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@#Abstract: Approximately 2 billion people worldwide are infected with helminths and the resulting helminthiasis is a heavy health burden for developing countries. Parasitic helminths are a class of multicellular parasites, mainly including trematodes, tapeworms and nematodes, wtih complex life cycle involving multiple developmental stages and typically one or more hosts. Understanding the growth, development, pathogenesis and transmission of these parasites at the molecular level is of great significance for the diagnosis and treatment of helminthiasis. Genetic manipulations, which alter the expression level of target genes, have greatly promoted the biomedical research. In recent years, with the release of genomic data of worms, genetic manipulation techniques, such as RNA interference and clustered regularly interspaced short palindromic repeats (CRISPR) gene editing, have been increasingly applied in the studies of parasitic helminths. This article reviews the progress of genetic manipulations in important medical worms, as well as the methods of genetic manipulations, which would be expected to inspire the future functional study in parasitic helminths.
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Genetic robustness refers to the fact that an organism has a buffer system that can maintain normal development, even if the existence of genetic mutations during biological development. Previous research on the underlying mechanism of genetic robustness mainly involves in genetic redundancy and distributed robustness, both of which are triggered at the protein level. Recently, a new genetic compensation response (GCR) mechanism has been discovered in zebrafish, which occurs in knockout rather than knockdown individuals and is triggered upstream of protein feedback regulation. Since there are many concepts related to genetic robustness, this review attempts to clarify these concepts from the types of compensation genes and triggering modes. Additionally, we aim to understand the latest discovered GCR mechanism and look forward to studying the function of specific genes based on functional compensation mechanisms.
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OBJECTIVES@#To explore the effect of acupuncture and moxibustion on intestinal flora in the rats with diarrhea-predominant irritable bowel syndrome (IBS-D) based on 16S rDNA technique.@*METHODS@#Ten rats were randomized from 58 SPF-grade male SD rats to be the blank group. The remained 48 rats were prepared to be IBS-D models by the modified method of acetic acid enema combined with binding tail-clip stress. Forty successfully-modeled rats were randomly divided into a model group, an acupuncture group, a moxibustion group and a western medication group, with 10 rats in each one. In the acupuncture group, the needle was inserted at bilateral "Zusanli" (ST 36) and remained for 15 min in each rat. In the moxibustion group, the suspending moxibustion was delivered at bilateral "Zusanli" (ST 36) for 15 min. The rats in the western medication group were given pinaverium bromide suspension (10 mL/kg) by intragastric administration. The above interventions were performed once daily for consecutive 14 days. The body mass and the score of fecal trait were compared before and after modeling, as well as after intervention in each group. Fecal water content, diarrhea index and colon transit time (CTT) were measured after modeling and intervention in the rats of each group separately. After intervention, the colonic morphology of rats in each group was observed, and using 16S rDNA technique, the intestinal flora was detected.@*RESULTS@#After modeling, compared with the blank group, the body mass and CTT were reduced (P<0.01); fecal trait scores, fecal water contents and diarrhea index increased (P<0.01) in the other 4 groups. After intervention, the body mass and CTT of the rats decreased (P<0.01), and fecal trait score, fecal water content and diarrhea index increased (P<0.01) in the model group compared with those in the blank group. In the acupuncture group, the moxibustion group and the western medication group, when compared with the model group, the body mass and CTT were elevated (P<0.01), while fecal trait scores, fecal water contents and diarrhea index declined (P<0.01). Compared with the western medication group, fecal water content decreased in the acupuncture group and the moxibustion group (P<0.05), while CTT increased in the acupuncture group (P<0.01), the body mass increased and fecal trait score was dropped in the moxibustion group (P<0.05). The colonic mucosa structure was clear and complete, and there was no obvious inflammatory cell infiltration in the blank group. The mild interstitial edema of intestinal mucosa was presented with the infiltration of few inflammatory cells in the model group. There was the infiltration of few inflammatory cells in the mucosa of the acupuncture group, the moxibustion group and the western medication group. Compared with the blank group, the indexes of Richness, Chao1, ACE and Shannon decreased in the model group (P<0.05). Indexes of Richness, Chao1 and ACE increased in the acupuncture group and the moxibustion group (P<0.05), and the Richness index in the western medication group increased (P<0.05) when compared with those in the model group. The relative abundance of Bacteroidetes, Proteobacteria and Prevotella increased (P<0.05), and that of Firmicutes and Muribaculaceae decreased (P<0.05) in the model group compared with those in the blank group. When compared with the model group, the relative abundance of Bacteroidetes, Proteobacteria and Prevotella was reduced (P<0.05), while that of Firmicutes and Muribaculaceae increased (P<0.05) in the acupuncture group, the moxibustion group and the western medication group; and that of Actinobacteria and Bifidobacterium increased in the acupuncture group and the moxibustion group (P<0.05). Compared with the blank group, the relative abundance of lipopolysaccharide (LPS) biosynthesis was elevated (P<0.05), and that of folate biosynthesis, lipoic acid metabolism, zeatin biosynthesis, ubiquinone and other terpenoid quinone biosynthesis decreased (P<0.05) in the model group. The relative abundance of LPS biosynthesis was dropped (P<0.05), and that of folate biosynthesis, lipoic acid metabolism, zeatin biosynthesis, ubiquinone and other terpenoid quinone biosynthesis increased (P<0.05) in the acupuncture group, the moxibustion group and the western medication group compared with those of the model group.@*CONCLUSIONS@#Either acupuncture or moxibustion can relieve the symptoms of IBS-D and protect intestinal mucosa, which may be associated with regulating the structure of intestinal flora and promoting nutrient metabolism and biosynthesis.
Subject(s)
Rats , Male , Animals , Irritable Bowel Syndrome/therapy , Moxibustion/methods , Rats, Sprague-Dawley , Gastrointestinal Microbiome , Lipopolysaccharides , Thioctic Acid , Ubiquinone , Zeatin , Acupuncture Therapy , Diarrhea/therapy , Terpenes , Water , Folic Acid , Acupuncture PointsABSTRACT
Abiotic stresses, predominately drought, heat, salinity, cold, and waterlogging, adversely affect cereal crops. They limit barley production worldwide and cause huge economic losses. In barley, functional genes under various stresses have been identified over the years and genetic improvement to stress tolerance has taken a new turn with the introduction of modern gene-editing platforms. In particular, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is a robust and versatile tool for precise mutation creation and trait improvement. In this review, we highlight the stress-affected regions and the corresponding economic losses among the main barley producers. We collate about 150 key genes associated with stress tolerance and combine them into a single physical map for potential breeding practices. We also overview the applications of precise base editing, prime editing, and multiplexing technologies for targeted trait modification, and discuss current challenges including high-throughput mutant genotyping and genotype dependency in genetic transformation to promote commercial breeding. The listed genes counteract key stresses such as drought, salinity, and nutrient deficiency, and the potential application of the respective gene-editing technologies will provide insight into barley improvement for climate resilience.
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【Objective】 To analyze the changes of microRNA (miRNA) expression profiles on day 1 and day 5 after storage with or without riboflavin and ultraviolet-B (UVB) light (VB2-PRT) treatment, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL) under VB2-PRT treatment. 【Methods】 20 apheresis platelet concentrates (5mL / sample), collected from voluntary donors, were split into two group after mixing and agitation. One was treated with riboflavin (final concentration 50 μmol/L) plus 6.24 J/mL UVB light(E group), and the other worked as a control group (C group) without any treatment. Both groups were subjected to agitated storage at (22±2) ℃ horizontally. The platelet concentrates were sampled on d1 and d5 (5mL) during storage, named as E1, E5, C1 and C5 groups, respectively, and sequenced by DNA nanoball (DNB) sequencing technology. The differentially expressed miRNAs between E and C groups were screened by using DEGseq and MA-plot analysis software, and GO function enrichment analysis and KEGG pathway enrichment analysis were further performed when the different expression between groups reached twofold and above. 【Results】 Compared with C1 group, 487 miRNAs with significantly different expression (P<0.01) were screened in E1 group, including 220 up-regulated miRNAs, such as miR-146a and let-7b, and 267 down regulated miRNAs, such as miR-7 and miR-1260. Compared with C5 group, 229 miRNAs with significantly different expression (P<0.01) were screened in E5, including 80 miRNAs with up-regulated expression, such as miR-423 and miR-378, and 149 down regulated miRNAs, such as miR-451 and miR-30.The target genes with differentially expressed miRNAs in E1 vs C1 groups and E5 vs C5 groups were similar in the numbers of enriched GO terms, including cell components, organelles, cell membrane and other cell structures, molecular functions such as adhesion, catalysis, molecular transformation, transportation, transcription factors and receptor activity, cell processing, metabolism, biological regulation, stress and other biological processes etc. Compared with E1 and C1 groups, E5 and C5 groups lacked of signal pathways related to environmental adaptation, translation and mucin synthesis, however, it increased inositol phosphate metabolism, phosphatidylinositol signaling system and chemokine signaling pathway. 【Conclusion】 The expression profiles of platelets miRNAs treated with VB2-PRT has changed significantly after storage for a period of time. Functional prediction suggests that these miRNAs might be involved in the regulation of platelet PSL induced by VB2-PRT.
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【Objective】 To investigate the changes of platelet microRNA (miRNA) expression profiles on d1 and d5 during storage with riboflavin and ultraviolet-B (UVB) light (VB2-PRT) treatment, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL) under VB2-PRT treatment. 【Methods】 20 apheresis platelet concentrates (5mL / sample) were collected from voluntary blood donors. After mixing and shaking, the samples was treated with riboflavin (final concentration 50 μmol/L) and 6.24J/mL UVB light for 8min, then split into two aliquots and agitated stored at (22±2) ℃. The concentrates were sampled (5mL) on d1 and d5, respectively, and sequenced by DNA nanoball (DNB) sequencing technology. The differentially expressed miRNAs between the two groups (at different storage periods) were screened by DEGseq and MA-plot analysis software. The miRNAs, reached more than 2 times different expression between groups, were considered significant different(P<0.01). The miRanda and TargetScan softwares were used to predict the target genes. Gene Ontology (GO) function enrichment analysis and Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed on the target genes of significant differentially expressed miRNAs. The expression of miRNAs was verified by real-time fluorescence quantitative PCR (qRT-PCR). 【Results】 miRNA expression profile: compared with d1 platelets, there were 590 miRNAs with significantly different expression (P< 0.01) in d5 group, including 255 up-regulated miRNAs (such as miR-99b, miR-7) and 335 down regulated miRNAs (such as miR-451a, miR-19b). Among the 272 known miRNAs, 112 were up-regulated and 160 were down regulated. There were 318 new miRNAs sequences. The enriched GO terms of target genes of differentially expressed miRNAs in d5 and d1 groups included cell components, organelles, cell membranes and other cellular structures, molecular functions such as adhesion, catalysis, molecular conversion, transportation, transcription factor and receptor activity, and biological processes such as cell processing, metabolism, biological regulation and stress. The corresponding pathways in the top 10 of KEGG enrichment were mainly secretion, glucose metabolism, signal transduction, membrane transport, translation, environmental adaptation and other signal pathways. The six randomly selected differentially expressed miRNAs verified by qRT-PCR was consistent with those of DNB sequencing. 【Conclusion】 The expression profiles of platelets miRNAs has changed significantly between d1- and d5-storage under VB2-PRT treatment. Functional prediction suggests that these miRNAs might be involved in the regulation of platelet PSL underVB2-PRT treatment.
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Triterpenoids are active constituents of many medicinal plants. The biosynthetic pathways of triterpenes have been fully studied and significant progress has been made. CYP450s are mainly involved in the post-modification process of triterpenoids biosynthesis, which plays a key role in the diversity of triterpenoids. Currently, a lot of CYP450s, which related to the biosynthesis of triterpenoids, have been cloned from many plants. In this paper, CYP450s involved in the biosynthesis of triterpenoids were reviewed, which provided a reference for the analysis of the biosynthetic pathway of triterpenoids and the functional study of CYP450s.
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Targeted replacement genome editing refers to DNA modification and engineering technology that could induce and achieve mutations of targeted gene replacement or knockin at a target gene or DNA region. In this review, the principles, implementation methods, factors that influence efficiency and accuracy, and applications of gene replacement editing were summarized and discussed. It provides the reference for gene functional characterization and genetic improvements through gene replacement strategies in higher plant especially crops.
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[ ABSTRACT] AIM:To detect the differentially expressed genes associated with ovarian serous cystadenocarcino -ma ( OV) by microarray and to analyze the participated signaling pathway .METHODS:We analyzed 16 datasets of Affy-metrix GeneChip Human Exon 1.0 ST Arrays from The Cancer Genome Atlas ( TCGA) , including 8 OV and 8 normal ovary samples.The function of differential genes was determined by pathway and gene ontology ( GO) analysis.The probable functions of the key genes were predicted according to intergenic signal transduction network .RESULTS:The 1 144 genes were identified as distinctively expressed in OV (P<0.05), 747 of which were up-regulated and 397 were down-regula-ted.The GO analysis results showed that the altered genes were involved in 362 up-regulated and 160 down-regulated sig-nificant functions (P<0.05) related to cell cycle, DNA replication, cell proliferation, cell apoptosis, cell adhesion, etc. The pathways of the different genes were involved in the 59 enrichment-related pathways (P<0.05), 45 of which were up-regulated and 14 were down-regulated.Among the 59 pathways, cell cycle, P53 signaling pathway, DNA replication, path-ways in cancer, PI3K-Akt signaling pathway, ECM-receptor signaling pathway, cell adhesion molecules and cell apoptosis were related to tumor genesis , development and metastasis .As a result, 229 genes with significant functions and pathways in GO and pathway analysis were selected to construct signal transduction network ( Signal-Net), 4 of which, CDK1, PLK1, MCM3 and PGK1, were found to play key roles in OV signal regulation network .CONCLUSION: The OV shows abundant differentially expressed genes that play key roles in cancer-related signal pathways .
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RNA interference (RNAi) is a gene-silencing technology by which small double-stranded RNAs are used to target the degradation of RNA with complementary sequence. RNAi is found in a wide variety of organisms (Caenorhabditis elegans, insects, plants, microorganisms and animals). With RNAi, we have harnessed the gene function to be explored, revolutionized our ability to perform large-scale genetic screens, and even therapeutic potential.
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Biology , Insecta , RNA , RNA Interference , RNA, Double-StrandedABSTRACT
AIM: To study the expression and function of novel gene AngRem104. METHODS: Northern blot was performed to detect the distribution of AngRem104 in human multiple normal tissues as well as the effect of AngⅡ and AT1R antagonist (losartan) on AngRem104 expression. The sense and antisense eukaryotic expression vectors of AngRem104 were constructed and transfected into human mesangial cells. RT-PCR was used to detect the expression of FN when AngRem104 was over-expressed. Primary sequence and motif analysis of AngRem104 protein were performed by on-line ExPasy predictive tools. RESULTS: AngRem104 was predicted to localize at the cellular nucleus. It was widely expressed in human heart, placenta, liver, muscle, kidney and pancreas. Moreover, the up-regulated expression of AngRem104 induced by AngⅡ was inhibited by losartan in a dose-dependent manner. CONCLUSION: AngRem104 is a novel nuclear protein related to the expression of fibronectin and could be up-regulated by AngⅡ in human MC.