内外增殖的抑制作用

为明确禽源α干扰素(IFN-α)对禽网状内皮组织增殖病病毒(REV)体内外增殖的抑制效果,本试验在细胞和动物水平上分别采用鸡胚成纤维(DF-1)细胞和海兰褐鸡作为模型,于REV接种前(预防)和接种后(治疗)分别添加不同浓度的禽IFN-α处理,分析禽IFN-α在体外和体内对REV增殖的影响。体外试验结果显示,与阳性对照组相比,禽IFN-α浓度为33 IU/mL的治疗组72 h细胞和上清中REV的增殖受到了显著抑制(P<0.05),禽IFN-α浓度为33 IU/mL的预防组72 h细胞中REV的增殖受到了显著抑制(P<0.05)。通过比较各组海兰褐鸡的平均体重、血液中REV阳性率和病毒载量、泄殖腔REV阳性率和排毒量、免疫器官指数和病毒载量系统评估禽IFN-α对REV体内增殖的影响,结果显示,与阳性对照组相比,禽IFN-α浓度为1 500 IU的治疗组海兰褐鸡21和28日龄时体重显著升高(P<0.05),14日龄时肝脏发育指数以及14和28日龄时的脾脏发育指数显著降低(P<0.05),14日龄时血液和脾脏中病毒载量显著降低(P<0.05),7和21日龄时泄殖腔排毒量显著降低(P<0.05)。本试验在体外细胞和体内动物2个层面均证实了禽IFN-α对REV增殖具有显著的抑制效果,这不仅为禽网状内皮组织增殖病的防控策略提供了新的思路和辅助手段,也为未来抗病毒药物的研发和应用提供了科学依据。

C57BL/6小鼠大脑皮层区与基底神经节隆起区神经元突触发育过程比较

目的:观察小鼠皮层区和基底神经节隆起(GE)区神经元突触发育过程,阐明兴奋性突触和抑制性突触在不同脑区的体内外发育差异。方法:C57BL/6雌鼠于妊娠第13.5~15.5天断颈处死后,经无菌操作取胚胎小鼠,显微镜下逐步分离获取胚胎小鼠脑组织皮层区和GE区。体外原代培养胚胎小鼠神经元,于培养3、7、14和21 d分别收集细胞样品,并将其作为培养3、7、14和21 d组。采用实时荧光定量PCR(RT-qPCR)法检测各组小鼠皮层区和GE区原代培养神经元中突触后表达蛋白突触后密度蛋白95(PSD95)及桥尾蛋白(Gephyrin) mRNA表达水平。免疫荧光法检测各组小鼠皮层区和GE区原代培养神经元中囊泡谷氨酸转运蛋白1(vGLUT1)、 PSD95、囊泡γ-氨基丁酸(GABA)转运蛋白(vGAT)及Gephyrin蛋白表达水平。免疫荧光法检测胚胎小鼠脑组织皮层区和GE区神经元中vGLUT1及vGAT蛋白表达水平。结果:与培养3 d组比较,培养14和21 d组小鼠皮层区和GE区原代培养神经元中PSD95及Gephyrin mRNA表达水平明显升高(P<0.01);与皮层区比较,培养14 d组小鼠GE区原代培养神经元中Gephyrin mRNA表达水平明显降低(P<0.01)。显微镜下观察,培养14 d组小鼠皮层区和GE区原代培养神经元中兴奋性突触及抑制性突触均初步发育,相关蛋白呈阳性表达;其中兴奋性突触相关蛋白阳性表达在皮层区神经元中更为明显,且突触前分子vGLUT1和突触后分子PSD95在皮层区神经元的胞体及突起部位均呈现共定位的特征;抑制性突触前分子vGAT蛋白和突触后分子Gephyrin蛋白在GE区神经元胞体及突起中也呈现共定位的特征,且突触前分子较相应的突触后分子蛋白阳性表达更明显。与皮层区比较,培养14 d组小鼠GE区原代培养神经元中vGLUT1和PSD95蛋白表达水平明显降低(P<0.01),vGAT和Gephyrin蛋白表达水平明显升高(P<0.01)。培养21 d组小鼠皮层区和GE区原代培养神经元突触相关蛋白阳性表达增加,兴奋性突触和抑制性突触进一步成熟并完善。皮层区和GE区原代培养神经元的胞体及突起部位均形成了丰富的突触前后对应的表达模式,突触结构逐步发育良好,且突触前分子较相应的突触后分子蛋白阳性表达更明显。与皮层区比较,培养21 d组小鼠GE区原代培养神经元中vGLUT1和PSD95蛋白表达水平均明显降低(P<0.01),vGAT和Gephyrin蛋白表达水平均明显升高(P<0.01)。与皮层区比较,胚胎小鼠脑组织GE区神经元中vGLUT1蛋白表达水平明显降低(P<0.01),vGAT蛋白表达水平明显升高(P<0.05)。结论:皮层区和GE区神经元的突触发育具有明显的差异性,皮层区兴奋性突触发育较早,GE区抑制性突触发育较早。突触的脑区特异性发育提示不同细胞类型的神经疾病可能具有不同的发育来源。

红小豆提取物对α-葡萄糖苷酶和胰脂肪酶的抑制作用

采用体积分数70%乙醇对红小豆进行提取,提取物经水混旋,然后依次采用石油醚、乙酸乙酯、正丁醇溶剂萃取。正丁醇萃取相浓缩干燥后经AB-8大孔树脂吸附,以不同体积分数乙醇洗脱,测定洗脱相浓缩液中总三萜皂苷和总黄酮的含量,并探究洗脱相浓缩液对α-葡萄糖苷酶和胰脂肪酶的抑制作用。结果显示:体积分数70%和95%乙醇洗脱组分中的总三萜皂苷含量高于其他组分,且体积分数70%乙醇洗脱组分的总黄酮含量最高,为(181.83±3.09)mg/g;体积分数95%、70%、50%乙醇洗脱组分对α-葡萄糖苷酶和胰脂肪酶均具有较好的抑制活性;体积分数95%乙醇洗脱组分对2种酶的抑制类型均为混合型抑制。

抗菌肽F1高产菌株筛选鉴定及其对多重耐药菌生物膜形成的抑制作用

该研究以L.paracasei subsp.Tolerans FX-6为出发菌株,经过^(60)Coγ射线辐照诱变,筛选出一株高产抗菌肽F1的突变菌株(菌26)。对菌26进行16S rDNA序列比较分析发现其与FX-6具有高度相似性。进一步对菌26的牛奶发酵粗提物进行分离纯化,成功富集抗菌肽F1。通过抑菌实验发现,菌26的牛奶发酵粗提物对E.coli的最低抑菌质量浓度(Minimum Inhibitory Concentration,MIC)为3.16 mg/mL,较FX-6牛奶发酵粗提物的MIC降低了75%,同时抗菌肽F1的产量也提高了3.03倍,初步推断抗菌肽F1质量浓度与菌株的抑菌活性之间可能存在正相关关系。前期研究已经发现抗菌肽F1对黏菌素耐药大肠杆菌SHP45的MIC为320.0μg/mL,该实验进一步研究表明当抗菌肽F1的质量浓度达到2×MIC时,可抑制50%以上的黏菌素耐药大肠杆菌SHP45生物膜的形成。基于以上研究结果表明,菌26比出发菌株具有更高的产抗菌肽F1能力,且抗菌肽F1对黏菌素耐药大肠杆菌SHP45的生物膜形成具有显著的抑制作用。该研究为高产抗菌肽菌株的获得提供研究思路,并有望为多重耐药细菌感染的防控提供物质基础。

自身免疫性肝炎患者血清巨噬细胞迁移抑制因子和白细胞介素-21水平变化及其对治疗应答的影响

目的探讨自身免疫性肝炎(AIH)患者血清巨噬细胞迁移抑制因子(MIF)和白细胞介素-21(IL-21)水平变化及其对治疗应答的影响。方法2017年12月~2022年11月我院诊治的AIH患者61例,均接受泼尼松联合硫唑嘌呤联合治疗。在治疗前,行肝穿刺检查,采用ELISA法检测血清MIF和IL-21水平。结果17例临床重度AIH患者血清MIF和IL-21水平分别(2.5±0.9)μg/L和(321.5±34.2)pg/mL,显著高于44例轻中度患者【分别为(1.5±0.5)μg/L和(174.7±20.5)pg/mL,P<0.05】;26例肝组织G3~G4炎症活动度患者血清MIF和IL-21水平分别(2.6±1.1)μg/L和(332.9±35.4)pg/mL,显著高于35例G1~G2患者【分别为(1.5±0.6)μg/L和(170.8±25.4)pg/mL,P<0.05】;经标准免疫抑制治疗,本组61例AIH患者生化学应答率为88.5%;9例超过6个月获得生化学应答患者血清MIF和IL-21水平分别(2.9±1.5)μg/L和(334.5±40.6)pg/mL,显著高于22例在6个月内应答患者【分别为(1.5±0.3)μg/L和(194.8±19.5)pg/mL,P<0.05】或23例在3个月内应答患者【分别为(0.9±0.2)μg/L和(100.6±9.2)pg/mL,P<0.05】。结论AIH患者血清MIF和IL-21水平升高,可能与病情重或对标准治疗应答差有关,值得进一步研究。

COPD急性加重期患者外周血单个核细胞SOCS-1、TLR4 mRNA及血清cTnT、尿酸水平变化分析

目的探讨慢性阻塞性肺疾病(COPD)急性加重期患者外周血单个核细胞中细胞因子信号抑制蛋白-1(SOCS-1)、Toll样受体4(TLR4)mRNA水平及血清心肌肌钙蛋白T(cTnT)、尿酸水平变化。方法收集COPD急性加重期(组)患者70例,COPD稳定期(组)患者40例,对照组健康志愿者40名。检测外周血单个核细胞SOCS-1、TLR4 mRNA水平及血清cTnT、尿酸浓度;行肺功能检查并记录相关指标(FEV1、FEV1%、FEV1/FVC%)。对COPD急性加重期患者进行1 a随访,分为预后不良组和预后良好组。对外周血单个核细胞SOCS-1、TLR4 mRNA水平、血清cTnT、尿酸浓度行Pearson相关性分析,并对COPD急性加重期患者预后评估价值进行ROC曲线分析。结果COPD急性加重期组SOCS-1 mRNA表达水平明显低于COPD稳定期组、对照组,TLR4 mRNA水平及血清cTnT、尿酸浓度明显高于COPD稳定期组和对照组(均P<0.01)。COPD急性加重期组FEV1、FEV1%、FEV1/FVC%明显低于COPD稳定期组和对照组(P<0.05)。COPD急性加重期患者FEV1/FVC%与外周血单个核细胞SOCS-1 mRNA表达水平呈正相关关系(P<0.01),与外周血单个核细胞TLR4 mRNA水平及血清cTnT、尿酸浓度呈负相关关系(P<0.01)。预后不良组SOCS-1 mRNA水平明显低于预后良好组,TLR4 mRNA水平及血清cTnT、尿酸浓度明显高于预后良好组(P<0.05)。外周血单个核细胞SOCS-1、TLR4 mRNA水平及血清cTnT、尿酸联合检测对COPD急性加重期患者预后具有较高的评估价值。结论COPD急性加重期患者SOCS-1低表达,TLR4、cTnT、尿酸高表达,且与肺功能水平密切相关,联合检测对患者预后具有较高的评估价值。

绿豆蛋白α-淀粉酶抑制肽的制备与鉴定

采用胃-胰蛋白酶水解绿豆蛋白及其分级蛋白,以水解物α-淀粉酶活性抑制率为主要指标,结合水解度、氨基酸组成及分子质量分析其抑制效果差异及原因。结果表明,绿豆蛋白肽对α-淀粉酶活性抑制率最高,为16.51%;与绿豆分级蛋白相比,绿豆蛋白的疏水氨基酸含量和水解度最高,分别为32.68%和6.28%,水解物的肽分子质量最小,均小于20kDa,因此选用绿豆蛋白制备α-淀粉酶抑制肽。然后分离鉴定绿豆蛋白肽,新发现了17条有潜在α-淀粉酶抑制效果的肽。本研究表明绿豆蛋白较其分级蛋白具有更强的α-淀粉酶抑制效果,能够用于降血糖的功能性食品或药物中。

抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用

目的探讨敲低长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)抑制核苷酸结合寡聚结构域样受体蛋白1(NLRP1)炎症小体在缓解阿尔茨海默病进展中的作用。方法选取9~10周龄遗传背景为C57/BL6的野生型小鼠(WT组,10只)或淀粉样前体蛋白(APP)/早老素1(PS1)转基因小鼠(30只)。APP/PS1转基因小鼠随机分为模型(model)组,模型+敲低lncRNA TUG1组[model+lncRNA TUG1短发夹RNA(shRNA)组]和model+shRNA非靶标(NT)组,每组10只。分别采集12周龄第1天(3月龄)和32周龄第1天(8月龄)小鼠外周血和脑皮质组织,并分离皮质中的原代小胶质细胞和原代星形胶质细胞,每个时间点每组5只小鼠。Real-time PCR分别测定3月龄和8月龄上述4个分组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和巨噬细胞移动抑制因子(MIF)mRNA的水平,以及原代星形胶质细胞中补体蛋白C1r和C1s mRNA的水平。ELISA法测定其外周血浆中MIF含量。对3月龄和8月龄小鼠脑皮质原代小胶质细胞和原代星形胶质细胞共培养。CCK-8法测定上述2种细胞的增殖能力。Western blotting分别测定3月龄和8月龄上述4个分组小鼠脑皮质组织中MIF、白细胞介素1β前体(pro-IL-1β)、凋亡相关斑点样蛋白(ASC)、Caspase-1(p20)、Caspase-1(full)、NLRP1及NLRP3蛋白的表达水平。采用免疫荧光染色法测定8月龄各分组小鼠脑皮质组织中β淀粉样蛋白(Aβ)表达。结果3月龄和8月龄时,与WT组小鼠相比,model组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF相对表达水平显著上调,原代小胶质细胞和原代星形胶质细胞增殖能力增强(P<0.05)。与model组相比,model+lncRNA TUG1 shRNA组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF的相对表达水平显著降低,原代小胶质细胞和原代星形胶质细胞增殖能力降低(P<0.05)。与WT组相比,model组小鼠外周血浆中MIF含量显著升高;小鼠脑皮质组织中pro-IL-1β、ASC、Caspase-1(p20)、Caspase-1(full)、NLRP1以及NLRP3的蛋白表达水平显著升高;Aβ免疫荧光强度明显增强(P<0.05)。与model组相比,model+lncRNA TUG1 shRNA组小鼠外周血浆中MIF含量显著降低;小鼠脑皮质组织中pro-IL-1β、ASC、Caspase-1(p20)、Caspase-1(full)和NLRP1的蛋白表达水平显著降低,Aβ免疫荧光强度明显降低(P<0.05),而NLRP3蛋白质的表达水平无明显变化(P>0.05)。与model组相比,model+shRNA NT组小鼠上述所有检测指标差异均无显著性(P>0.05)。结论APP/PS1转基因小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF因子表达上调与脑皮质内NLRP1炎症小体激活成正相关,敲低lncRNA TUG1可缓解阿尔茨海默病的进展。