Biomimetic natural biomaterials for tissue engineering and regenerative medicine:new biosynthesis methods,recent advances,and emerging applications

Biomimetic materials have emerged as attractive and competitive alternatives for tissue engineering(TE)and regenerative medicine.In contrast to conventional biomaterials or synthetic materials,biomimetic scaffolds based on natural biomaterial can offer cells a broad spectrum of biochemical and biophysical cues that mimic the in vivo extracellular matrix(ECM).Additionally,such materials have mechanical adaptability,micro-structure interconnectivity,and inherent bioactivity,making them ideal for the design of living implants for specific applications in TE and regenerative medicine.This paper provides an overview for recent progress of biomimetic natural biomaterials(BNBMs),including advances in their preparation,functionality,potential applications and future challenges.We highlight recent advances in the fabrication of BNBMs and outline general strategies for functionalizing and tailoring the BNBMs with various biological and physicochemical characteristics of native ECM.Moreover,we offer an overview of recent key advances in the functionalization and applications of versatile BNBMs for TE applications.Finally,we conclude by offering our perspective on open challenges and future developments in this rapidly-evolving field.

OsFK1 encodes C-14 sterol reductase,which is involved in sterol biosynthesis and affects premature aging of leaves in rice

The enzyme C-14 sterol reductase is involved in biosynthesis of brassinosteroids(BR)and sterols,as well as plant development.OsFK1,a member of the sterol biosynthesis pathway located in the endoplasmic reticulum(ER),encodes C-14 sterol reductase.However,there is little research on the function of C-14 sterol reductase in rice.Compared with the wild type,an osfk1 mutant showed dwarf phenotype and premature aging in the second leaf during the trefoil stage,and abnormal development of leaf veins during the tillering stage.The osfk1 mutant showed signs of aberrant PCD,as evidenced by TUNEL staining.This suggested that high ROS buildup caused DNA damage and ROS-mediated cell death in the mutant.The osfk1 mutant also showed decreased chlorophyll content and aberrant chloroplast structure.Sequencing of the osfk1 mutant allele revealed a non-synonymous G to A mutation in the final intron,leading to early termination.Here,we identified the OsFK1 allele,cloned it by Mutmap sequencing,and verified it by complementation.HPLC-MS/MS assays demonstrated that the osfk1 mutation caused lower phytosterol levels.These findings showed that the OsFK1 allele encoding C-14 sterol reductase is involved in phytosterol biosynthesis and mediates normal development of rice plants.

Chain Elongation Using Native Soil Inocula:Exceptional n-Caproate Biosynthesis Performance and Microbial Mechanisms

This study demonstrates the feasibility and effectiveness of utilizing native soils as a resource for inocula to produce n-caproate through the chain elongation(CE)platform,offering new insights into anaerobic soil processes.The results reveal that all five of the tested soil types exhibit CE activity when supplied with high concentrations of ethanol and acetate,highlighting the suitability of soil as an ideal source for n-caproate production.Compared with anaerobic sludge and pit mud,the native soil CE system exhibited higher selectivity(60.53%),specificity(82.32%),carbon distribution(60.00%),electron transfer efficiency(165.00%),and conductivity(0.59 ms∙cm^(-1)).Kinetic analysis further confirmed the superiority of soil in terms of a shorter lag time and higher yield.A microbial community analysis indicated a positive correlation between the relative abundances of Pseudomonas,Azotobacter,and Clostridium and n-caproate production.Moreover,metagenomics analysis revealed a higher abundance of functional genes in key microbial species,providing direct insights into the pathways involved in n-caproate formation,including in situ CO_(2)utilization,ethanol oxidation,fatty acid biosynthesis(FAB),and reverse beta-oxidation(RBO).The numerous functions in FAB and RBO are primarily associated with Pseudomonas,Clostridium,Rhodococcus,Stenotrophomonas,and Geobacter,suggesting that these genera may play roles that are involved or associated with the CE process.Overall,this innovative inoculation strategy offers an efficient microbial source for n-caproate production,underscoring the importance of considering CE activity in anaerobic soil microbial ecology and holding potential for significant economic and environmental benefits through soil consortia exploration.

Rational Engineering of Secondary Metabolic Pathways in a Heterologous Host to Enable the Biosynthesis of Hibarimicin Derivatives with Enhanced Anti-Melanomic Activity

A 61-kb biosynthetic gene cluster(BGC),which is accountable for the biosynthesis of hibarimicin(HBM)B from Microbispora rosea subsp.hibaria TP-A0121,was heterologously expressed in Streptomyces coelicolor M1154,which generated a trace of the target products but accumulated a large amount of shunt products.Based on rational analysis of the relevant secondary metabolism,directed engineering of the biosynthetic pathways resulted in the high production of HBM B,as well as new HBM derivates with improved antitumor activity.These results not only establish a biosynthetic system to effectively synthesize HBMs-a class of the largest and most complex Type-Ⅱpolyketides,with a unique pseudo-dimeric structure-but also set the stage for further engineering and deep investigation of this complex biosynthetic pathway toward potent anticancer drugs.

Wetting alternating with partial drying during grain filling increases lysine biosynthesis in inferior rice grain

Lysine content is a criterion of the nutritional quality of rice.Understanding the process of lysine biosynthesis in early-flowering superior grain(SG)and late-flowering inferior grain(IG)of rice would advance breeding and cultivation to improve nutritional quality.However,little information is available on differences in lysine anabolism between SG and IG and the underlying mechanism,and whether and how irrigation regimes affect lysine anabolism in these grains.A japonica rice cultivar was grown in the field and two irrigation regimes,continuous flooding(CF)and wetting alternating with partial drying(WAPD),were imposed from heading to the mature stage.Lysine content and activities of key enzymes of lysine biosynthesis,and levels of brassinosteroids(BRs)were lower in the IG than in the SG at the early grainfilling stage but higher at middle and late grain-filling stages.WAPD increased activities of these key enzymes,BR levels,and contents of lysine and total amino acids in IG,but not SG relative to CF.Application of 2,4-epibrassinolide to rice panicles in CF during early grain filling reproduced the effects of WAPD,but neither treatment altered the activities of enzymes responsible for lysine catabolism in either SG or IG.WAPD and elevated BR levels during grain filling increased lysine biosynthesis in IG.Improvement in lysine biosynthesis in rice should focus on IG.

Transcriptome analysis reveals steroid hormones biosynthesis pathway involved in abdominal fat deposition in broilers

Excessive abdominal fat deposition reduces the feed efficiency and increase the cost of production in broilers.Therefore,it is an important task for poultry breeders to breed broilers with low abdominal fat.Abdominal fat deposition is a highly complex biological process,and its molecular basis remains elusive.In this study,we performed transcriptome analysis to compare gene expression profiles at different stages of abdominal fat deposition to identify the key genes and pathways involved in abdominal fat accumulation.We found that abdominal fat weight(AFW)increased gradually from day 35(D35)to 91(D91),and then decreased at day 119(D119).Accordingly,after detecting differentially expressed genes(DEGs)by comparing gene expression profiles at D35 vs.D63 and D35 vs.D91,and identifying gene modules associated with fat deposition by weighted gene co-expression network analysis(WGCNA),we performed intersection analysis of the detected DEGs and WGCNA gene modules and identified 394 and 435 intersecting genes,respectively.The results of the Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses showed that the steroid hormone biosynthesis and insulin signaling pathways were co-enriched in all intersecting genes,steroid hormones have been shown that regulated insulin signaling pathway,indicating the importance of the steroid hormone biosynthesis pathway in the development of broiler abdominal fat.We then identified 6 hub genes(ACTB,SOX9,RHOBTB2,PDLIM3,NEDD9,and DOCK4)related to abdominal fat deposition.Further analysis also revealed that there were direct interactions between 6 hub genes.SOX9 has been shown to bind to proteins required for steroid hormone receptor binding,and RHOBTB2 indirectly regulates the steroid hormones biosynthesis through cyclin factor,and ultimately affect fat deposition.Our results suggest that the genes RHOBTB2 and SOX9 play an important role in fat deposition in broilers,by regulating steroid hormone synthesis.These findings provide new targets and directions for further studies on the mechanisms of fat deposition in chicken.

Identification and Molecular Characterization of the Alkaloid Biosynthesis Gene Family in Dendrobium catenatum

As one of the main active components of Dendrobium catenatum, alkaloids have high medicinal value. The physicochemicalproperties, conserved domains and motifs, phylogenetic analysis, and cis-acting elements of the genefamily members in the alkaloid biosynthesis pathway of D. catenatum were analyzed by bioinformatics, and theexpression of the genes in different years and tissues was analyzed by qRT-PCR. There are 16 gene families,including 25 genes, in the D. catenatum alkaloid biosynthesis pathway. The analysis of conserved domains andmotifs showed that the types, quantities, and orders of domains and motifs were similar among members ofthe same family, but there were significant differences among families. Phylogenetic analysis indicated that thegene family members showed some evolutionary conservation. Cis-acting element analysis revealed that therewere a large number of light-responsive elements and MYB (v-myb avian myeloblastosis viral oncogene homolog)-related elements in these genes. qRT-PCR showed that expressions of gene family members involved in alkaloidsynthesis were different in different years and tissues of D. catenatum. This study provides a theoretical basisfor further exploration of the regulatory mechanisms of these genes in the alkaloid biosynthesis of D. catenatum.

Research Progress on Functions and Biosynthesis of D-Psicose

D-Psicose,a naturally occurring rare sugar,exhibits a sweetness approximately 70%that of sucrose.It possesses high solubility,antioxidant activity,anti-inflammatory properties,and the ability to regulate cholesterol levels and enhance insulin sensitivity.However,D-psicose is relatively scarce in nature,making large-scale extraction and utilization impractical.Consequently,the development of cost-effective synthetic strategies for D-psicose is pivotal for its industrial application.In recent years,the Izumoring strategy has emerged as an efficient alternative to chemical synthesis for producing D-psicose.Nonetheless,limitations in the biotransformation of D-psicose,primarily governed by the conversion rate of D-psicose 3-epimerase(DPEase)and enzyme yield,continue to pose challenges in achieving economically viable production.Enzyme engineering and the establishment of high-level expression systems remain crucial avenues for reducing the overall biosynthesis costs.

The biosynthesis of alarm pheromone in the wheat aphid Rhopalosiphum padi is regulated by hormones via fatty acid metabolism

Aphids are major insect pests in agriculture and forestry worldwide.Following attacks by natural enemies,many aphids release an alarm pheromone to protect their population.In most aphids,the main component of the aphid alarm pheromone(AAP)is the sesquiterpene hydrocarbon(E)-β-farnesene(EβF).However,the mechanisms behind its biosynthesis and regulation remain poorly understood.In this study,we used the bird cherry–oat aphid Rhopalosiphum padi,which is an important wheat aphid,to investigate the regulatory mechanisms of EβF biosynthesis.Our results showed that EβF biosynthesis occurs during the mature embryo period and the molting period of the 1st-and 2nd-instar nymphs.Triglycerides provide the prerequisite material for EβF production and release.Based on transcriptome sequencing,RNAi analysis,hormone treatments,and quantitative measurements,we found that the biosynthesis of EβF utilizes acetyl coenzyme A produced from fatty acid degradation,which can be suppressed by juvenile hormone but it is promoted by 20-hydroxyecdysone through the modulation of fatty acid metabolism.This is the first systemic study on the modulation of EβF production in aphids.The results of our study provide insights into the molecular regulatory mechanisms of AAP biosynthesis,as well as valuable information for designing potential aphid control strategies.

Biosynthesis of Silver Nanoparticles with Clerodendron phlomoides Leave Extract:Particle Morphology,Antimicrobial Potential and Application

Silver nanoparticles are versatile nanomaterials that have found numerous applications in various fields.The use of plant extract for the synthesis of silver is a green and sustainable approach.Clerodendron phlomoides leaves extract has been found to contain various phytochemicals,such as phenols,flavonoids,tannins,and alkaloids,which possess reducing and stabilizing properties that can aid the production of silver particles.In this paper,morphological and topographical analyses were performed on silver nanoparticles.The biosynthesized silver nanoparticles showed antimicrobial potential against wound pathogens.SEM and TEM micrographs revealed that the particles were sphere and nanosized,which makes them suitable for various biomedical applications.

基于环境DNA技术的珠江中下游鱼类多样性初步研究

通过环境DNA技术(Environmental DNA,eDNA)检测珠江中下游鱼类生物多样性,探索珠江中下鱼类多样性监测和保护的新途径。2023年2月在珠江中下游设置了桂平、藤县、封开、德庆、肇庆和九江共6个采样点,通过水样采集及过滤、eDNA提取、遗传标记扩增及测序和数据库比对分析等流程检测鱼类多样性。结果表明,6个采样点共检测出30种鱼类,隶属于4目10科27属,其中土著鱼类26种,外来种4种。较已有传统调查数据新检出2种鱼类:美丽沙鳅(Botia pulchra)和齐氏罗非鱼(Oceochromis zillii)。鱼类优势种为子陵吻鰕虎鱼(Rhinogobius giurinus)、瓦氏黄颡鱼(Pelteobagrus vachellii)、鲢(Hypophthalmichthys molitrix)、尼罗罗非鱼(O.nilotica)、齐氏罗非鱼、南方波鱼(Rasbora steineri)和鲤(Cyprinus carpio)。根据Shannon指数和Simpson指数显示,eDNA检测九江和桂平站点的鱼类多样性最高,藤县的最低。作为一种新的检测方法,eDNA技术可用于快速检测珠江中下游鱼类的多样性及分布,在实际应用中可将eDNA技术与传统的监测方法相结合,以提供更全面的鱼类生物多样性数据信息。

HBsAg ELISA+/HBV DNA NAT-献血者血清学与分子生物学特征分析

目的 了解西安地区无偿献血人群HBsAg ELISA检测结果与HBV DNA检测结果不一致的标本相关血清学标志物的分布情况。方法 收集2022年11月1日—2023年4月30日陕西省血液中心HBsAg ELISA+/HBV DNA NAT-(ELISA+/NAT-)标本共计71份,对其采用电化学发光法检测乙肝血清学标志物,同时复检巢式PCR扩增HBV S区和C区基因片段。结果 双ELISA+/NAT-标本(n=30)巢式PCR检测阳性率远高于单ELISA+/NAT-标本(n=41)(60%vs 24.40%,P<0.05)。前者献血者100%为初次献血者,血清抗-HBc阳性率100%,血清学模式以1、4、5此3项阳性(80%)为主;后者献血者中31.7%为重复献血者,血清抗-HBc阳性率仅为19.51%,血清学模式以单2项阳性(43.90%)和全阴(36.58%)为主。结论 单ELISA+结果存在较多假阳性,导致不必要的血液报废;而NAT-标本可能存在低水平的HBV DNA,产生漏检风险。建议针对单HBsAg ELISA+/NAT-献血者,采用多套系统多种方法追溯检测,提高献血者HBV筛查的准确度,减少不必要的血液浪费。

中年男性精子DNA碎片率对体外受精-胚胎移植妊娠结局的影响

目的探讨中年男性精子DNA碎片率(DFI)与精液质量和体外受精-胚胎移植妊娠结局的关系。方法共收集180例接受常规体外受精-胚胎移植治疗且男性年龄>38岁的精液标本。根据DFI的阈值分成(<30%和≥30%)两组。主要测量指标包括:常规精液参数、激素水平、DFI、受精率、优质胚胎率、临床妊娠率等。结果通过比较分析发现,DFI与男性卵泡刺激素(FSH)、精子活力、精子形态密切相关,且精子活力随着DFI水平的升高而下降(P<0.05);当DFI≥30%时,优质胚胎率下降(P<0.05),但两组的临床妊娠率差异无显著性(P>0.05)。结论DFI可以作为中年男性精液常规分析的重要参考指标,虽然DFI影响优质胚胎率,但与辅助生殖治疗的临床妊娠结局无关。

完带汤防治脾虚湿盛型外阴阴道假丝酵母菌病的临床疗效及对DNA损伤的影响

目的探讨完带汤治疗脾虚湿盛型外阴阴道假丝酵母菌病(Vulvovaginal candidiasis,VVC)的临床疗效及对DNA损伤的影响。方法将符合纳入标准的70例脾虚湿盛VVC患者随机分为完带汤组和氟康唑组各35例,治疗期间2组各脱落5例。氟康唑组采用150 mg氟康唑口服一次;完带汤组采用完带汤口服14 d。治疗后比较2组患者中医证候积分变化情况;评估2组患者临床治愈(Test of cure,TOC)率及临床缓解(Clinical improvement,CI)率;比色法检测阴道灌洗液中8-羟基脱氧鸟苷(8-hydroxydeoxyguanosine,8-OHDG)水平变化以评估DNA损伤情况;治疗后3月评估患者访视临床完全缓解(Follow up,FU)、真菌学转阴及复发率。结果治疗后,2组患者中医证候积分均呈现不同程度改善(P<0.05,P<0.01);完带汤组优于氟康唑组(P<0.05,P<0.01);完带汤组TOC、CI、真菌学转阴率与氟康唑组未见明显差异(P>0.05);但完带汤组FU及复发率均显著优于氟康唑组(P<0.05,P<0.01)。治疗后,氟康唑组阴道灌洗液中8-OHDG表达显著增加(P<0.001),完带汤组未见明显变化,显著低于氟康唑组(P<0.001)。结论完带汤总体临床疗效与氟康唑相当,但在改善中医证候、防止复发方面优于氟康唑,同时具有不加剧阴道细胞DNA损伤的优势。

不同运动方式对人体DNA损伤、DNA甲基化和端粒长度的影响

背景:运动不仅是改善身体健康和心理健康的有效手段,还对代谢性和心脑血管等疾病的发生、发展具有良好的干预效果,其原因与表观遗传因素有关。目的:总结不同运动方式对人体DNA损伤、DNA甲基化和端粒长度的影响,并分析运动调控表观遗传修饰的可能机制,以期为运动改善机体功能提供参考。方法:以“运动,有氧训练,急性运动,无氧训练,抗阻训练,DNA损伤,DNA甲基化,端粒”为中文检索词,以“exercise,sport,aerobic exercise,anaerobic exercise,resistance training,acute exercise,DNA methylation,DNA damage,telomere”为英文检索词。在PubMed、Embase、Web of Science、中国知网数据库中进行检索,并根据纳入与排除标准筛选文献,最终纳入70篇文献。结果与结论:①长期有氧、抗阻和无氧运动均能改善DNA损伤,其原因与运动可以提高机体的抗氧化能力有关。而急性运动则会通过上调活性氧和活性氮氧化物的表达进而加剧DNA损伤程度;②急性运动、长期抗阻运动和无氧运动在降低DNA甲基化方面具有积极作用,其关键机制可能是运动诱导的活性氧使氧化型谷胱甘肽/还原型谷胱甘肽比值和DNA甲基化转移酶、10-11易位酶的表达发生了改变,进而对DNA甲基化产生调控作用;③与其他运动形式相比,长时间有氧运动可能更具有增加端粒长度的潜在价值,其中的生物学机制涉及炎症、氧化应激、DNA甲基化和微小RNA的表达调控;④基于当前文献可知,有氧运动持续2年以上可以增加端粒长度,未来的研究也应进一步明确最佳的运动持续时间。

基于混沌理论与DNA动态编码的卫星图像加密算法

针对卫星图像在传输、存储过程中涉及的信息安全问题,该文提出一种新型的基于混沌理论与DNA动态编码的卫星图像加密算法。首先,提出一种改进型无限折叠混沌映射,拓宽了原有无限折叠混沌映射的混沌区间。之后,结合改进型Chebyshev混沌映射与SHA-256哈希算法,生成加密算法的密钥流,提升算法的明文敏感性。然后,利用混沌系统的状态值对Hilbert局部置乱后的像素进行DNA编码,实现DNA动态编码,解决了DNA编码规则较少所带来的容易受到暴力攻击的弱点。最后,使用混沌序列完成进一步混沌加密,从而彻底混淆原始像素信息,增加加密算法的随机性与复杂性,得到密文图像。实验结果表明,该算法具有较好的加密效果和应对各种攻击的能力。

环境DNA技术发展及其在长江流域水生生态学领域的应用研究进展

长江流域鱼类资源丰富、生物多样性高。近年来,受人为干扰、环境变化等因素影响,鱼类资源急剧衰退,全面了解该流域水生生态学信息、进行长江大保护迫在眉睫。随着分子生物学技术的发展,环境DNA技术应运而生,其相比于传统调查方式更加高效、灵敏,应用领域更广;该技术的灵敏性使其非常适合于检测濒危物种、低密度物种入侵、瞬时和隐秘物种的存在,特别是当检测低密度物种的采样工作难以控制时,其敏感性、简便性和降低危害性的优势愈加显现出来。因此,该技术已被广泛应用于食品微生物、生物监测、群落生态学、古环境、保护生物学和生物入侵等领域的研究。介绍了环境DNA定义、发展史、研究方法与优劣势,在此基础上概述了其在长江流域水生生态学领域的应用研究进展,最后展望了环境DNA技术与环境RNA技术相结合的技术革新以及新一代测序手段、大数据及机器智能技术多技术结合助力该领域研究的前景,以期为长江流域持续性生态学监测提供借鉴和参考。

DNA条形码技术在蛇类鉴别中的应用

从NCBI中GenBank数据库下载蛇类细胞色素b基因(Cytb)序列7329条,以MT765098.1序列为标准进行对比和修剪,获得蛇类Cytb序列4665条。对蛇类Cytb序列进行核苷酸饱和度、遗传多样性、种内和种间遗传距离计算,构建系统发育树。结果表明,基于Kimura–2–Parameter模型,蛇类平均种内遗传距离为3.3%,普遍小于6.7%,而平均种间遗传距离为19.96%,普遍高于9.3%,说明蛇类物种间遗传距离存在较大差异。根据蛇类物种间遗传距离,识别出蛇类23个物种的亚种,Pareas和Hydrophis属物种含有复合体,Atractus dunni、A.iridescen和A.occidentali互为姐妹物种。

围绕FFPE样本特性的DNA纯化纳米磁珠优化

目的 探讨围绕福尔马林固定石蜡包埋(formalin fixed paraffin embedded, FFPE)样本特性获取更高质量/得率的纳米磁珠核酸提取方案,改进分子病理技术。方法 合成4大类15个小类的备选磁珠,以FFPE样本为中心,筛选高质量/得率磁珠。模拟常规组织、粗针穿刺(肝脏)、纤维支气管镜样本(肺),装管相同张数连片。采用筛选的最佳磁珠与市场出售的常见磁珠试剂提取核酸,横向对比纯化总量、片段大小等质量参数。应用PCR和Sanger验证核酸的下游应用。结果 以FFPE样本DNA为中心筛选自制纳米磁珠,获得最佳性能纳米磁珠总回收率为58.5%±1.58%,5种市售商品化磁珠和3种国产磁珠总回收率为18.68%~40.71%。相同组织量(连片)提取的DNA总量,在模拟常规组织、粗针穿刺和纤维支气管镜样本核酸得率比市售试剂盒提高39.49%~181.72%(P<0.05)。模拟纤维支气管镜样本1张(4μm)总量可达100 ng以上、5张总量可达400 ng以上。结论 以FFPE样本DNA为中心筛选的DNA纯化纳米磁珠,与商品化磁珠相比有较大提升,为临床分子病理检测的质量保证、自动化检测和项目拓展提供空间。

Taq DNA聚合酶的分子改造及其在探针法qPCR直扩体系中的应用

Taq DNA聚合酶作为实时荧光定量聚合酶链式反应(qPCR)技术的核心组分,其性能优劣直接影响qPCR技术的进一步发展。然而,野生型Taq DNA聚合酶的耐抑制剂性能差、延伸性能不足。为获得具有高性能的Taq DNA聚合酶,采用基因工程技术将双链DNA结合蛋白Sso7d或Sto7d融合在野生型Taq DNA聚合酶的N端或C端,构建了4个均可溶表达的改造体,再经过耐受性测试筛选较优的改造体,结果显示:改造体Taq-Sto的耐受性最高,其热稳定性不受影响,且在1 s/kbp的延伸条件下能成功扩增靶标,表明Taq-Sto具有增强的延伸性能,在TaqMan探针法qPCR体系中对腐殖酸、单宁酸、全血等抑制剂同样表现出良好的耐受性。EMSA实验发现:Taq-Sto对DNA模板的结合亲和力有所提高,有利于增强Taq-Sto对模板的竞争力;将Taq-Sto应用于非洲猪瘟病毒(ASFV)的TaqMan探针法qPCR检测,与商品化试剂相比,Taq-Sto具有更低的ASFV检出限,且在体积分数为2%~6%的猪粪便样本或猪肉样本中的检测灵敏度分别为100.0%和85.4%,说明Taq-Sto在直扩qPCR检测领域更具有优势。