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italic>Rehmannia glutinosa belongs to the Scrophulariaceae family with important medicinal value. In order to effectively explore the transcriptome information of R. glutinosa and identify the genes encoding enzymes involved in phenylethanol glycoside (PhGs) biosynthesis, the leaves, stems and tuberous roots of R. glutinosa were used for transcriptome sequencing using Pacific Biosiences RS II platform. A total of 27 773 transcripts were generated with an average length of 2 380 bp, and 27 236 coding sequences (CDS) were predicted. Using BLAST software, non-redundant transcript sequences were annotated with NR, NT, GO, COG, KEGG, SwissProt and Interpro databases and a total of 27 399 annotated genes were obtained. Among them, the number of genes related to Sesamum indicum in the NR database was the highest (81.44%), which is consistent with their evolutionary relationship. Enzymes likely involved in the biosynthesis of isoacteoside, echinacoside, cistanosides A, cistanosides F, 2′-acetylacteoside and leonoside F were identified, and 143 genes were identified in R. glutinosa full-length transcriptome. The expression levels of 19 genes correlated with acteoside content in twelve tissues of R. glutinosa, and most showed higher expression levels in leaf tissues and floral organs. This study provides more reliable transcriptome data for screening R. glutinosa for functional genes and provides a foundation for the study of the molecular mechanisms of PhGs biosynthesis.
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Objective:To study the expression characteristics of myocardial strain index after the abnormal origin of the left coronary artery of the pulmonary artery in children was repaired.Methods:The data of 30 children (study group) with abnormal origin of pulmonary artery left coronary artery repair from August 2017 to August 2021 were analyzed. In addition, healthy children during the same period were selected as the control group, and the study group was compared before and after treatment and the control group. Circumferential and radial peak myocardial strain index, post-contraction strain index.Results:The longitudinal, circumferential, and radial overall peak strain indexes of the study group before and after treatment were significantly lower than those of the control group, and the longitudinal, circumferential, and radial overall peak strain indexes of the study group after treatment were significantly higher than those before treatment ( P<0.05); The longitudinal, circumferential, and radial peak strain indexes of the study group before treatment were significantly lower than those of the control group. After treatment in the study group, the middle section of the longitudinal inferior wall, the middle section of the anterior wall, the basal section of the anterior wall, the apex, and the circumferential direction were significantly lower The peak strain index of the basal segment of the inferior wall and the middle segment of the inferior wall was significantly lower than that of the control group; and the longitudinal, circumferential, and radial peak strain indexes of the study group after treatment were significantly higher than those before treatment ( P<0.05); the study group children before treatment Longitudinal, circumferential, and radial PSI indexes were significantly lower than those of the control group. After treatment, the study group was treated in the longitudinal inferior wall, septal apical segment, circumferential inferior wall basal segment, inferior wall middle segment, and radial PSI anterior wall basal segment, apex. The part was significantly higher than that of the control group; and the longitudinal, circumferential, and radial PSI of the study group after treatment were significantly lower than before treatment ( P<0.05). Conclusion:After ALCAPA repair, the overall and regional strain and overall synchronization are improved, indicating that the resting myocardium has recovered, but the strain of certain segments supplied by the abnormal left coronary artery fails to normalize after ALCAPA repair. Persistent myocardial injury is consistent, which can provide some guidance for the prognosis assessment of children with ALCAPA.
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Glutamine synthetase is a key enzyme in plant nitrogen assimilation. To study the structure of wheat glutamine synthetase isoenzymes, GS1, GSr, GSe, GS2 and GS2p of wheat were cloned into pET-21a, and the expression condition was optimized. Although wheat glutamine synthetase isoenzymes had 70%-80% amino acid sequence homology, the isoforms expressed with different characteristics. Induced at 30 °C, the most expression level of GSr, GSe and GS2 was after 3 h, and of GS1 was at the 7 h whereas no GS2p was expressed, and the GS isoenzymes showed different expression level, with the order of GS1 (22%)>GSr (15%)>GS2 (12%)>GSe (5%). GSe expressed as soluble protein, and GS1 expressed mainly as soluble protein whereas GSr and GS2 expressed as insoluble proteins. Induced at 30 °C for 3 h, mRNA transcript levels of GS isoforms were different, with the order of GSr (7.59)>GS2 (1.84)>GS2p (1.66)>GSe (1.46)>GS1 (1.00). The levels of mRNA transcription were not consistent with the level of the protein translation. The analysis of mRNA secondary structure showed the free energy of translation initiation region of glutamine synthetase isoforms was different, with the order of GS1 (14.4)<GSr (17.2)<GS2 (22.6) <GSe (25.4) <GS2p (31.6), the smaller freed energy, the more unstable mRNA secondary structure of translation initiation region and the higher level of protein expression. Soluble expression condition of glutamine synthetase isozymes was also different, with GS1, GSr, GSe and GS2 induced at 30 °C for 5 h, 16 °C for 15 h, 37 °C for 5 h, and 25 °C for 7 h respectively. The soluble protein showed different expression level with GS1 (20%)>GSr (13%)>GS2 (10%)>GSe (7%), and different activities with GS1>GSe>GS2, and the activity of GSr was not detected. The gene sequence of glutamine synthetase isoenzymes determines the amount, status and activity of proteins expressed in prokaryotic cells.
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To study the effects of Acaí on biological expression characteristics in rats with deficiency-heat and deficiency-cold syndromes, SD rats were divided into blank group, deficiency-heat model group, deficiency-heat+Phellodendri Chinensis Cortex group, deficiency-heat+Acaí high dose and low dose groups, deficiency-cold model group, deficiency-cold+Cinnamomi Cortex group, deficiency-cold+Acaí high dose and low dose groups. The rats were treated with intramuscular injection of hydrocortisone (20 mg•kg⁻¹) or dexamethasone sodium phosphate (0.35 mg•kg⁻¹) for 21 days to set up deficiency-heat model and deficiency-cold models. The levels of cAMP, cGMP, T3, T4 and rT3 were detected by radioimmunoassay. The levels of TP, UA, TC, TG and ALB were detected by colorimetry. The level of cAMP, cAMP/cGMP in serum were reduced in Acaí high dose group (P<0.05, P<0.001). The levels of T3, T4 and rT3 were significantly reduced in the Acaí high dose group (P<0.01, P<0.001, P<0.05). The levels of TP, UA, TC, TG and ALB were significantly reduced in the Acaí high dose group (P<0.001, P<0.05, P<0.05, P<0.05, P<0.01). However, Acaí had no obvious effects on deficiency-cold models. Acaí showed the same effect with Phellodendri Chinensis Cortex in adjusting the levels of deficiency-heat rats; but unlike Cinnamomi Cortex, Acaí showed no obvious effects in adjusting the levels of deficiency-cold rats.
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Objective: To screen the reference genes of Lonicera macranthoides for gene expression analysis and to study the spatio-temporal expression characteristics of LmAGL15 which was a member of Mads-Box family. Methods: In this study, 18 S rRNA, Ubiquilin, Actin and Efl-β of L. macranthoides were cloned and the stabilities of the four housekeeping genes were evaluated in different positions (leaves, stems, and buds) and different periods of bud development. In addition, the spatio-temporal expression of LmAGL15 gene was analyzed. Results: 18 S rRNA was the most suitable reference gene for spatio-temporal expression analysis in L. macranthoides; The relative expression of LmAGL15 was low in leaves and stems, and that in buds was higher. Conclusion: 18 S rRNA is the most suitable reference gene in L. macranthoides. The relative expression of LmAGL15 changes significantly in leaves, stems, and buds.
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Objective: To screen the reference genes of Siraitia grosvenorii for gene expression analysis, and to study the spatio-temporal expression characteristics of 3-hydroxy-3-methylglutaryl coenzyme A (HMGR) which was the key enzyme of mogroside V biosynthesis. Methods: In this study, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), microtubule associated protein (α-tubulin), actin (β-actin), and ubiquitin (UBQ5) fragments of S. grosvenorii were cloned, and the stabilities of the four housekeeping genes were evaluated in different positions (leaves, stems, and fruits) and different periods of fruit development. In addition, the spatio-temporal expression of HMGR gene was analyzed. Results: UBQ5 was the most suitable reference gene for spatio-temporal expression analysis in S. grosvenorii; The relative expression of HMGR was low in leaves, and that in stems and fruits was higher; The relative expression quantity of HMGR in fruits showed the fluctuation changes that increased firstly and then decreased, then increased and decreased again; The highest expression of HMGR was at 70 d of fruit development period, followed by 5, 30, 10, and 50 d. Conclusion: UBQ5 is the most suitable reference gene in S. grosvenorii. The relative expression of HMGR changes significantly in leaves, stems, and fruits. The relative expression quantity of HMGR in fruits shows the fluctuation changes, which is similar to synthetic accumulation pattern of mogroside V.