MIR396-GRF/GIF enhances in planta shoot regeneration of Dendrobium catenatum.

Recent studies on co-transformation of the growth regulator, TaGRF4-GIF1 chimera (Growth Regulating Factor 4-GRF Interacting Factor 1), in cultivated wheat varieties (Triticum aestivum), showed improved regeneration efficiency, marking a significant breakthrough. Here, a simple and reproducible protocol using the GRF4-GIF1 chimera was established and tested in the medicinal orchid Dendrobium catenatum, a monocot orchid species. TaGRF4-GIF1 from T. aestivum and DcGRF4-GIF1 from D. catenatum were reconstructed, with the chimeras significantly enhancing the regeneration efficiency of D. catenatum through in planta transformation. Further, mutating the microRNA396 (miR396) target sites in TaGRF4 and DcGRF4 improved regeneration efficiency. The target mimicry version of miR396 (MIM396) not only boosted shoot regeneration but also enhanced plant growth. Our methods revealed a powerful tool for the enhanced regeneration and genetic transformation of D. catenatum.

Whole Genome Duplication Events Likely Contributed to the Aquatic Adaptive Evolution of Parkerioideae.

As the only aquatic lineage of Pteridaceae, Parkerioideae is distinct from many xeric-adapted species of the family and consists of the freshwater Ceratopteris species and the only mangrove ferns from the genus Acrostichum. Previous studies have shown that whole genome duplication (WGD) has occurred in Parkerioideae at least once and may have played a role in their adaptive evolution; however, more in-depth research regarding this is still required. In this study, comparative and evolutionary transcriptomics analyses were carried out to identify WGDs and explore their roles in the environmental adaptation of Parkerioideae. Three putative WGD events were identified within Parkerioideae, two of which were specific to Ceratopteris and Acrostichum, respectively. The functional enrichment analysis indicated that the lineage-specific WGD events have played a role in the adaptation of Parkerioideae to the low oxygen concentrations of aquatic habitats, as well as different aquatic environments of Ceratopteris and Acrostichum, such as the adaptation of Ceratopteris to reduced light levels and the adaptation of Acrostichum to high salinity. Positive selection analysis further provided evidence that the putative WGD events may have facilitated the adaptation of Parkerioideae to changes in habitat. Moreover, the gene family analysis indicated that the plasma membrane H+-ATPase (AHA), vacuolar H+-ATPase (VHA), and suppressor of K+ transport growth defect 1 (SKD1) may have been involved in the high salinity adaptation of Acrostichum. Our study provides new insights into the evolution and adaptations of Parkerioideae in different aquatic environments.

Neottiamaolanensis, a replacement name for Neottiabifida M.N.Wang (Orchidaceae).

According to Articles 53.1 of the International Code of Nomenclature for Algae, Fungi, and Plants (Shenzhen Code), Neottiabifida M.N.Wang (as 'bifidus'; PhytoKeys 229: 222, 2023) is an illegitimate name, and hence a new name Neottiamaolanensis M. N. Wang is proposed here.

Impeding DNA Polymerase ß Activity by Oleic Acid to Inhibit Base Excision Repair and Induce Mitochondrial Dysfunction in Hepatic Cells.

Free fatty acids (FFAs) hepatic accumulation and the resulting oxidative stress contribute to several chronic liver diseases including nonalcoholic steatohepatitis. However, the underlying pathological mechanisms remain unclear. In this study, we propose a novel mechanism whereby the toxicity of FFAs detrimentally affects DNA repair activity. Specifically, we have discovered that oleic acid (OA), a prominent dietary free fatty acid, inhibits the activity of DNA polymerase ß (Pol ß), a crucial enzyme involved in base excision repair (BER), by actively competing with 2'-deoxycytidine-5'-triphosphate. Consequently, OA hinders the efficiency of BER, leading to the accumulation of DNA damage in hepatocytes overloaded with FFAs. Additionally, the excessive presence of both OA and palmitic acid (PA) lead to mitochondrial dysfunction in hepatocytes. These findings suggest that the accumulation of FFAs hampers Pol ß activity and contributes to mitochondrial dysfunction, shedding light on potential pathogenic mechanisms underlying FFAs-related diseases.

Neottiabifidus (Orchidaceae, Epidendroideae, Neottieae), a new mycoheterotrophic species from Guizhou, China.

Neottiabifidus, a new mycoheterotrophic orchid, found in Maolan National Nature Reserve in Guizhou Province, China, is described and illustrated here. The new species is close to N.nidus-avis, N.kiusiana and N.papilligera but differs in having a finely pubescent rachis with fewer flowers, a finely pubescent pedicel, and a fishtail-shaped lip that is deeply bilobed to the middle of the lip, with the lobes diverging at an acute angle (45°) to each other and mesochile with many papillae. Additionally, N.bifidus is well supported as a new species by molecular phylogenetic results based on ITS and chloroplast genome. The chloroplast genome of the novelty, which contains an LSC region of 33,819 bp, SSC region of 5,312 bp and IRs of 46,762 bp was assembled and annotated. A key to mycoheterotrophic Neottia species in China is also provided.

Metabolic Pathway Engineering Improves Dendrobine Production in Dendrobium catenatum.

The sesquiterpene alkaloid dendrobine, widely recognized as the main active compound and a quality control standard of medicinal orchids in the Chinese Pharmacopoeia, demonstrates diverse biological functions. In this study, we engineered Dendrobium catenatum as a chassis plant for the production of dendrobine through the screening and pyramiding of key biosynthesis genes. Initially, previously predicted upstream key genes in the methyl-D-erythritol 4-phosphate (MEP) pathway for dendrobine synthesis, including 4-(Cytidine 5'-Diphospho)-2-C-Methyl-d-Erythritol Kinase (CMK), 1-Deoxy-d-Xylulose 5-Phosphate Reductoisomerase (DXR), 2-C-Methyl-d-Erythritol 4-Phosphate Cytidylyltransferase (MCT), and Strictosidine Synthase 1 (STR1), and a few downstream post-modification genes, including Cytochrome P450 94C1 (CYP94C1), Branched-Chain-Amino-Acid Aminotransferase 2 (BCAT2), and Methyltransferase-like Protein 23 (METTL23), were chosen due to their deduced roles in enhancing dendrobine production. The seven genes (SG) were then stacked and transiently expressed in the leaves of D. catenatum, resulting in a dendrobine yield that was two-fold higher compared to that of the empty vector control (EV). Further, RNA-seq analysis identified Copper Methylamine Oxidase (CMEAO) as a strong candidate with predicted functions in the post-modification processes of alkaloid biosynthesis. Overexpression of CMEAO increased dendrobine content by two-fold. Additionally, co-expression analysis of the differentially expressed genes (DEGs) by weighted gene co-expression network analysis (WGCNA) retrieved one regulatory transcription factor gene MYB61. Overexpression of MYB61 increased dendrobine levels by more than two-fold in D. catenatum. In short, this work provides an efficient strategy and prospective candidates for the genetic engineering of D. catenatum to produce dendrobine, thereby improving its medicinal value.

Study on the Hydration Reaction of Typical Clay Minerals under Alkali and Sulfate Compound Activation.

Sand, stone, tailings and other aggregates often contain a small amount of clay mineral and their hydration activity is low, thereby lowering concrete performance indexes while negatively affecting their resource utilisation. In this study, clay minerals, calcium hydroxide and desulfurised gypsum were used to prepare cementitious materials to examine kaolinite, montmorillonite, illite and chlorite clay mineral contents under compound activation. The effects of curing temperature and water reducer on clay samples were analysed. The results showed that the compressive strength of kaolinite samples cured at 25 °C and 55 °C reached 1.09 and 4.93 MPa in 28 days and increased by 43% and 12%, respectively, after adding a 0.3% water reducer. Montmorillonite was activated and its compressive strength reached 5.33 MPa after curing at 55 °C in 28 days. Illite exhibited some activity and its compressive strength reached 1.43 MPa after curing at 55 °C in 28 days and the strength increased slightly after adding a water reducer. The chlorite sample had no strength after activation under the same conditions. Furthermore, X-ray diffraction and scanning electron microscope and energy-dispersive spectroscopy microstructure analyses showed that after alkali and sulfate activation, the hydration products of activated clay minerals mainly included ettringite, hydrated calcium aluminate and hydrated calcium silicate. The increase in curing temperature accelerated the reaction speed and improved the early strength. However, the effect on chlorite minerals was not obvious.

Risk factors for delayed postoperative hyponatremia in patients with non-functioning pituitary adenomas undergoing transsphenoidal surgery: A single-institution study.

Purpose: We aimed to assess factors influencing the occurrence of delayed hyponatremia after transsphenoidal surgery (TSS) in patients with a non-functional pituitary adenoma (NFPA). Methods: We retrospectively collected the clinical data of patients who underwent TSS for NFPA between January 2016 and January 2021. The pituitary region was preoperatively scanned with 3.0 T magnetic resonance imaging. The risk factors for delayed postoperative hyponatremia for NFPA were identified by univariate and multivariable logistic regression analysis. Results: We selected 166 patients with NFPA who fulfilled the inclusion criteria. Delayed postoperative hyponatremia occurred in 28 patients and did not in 138. Multivariable logistic regression analyses demonstrated that higher odds of developing delayed postoperative hyponatremia were independently associated with larger craniocaudal dimension (OR = 1.128, P = 0.034), as well as preoperative hyperprolactinemia (OR = 2.618, P = 0.045) and larger preoperative pituitary stalk deviation angle (OR = 3.033, P = 0.022). Conclusion: We identified the independent risk factors for delayed hyponatremia after TSS for NFPA; these included preoperative hyperprolactinemia, craniocaudal diameter, and preoperative pituitary stalk deviation angle.

Genome-wide identification and characterization of active ingredients related ß-Glucosidases in Dendrobium catenatum.

BACKGROUND: Dendrobium catenatum/D. officinale (here after D. catenatum), a well-known economically important traditional medicinal herb, produces a variety of bioactive metabolites including polysaccharides, alkaloids, and flavonoids with excellent pharmacological and clinical values. Although many genes associated with the biosynthesis of medicinal components have been cloned and characterized, the biosynthetic pathway, especially the downstream and regulatory pathway of major medicinal components in the herb, is far from clear. ß-glucosidases (BGLUs) comprise a diverse group of enzymes that widely exist in plants and play essential functions in cell wall modification, defense response, phytohormone signaling, secondary metabolism, herbivore resistance, and scent release by hydrolyzing ß-D-glycosidic bond from a carbohydrate moiety. The recent release of the chromosome-level reference genome of D. catenatum enables the characterization of gene families. Although the genome-wide analysis of the BGLU gene family has been successfully conducted in various plants, no systematic analysis is available for the D. catenatum. We previously isolated DcBGLU2 in the BGLU family as a key regulator for polysaccharide biosynthesis in D. catenatum. Yet, the exact number of DcBGLUs in the D. catenatum genome and their possible roles in bioactive compound production deserve more attention. RESULTS: To investigate the role of BGLUs in active metabolites production, 22 BGLUs (DcBGLU1-22) of the glycoside hydrolase family 1 (GH1) were identified from D. catenatum genome. Protein prediction showed that most of the DcBGLUs were acidic and phylogenetic analysis classified the family into four distinct clusters. The sequence alignments revealed several conserved motifs among the DcBGLU proteins and analyses of the putative signal peptides and N-glycosylation site revealed that the majority of DcBGLU members dually targeted to the vacuole and/or chloroplast. Organ-specific expression profiles and specific responses to MeJA and MF23 were also determined. Furthermore, four DcBGLUs were selected to test their involvement in metabolism regulation. Overexpression of DcBGLU2, 6, 8, and 13 significantly increased contents of flavonoid, reducing-polysaccharide, alkaloid and soluble-polysaccharide, respectively. CONCLUSION: The genome-wide systematic analysis identified candidate DcBGLU genes with possible roles in medicinal metabolites production and laid a theoretical foundation for further functional characterization and molecular breeding of D. catenatum.

Polß modulates the expression of type I interferon via STING pathway.

DNA Polymerase ß (Polß) is a key enzyme in base excision repair (BER), which is very important in maintaining the stability and integrity of the genome. Mutant Polß is closely associated with carcinogenesis. However, Polß is highly expressed in most cancers, but the underlying mechanism is not well understood. Here, we found that breast cancer cells MCF-7 with Polß knockdown exhibited high levels of type I interferon and were easily eliminated by natural killer (NK) cells.Similarly, Polß-mutant (R137Q) mice exhibited chronic inflammation symptoms in multiple organs and upregulated type I interferon levels. Further results showed that Polß deficiency caused more DNA damage accumulation in cells and triggered the leakage of damaged DNA into the cytoplasm, which activated the STING/IRF3 pathway, promoted phosphorylated IRF3 translocating into the nucleus and enhanced the expression of type I interferon and proinflammatory cytokines. In addition, this effect could be eliminated by Polß overexpression, STING inhibitor or STING knockdown. Taken together, our findings provide mechanistic insight into the role of Polß in cancers by linking DNA repair and the inflammatory STING pathway.

When immunotherapy meets surgery in non-small cell lung cancer.

The results of the most recent Checkmate-816 trial in The New England Journal of Medicine using combination neoadjuvant immunotherapy with platinum-based chemotherapy in resectable non-small cell lung cancer demonstrate the effectiveness of neoadjuvant immunotherapy and provide further support that biology and personalized therapy represent the foundation of lung cancer treatment.

Microscopic Transsphenoidal Resection of Giant Pituitary Adenomas: Analysis of the Factors Limiting the Degree of Resection in 73 Cases.

Purpose: To analyze the risk factors affecting the gross-total resection of giant pituitary adenomas using a transsphenoidal approach under a microscope to provide a reference basis for formulating an appropriate surgical strategy. Methods: The clinical data of patients who underwent microscopic transsphenoidal resection of giant pituitary adenomas in a single center from January 2011 to December 2020 were retrospectively analyzed. Based on magnetic resonance imaging and surgical records, the predictive factors affecting the gross-total resection of giant pituitary adenomas under microscopy were determined through univariate and multivariate analyses. Results: A total of 73 patients with giant pituitary adenomas underwent transsphenoidal microsurgery. Gross-total resection was performed in 19 cases (26%), subtotal resection in 31 cases (42%), partial resection in 21 cases (29%), and the degree of resection was <50% in only two cases (3%). After binary logistic analysis, it was found that it was more difficult to completely remove giant pituitary adenomas with a Knosp grade 3-4 [odds ratio (OR) = 0.214, 95% confidence interval (CI): 0.05-0.917; P = 0.038], greater proportion of tumor suprasellar volume (odds ratio = 0.937, 95% confidence interval: 0.898-0.978; P = 0.003), and intraoperative evidence of invasion of the cavernous sinus (odds ratio = 0.187, 95% CI: 0.039-0.898; P = 0.036). Conclusion: It is difficult to remove a giant pituitary adenoma invading the cavernous sinus completely with a higher degree of invasion of the suprasellar region using microscopic transsphenoidal surgery. The combined application of multiple surgical methods can help to improve the degree of resection during a single operation.

[Effects of landscape complexity and local management on bee pollinator diversity in apple orchards in Changping District, Beijing, China.] / åŒ—äº¬æ˜Œå¹³åŒºæ™¯è§‚å¤æ‚åº¦å’Œå±€åœ°ç®¡ç†å¯¹è‹¹æžœå›­ä¼ ç²‰èœ‚å¤šæ ·æ€§çš„å½±å“.

Pollinators provide important ecosystem services for crop production and food security. With the development of agricultural economy and the increasing intensity of land-use, a large number of natural or semi-natural habitats have been converted to croplands. Landscape homogenization and intensive management lead to the decline of wild bee diversity and threaten the sustainable agricultural production. In this study, we investigated the effects of landscape complexity (proportion of semi-natural habitats), local management practices (local flowering plant diversity and soil total nitrogen), and their interactions on diversity of bee pollinators in apple orchard in Changping District, Beijing. A total of 8642 bee individuals were captured, including 5125 honey bees and 3517 wild bees from 5 families, 14 genera, and 49 species. The optimal landscape scale for the response of bee diversity to landscape complexity and local management intensity was 500 m. Within 500 m radius of the site, the abundance of overall bees and wild bees significantly increased with increasing proportion of semi-natural habitats. The landscape complexity interacting with local flowering plant diversity significantly affected the richness of overall bee and wild bee. When the proportion of semi-natural habitats surrounding the apple orchards was low (≤29.9%), we found a positive effect of flowering plant diversity on the richness of overall bee and wild bee, whereas a reversed trend was found when the proportion of semi-natural habitats surrounding the apple orchards was high (>29.9%). In addition, the abundance of honey bees significantly increased with the increase of local flowering plant diversity and soil total nitrogen. The soil total nitrogen interacting with local flowering plant diversity significantly affected the honey bee abundance. At low levels of soil total nitrogen (≤1.9 g·kg-1), there was a positive effect of flowering plant diversity on honey bee abundance; whereas this trend was reversed at high levels of soil total nitrogen (>1.9 g·kg-1). Increasing the proportion of semi-natural habitats in agricultural landscape was beneficial to the increase of wild bee abundance, and flowering plant diversity could promote bee diversity but depending on landscape scale (proportion of semi-natural habitats) and local scale (nitrogen application). Therefore, multi-scale factors should be considered to develop conservation strategies to maintain the diversity of wild bees in agricultural landscape. Maintaining a higher proportion of cultivated land as much as possible is still a long-term requirement for production, while maintaining intermediate landscape complexity, increasing the diversity of flowering plants on the ground, and reducing the application of nitrogen fertilizer would be effective ways to promote the diversity of pollinating bees in apple orchards.

Toward personalized treatment approaches for non-small-cell lung cancer.

Worldwide, lung cancer is the most common cause of cancer-related deaths. Molecular targeted therapies and immunotherapies for non-small-cell lung cancer (NSCLC) have improved outcomes markedly over the past two decades. However, the vast majority of advanced NSCLCs become resistant to current treatments and eventually progress. In this Perspective, we discuss some of the recent breakthrough therapies developed for NSCLC, focusing on immunotherapies and targeted therapies. We highlight our current understanding of mechanisms of resistance and the importance of incorporating genomic analyses into clinical studies to decipher these further. We underscore the future role of neoadjuvant and maintenance combination therapy approaches to potentially cure early disease. A major challenge to successful development of rational combination therapies will be the application of robust predictive biomarkers for clear-cut patient stratification, and we provide our views on clinical research areas that could influence how NSCLC will be managed over the coming decade.

Agonistic CD40 Antibodies in Cancer Treatment.

CD40 is expressed on a variety of antigen-presenting cells. Stimulation of CD40 results in inflammation by upregulation of other costimulatory molecules, increased antigen presentation, maturation (licensing) of dendritic cells, and activation of CD8+ T cells. Here we analyzed gene expression data from The Cancer Genome Atlas in melanoma, renal cell carcinoma, and pancreatic adenocarcinoma and found correlations between CD40 and several genes involved in antigen presentation and T cell function, supporting further exploration of CD40 agonists to treat cancer. Agonist CD40 antibodies have induced anti-tumor effects in several tumor models and the effect has been more pronounced when used in combination with other treatments (immune checkpoint inhibition, chemotherapy, and colony-stimulating factor 1 receptor inhibition). The reduction in tumor growth and ability to reprogram the tumor microenvironment in preclinical models lays the foundation for clinical development of agonistic CD40 antibodies (APX005M, ChiLob7/4, ADC-1013, SEA-CD40, selicrelumab, and CDX-1140) that are currently being evaluated in early phase clinical trials. In this article, we focus on CD40 expression and immunity in cancer, agonistic human CD40 antibodies, and their pre-clinical and clinical development. With the broad pro-inflammatory effects of CD40 and its ligand on dendritic cells and macrophages, and downstream B and T cell activation, agonists of this pathway may enhance the anti-tumor activity of other systemic therapies.

Two new dibenzyl derivatives from the stems of Dendrobium catenatum.

Two new dibenzyl derivatives, dendrocandins V-W (1-2), together with six known compounds (3-8), have been isolated from the dried stems of Dendrobium catenatum. Their structures were mainly elucidated on the basis of HRESIMS, one- and two-dimensional NMR techniques. The isolated compounds 5-8 were evaluated in vitro for their antioxidant and hypoglycemic activities. Compound 8 showed moderate potent DPPH scavenging activity with IC50 value of 34.45 ± 1.07 µM. And compounds 5, 7-8 exhibited significant ABTS radical scavenging activities with IC50 values of 10.03 ± 0.88, 5.32 ± 1.13 and 9.01 ± 1.39 µM. Compounds 6-7 showed potent α-glucosidase inhibitory activities with IC50 values of 36.05 ± 0.67 and 159.59 ± 0.86 µM.

Cytological, Biochemical, and Transcriptomic Analyses of a Novel Yellow Leaf Variation in a Paphiopedilum (Orchidaceae) SCBG COP15.

The genus Paphiopedilum, belonging to the Orchidaceae, has high ornamental value. Leaf variations can considerably improve the economic and horticultural value of the orchids. In the study, a yellow leaf mutant of a Paphiopedilum hybrid named P. SCBG COP15 was identified during the in vitro plant culture process; however, little is known about their molecular mechanisms. For this, RNA-seq libraries were created and used for the transcriptomic profiling of P. SCBG COP15 and the yellow mutant. The Chl a, Chl b, and carotenoid contents in the yellow leaves decreased by approximately 75.99%, 76.92%, and 56.83%, respectively, relative to the green leaves. Decreased chloroplasts per cell and abnormal chloroplast ultrastructure were observed by electron microscopic investigation in yellowing leaves; photosynthetic characteristics and Chl fluorescence parameters were also decreased in the mutant. Altogether, 34,492 unigenes were annotated by BLASTX; 1,835 DEGs were identified, consisting of 697 upregulated and 1138 downregulated DEGs. HEMA, CRD, CAO, and CHLE, involved in Chl biosynthesis, were predicted to be key genes responsible for leaf yellow coloration. Our findings provide an essential genetic resource for understanding the molecular mechanism of leaf color variation and breeding new varieties of Paphiopedilum with increased horticultural value.

Gigantol Attenuates the Metastasis of Human Bladder Cancer Cells, Possibly Through Wnt/EMT Signaling.

BACKGROUND: Bladder cancer has long been recognized as one of the most common and aggressive human malignant carcinomas due to the increased invasiveness and metastasis. The discovery and development of natural compounds from Dendrobium species for cancer therapy have garnered increasing attention in recent years. Among those natural elements, the bibenzyl compound gigantol has promising therapeutic potential against several cancer cell lines; however, its roles on bladder tumor metastasis have not been investigated. MATERIALS AND METHODS: Here in this in vitro study, we utilized viability tests, cell migration, cell invasion and apoptosis assays to evaluate the anti-tumor activity of gigantol on three human bladder cancer cell lines (SW780, 5637, and T24) and a normal human bladder cell line (SVHUC-1). Cells were treated with different concentrations of gigantol (0, 40, 80, and 160 µM) for 24, 48 and 72 h. RESULTS: Here in this study, we showed that gigantol suppressed cancer cell proliferation but not normal SVHUC-1 cells. The inhibitory effect of the compound on cell migration and invasion was also exhibited in the cancer cell lines. Cell apoptosis assay by flow cytometry revealed enhanced apoptotic effects of gigantol on cancer cells. Gene expression analysis revealed that Wnt/EMT signaling might involve in the response of bladder cancer cells to gigantol. CONCLUSION: Therefore, the present data demonstrate gigantol as a strong anticancer reagent against bladder cancer possibly through Wnt/EMT signaling.