Transcriptomic Insights and Cytochrome P450 Gene Analysis in Kadsura coccinea for Lignan Biosynthesis.

Kadsura coccinea is a medicinal plant from the Schisandraceae family that is native to China and has great pharmacological potential due to its lignans. However, there are significant knowledge gaps regarding the genetic and molecular mechanisms of lignans. We used transcriptome sequencing technology to analyze root, stem, and leaf samples, focusing on the identification and phylogenetic analysis of Cytochrome P450 (CYP) genes. High-quality data containing 158,385 transcripts and 68,978 unigenes were obtained. In addition, 36,293 unigenes in at least one database, and 23,335 across five databases (Nr, KEGG, KOG, TrEMBL, and SwissProt) were successfully annotated. The KEGG pathway classification and annotation of these unigenes identified 10,825 categorized into major metabolic pathways, notably phenylpropanoid biosynthesis, which is essential for lignan synthesis. A key focus was the identification and phylogenetic analysis of 233 Cytochrome P450 (CYP) genes, revealing their distribution across 38 families in eight clans, with roots showing specific CYP gene expression patterns indicative of their role in lignan biosynthesis. Sequence alignment identified 22 homologous single genes of these CYPs, with 6 homologous genes of CYP719As and 1 of CYP81Qs highly expressed in roots. Our study significantly advances the understanding of the biosynthesis of dibenzocyclooctadiene lignans, offering valuable insights for future pharmacological research and development.

Embryological observations on seed abortion in Hibiscus syriacus L. and physiological studies on nutrients, enzyme activity and endogenous hormones.

Under natural conditions, most Hibiscus syriacus L. individuals form very few mature seeds or the mature seeds that do form are of poor quality. As a result, seed yield is poor and seeds have low natural germinability. These phenomena strongly hinder utilization of the excellent germplasm resources of H. syriacus. The study has shown that pollen activity and stigma receptivity were high on the day of anthesis, and the pistils and stamens were fertile. Pollen release and stigma receptivity were synchronous. But in styles following self and cross-pollination, pollen tube abnormalities (distortion and twisting of the pollen tubes) and callose deposition were observed. Cross-pollinated pollen tubes elongated faster and fewer pollen tube abnormalities were observed compared with self-pollinated pollen tubes. And during embryo development, abnormalities during the heart-shaped embryo stage led to embryo abortion. Imbalance in antioxidant enzyme activities and low contents of auxin and cytokinin during early stages of embryo development may affect embryo development. Therefore, a low frequency of outcrossing and mid-development embryo abortion may be important developmental causes of H. syriacus seed abortion. Nutrient deficiencies, imbalance in antioxidant enzyme activities, and a high content of abscisic acid at advanced stages of seed development may be physiological causes of seed abortion.

Transcriptomic Analysis Reveals Key Candidate Genes Related to Seed Abortion in Chinese Jujube (Ziziphus jujuba Mill).

Background: Seed abortion is a common phenomenon in Chinese jujube that seriously hinders the process of cross-breeding. However, the molecular mechanisms of seed abortion remain unclear in jujube. Methods: Here, we performed transcriptome sequencing using eight flower and fruit tissues at different developmental stages in Ziziphus jujuba Mill. 'Zhongqiusucui' to identify key genes related to seed abortion. Histological analysis revealed a critical developmental process of embryo abortion after fertilization. Results: Comparisons of gene expression revealed a total of 14,012 differentially expressed genes. Functional enrichment analyses of differentially expressed genes between various sample types uncovered several important biological processes, such as embryo development, cellular metabolism, and stress response, that were potentially involved in the regulation of seed abortion. Furthermore, gene co-expression network analysis revealed a suite of potential key genes related to ovule and seed development. We focused on three types of candidate genes, agamous subfamily genes, plant ATP-binding cassette subfamily G transporters, and metacaspase enzymes, and showed that the expression profiles of some members were associated with embryo abortion. Conclusion: This work generates a comprehensive gene expression data source for unraveling the molecular mechanisms of seed abortion and aids future cross-breeding efforts in jujube.

3D printable Sodium alginate-Matrigel (SA-MA) hydrogel facilitated ectomesenchymal stem cells (EMSCs) neuron differentiation.

Ectomesenchymal stem cells (EMSCs) are typical adult stem cells obtained from the cranial neural crest. They have the potential to differentiate into various cell types, such as osseous cells, neurons and glial cells. Three-dimensional (3 D) printing is a novel method to construct biological structures by rapid prototyping. Previously, our group reported on the stemness and multi-lineage differentiation potential of EMSCs on gels. However, the exploration of EMSCs in 3 D printing and then evaluation of the growth and neuronal differentiation of EMSCs on extruded 3 D printable hybrid hydrogels has not been reported. Therefore, the current study explored the novel hybrid Sodium alginate-Matrigel (SA-MA) hydrogel extruded 3 D printing to design an in vitro scaffold to promote the differentiation and growth of EMSCs. In addition, the physical properties of the hydrogel were characterized and its drug-releasing property determined. Notably, the results showed that the construct exhibited a sustain-released effect of growth factor BDNF in accordance with the Higuchi equation. Moreover, the cell survival rate on the 3 D printed scaffold was 88.22 ± 1.13% with higher neuronal differentiation efficiency compared with 2 D culture. Thus, SA-MA's ability to enhanced EMSCs neuronal differentiation offers a new biomaterial for neurons regeneration in the treatment of spinal cord injury.

EMSCs Build an All-in-One Niche via Cell-Cell Lipid Raft Assembly for Promoted Neuronal but Suppressed Astroglial Differentiation of Neural Stem Cells.

The therapeutic efficiency of allogenic/intrinsic neural stem cells (NSCs) after spinal cord injury is severely compromised because the hostile niche at the lesion site incurs massive astroglial but not neuronal differentiation of NSCs. Although many attempts are made to reconstruct a permissive niche for nerve regeneration, solely using a living cell material to build an all-in-one, multifunctional, permissive niche for promoting neuronal while inhibiting astroglial differentiation of NSCs is not reported. Here, ectomesenchymal stem cells (EMSCs) are reported to serve as a living, smart material that creates a permissive, all-in-one niche which provides neurotrophic factors, extracellular matrix molecules, cell-cell contact, and favorable substrate stiffness for directing NSC differentiation. Interestingly, in this all-in-one niche, a corresponding all-in-one signal-sensing platform is assembled through recruiting various niche signaling molecules into lipid rafts for promoting neuronal differentiation of NSCs, and meanwhile, inhibiting astrocyte overproliferation through the connexin43/YAP/14-3-3θ pathway. In vivo studies confirm that EMSCs can promote intrinsic NSC neuronal differentiation and domesticating astrocyte behaviors for nerve regeneration. Collectively, this study represents an all-in-one niche created by a single-cell material-EMSCs for directing NSC differentiation.

Porphyra polysaccharide-derived carbon dots for non-viral co-delivery of different gene combinations and neuronal differentiation of ectodermal mesenchymal stem cells.

In this study, multifunctional fluorescent carbon dots (CDs) were synthesized using a one-pot hydrothermal carbonization reaction, with the naturally-occurring porphyra polysaccharide (PPS) serving as a single carbon source for the first time and ethylenediamine (Ed) acting as the surface passivation agent. The resulting CDs enjoyed a high quantum yield (56.3%), excitation-dependent fluorescence, small size (<10 nm), spherical shape, uniform distribution, positive surface charge, low cytotoxicity and excellent ability to condense macromolecular plasmid DNA. The synthesized CDs were employed for neuronal induction from ectodermal mesenchymal stem cells for the first time via highly efficient non-viral gene delivery. The optimal combination of factors (Ascl1 and Brn2) was selected from seven different combinations out of Ascl1, Brn2 and Sox2 according to the expression of neuronal markers (Tuj1, Map2 and Tau). The results of qRT-PCR demonstrated that the CDs possessed a significantly higher transfection efficiency than the commercially available transfection reagents PEI (25 kDa) and Lipofectamine2000. Moreover, the CDs/pDNA nanoparticles exhibited more efficient neuronal differentiation of the EMSCs than the AT-RA-containing induction medium. Furthermore, the CDs/pDNA nanoparticles could enter cells via both caveolae- and clathrin-mediated endocytosis. Taken together, the natural polysaccharide PPS-derived CDs enriched the current application of CDs by employing the CDs as a novel non-viral gene carrier for neuronal differentiation of adult stem cells, which held great promise in tissue engineering and bioimaging.

A pilot study of perioperative esmolol for myocardial protection during on-pump cardiac surgery.

The protective effects of preprocedural esmolol on myocardial injury and hemodynamics have not, to date, been investigated in patients who were scheduled for cardiac surgeries under a cardiopulmonary bypass (CPB). A pilot randomized controlled trial was performed at The First Affiliated Hospital of Dalian Medical University (Dalian, China). Patients scheduled for elective open-heart surgeries under CBP were included, and were randomized to esmolol and control groups. For patients in the esmolol groups, intravenous esmolol (70 µg/kg/min) was administered at the time of incision until CPB was performed. For patients assigned to the control group, equal volumes of 0.9% saline were administered. Markers of myocardial injury and hemodynamic parameters were observed until 12 h post surgery. A total of 24 patients were included in the present study. No significant differences in hemodynamic parameters, including the central venous pressure and heart rate, were detected between patients in the two groups during the perioperative period or within the first 12 h post-surgery (P>0.05), except for the mean arterial pressure, which was higher in the esmolol group compared with the control group at 5 and 12 h post-surgery (P<0.05). However, the serum level of cardiac troponin I was higher in patients of the control group compared with those of the esmolol group during the preoperative period (P<0.05). Although creatinine kinase was significantly different at T2 between the two groups, its MB isoenzyme was not significantly different between the groups (P>0.05). In addition, administration of esmolol was not associated with an increased risk for severe complications and adverse events in these patients. In conclusion, preoperative esmolol may be an effective and safe measure of myocardial protection for patients who undergo elective cardiac surgeries under CBP.

[Cloning and characterization of a transcription factor ZmNAC1 in maize (Zea mays)].

NAC transcription factors are a family of functionally diverse proteins. They are unique to plants and play an important role in regulation of plant growth and development, hormone regulation and responses to various stresses. A cDNA encoding the NAC-like gene homologue was isolated from maize (Zea mays L.) by RT-PCR and designated ZmNAC1 (GenBank Accession No. EU224278). Sequence analysis showed that cDNA of ZmNAC1 was 1,029 bp long and contained a single open reading frame (ORF, 26 to approximately 907 bp). The predicted ZmNAC1 protein has 293 amino acids with an estimated molecular mass of 32.3 kDa and an isoelectric point of 8.65. RT-PCR analysis showed that the expression of ZmNAC1 was induced by low temperature, PEG, salt, and ABA, respectively. These results suggest that ZmNAC1 may play important roles in biotic and abiotic resistance pathways. This is the first NAC-like gene reported in maize.