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OBJECTIVE To investigate the improvement effect and mechanism of calycosin （CA） on acute inflammatory injury secondary to intracerebral hemorrhage. METHODS Male C57BL/6 mice were injected with type Ⅶ collagenase into the basal ganglia to establish an intracerebral hemorrhage model， which were divided into sham-operation group（phosphate buffered saline instead of collagenase）， model group， and different CA dose groups（15，30，60，120 mg/kg）. Based on the modified neurological severity score （mNSS） to screen the intervention doses， the volume of intracerebral hemorrhage， brain water content， the expressions of ionized calcium-binding adaptor molecule 1 （Iba1） in brain tissue， Toll-like receptor 4 （TLR4） and its downstream inflammatory factors [tumor necrosis factor-α （TNF-α）， inducible nitric-oxide synthase （iNOS）， interleukin-1β （IL- 1β）] in brain tissue， and the apoptosis of cells in brain tissue were detected. Primary microglia were cultured in vitro， and the expressions of TLR4 and its downstream inflammatory factors were detected. Primary neurons and primary microglia were co- cultured in vitro， and the apoptosis of neurons was detected. RESULTS The doses of 30 mg/kg and 60 mg/kg were selected as intervention doses of CA for subsequent experiments. Compared with the sham-operation group， the mice in the model group had cerebral hemorrhage， the volume of cerebral hemorrhage and brain water content were significantly increased （P＜0.05）； the positive expression rate of Iba1 protein in brain tissue was significantly increased， and the relative expression levels of TLR4， TNF-α， IL-1β and iNOS protein in brain tissue were up-regulated significantly. The apoptosis rate also increased significantly （P＜0.05）. Compared with model group， the above indexes of the mice in the 30 and 60 mg/kg CA groups were significantly improved （P＜0.05）. CA significantlyreduced the relative expression levels of TLR4 and its downstream inflammatory factors in microglia， and reduced the apoptosis of neurons in the co-culture system of primary neurons and primary microglia （P＜0.05）. CONCLUSIONS CA can exert a protective effect on the brain， which may be related to relieving the secondary acute inflammatory injury after intracerebral hemorrhage by inhibiting TLR4-mediated inflammatory response.
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The endangered medicinal plant Glehnia littoralis is one of the important natural source of furanocoumarin, which has been used as mucolytic, antitussive, antitumour and antibacterial. However, the genetic information of furanocoumarin biosynthesis in G. littoralis is scarce at present. The objective of this study was to mine the putative candidate genes involved in the biosynthesis pathway of furanocoumarin and provide references for gene identification, and functional genomics of G. littoralis. We carried out the transcriptome analysis of leaves and roots in G. littoralis, which provided a dataset for gene mining. Psoralen, imperatorin and isoimperatorin were detected in G. littoralis by high performance liquid chromatography analysis. Candidate key genes were mined based on the annotations and local BLAST with homologous sequences using BioEdit software. The relative expression of genes was analysed using quantitative real-time polymerase chain reaction. Further, the CYP450 genes were mined using phylogenetic analyses using MEGA 6.0 software. A total of 156,949 unigenes were generated, of which 9021 were differentially-expressed between leaves and roots. A total of 82 unigenes encoding eight enzymes in furanocoumarin biosynthetic pathway were first obtained. Seven genes that encoded key enzymes in the downstream furanocoumarin biosynthetic pathway and expressed more in roots than leaves were screened. Twenty-six candidate CYP450 unigenes expressed abundantly in roots and were chiefly concentrated in CYP71, CYP85 and CYP72 clans. Finally, we filtered 102 differentially expressed transcription factors (TFs) unigenes. The transcriptome of G. littoralis was characterized which would help to elucidate the furanocoumarin biosynthetic pathway in G. littoralis and provide an invaluable resource for further study of furanocoumarin.
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Scutellaria baicalensis (S. baicalensis) and Scutellaria barbata (S. barbata) are common medicinal plants of the Lamiaceae family. Both produce specific flavonoid compounds, including baicalein, scutellarein, norwogonin, and wogonin, as well as their glycosides, which exhibit antioxidant and antitumor activities. Here, we report chromosome-level genome assemblies of S. baicalensis and S. barbata with quantitative chromosomal variation (2n = 18 and 2n = 26, respectively). The divergence of S. baicalensis and S. barbata occurred far earlier than previously reported, and a whole-genome duplication (WGD) event was identified. The insertion of long terminal repeat elements after speciation might be responsible for the observed chromosomal expansion and rearrangement. Comparative genome analysis of the congeneric species revealed the species-specific evolution of chrysin and apigenin biosynthetic genes, such as the S. baicalensis-specific tandem duplication of genes encoding phenylalanine ammonia lyase and chalcone synthase, and the S. barbata-specific duplication of genes encoding 4-CoA ligase. In addition, the paralogous duplication, colinearity, and expression diversity of CYP82D subfamily members revealed the functional divergence of genes encoding flavone hydroxylase between S. baicalensis and S. barbata. Analyzing these Scutellaria genomes reveals the common and species-specific evolution of flavone biosynthetic genes. Thus, these findings would facilitate the development of molecular breeding and studies of biosynthesis and regulation of bioactive compounds.
Subject(s)
Evolution, Molecular , Flavonoids/biosynthesis , Genome, Plant , Plant Extracts/genetics , Scutellaria/metabolism , Whole Genome SequencingABSTRACT
Objective@#To investigate the application value of CTDIvol and size-specific dose estimate(SSDE) in evaluating the radiation dose of adult chest CT.@*Methods@#A retrospective analysis was made on the CTDIvoland SSDE of 128 patients who underwent chest CT scanning in the First Affiliated Hospital of Zhejiang Chinese Medical University from March to April 2017 and all images are adequate for diagnosis. The subjects were divided into three groups according to body mass index (BMI): Group A, 38 cases with 16≤BMI<21.1 kg/m2; Group B, 53 cases with 21.1≤BMI<23.9 kg/m2; Group C, 37 cases with 23.9≤BMI<34.1 kg/m2. The diameters of anterior-posterior (AP) and left-right (LAT) of each patient were measured in the slice of nipple level, and CTDIvol, effective diameter (ED), conversion factor (fsize) and SSDE of each patient were recorded and calculated. Meanwhile, the differences between CTDIvol and SSDE in different BMI groups were compared.@*Results@#SSDE of each group was higher than CTDIvol, and increased by 50.13%, 42.83% and 33.68% in group A, B and C, respectively. There were significant differences in radiation dose between CTDIvol and SSDE in the three groups (t=-48.873, -57.001, -32.651, P<0.05). There were significant differences in ED, fsize, CTDIvol and SSDE among the three groups (F=51.456, 47.749, 113.916, 106.449, P<0.05).@*Conclusions@#SSDE can evaluate the radiation dose absorbed by patients with different body types in chest CT, while CTDIvol underestimated the radiation dose.The lower BMI, the greater the underestimated dose value, the more radiation dose absorbed.
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Objective To investigate the application value of CTDIvol and size-specific dose estimate( SSDE) in evaluating the radiation dose of adult chest CT. Methods A retrospective analysis was made on the CTDIvol and SSDE of 128 patients who underwent chest CT scanning in the First Affiliated Hospital of Zhejiang Chinese Medical University from March to April 2017 and all images are adequate for diagnosis. The subjects were divided into three groups according to body mass index ( BMI):Group A,38 cases with 16≤BMI<21. 1 kg/m2; Group B,53 cases with 21. 1≤BMI<23. 9 kg/m2; Group C,37 cases with 23. 9≤BMI<34. 1 kg/m2. The diameters of anterior-posterior (AP) and left-right (LAT) of each patient were measured in the slice of nipple level, and CTDIvol , effective diameter ( ED) , conversion factor ( fsize ) and SSDE of each patient were recorded and calculated. Meanwhile, the differences between CTDIvol and SSDE in different BMI groups were compared. Results SSDE of each group was higher than CTDIvol , and increased by 50. 13％, 42. 83％ and 33. 68％ in group A, B and C, respectively. There were significant differences in radiation dose between CTDIvol and SSDE in the three groups ( t=-48. 873,-57. 001, -32. 651, P<0. 05) . There were significant differences in ED, fsize , CTDIvol and SSDE among the three groups ( F=51. 456, 47. 749, 113. 916, 106. 449, P<0. 05) . Conclusions SSDE can evaluate the radiation dose absorbed by patients with different body types in chest CT, while CTDIvol underestimated the radiation dose. The lower BMI, the greater the underestimated dose value, the more radiation dose absorbed.
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Tanshinones are a class of bioactive components in the traditional Chinese medicine , and their biosynthesis and regulation have been widely studied. Current studies show that basic leucine zipper (bZIP) proteins regulate plant secondary metabolism, growth and developmental processes. However, the bZIP transcription factors involved in tanshinone biosynthesis are unknown. Here, we conducted the first genome-wide survey of the bZIP gene family and analyzed the phylogeny, gene structure, additional conserved motifs and alternative splicing events in A total of 70 SmbZIP transcription factors were identified and categorized into 11 subgroups based on their phylogenetic relationships with those in . Moreover, seventeen genes underwent alternative splicing events. According to the transcriptomic data, the genes that were highly expressed in the Danshen root and periderm were selected. Based on the prediction of bZIP binding sites in the promoters and the co-expression analysis and co-induction patterns in response to Ag treatment quantitative real-time polymerase chain reaction (qRT-PCR), we concluded that and potentially participate in the regulation of tanshinone biosynthesis. These results provide a foundation for further functional characterization of the candidate genes, which have the potential to increase tanshinone production.
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Global concerns have been paid to the potential hazard of traditional herbal medicinal products (THMPs). Substandard and counterfeit THMPs, including traditional Chinese patent medicine, health foods, dietary supplements, etc. are potential threats to public health. Recent marketplace studies using DNA barcoding have determined that the current quality control methods are not sufficient for ensuring the presence of authentic herbal ingredients and detection of contaminants/adulterants. An efficient biomonitoring method for THMPs is of great needed. Herein, metabarcoding and single-molecule, real-time (SMRT) sequencing were used to detect the multiple ingredients in Jiuwei Qianghuo Wan (JWQHW), a classical herbal prescription widely used in China for the last 800 years. Reference experimental mixtures and commercial JWQHW products from the marketplace were used to confirm the method. Successful SMRT sequencing results recovered 5416 and 4342 circular-consensus sequencing (CCS) reads belonging to the ITS2 and regions. The results suggest that with the combination of metabarcoding and SMRT sequencing, it is repeatable, reliable, and sensitive enough to detect species in the THMPs, and the error in SMRT sequencing did not affect the ability to identify multiple prescribed species and several adulterants/contaminants. It has the potential for becoming a valuable tool for the biomonitoring of multi-ingredient THMPs.
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Drought stress could significantly affect the growth of medical plants, while moderate drought stress can effectively improve the quality of medicinal plants. Recently Herbgenomics promotes innovative research on the molecular mechanism for the quality of medicinal plants affected by drought stress. Here, we summarized the effects of drought stress on active compounds including alkaloids, flavonoids, terpenoids, phenols and saponins, and biomass of medicinal plants. Moreover, the studies on the quality of medicinal plants affected by drought stress using Herbgenomic technologies, the reactive oxygen species metabolism, and ABA signaling pathway, were reviewed. It is proposed to reveal the molecular mechanism for the quality of medicinal plants affected by drought stress, and to explore molecular markers closely related to drought-tolerance traits using medicinal model plant system based on Herbgenomics. This review provides a theoretical basis for improving the quality of medicinal plants, breeding of drought-tolerant medicinal plants and guiding agricultural production of traditional Chinese medicine.
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To develop monoclonal antibodies (McAbs) against chicken interleukin 4 (chIL-4), we subcloned the mature chIL-4 gene into prokaryotic expression vectors pET-28a and pGEX-6P-1, then expressed and purified the recombinant proteins. We immunized BALB/c mice with the purified His-chIL-4 protein and fused the murine splenocytes with SP2/0 after 4 times of immunization. We used the GST-chIL-4 protein as a coating antigen to establish an indirect ELISA to screen positive clones. After screening and 3 rounds of cloning process, we obtained 3 hybridomas that stably secreted McAbs against chIL-4, and named 1G11-3B, 2E5-3D, and 1G11-5H. The isotypes of these McAbs were all IgG1 and the dissociation constant (Kd) of these McAbs were 1.79×10⁻⁹, 1.61×10⁻⁹, and 2.36×10⁻⁹, respectively. These McAbs specifically bound to chIL-4 expressed by either prokaryotic or eukaryotic system as determined by Western blotting and indirect immunofluorescence assay. The binding domains of chIL-4 recognized by 1G11-3B, 2E5-3D, and 1G11-5H were located between aa 1-40, 80-112, and 40-80, respectively, as determined by Western blotting. These McAbs would help to detect chIL-4 and to elucidate the biological roles of chIL-4 in immune responses.
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Transcription factor is one of the key factors in the regulation of gene expression at the transcriptional level. It plays an important role in plant growth, active components biosynthesis and response to environmental change. This paper summarized the structure and classification of bHLH transcription factors and elaborated the research progress of bHLH transcription factors which regulate the active components in plants, such as flavonoids, alkaloids, and terpenoids. In addition, the possibility of increasing the concentration of active substances by bHLH in medicinal plants was assessed. The paper emphasized great significance of model plants and multidisciplinary research fields including modern genomics, transcriptomics, metabolomics and bioinformatics, providing the contribution to improve the discovery and function characterization of bHLH transcription factors. Accelerating the research in the mechanism of bHLH transcription factors on the regulation of active components biosynthesis will promote the development of breeding and variety improvement of Chinese medicinal materials, also ease the pressure of resources exhaustion of traditional Chinese medicine home and abroad.
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The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.
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@#Spinal cord compression injury (SCCI) is common in orthopedics manifested as system dysfunction, and even Results in disability and death. This article would review the recent researches for SCCI.