Landscape of Sequence Variations in Homologous Copies of FAD2 and FAD3 in Rapeseed(Brassica napus L.)Germplasm with High/Low Linolenic Acid Trait

Genetic manipulation(either restraint or enhancement)of the biosynthesis pathway ofα-linolenic acid(ALA)in seed oil is an important goal in Brassica napus breeding.B.napus is a tetraploid plant whose genome often har-bors four and six homologous copies,respectively,of the two fatty acid desaturases FAD2 and FAD3,which con-trol the last two steps of ALA biosynthesis during seed oil accumulation.In this study,we compared their promoters,coding sequences,and expression levels in three high-ALA inbred lines 2006L,R8Q10,and YH25005,a low-ALA line A28,a low-ALA/high-oleic-acid accession SW,and the wildtype ZS11.The expression levels of most FAD2 and FAD3 homologs in the three high-ALA accessions were higher than those in ZS11 and much higher than those in A28 and SW.The three high-ALA accessions shared similar sequences with the pro-moters and CDSs of BnFAD3.C4 and BnFAD3.A3.In A28 and SW,substitution of three amino acid residues in BnFAD2.A5 and BnFAD2.C5,an absence of BnFAD2.C1 locus,and a 549 bp long deletion on the BnFAD3.A3 promoter were detected.The profile of BnFAD2 mutation in the two low-ALA accessions A28 and SW is different from that reported in previous studies.The mutations in BnFAD3 in the high-ALA accessions are reported for thefirst time.In identifying the sites of these mutations,we provide detailed information to aid the design of mole-cular markers for accelerated breeding schemes.

Preparation of High-Purity Linolenic Acid from Oil of Lithospermum Erythrorhizon by Urea Inclusion and Column Chromatography

Aim To separate high purity linolenic acid from the oil of Lithospermumerythrorhizon growing in the Northeast of China. Methods Urea inclusion and column chromatographywere used. Results Unsaturated fatty acid was separated, with a purity of 99.30 wt% of linolenicacid. Conclusion The experiment shows excellent reproducibility and high feasibility for industrialproduction.

Preparation and Charaterization of Self-assembled Nanoparticles Based on Linolenic-acid Modified Chitosan

Chitosan was modified by conjugating coupling with linolenic acid through the 1-ethyl-3-（3-dimethylami- nopropyyl） carbodiimide （EDC）-mediated reaction. The degree of substitution 1.8% （ i.e. 1.8 linolenic acid group per 100 anhydroglucose units） was measured by ^1H NMR. The critical aggregation concentration （CAC） of the self-aggregate of hydrophobically modified chitosan was determined by measuring the fluorescence intensity of the pyrene as a fluorescent probe. The CAC value in phosphate-buffered saline （PBS） solution （pH 7.4） was 5 × 10^-2 mg mL^-1. The average particle size of selfaggregates of hydrophobically modified chitosan in PBS solution （pH7.4） was 210.8 nm with a unimodal size distribution ranging from 100 to 500 nm. Transmission electron microscopy （TEM） study showed that the formation of near spherical shape nanoparticles has enough structural integrity. The loading ability of hydrophibically modified chitosan （LA-chitosan） was investigated by using bovine serum albumin （BSA） as the model. The loading capacity of self-aggregated nanoparticles increases （ 19.85 % ± 0.04 % to 37.57 % ± 0.25 % ） with the concentration of BSA （0.1-0.5 mg mL^-1 ）.

Evaluation of anti-tubercular activity of linolenic acid and conjugatedlinoleic acid as effective inhibitors against Mycobacterium tuberculosis

Objective: To evaluate a new pharmacological activity/effect of linolenic acid(α- and γ-form) and conjugated-linoleic acid(CLA) causing antibacterial activity against Mycobacterium tuberculosis(Mtb). Methods: The anti-Mtb activity/effect of linolenic acid and CLA were determined using different anti-Mtb indicator methods such as resazurin microtiter assay(REMA) and MGIT 960 system assay. The Mtb was incubated with various concentrations(12.5–200) μg/m L of the compounds and anti-Mtb first-line drugs for 5 d in the REMA, and for 3 wk in MGIT 960 system assay. Results: Linolenic acid and CLA obviously indicated their anti-Mtb activity/effect by strongly inhibiting the growth/proliferation of Mtb in a dosedependent manner in the REMA and the MGIT 960 system assay. Interestingly, linolenic acid and CLA consistently induced anti-Mtb activity/effect by effectively inhibiting the growth/proliferation of Mtb in MGIT 960 system for 21 d with a single-treatment, and their minimum inhibitory concentrations were measured as 200 μg/m L respectively. Conclusions: These results demonstrate that linolenic acid and CLA not only have effective anti-Mtb activity/properties, but also induce the selective-anti-Mtb effects by strongly inhibiting and blocking the growth/proliferation of Mtb through a new pharmacological activity/action. Therefore, this study provides novel perspectives for the effective use of them and the potential that can be used as potent anti-Mtb candidate drugs, as well as suggests the advantage of reducing the cost and/or time for developing a new/substantive drug by effectively repurposing the existing drugs or compounds as one of new strategies for the global challenge of tuberculosis.

Formulation ofα-linolenic acid microemulsion free of co-surfactant

A novel class ofα-linolenic acid-in-water microemulsion free of co-surfactant was investigated as potential food delivery systems.Rough demarcation within the transparent region was deduced from the results of conductivity and polarizing optical microscopy.The microemulsion mean hydrodynamic diameter and characterization were determined by dynamic light scattering and negative-staining TEM.The location of ALA molecules in the microemulsion formulations was determined by ~1H NMR spectroscopy.

γ-亚麻酸通过下调氧化应激反应抑制NLRP3介导的焦亡并改善脑梗塞小鼠认知障碍

目的:探究γ-亚麻酸(GLA)对脑梗塞小鼠认知障碍的影响及机制。方法:选取18只雄性小鼠,分为对照组6只、模型组6只和GLA组6只,对模型组以及GLA组以大脑中动脉闭塞术进行脑栓塞小鼠认知障碍模型制作,对照组进行假手术,即手术过程相同但不插入线栓。对照组和模型组每日以1mg/kg生理盐水灌胃,GLA组每日以1mg/kg GLA灌胃14d。以Morris水迷宫评估小鼠的认知障碍情况。处死小鼠并取小鼠大脑组织,以HE染色观察小鼠脑梗死面积,以TUNEL染色观察小鼠神经元凋亡情况,以Western Blot实验检测小鼠海马体中P22、P47、NLRP3、IL-1β、GSDMD以及Caspase-1蛋白表达水平。结果:模型组小鼠的逃逸潜伏期短于GLA组以及对照组,GLA组则短于对照组,模型组的穿越环次数最多,GLA组次之,对照组最少;模型组大脑梗死面积以及神经元凋亡数目最多,GLA组次之,对照组大脑无梗死且无神经元凋亡;模型组P22、P47、NLRP3、IL-1β、GSDMD以及Caspase-1蛋白表达高于GLA组和对照组,GLA组的蛋白表达水平则高于对照组。结论:GLA通过下调氧化应激反应抑制NLRP3介导的焦亡并改善脑梗塞导致的小鼠认知障碍。

γ-亚麻酸对糖尿病小鼠记忆衰退的影响及机制

目的 探究γ-亚麻酸(GLA)对糖尿病小鼠记忆衰退的影响及机制。方法 选取16只8周龄野生型小鼠,雌雄各半,分成GLA组及对照组,对所有小鼠使用1 g/kg D-果糖加10%饮用水饮食(持续40 d)诱导糖尿病小鼠记忆衰退模型。对照组小鼠给予每日1 mg/kg生理盐水灌胃,GLA组小鼠给予每日1 mg/kg GLA灌胃。干预7 d后,以Morris水迷宫评估小鼠的学习和记忆障碍情况,处死小鼠取大脑皮层及海马组织,以HE染色检查二者病理改变,以透射电镜观察小鼠海马神经元超微结构变化,以Western Blot实验检测小鼠海马组织中GRP78、GRP94、NOX2和NOX4蛋白表达水平。结果 GLA组小鼠的逃逸潜伏期短于对照组,穿越环次数高于对照组(P<0.05)。GLA组小鼠大脑皮层和海马组织病理变化以及海马神经元超微结构损伤程度均较对照组轻。GLA组小鼠海马组织GRP78、GRP94、NOX2和NOX4蛋白表达水平低于对照组(P<0.05)。结论 GLA能通过抑制海马神经元内质网应激反应,进而改善糖尿病小鼠记忆衰退。

6种富含α-亚麻酸食用油脂的主要组成成分及消化特征研究进展

α-亚麻酸(ALA)是人体必需脂肪酸,在维持人体正常生理活动中具有多重作用。旨在确定ALA在食用油脂中的物质基础与其功能的构效关系,实现精准的膳食脂质营养,对6种富含ALA食用油脂的总脂肪酸组成、甘油三酯中脂肪酸的位置分布情况、甘油三酯分子构成、脂质伴随物和消化特性等研究内容进行了总结和分析。6种富含ALA食用油脂主要脂肪酸组成为棕榈酸、硬脂酸、油酸、亚油酸、ALA,其中:紫苏籽油、亚麻籽油和牡丹籽油的ALA含量较高;不同富含ALA食用油脂甘油三酯中脂肪酸的位置分布存在显著差异,蚕蛹油的ALA主要分布在sn-2位,牡丹籽油和火麻仁油的ALA主要分布在sn-1,3位;不同富含ALA食用油脂甘油三酯的分子结构类型存在显著差异;富含ALA食用油脂的脂质伴随物主要为维生素E和甾醇,其中沙棘籽油的维生素E和甾醇含量均高于其余5种食用油脂;富含ALA食用油脂的消化程度和消化速率均低于非富含ALA食用油脂。

18β-glycyrrhetinic acid inhibits proliferation of gastric cancer cells through regulating the miR-345-5p/TGM2 signaling pathway

BACKGROUND Gastric cancer(GC)is a common gastrointestinal malignancy worldwide.Based on cancer-related mortality,the current prevention and treatment strategies for GC still show poor clinical results.Therefore,it is important to find effective drug treatment targets.AIM To explore the molecular mechanism of 18β-glycyrrhetinic acid(18β-GRA)regulating the miR-345-5p/TGM2 signaling pathway to inhibit the proliferation of GC cells.METHODS CCK-8 assay was used to determine the effect of 18β-GRA on the survival rate of GES-1 cells and AGS and HGC-27 cells.Cell cycle and apoptosis were detected by flow cytometry,cell migration was detected by a wound healing assay,the effect of 18β-GRA on subcutaneous tumor growth in BALB/c nude mice was investigated,and the cell autophagy level was determined by MDC staining.TMT proteomic analysis was used to detect the differentially expressed autophagy-related proteins in GC cells after 18β-GRA intervention,and then the protein-protein interaction was predicted using STRING(https://string-db.org/).MicroRNAs(miRNAs)transcriptome analysis was used to detect the miRNA differential expression profile,and use miRBase(https://www.mirbase/)and TargetScan(https://www.targetscan.org/)to predict the miRNA and complementary binding sites.Quantitative real-time polymerase chain reaction was used to detect the expression level of miRNA in 18β-GRA treated cells,and western blot was used to detect the expression of autophagy related proteins.Finally,the effect of miR-345-5p on GC cells was verified by mir-345-5p overexpression.RESULTS 18β-GRA could inhibit GC cells viability,promote cell apoptosis,block cell cycle,reduce cell wound healing ability,and inhibit the GC cells growth in vivo.MDC staining results showed that 18β-GRA could promote autophagy in GC cells.By TMT proteomic analysis and miRNAs transcriptome analysis,it was concluded that 18β-GRA could down-regulate TGM2 expression and up-regulate miR-345-5p expression in GC cells.Subsequently,we verified that TGM2 is the target of miR-345-5p,and that overexpression of miR-345-5p significantly inhibited the protein expression level of TGM2.Western blot showed that the expression of autophagy-related proteins of TGM2 and p62 was significantly reduced,and LC3II,ULK1 and AMPK expression was significantly increased in GC cells treated with 18β-GRA.Overexpression of miR-345-5p not only inhibited the expression of TGM2,but also inhibited the proliferation of GC cells by promoting cell apoptosis and arresting cell cycle.CONCLUSION 18β-GRA inhibits the proliferation of GC cells and promotes autophagy by regulating the miR-345-5p/TGM2 signaling pathway.

Chlorogenic acid alleviates hypoxic-ischemic brain injury in neonatal mice

Recent studies have shown that chlorogenic acid(CGA),which is present in coffee,has protective effects on the nervous system.However,its role in neonatal hypoxic-ischemic brain injury remains unclear.In this study,we established a newborn mouse model of hypoxic-ischemic brain injury using a modified Rice-Vannucci method and performed intraperitoneal injection of CGA.We found that CGA intervention effectively reduced the volume of cerebral infarct,alleviated cerebral edema,restored brain tissue structure after injury,and promoted axon growth in injured brain tissue.Moreover,CGA pretreatment alleviated oxygen-glucose deprivation damage of primary neurons and promoted neuron survival.In addition,changes in ferroptosis-related proteins caused by hypoxic-ischemic brain injury were partially reversed by CGA.Furthermore,CGA intervention upregulated the expression of the key ferroptosis factor glutathione peroxidase 4 and its upstream glutamate/cystine antiporter related factors SLC7A11 and SLC3A2.In summary,our findings reveal that CGA alleviates hypoxic-ischemic brain injury in neonatal mice by reducing ferroptosis,providing new ideas for the treatment of neonatal hypoxic-ischemic brain injury.

富含α-亚麻酸的中长链脂肪酸结构脂的酶法合成与理化性质分析

旨在为富含多不饱和脂肪酸(PUFA)的中长链脂肪酸甘油三酯(MLCT)产品的应用提供参考,采用脂肪酶Lipozyme TL IM催化精炼桑蚕蛹油(SPO)与三辛酸甘油酯(TRI)进行酯交换反应合成富含α-亚麻酸(ALA)的MLCT(ALA-MLCT),考察了反应时间对ALA-MLCT合成的影响,并对合成的ALA-MLCT的甘油三酯组成、质量指标、氧化稳定性、乳液性能和体外消化特性进行了分析。结果表明:Lipozyme TL IM催化效率高,反应10 h TRI的转化率达到98%以上,反应12 h合成的产物ALA-MLCT中含有ALA的MLCT含量可以达到40.41%,且主要为LnCaCa (10.95%)、PLnCa (7.69%)和OLnCa (6.21%),其质量指标符合中长链脂肪酸食用油标准征求意见稿2022中的一级食用油标准;与SPO和酯交换反应底物物理混合物(SPO与TRI质量比1∶4,SPO+TRI)相比,产物ALA-MLCT氧化稳定性显著提高,其乳液粒径较小且分布均匀,具有更好的物理稳定性;体外模拟消化实验表明,与SPO+TRI相比,ALA-MLCT更快达到消化平衡,但最终二者消化率均可以达到90%以上。综上,合成的ALA-MLCT结构脂有助于提高ALA的生物可及性,有望作为功能性食用油脂。

γ-亚麻酸的富集及其生理功效研究进展

γ-亚麻酸(GLA)是含有三个双键的多不饱和脂肪酸,因其具有多种生物学活性而备受关注。GLA目前主要的植物来源为琉璃苣、月见草和黑加仑等,微生物来源大多为藻类和真菌。现有的GLA分离纯化技术各有优缺点,单一的富集方法无法满足高纯度GLA的要求,采用不同方法综合使用、结合现代化分离提取技术是未来制备GLA的方法。同时,多项研究表明,GLA在机体的物质代谢和生理调控上发挥着十分重要的作用,对血脂、抗血栓性心血管疾病、癌症和糖尿病等慢性疾病具有很好的预防效果,但具体的作用机理仍需进一步研究。综上所述,GLA是一种对人体有益的脂肪酸,但其人体内的代谢途径和生理功效仍然需要进一步挖掘,从而为GLA作为功能性保健食品的开发提供理论支撑。

Effects of Ce^(3+) on improvement of spectral characteristics and function of chloroplasts damaged by linolenic acid in spinach

Linolenic acid has great effects on the structure and function of chloroplast. We studied the effects of Ce3＋ on the improvement of chloroplast spectral characteristics and oxygen evolution damaged by linolenic acid in spinach. Results showed that Ce3＋ could decrease the light absorption increased by linolenic acid and promote the distribution of excitation energy to PS II and alleviate the decrease of PS Ⅱ fluo- rescence yield caused by linolenic acid. The linolenic acid treatments in various concentrations reduced the oxygen-evolving rate of chloroplasts, but the rate was accelerated since adding Ce3＋.

Effect of Altering Linoleic Acid and Linolenic Acid Dietary Ratios on the Performance and Tissue Fatty Acid Profiles of Prawn, Macrobrachium rosenbergii （de Man 1879） Post Larvae

A 90-day experiment was conducted to investigate the effects of different dietary linoleic acid （LA; 18：2n-6） and linolenic acid ratios （LNA; 18：3n-3） on growth induces, feed utilization and tissue fatty acid profile of freshwater prawn, Macrobrachium rosenbergii post-larvae （PL）. The experiment was conducted in cubic indoor fiberglass tanks, each holding 700 L in triplicate. Post-larvae with an average weight of 20.8 ± 0.20 mg were stocked at 80 PL m2. Five experimental isocaloric （15.06 MJ kgl digestible energy）, and isonitrogenous （30.45% digestible protein） diets were formulated by blending of soybean oil and linseed oil to containing five dietary LA/LNA ratios （7.80, 2.75, 1.28, 0.65 and 0.30）. The highest survival values were recorded for prawn PL fed diet containing 0.65 LA/LAN ratios. Growth indices of PL significantly increased （P 〈 0.05） with decreased dietary LA/LAN ratios to 0.65. The same trend was observed for the highest （P ≤ 0.05） protein efficiency ratio, protein productive value, fat retention, energy retention and best feed conversion ratio. The total whole tissue polyunsaturated fatty acid composition of M. rosenbergii PL was dominated by LA followed by LAN. Post larvae fed the diets containing higher LA/LNA ratios showed a higher tissue LA/LNA ratio. The obtained findings revealed that fatty acid patterns ofM. rosenbergii PL were influenced by fatty acid profiles of diets. The diet containing 0.65 LA/LNA ratio is recommended to obtaining optimum growth performance and feed utilization for M. rosenbergii PL.

Alleviation effects of Ce^(3+) on inhibition of photochemical activity caused by linolenic acid in spinach chloroplast

Linolenic acid has great effects on the structure and function of chloroplast. The function of Ce^3＋ on the improvement of chloroplast photoreduction activity and oxygen evolution damaged by linolenic acid in spinach by in vitro investigation was studied. Results showed that adding Ce^3＋ to the linolenic acid treated chloroplast could greatly decrease the reduction linolenic acid exerted on the whole chain electron transport rate and the photoreduction activity of photosystem Ⅱ （PSII） and photosystem Ⅰ （PSI） as well as the oxygen evolution rate of chloroplast. It indicated that Ce^3＋ had the ability to relieve the inhibition of the photochemical reaction of chloroplast caused by linolenic acid to some extent.

MicroRNA-502-3p regulates GABAergic synapse function in hippocampal neurons

Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.

日粮添加α-亚麻酸对育肥猪肌肉脂肪酸组成与肠道菌群的影响

[目的]本试验旨在研究日粮添加α-亚麻酸对育肥猪肌肉脂肪酸组成与肠道菌群的影响。[方法]选取36头120日龄杜×长×大三元猪进行分栏喂养,根据体重相近原则随机分为2组,每组3个重复,每重复6头,分别为对照组(CON)和添加α-亚麻酸组(ALA)。CON组饲喂基础日粮,ALA组饲喂含5 g·kg^(-1) α-亚麻酸的基础日粮。饲喂至162日龄时在每个重复中随机选取6头猪屠宰并采样分析。[结果]相较于CON组,ALA组猪平均终末重、日增重、日采食量及料重比均未见显著差异,但料重比降低7.7%,ALA组猪背最长肌肉色、滴水损失无显著变化。在血液生化指标中,ALA组总胆固醇和游离脂肪酸含量相较于CON组显著降低(P<0.05)。对背最长肌脂肪酸组成分析结果显示,与CON组相比,ALA组中饱和脂肪酸(saturated fatty acid, SFA)含量和n-6多不饱和脂肪酸(omega-6 polyunsaturated fatty acid, PUFA)/n-3 PUFA的比值显著降低(P<0.05),C18:3n-3(ALA)、C20:5n-3(DPA)含量显著升高(P<0.05),n-3 PUFA总含量也显著升高(P<0.05)。与CON组相比,ALA组结肠内容物中瘤胃球菌科、毛螺菌科、韦荣氏菌科、丁酸球菌科的丰度均显著增加(P<0.05),而氨基酸球菌科、产粪甾醇真细菌、脱硫弧菌、消化链球菌科、坦纳菌科、梭状芽胞杆菌科和克里斯滕森菌科的丰度均显著降低(P<0.05)。[结论]日粮添加5 g·kg^(-1) α-亚麻酸改善了育肥猪肌肉脂肪酸组成,降低了肌肉中饱和脂肪酸(SFA)含量,提高了n-3 PUFA含量并降低了n-6 PUFA/n-3 PUFA的比值,显著促进了育肥猪肠道中瘤胃球菌科、毛螺菌科、韦荣氏菌科等常见的短链脂肪酸(SCFA)产生菌的增殖。

18β-glycyrrhetinic acid promotes gastric cancer cell autophagy and inhibits proliferation by regulating miR-328-3p/signal transducer and activator of transcription 3

BACKGROUND Gastric cancer(GC)is one of the most common cancer types worldwide,and its prevention and treatment methods have garnered much attention.As the active ingredient of licorice,18β-glycyrrhetinic acid(18β-GRA)has a variety of pharmacological effects.The aim of this study was to explore the effective target of 18β-GRA in the treatment of GC,in order to provide effective ideas for the clinical prevention and treatment of GC.AIM To investigate the mechanism of 18β-GRA in inhibiting cell proliferation and promoting autophagy flux in GC cells.METHODS Whole transcriptomic analyses were used to analyze and screen differentially expressed microRNAs(miRNAs)in GC cells after 18β-GRA intervention.Lentivirus-transfected GC cells and the Cell Counting Kit-8 were used to detect cell proliferation ability,cell colony formation ability was detected by the clone formation assay,and flow cytometry was used to detect the cell cycle and apoptosis.A nude mouse transplantation tumor model of GC cells was constructed to verify the effect of miR-328-3p overexpression on the tumorigenicity of GC cells.Tumor tissue morphology was observed by hematoxylin and eosin staining,and microtubule-associated protein light chain 3(LC3)expression was detected by immunohistochemistry.TransmiR,STRING,and miRWalk databases were used to predict the relationship between miR-328-3p and signal transducer and activator of transcription 3(STAT3)-related information.Expression of STAT3 mRNA and miR-328-3p was detected by quantitative polymerase chain reaction(qPCR)and the expression levels of STAT3,phosphorylated STAT3(p-STAT3),and LC3 were detected by western blot analysis.The targeted relationship between miR-328-3p and STAT3 was detected using the dual-luciferase reporter gene system.AGS cells were infected with monomeric red fluorescent protein-green fluorescent protein-LC3 adenovirus double label.LC3 was labeled and autophagy flow was observed under a confocal laser microscope.RESULTS The expression of miR-328-3p was significantly upregulated after 18β-GRA intervention in AGS cells(P=4.51E-06).Overexpression of miR-328-3p inhibited GC cell proliferation and colony formation ability,arrested the cell cycle in the G0/G1 phase,promoted cell apoptosis,and inhibited the growth of subcutaneous tumors in BALB/c nude mice(P<0.01).No obvious necrosis was observed in the tumor tissue in the negative control group(no drug intervention or lentivirus transfection)and vector group(the blank vector for lentivirus transfection),and more cells were loose and necrotic in the miR-328-3p group.Bioinformatics tools predicted that miR-328-3p has a targeting relationship with STAT3,and STAT3 was closely related to autophagy markers such as p62.After overexpressing miR-328-3p,the expression level of STAT3 mRNA was significantly decreased(P<0.01)and p-STAT3 was downregulated(P<0.05).The dual-luciferase reporter gene assay showed that the luciferase activity of miR-328-3p and STAT33’untranslated regions of the wild-type reporter vector group was significantly decreased(P<0.001).Overexpressed miR-328-3p combined with bafilomycin A1(Baf A1)was used to detect the expression of LC3 II.Compared with the vector group,the expression level of LC3 II in the overexpressed miR-328-3p group was downregulated(P<0.05),and compared with the Baf A1 group,the expression level of LC3 II in the overexpressed miR-328-3p+Baf A1 group was upregulated(P<0.01).The expression of LC3 II was detected after intervention of 18β-GRA in GC cells,and the results were consistent with the results of miR-328-3p overexpression(P<0.05).Additional studies showed that 18β-GRA promoted autophagy flow by promoting autophagosome synthesis(P<0.001).qPCR showed that the expression of STAT3 mRNA was downregulated after drug intervention(P<0.05).Western blot analysis showed that the expression levels of STAT3 and p-STAT3 were significantly downregulated after drug intervention(P<0.05).CONCLUSION 18β-GRA promotes the synthesis of autophagosomes and inhibits GC cell proliferation by regulating the miR-328-3p/STAT3 signaling pathway.

Fixed-Bed Column Adsorption Modeling of MnO4- Ions from Acidic Aqueous Solutions on Activated Carbons Prepared with the Biomass

Activated carbons calcined at 400˚C and 600˚C (AC-400 and AC-600), prepared using palm nuts, collected in the town of Franceville in Gabon, were used to study the dynamic adsorption of MnO4- ions in acidic media on fixed bed column and on the kinetic modeling of experimental data of breakthrough curves of MnO4- ions obtained. Results on the adsorption of MnO4- ions in fixed-bed dynamics obtained on AC-400 and AC-600 adsorbents beds indicated that the AC-400 bed appears to be the most efficient in removing MnO4- ions in acidic media. Indeed, the adsorbed amounts, the adsorbed capacities at saturation and the elimination percentage of MnO4- ions obtained with AC-400 (31.24 mg;52.06 mg·g-1 and 41.65% respectively) were higher compared to those obtained with AC-600 (9.87 mg;16.45 mg·g-1 and 17.79% respectively). The breakthrough curves kinetic modeling revealed that the Thomas model and the pseudo-first-order kinetic model were the most suitable models to describe the adsorption of MnO4- ions on adsorbents studied in our experimental conditions. The results of the intraparticle diffusion model showed that intraparticle diffusion was involved in the adsorption mechanism of MnO4- ions on investigated adsorbents and was not the limiting step and the only process controlling MnO4- ions adsorption. In contrast to AC-400, the intraparticle diffusion on AC-600 bed plays an important role in the adsorption mechanism of MnO4- ions.