Jingyi Tu, Changqing Shen, Ruiling Lei, Jie Yang, Shicheng Wang, Siqi Peng, Lang Li, Xiaoyan Qiu
Embryo implantation involves a complex interaction between the embryo and the endometrium of the mother, the study of which faces a variety of problems. The modeling of endometrial epithelial organoids and endometrial assembloids provides a new way to study the process of embryo implantation in vitro. This paper summarized the latest research progress in embryo implantation, the regulation mechanism of endometrial receptivity by estrogen- progesterone coordination and embryo-derived signals, the establishment of endometrial organoids, and the development and application of endometrial assembloids in the research on mother-embryo interaction, providing new strategies for studying the communication between embryo and maternal uterus during implantation.
{"title":"[Research progress in animal embryo implantation and endometrial organoids].","authors":"Jingyi Tu, Changqing Shen, Ruiling Lei, Jie Yang, Shicheng Wang, Siqi Peng, Lang Li, Xiaoyan Qiu","doi":"10.13345/j.cjb.240128","DOIUrl":"https://doi.org/10.13345/j.cjb.240128","url":null,"abstract":"<p><p>Embryo implantation involves a complex interaction between the embryo and the endometrium of the mother, the study of which faces a variety of problems. The modeling of endometrial epithelial organoids and endometrial assembloids provides a new way to study the process of embryo implantation <i>in vitro</i>. This paper summarized the latest research progress in embryo implantation, the regulation mechanism of endometrial receptivity by estrogen- progesterone coordination and embryo-derived signals, the establishment of endometrial organoids, and the development and application of endometrial assembloids in the research on mother-embryo interaction, providing new strategies for studying the communication between embryo and maternal uterus during implantation.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4452-4466"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Argonaute proteins are active throughout the lifetime in a variety of organisms and they bind to small RNAs (sRNAs) to regulate gene expression. The Argonaute proteins of vertebrates can be classified into two clades: the Ago clade and the Piwi clade. Both clades have N, L1, L2, PAZ, MID and PIWI domains. The N domain is involved in the loading of sRNAs. L1 and L2 domains facilitate the linking between domains. The PAZ and MID domains exert functions by anchoring sRNAs. The PIWI domain of some Argonaute proteins has RNase H-like structure and exerts the endonuclease function. Ago proteins regulate gene expression at transcriptional and post-transcriptional levels. Piwi proteins mainly exist in the germ cells, silencing transposons in different ways to keep genome integrality and regulating gene expression. In recent years, great progress has been made in Argonaute proteins in terms of the crystal structures, functions, and expression patterns. By reviewing the relevant studies, we elaborate on the structures, sRNA dependence, gene expression regulation, and biological roles of the Ago and Piwi proteins in vertebrates, aiming to clarify the roles of Argonaute proteins in epigenetic regulation and provide a reference for further research and application of these proteins.
Argonaute蛋白在多种生物体的整个生命周期中都很活跃,它们与小rna (sRNAs)结合以调节基因表达。脊椎动物的Argonaute蛋白可分为两个分支:Ago分支和Piwi分支。两个分支都有N、L1、L2、PAZ、MID和PIWI结构域。N结构域参与srna的装载。L1和L2域促进了域之间的连接。PAZ和MID结构域通过锚定srna发挥功能。一些Argonaute蛋白的PIWI结构域具有RNase h -样结构并发挥核酸内切酶功能。Ago蛋白在转录和转录后水平调控基因表达。Piwi蛋白主要存在于生殖细胞中,通过不同方式沉默转座子,保持基因组完整性,调节基因表达。近年来,对Argonaute蛋白在晶体结构、功能和表达模式等方面的研究取得了很大进展。本文通过对相关研究的回顾,阐述了Ago和Piwi蛋白在脊椎动物中的结构、sRNA依赖性、基因表达调控及其生物学作用,旨在阐明Argonaute蛋白在表观遗传调控中的作用,为这些蛋白的进一步研究和应用提供参考。
{"title":"[Research progress in Argonaute proteins of vertebrates].","authors":"Xianhua Xie, Meng Ru, Jianling Peng, Linjian Weng, Jianzhen Huang","doi":"10.13345/j.cjb.240026","DOIUrl":"https://doi.org/10.13345/j.cjb.240026","url":null,"abstract":"<p><p>Argonaute proteins are active throughout the lifetime in a variety of organisms and they bind to small RNAs (sRNAs) to regulate gene expression. The Argonaute proteins of vertebrates can be classified into two clades: the Ago clade and the Piwi clade. Both clades have N, L1, L2, PAZ, MID and PIWI domains. The N domain is involved in the loading of sRNAs. L1 and L2 domains facilitate the linking between domains. The PAZ and MID domains exert functions by anchoring sRNAs. The PIWI domain of some Argonaute proteins has RNase H-like structure and exerts the endonuclease function. Ago proteins regulate gene expression at transcriptional and post-transcriptional levels. Piwi proteins mainly exist in the germ cells, silencing transposons in different ways to keep genome integrality and regulating gene expression. In recent years, great progress has been made in Argonaute proteins in terms of the crystal structures, functions, and expression patterns. By reviewing the relevant studies, we elaborate on the structures, sRNA dependence, gene expression regulation, and biological roles of the Ago and Piwi proteins in vertebrates, aiming to clarify the roles of Argonaute proteins in epigenetic regulation and provide a reference for further research and application of these proteins.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4418-4438"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Epigallocatechin gallate (EGCG), the predominant polyphenol in green tea, exerts a spectrum of physiological activities, including antioxidant, anticancer, and anti-inflammatory effects. Emerging research underscores the significance of EGCG in modulating oocyte aging. EGCG can enhance antioxidant defenses, improve mitochondrial functions, and inhibit apoptotic pathways, thereby retarding the aging of oocytes. This review delineates the main molecular features of EGCG and expounds its regulatory mechanisms concerning oocyte aging, enriching the knowledge on the role of EGCG in the amelioration of oocyte aging.
{"title":"[Research progress in biological activities and oocyte aging-regulating effect of EGCG].","authors":"Weiying Zhang, Huizhu Zhang, Yujun Li, Daoliang Lan, Xianrong Xiong, Yaying Wang, Jian Li, Honghong He","doi":"10.13345/j.cjb.240192","DOIUrl":"https://doi.org/10.13345/j.cjb.240192","url":null,"abstract":"<p><p>Epigallocatechin gallate (EGCG), the predominant polyphenol in green tea, exerts a spectrum of physiological activities, including antioxidant, anticancer, and anti-inflammatory effects. Emerging research underscores the significance of EGCG in modulating oocyte aging. EGCG can enhance antioxidant defenses, improve mitochondrial functions, and inhibit apoptotic pathways, thereby retarding the aging of oocytes. This review delineates the main molecular features of EGCG and expounds its regulatory mechanisms concerning oocyte aging, enriching the knowledge on the role of EGCG in the amelioration of oocyte aging.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4382-4395"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jing Hong, Yongyong Dai, Qijun Nie, Zhiqiang Liao, Liangcai Peng, Dan Sun
The rapid development of modern industries is accompanied with the aggravating water heavy metal pollution, which poses a potential threat to the aquatic environment and the health of local populations. As an efficient and economical adsorbent, biochar demonstrates the adsorption capacity for heavy metal ions and its adsorption capacity is significantly enhanced after modification. Therefore, biochar can effectively mitigate environmental pollution. By reviewing the existing studies, we summarize the modification methods of biochar, compare the advantages and disadvantages of physical, biological, and chemical modification methods, analyze the effects of modification on the adsorption capacity of biochar for heavy metal ions, and expound the modification mechanism of biochar. On this basis, this article puts forward the future research directions of the application of biochar in treating coexisting pollutants, aiming to provide a reference for the application of biochar in the purification of heavy metal-containing wastewater.
{"title":"[Research progress in the adsorption of heavy metal ions from wastewater by modified biochar].","authors":"Jing Hong, Yongyong Dai, Qijun Nie, Zhiqiang Liao, Liangcai Peng, Dan Sun","doi":"10.13345/j.cjb.240263","DOIUrl":"https://doi.org/10.13345/j.cjb.240263","url":null,"abstract":"<p><p>The rapid development of modern industries is accompanied with the aggravating water heavy metal pollution, which poses a potential threat to the aquatic environment and the health of local populations. As an efficient and economical adsorbent, biochar demonstrates the adsorption capacity for heavy metal ions and its adsorption capacity is significantly enhanced after modification. Therefore, biochar can effectively mitigate environmental pollution. By reviewing the existing studies, we summarize the modification methods of biochar, compare the advantages and disadvantages of physical, biological, and chemical modification methods, analyze the effects of modification on the adsorption capacity of biochar for heavy metal ions, and expound the modification mechanism of biochar. On this basis, this article puts forward the future research directions of the application of biochar in treating coexisting pollutants, aiming to provide a reference for the application of biochar in the purification of heavy metal-containing wastewater.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4467-4479"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Dan Chen, Jiani Yuan, Xiaojun Deng, Lei Ding, Zhongwei An, Wen Luo
This study aims to optimize the process for preparing chitosan-based ultrasound-coupled hydrogel pads and investigate their application potential in ultrasonography. Chitosan, 2-acrylamido-2-methylpropanesulfonic acid, and N-isopropylacrylamide were used as the main materials to prepare chitosan-based ultrasound-coupled hydrogel pads. The free-radical polymerization conditions were optimized by a three-factor, three-level orthogonal test with the tensile strength and ultrasound image quality of the hydrogel pads as evaluation indicators. The optimal prescription was selected by optimizing three factors of raw material ratio, polymerization temperature, and freeze-drying time. The structure and performance of the hydrogel pads were characterized by a scanning electron microscope, a universal testing machine, and an ultrasonic diagnostic instrument. The results showed that the optimal prescription was as follows: the chitosan: 2-acrylamide-2-methylpropanesulfonic acid: N-isopropylacrylamide ratio of 2:0.55:17.27, the polymerization temperature of 25 ℃, and the freeze-drying time of 48 h. The ultrasonically-coupled hydrogel pads prepared under these conditions were transparent, with a porous structure, good adhesion, and high tensile strength. The hydrogel pads had good swelling properties and the swelling degree decreased slowly on day 10. The quality of the ultrasound images obtained via chitosan-based hydrogel pads was not significantly different from that obtained via medical ultrasound coupling agent. In this study, we analyzed the effects of different preparation processes on the gel formation of chitosan-based ultrasound-coupled hydrogel pads. The hydrogel pads were transparent and mild and non-irritating to the human body, serving as an ultrasound transmission material for ultrasonography.
{"title":"[Preparation and methodological analysis of chitosan-based ultrasound-coupled hydrogel pads].","authors":"Dan Chen, Jiani Yuan, Xiaojun Deng, Lei Ding, Zhongwei An, Wen Luo","doi":"10.13345/j.cjb.240586","DOIUrl":"https://doi.org/10.13345/j.cjb.240586","url":null,"abstract":"<p><p>This study aims to optimize the process for preparing chitosan-based ultrasound-coupled hydrogel pads and investigate their application potential in ultrasonography. Chitosan, 2-acrylamido-2-methylpropanesulfonic acid, and N-isopropylacrylamide were used as the main materials to prepare chitosan-based ultrasound-coupled hydrogel pads. The free-radical polymerization conditions were optimized by a three-factor, three-level orthogonal test with the tensile strength and ultrasound image quality of the hydrogel pads as evaluation indicators. The optimal prescription was selected by optimizing three factors of raw material ratio, polymerization temperature, and freeze-drying time. The structure and performance of the hydrogel pads were characterized by a scanning electron microscope, a universal testing machine, and an ultrasonic diagnostic instrument. The results showed that the optimal prescription was as follows: the chitosan: 2-acrylamide-2-methylpropanesulfonic acid: N-isopropylacrylamide ratio of 2:0.55:17.27, the polymerization temperature of 25 ℃, and the freeze-drying time of 48 h. The ultrasonically-coupled hydrogel pads prepared under these conditions were transparent, with a porous structure, good adhesion, and high tensile strength. The hydrogel pads had good swelling properties and the swelling degree decreased slowly on day 10. The quality of the ultrasound images obtained <i>via</i> chitosan-based hydrogel pads was not significantly different from that obtained <i>via</i> medical ultrasound coupling agent. In this study, we analyzed the effects of different preparation processes on the gel formation of chitosan-based ultrasound-coupled hydrogel pads. The hydrogel pads were transparent and mild and non-irritating to the human body, serving as an ultrasound transmission material for ultrasonography.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4586-4593"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897263","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The fiber type transition of skeletal muscle is an intricate and essential physiological process in the body, significantly influencing both the function and metabolism of skeletal muscle. This phenomenon is not only affected by external environmental changes but also intricately regulated by internal physiological mechanisms. Therefore, exploring the physiological process of muscle fiber type transition holds considerable significance for the treatment of human neuromuscular disorders and the improvement of meat quality in livestock and poultry. It has been discovered that the cytokines secreted by skeletal muscle, i.e., myokines, play a role in the fiber type transition of skeletal muscle. Myokines mainly act on skeletal muscle in autocrine and paracrine forms to participate in signal transduction and regulate the fiber type transition of skeletal muscle. This paper reviews the functional differences among various muscle fiber types, expounds the effects and mechanisms of myokines in regulating the transition processes of these fiber types, and prospects the future research directions in this field. This review is expected to provide theoretical support for enhancing the meat quality of livestock and poultry and treating skeletal muscle-related diseases.
{"title":"[Progress and prospects of the effects and mechanisms of myokines in regulating fiber type transition of skeletal muscle].","authors":"Boyu Huang, Ziyi Zhang, Weijun Pang","doi":"10.13345/j.cjb.240729","DOIUrl":"https://doi.org/10.13345/j.cjb.240729","url":null,"abstract":"<p><p>The fiber type transition of skeletal muscle is an intricate and essential physiological process in the body, significantly influencing both the function and metabolism of skeletal muscle. This phenomenon is not only affected by external environmental changes but also intricately regulated by internal physiological mechanisms. Therefore, exploring the physiological process of muscle fiber type transition holds considerable significance for the treatment of human neuromuscular disorders and the improvement of meat quality in livestock and poultry. It has been discovered that the cytokines secreted by skeletal muscle, <i>i</i>.<i>e</i>., myokines, play a role in the fiber type transition of skeletal muscle. Myokines mainly act on skeletal muscle in autocrine and paracrine forms to participate in signal transduction and regulate the fiber type transition of skeletal muscle. This paper reviews the functional differences among various muscle fiber types, expounds the effects and mechanisms of myokines in regulating the transition processes of these fiber types, and prospects the future research directions in this field. This review is expected to provide theoretical support for enhancing the meat quality of livestock and poultry and treating skeletal muscle-related diseases.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4365-4381"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897267","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lipid nanoparticles serve as a promising drug delivery system due to the good biocompatibility, non-immunogenicity, and high drug loading efficiency. However, unmodified lipid nanoparticles have limitations such as poor stability, easy hydrolysis, and rapid removal. To overcome these shortcomings, researchers have developed peptide modification, antibody modification, ligand modification, nucleic acid aptamer modification, and polysaccharide modification for lipid nanoparticles. Polysaccharides are a class of natural polymers, and the polysaccharide-modified lipid nanoparticles exhibit good biocompatibility, precise targeting, and low toxicity. Therefore, polysaccharide-modified lipid nanoparticles demonstrate great potential in clinical treatment. This review summarizes the preparation and application of polysaccharide-modified lipid nanoparticles, aiming to provide a reference for further research and development of new lipid nanoparticles.
{"title":"[Research progress in polysaccharide-modified lipid nanoparticles for drug delivery].","authors":"Yuqing Ma, Haiyun Liu, Xiaoqiang Wang, Shiqi Sun, Huichen Guo","doi":"10.13345/j.cjb.240188","DOIUrl":"https://doi.org/10.13345/j.cjb.240188","url":null,"abstract":"<p><p>Lipid nanoparticles serve as a promising drug delivery system due to the good biocompatibility, non-immunogenicity, and high drug loading efficiency. However, unmodified lipid nanoparticles have limitations such as poor stability, easy hydrolysis, and rapid removal. To overcome these shortcomings, researchers have developed peptide modification, antibody modification, ligand modification, nucleic acid aptamer modification, and polysaccharide modification for lipid nanoparticles. Polysaccharides are a class of natural polymers, and the polysaccharide-modified lipid nanoparticles exhibit good biocompatibility, precise targeting, and low toxicity. Therefore, polysaccharide-modified lipid nanoparticles demonstrate great potential in clinical treatment. This review summarizes the preparation and application of polysaccharide-modified lipid nanoparticles, aiming to provide a reference for further research and development of new lipid nanoparticles.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4339-4350"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The effects of host factors ADP-ribosylation factor 4 (ARF4) and ADP-ribosylation factor 5 (ARF5) upon Zika virus (ZIKV) infection in vivo were characterized by construction of gene knockout mice via CRISPR-Cas9. Firstly, ARF5 and ARF4 genes were modified by the CRISPR-Cas9 system and then microinjected into the fertilized eggs of C57BL/6JGpt mice. Fertilized eggs were transplanted to obtain ARF4 or ARF5 knockout (ARF4KO or ARF5KO) mice, and ARF4/5 double knockout mice were achieved by the mating between ARF4KO and ARF5KO mice (ARF4KO/ARF5KO). Then, the mouse genotypes were identified by PCR to identify the positive knockout mice, and RT-qPCR was employed to examine the knockout efficiency. The mice were then infected with ZIKV and the blood and tissue samples were collected after 2, 4, and 6 days. RT-qPCR was then employed to determine the virus load, and hematoxylin-eosin staining was employed to observe the pathological changes in the tissue. The results showed that expected PCR bands were detected from ARF4KO-/+, ARF5KO-/-, and ARF4KO-/+/ARF5KO-/- mice, respectively. The results of mRNA transcription measurement indicated the significant knockdown of ARF4 by 37.8%-50.0% but not ARF5 in ARF4KO-/+ compared with the wild-type mice. Meanwhile, complete knockout of ARF5 and no changes in ARF4 were observed in ARF5KO-/- mice. Additionally, completed knockout of ARF5 and down-regulated mRNA level of ARF4 in the lung, kidney, and testis were detected in ARF4KO-/+/ARF5KO-/-mice in comparison with the wild-type mice. The virus load in the serum decreased in ARF4KO-/+ mice, while it showed no significant change in ARF5KO-/- or ARF4KO-/+/ARF5KO-/- mice compared with that in the wild type. Meanwhile, ARF4KO-/+ mice showcased no significant difference in virus load in various tissues but attenuated pathological changes in the brain and testis compared with the wild-type mice. We successfully constructed ARF4KO and ARF5KO mice by CRISPR-Cas9 in this study. ARF4 rather than ARF5 is essential for ZIKV infection in vivo. This study provided animal models for studying the roles of ARF4 and ARF5 in ZIKV infection and developing antivirals.
{"title":"[Characterization of host factors <i>ARF4</i> and <i>ARF5</i> upon Zika virus infection <i>in vivo</i> by construction of gene knockout mice].","authors":"Kao Deng, Mingyuan Li, Huiying Zhang, Yongqiang Deng, Yuan Qin, Chengfeng Qin","doi":"10.13345/j.cjb.240307","DOIUrl":"https://doi.org/10.13345/j.cjb.240307","url":null,"abstract":"<p><p>The effects of host factors ADP-ribosylation factor 4 (ARF4) and ADP-ribosylation factor 5 (ARF5) upon Zika virus (ZIKV) infection <i>in vivo</i> were characterized by construction of gene knockout mice via CRISPR-Cas9. Firstly, <i>ARF5</i> and <i>ARF4</i> genes were modified by the CRISPR-Cas9 system and then microinjected into the fertilized eggs of C57BL/6JGpt mice. Fertilized eggs were transplanted to obtain <i>ARF4</i> or <i>ARF5</i> knockout (ARF4KO or ARF5KO) mice, and <i>ARF4</i>/<i>5</i> double knockout mice were achieved by the mating between ARF4KO and ARF5KO mice (ARF4KO/ARF5KO). Then, the mouse genotypes were identified by PCR to identify the positive knockout mice, and RT-qPCR was employed to examine the knockout efficiency. The mice were then infected with ZIKV and the blood and tissue samples were collected after 2, 4, and 6 days. RT-qPCR was then employed to determine the virus load, and hematoxylin-eosin staining was employed to observe the pathological changes in the tissue. The results showed that expected PCR bands were detected from ARF4KO<sup>-/+</sup>, ARF5KO<sup>-/-</sup>, and ARF4KO<sup>-/+</sup>/ARF5KO<sup>-/-</sup> mice, respectively. The results of mRNA transcription measurement indicated the significant knockdown of <i>ARF4</i> by 37.8%-50.0% but not <i>ARF5</i> in ARF4KO<sup>-/+</sup> compared with the wild-type mice. Meanwhile, complete knockout of <i>ARF5</i> and no changes in <i>ARF4</i> were observed in ARF5KO<sup>-/-</sup> mice. Additionally, completed knockout of <i>ARF5</i> and down-regulated mRNA level of <i>ARF4</i> in the lung, kidney, and testis were detected in ARF4KO<sup>-/+</sup>/ARF5KO<sup>-/-</sup>mice in comparison with the wild-type mice. The virus load in the serum decreased in ARF4KO<sup>-/+</sup> mice, while it showed no significant change in ARF5KO<sup>-/-</sup> or ARF4KO<sup>-/+</sup>/ARF5KO<sup>-/-</sup> mice compared with that in the wild type. Meanwhile, ARF4KO<sup>-/+</sup> mice showcased no significant difference in virus load in various tissues but attenuated pathological changes in the brain and testis compared with the wild-type mice. We successfully constructed ARF4KO and ARF5KO mice by CRISPR-Cas9 in this study. ARF4 rather than ARF5 is essential for ZIKV infection <i>in vivo</i>. This study provided animal models for studying the roles of ARF4 and ARF5 in ZIKV infection and developing antivirals.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4605-4615"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To construct a recombinant Bacillus subtilis strain expressing SpaA and CbpB of Erysipelothrix rhusiopathiae for oral administration, we constructed the recombinant plasmid pDG1730-CBJA by fusion PCR and seamless cloning. The plasmid was introduced into B. subtilis KC strain by natural transformation, and the recombinant strain KC-spaA-cbpB was screened out on the plate containing spectinomycin (sper) and confirmed by PCR and starch degradation test. The SpaA and CbpB expressed by KC-spaA-cbpB were detected by Western blotting and indirect immunofluorescence assay, and the genetic stability of the recombinant strain in mice was determined. The plasmid pMAD-∆sper with knockout of sper was constructed and transformed into KC-spaA-cbpB. The sper-deleted mutant strain KC-spaA-cbpB: : ∆sper was screened and identified, and its immunogenicity in a mouse model was evaluated by oral immunization. The results showed that the recombinant strain KC-spaA-cbpB was stable in mice, expressing SpaA on the cell surface and CbpB on the spore surface. KC-spaA-cbpB: : ∆sper expressed SpaA and CbpB. The mice vaccinated with the spores of KC-spaA-cbpB: : ∆sper had higher levels of SpaA and CbpB-specific IgG in the serum that those vaccinated with the wild-type spores 42 days after vaccination by gavage (P < 0.01). The protective rate of mice immunized with the recombinant spores was 67.5%. The results indicated that a recombinant B. subtilis strain expressing SpaA and CbpB of E. rhusiopathiae was successfully constructed, and the recombinant strain laid a foundation for the development of oral live vector vaccines for swine erysipelas.
{"title":"[Construction of a recombinant <i>Bacillus subtilis</i> strain expressing SpaA and CbpB of <i>Erysipelothrix rhusiopathiae</i> and evaluation of the strain immunogenicity in a mouse model].","authors":"Zhonglin Cheng, Hao Huang, Siyi Cao, Huahui Shi, Jiye Gao, Jixiang Li","doi":"10.13345/j.cjb.240456","DOIUrl":"https://doi.org/10.13345/j.cjb.240456","url":null,"abstract":"<p><p>To construct a recombinant <i>Bacillus subtilis</i> strain expressing SpaA and CbpB of <i>Erysipelothrix rhusiopathiae</i> for oral administration, we constructed the recombinant plasmid pDG1730-CBJA by fusion PCR and seamless cloning. The plasmid was introduced into <i>B</i>. <i>subtilis</i> KC strain by natural transformation, and the recombinant strain KC-<i>spaA</i>-<i>cbpB</i> was screened out on the plate containing spectinomycin (<i>spe</i><sup><i>r</i></sup>) and confirmed by PCR and starch degradation test. The SpaA and CbpB expressed by KC-<i>spaA</i>-<i>cbpB</i> were detected by Western blotting and indirect immunofluorescence assay, and the genetic stability of the recombinant strain in mice was determined. The plasmid pMAD-∆<i>spe</i><sup>r</sup> with knockout of <i>spe</i><sup><i>r</i></sup> was constructed and transformed into KC-<i>spaA-cbpB</i>. The <i>spe</i><sup><i>r</i></sup>-deleted mutant strain KC-<i>spaA</i>-<i>cbpB</i>: : ∆<i>spe</i><sup>r</sup> was screened and identified, and its immunogenicity in a mouse model was evaluated by oral immunization. The results showed that the recombinant strain KC-<i>spaA</i>-<i>cbpB</i> was stable in mice, expressing SpaA on the cell surface and CbpB on the spore surface. KC-<i>spaA</i>-<i>cbpB</i>: : ∆<i>spe</i><sup>r</sup> expressed SpaA and CbpB. The mice vaccinated with the spores of KC-<i>spaA</i>-<i>cbpB</i>: : ∆<i>spe</i><sup>r</sup> had higher levels of SpaA and CbpB-specific IgG in the serum that those vaccinated with the wild-type spores 42 days after vaccination by gavage (<i>P</i> < 0.01). The protective rate of mice immunized with the recombinant spores was 67.5%. The results indicated that a recombinant <i>B</i>. <i>subtilis</i> strain expressing SpaA and CbpB of <i>E</i>. <i>rhusiopathiae</i> was successfully constructed, and the recombinant strain laid a foundation for the development of oral live vector vaccines for swine erysipelas.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4521-4532"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Anqi Huang, Yinfeng Liang, Sirui Wang, Runrun She, Jin Yan, Yingyu Wang, Luyao Zhang, Mingchun Liu
Mastoparans (MP), a class of α-helix cationic insect-derived antimicrobial peptides, have a broad spectrum of biological activities including inhibiting bacteria, fungi, viruses, and parasites. Amino acid substitution, peptide modification, peptide chain cyclization, and dosage form modification can enhance the biological activities and target and reduce the toxicity of mastoparans. In this review, we summarize the structure, biological function and modification methods of mastoparans, and prospect the development of antibacterial drugs based on mastoparans, so as to provide reference for the research of mastoparans as a new antibacterial drug.
{"title":"[Research progress in mastoparans].","authors":"Anqi Huang, Yinfeng Liang, Sirui Wang, Runrun She, Jin Yan, Yingyu Wang, Luyao Zhang, Mingchun Liu","doi":"10.13345/j.cjb.240482","DOIUrl":"https://doi.org/10.13345/j.cjb.240482","url":null,"abstract":"<p><p>Mastoparans (MP), a class of α-helix cationic insect-derived antimicrobial peptides, have a broad spectrum of biological activities including inhibiting bacteria, fungi, viruses, and parasites. Amino acid substitution, peptide modification, peptide chain cyclization, and dosage form modification can enhance the biological activities and target and reduce the toxicity of mastoparans. In this review, we summarize the structure, biological function and modification methods of mastoparans, and prospect the development of antibacterial drugs based on mastoparans, so as to provide reference for the research of mastoparans as a new antibacterial drug.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"40 12","pages":"4408-4417"},"PeriodicalIF":0.0,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142897290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}