Biomimetic Nanovehicle-Enabled Targeted Depletion of Intratumoral Fusobacterium nucleatum Synergizes with PD-L1 Blockade against Breast Cancer.

Immune checkpoint blockade (ICB) therapy has been approved for breast cancer (BC), but clinical response rates are limited. Recent studies have shown that commensal microbes colonize a variety of tumors and are closely related to the host immune system response. Here, we demonstrated that Fusobacterium nucleatum (F.n), which is prevalent in BC, creates an immunosuppressive tumor microenvironment (ITME) characterized by a high-influx of myeloid cells that hinders ICB therapy. Administering the antibiotic metronidazole in BC can deplete F.n and remodel the ITME. To prevent an imbalance in the systemic microbiota caused by antibiotic administration, we designed a biomimetic nanovehicle for on-site antibiotic delivery inspired by F.n homing to BC. Additionally, ferritin-nanocaged doxorubicin was coloaded into this nanovehicle, as immunogenic chemotherapy has shown potential for synergy with ICB. It has been demonstrated that this biomimetic nanovehicle can be precisely homed to BC and efficiently eliminate intratumoral F.n without disrupting the diversity and abundance of systemic microbiota. This ultimately remodels the ITME, improving the therapeutic efficacy of the PD-L1 blocker with a tumor inhibition rate of over 90% and significantly extending the median survival of 4T1 tumor-bearing mice.

Enhancement of the anthocyanin contents of Caladium leaves and petioles via metabolic engineering with co-overexpression of AtPAP1 and ZmLc transcription factors.

Introduction: Metabolic engineering of anthocyanin synthesis is an active research area for pigment breeding and remains a research hotspot involving AtPAP1 and ZmLc transcription factors. Caladium bicolor is a desirable anthocyanin metabolic engineering receptor, with its abundant leaf color and stable genetic transformation system. Methods: We transformed C. bicolor with AtPAP1 and ZmLc and successfully obtained transgenic plants. We then used a combination of metabolome, transcriptome, WGCNA and PPI co-expression analyses to identify differentially expressed anthocyanin components and transcripts between wild-type and transgenic lines. Results: Cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside and peonidin-3-O-rutinoside are the main components of anthocyanins in the leaves and petioles of C. bicolor. Exogenous introduction of AtPAP1 and ZmLc resulted in significant changes in pelargonidins, particularly pelargonidin-3-O-glucoside and pelargonidin-3-O-rutinoside in C. bicolor. Furthermore, 5 MYB-TFs, 9 structural genes, and 5 transporters were found to be closely associated with anthocyanin synthesis and transport in C. bicolor. Discussion: In this study, a network regulatory model of AtPAP1 and ZmLc in the regulation of anthocyanin biosynthesis and transport in C. bicolor was proposed, which provides insights into the color formation mechanisms of C. bicolor, and lays a foundation for the precise regulation of anthocyanin metabolism and biosynthesis for economic plant pigment breeding.

An exotic plant successfully invaded as a passenger driven by light availability.

Invasive exotic plant species (IEPs) are widely distributed across the globe, but whether IEPs are drivers or passengers of habitat change in the invaded spaces remains unclear. Here, we carried out a vegetation and soil survey in 2018 and two independent field experiments (Pedicularis kansuensis removal in 2014 and 2015, and fertilization experiment since 2012) and found that the invasive annual P. kansuensis was at a disadvantage in light competition compared with perennial native grasses, but the successful invasion of P. kansuensis was due to the sufficient light resources provided by the reduced coverage of the native species. Conversely, nitrogen enrichment can effectively inhibit P. kansuensis invasion by increasing the photocompetitive advantage of the native species. sP. kansuensis invasion did not reduce species richness, but did increase plant community coverage, productivity and soil nutrients. Furthermore, the removal of P. kansuensis had little effect on the plant community structure and soil properties. Our results suggest that the passenger model perfectly explains the benign invasive mechanism of P. kansuensis. The invasion "ticket" of P. kansuensis is a spare ecological niche for light resources released by overgrazing.

Herpesvirus-Mimicking DNAzyme-Loaded Nanoparticles as a Mitochondrial DNA Stress Inducer to Activate Innate Immunity for Tumor Therapy.

Virus-based immunotherapy is a promising approach to treat tumor. Closely mimicking the structure and sequential infection processes of natural viruses is highly desirable for effective tumor immunotherapy but remains challenging. Here, inspired by the robust innate immunity induced by herpesvirus, a herpesvirus-mimicking nanoparticle (named Vir-ZM@TD) is engineered for tumor therapy by mimicking the structure and infection processes of herpesvirus. In this biomimetic system, DNAzyme-loaded manganese-doped zeolitic imidazolate framework-90 (ZIF-90) nanoparticles (ZM@TD) mimic the virus nucleocapsid containing the genome; the erythrocyte membrane mimics the viral envelope; and two functional peptides, RGD and HA2 peptides, resemble the surface glycoprotein spikes of herpesvirus. Vir-ZM@TD can both effectively evade rapid clearance in the blood circulation and closely mimic the serial infection processes of herpesvirus, including specific tumor targeting, membrane fusion-mediated endosomal escape, and TFAM (transcription factor A, mitochondrial) deficiency-triggered mitochondrial DNA stress, as well as the release of manganese ions (Mn2+ ) from organelles into the cytosol, ultimately effectively priming cGAS-STING pathway-mediated innate immunity with 68% complete regression of primary tumors and extending by 32 days the median survival time of 4T1-tumor-bearing mice.

A Legume Host Benefits More from Arbuscular Mycorrhizal Fungi Than a Grass Host in the Presence of a Root Hemiparasitic Plant.

In nature, most plants parasitized by root hemiparasites are also colonized by mutualistic arbuscular mycorrhizal (AM) fungi, highlighting the prevalence of this tripartite interaction. AM colonization is generally found to improve the growth of parasitized legumes but has little impact on grass hosts parasitized by root hemiparasites, and the underlying mechanisms are still unclear. In this study, we conducted a pot experiment to test the influence of AM fungus (Glomus mosseae) on the growth and photosynthesis of leguminous Trifolium repens and gramineous Elymus nutans in the presence of a root hemiparasitic plant (Pedicularis kansuensis). The results showed that inoculation with AM fungi significantly improved the growth performance of parasitized legumes via enhancing their nutrient status and photosynthetic capacity, even though a larger P. kansuensis parasitized the legume host in the AM treatment. In contrast, AM colonization slightly improved the shoot DW of grass hosts by suppressing haustoria formation and the growth of P. kansuensis. Our results demonstrated that legume hosts benefit more from AM inoculation than grass hosts in the presence of hemiparasitic plants, and set out the various mechanisms. This study provides new clues for parsing the tritrophic interaction of AM fungi, parasitic plants, and host plants.

Preparation of formate-free PMA@MOF-808 catalysts for deep oxidative desulfurization of model fuels.

Due to the increasingly serious environmental problems caused by the combustion of sulfides in fuel, deep desulfurization of fuel became particularly urgent. Herein, the catalyst (PMA@MOF-808) of the Zr-based metal-organic framework (MOF-808) encapsulating phosphomolybdic acid (PMA) was prepared via a one-pot hydrothermal method. Besides, the formate ions of PMA@MOF-808 were removed by posttreatment with methanol, resulting in formate-free PMA@MOF-808-H catalysts with unsaturated open metal sites. The as-synthesized catalysts were systematically characterized by XRD, FT-IR, SEM, BET, TGA, 1H NMR and XPS. The catalysts were also applied in catalytic oxidation desulfurization of fuel. The results indicated that the introduction of PMA and the removal of formate ions can improve the desulfurization performance of catalysts. Formate-free 0.2-PMA@MOF-808-H catalyst can reach 100% desulfurization rate for DBT. Besides, the kinetic properties were studied, and the apparent activation energy was 29.34 kJ/mol.

Genomes shed light on the evolution of Begonia, a mega-diverse genus.

Clarifying the evolutionary processes underlying species diversification and adaptation is a key focus of evolutionary biology. Begonia (Begoniaceae) is one of the most species-rich angiosperm genera with c. 2000 species, most of which are shade-adapted. Here, we present chromosome-scale genome assemblies for four species of Begonia (B. loranthoides, B. masoniana, B. darthvaderiana and B. peltatifolia), and whole genome shotgun data for an additional 74 Begonia representatives to investigate lineage evolution and shade adaptation of the genus. The four genome assemblies range in size from 331.75 Mb (B. peltatifolia) to 799.83 Mb (B. masoniana), and harbor 22 059-23 444 protein-coding genes. Synteny analysis revealed a lineage-specific whole-genome duplication (WGD) that occurred just before the diversification of Begonia. Functional enrichment of gene families retained after WGD highlights the significance of modified carbohydrate metabolism and photosynthesis possibly linked to shade adaptation in the genus, which is further supported by expansions of gene families involved in light perception and harvesting. Phylogenomic reconstructions and genomics studies indicate that genomic introgression has also played a role in the evolution of Begonia. Overall, this study provides valuable genomic resources for Begonia and suggests potential drivers underlying the diversity and adaptive evolution of this mega-diverse clade.

Boosting 5-ALA-based photodynamic therapy by a liposomal nanomedicine through intracellular iron ion regulation.

5-Aminolevulinic acid (5-ALA) has been approved for clinical photodynamic therapy (PDT) due to its negligible photosensitive toxicity. However, the curative effect of 5-ALA is restricted by intracellular biotransformation inactivation of 5-ALA and potential DNA repair of tumor cells. Inspired by the crucial function of iron ions in 5-ALA transformation and DNA repair, a liposomal nanomedicine (MFLs@5-ALA/DFO) with intracellular iron ion regulation property was developed for boosting the PDT of 5-ALA, which was prepared by co-encapsulating 5-ALA and DFO (deferoxamine, a special iron chelator) into the membrane fusion liposomes (MFLs). MFLs@5-ALA/DFO showed an improved pharmaceutical behavior and rapidly fused with tumor cell membrane for 5-ALA and DFO co-delivery. MFLs@5-ALA/DFO could efficiently reduce iron ion, thus blocking the biotransformation of photosensitive protoporphyrin IX (PpIX) to heme, realizing significant accumulation of photosensitivity. Meanwhile, the activity of DNA repair enzyme was also inhibited with the reduction of iron ion, resulting in the aggravated DNA damage in tumor cells. Our findings showed MFLs@5-ALA/DFO had potential to be applied for enhanced PDT of 5-ALA.

The bright side of parasitic plants: what are they good for?

Parasitic plants are mostly viewed as pests. This is caused by several species causing serious damage to agriculture and forestry. There is however much more to parasitic plants than presumed weeds. Many parasitic plans exert even positive effects on natural ecosystems and human society, which we review in this paper. Plant parasitism generally reduces the growth and fitness of the hosts. The network created by a parasitic plant attached to multiple host plant individuals may however trigger transferring systemic signals among these. Parasitic plants have repeatedly been documented to play the role of keystone species in the ecosystems. Harmful effects on community dominants, including invasive species, may facilitate species coexistence and thus increase biodiversity. Many parasitic plants enhance nutrient cycling and provide resources to other organisms like herbivores or pollinators, which contributes to facilitation cascades in the ecosystems. There is also a long tradition of human use of parasitic plants for medicinal and cultural purposes worldwide. Few species provide edible fruits. Several parasitic plants are even cultivated by agriculture/forestry for efficient harvesting of their products. Horticultural use of some parasitic plant species has also been considered. While providing multiple benefits, parasitic plants should always be used with care. In particular, parasitic plant species should not be cultivated outside their native geographical range to avoid the risk of their uncontrolled spread and the resulting damage to ecosystems.

Non-invasive red-light optogenetic control of Drosophila cardiac function.

Drosophila is a powerful genetic model system for cardiovascular studies. Recently, optogenetic pacing tools have been developed to control Drosophila heart rhythm noninvasively with blue light, which has a limited penetration depth. Here we developed both a red-light sensitive opsin expressing Drosophila system and an integrated red-light stimulation and optical coherence microscopy (OCM) imaging system. We demonstrated noninvasive control of Drosophila cardiac rhythms using a single light source, including simulated tachycardia in ReaChR-expressing flies and bradycardia and cardiac arrest in halorhodopsin (NpHR)-expressing flies at multiple developmental stages. By using red excitation light, we were able to pace flies at higher efficiency and with lower power than with equivalent blue light excitation systems. The recovery dynamics after red-light stimulation of NpHR flies were observed and quantified. The combination of red-light stimulation, OCM imaging, and transgenic Drosophila systems provides a promising and easily manipulated research platform for noninvasive cardiac optogenetic studies.

FlyNet 2.0: drosophila heart 3D (2D + time) segmentation in optical coherence microscopy images using a convolutional long short-term memory neural network.

A custom convolutional neural network (CNN) integrated with convolutional long short-term memory (LSTM) achieves accurate 3D (2D + time) segmentation in cross-sectional videos of the Drosophila heart acquired by an optical coherence microscopy (OCM) system. While our previous FlyNet 1.0 model utilized regular CNNs to extract 2D spatial information from individual video frames, convolutional LSTM, FlyNet 2.0, utilizes both spatial and temporal information to improve segmentation performance further. To train and test FlyNet 2.0, we used 100 datasets including 500,000 fly heart OCM images. OCM videos in three developmental stages and two heartbeat situations were segmented achieving an intersection over union (IOU) accuracy of 92%. This increased segmentation accuracy allows morphological and dynamic cardiac parameters to be better quantified.

US-triggered ultra-sensitive "thrombus constructor" for precise tumor therapy.

Embolization therapy is an attractive strategy for antitumor therapy, especially for solid tumors. In vivo self-coagulation behavior holds great potential in a new type of tumor embolization therapy. However, spatiotemporal controllable in situ formation of thrombus in tumor is a challenge. Herein, an ultrasound (US)-responsive ultra-sensitive "thrombus constructor" (UUNC), which was prepared by loading thrombin into a nanobubble, and modified with NGR peptide on its surface, is rational designed for tumor embolization therapy. Benefiting from the targeting ability of NGR peptides to tumor neovascularization, UUNC efficiently enriched in tumor vessels, and then released thrombin rapidly to form thrombi in situ of tumor blood vessels in the presence of US. In vivo antitumor experiments demonstrated that UUNC could significantly lead to tumor cell apoptosis and necrosis, and the tumor growth inhibition rate (TGI) was 85.3% with a transient US in tumor, while maintain high stability, and no obvious thrombus was observed in normal tissues. UUNC holds an attractive potential for tumor embolization therapy via spatiotemporal controllable thrombus construct strategy.

Sub-3 nm Rh nanoclusters confined within a metal-organic framework for enhanced hydrogen generation.

Ultrafine sub-3 nm Rh nanoclusters confined within a metal-organic framework (UIO-66) were constructed through a double-solvent host-guest strategy and achieved superior hydrogen generation via ammonia borane hydrolysis compared with Rh nanoclusters dispersed on the exterior walls.

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila.

Dendrites are the branched projections of a neuron, and dendritic morphology reflects synaptic organization during the development of the nervous system. Drosophila larval neuronal dendritic arborization (da) is an ideal model for studying morphogenesis of neural dendrites and gene function in the development of nervous system. There are four classes of da neurons. Class IV is the most complex with a branching pattern that covers almost the entire area of the larval body wall. We have previously characterized the effect of silencing the Drosophila ortholog of SOX5 on class IV neuronal dendritic arborization complexity (NDAC) using four parameters: the length of dendrites, the surface area of dendrite coverage, the total number of branches, and the branching structure. This protocol presents the workflow of NDAC quantitative analysis, consisting of larval dissection, confocal microscopy, and image analysis procedures using ImageJ software. Further insight into da neuronal development and its underlying mechanisms will improve the understanding of neuronal function and provide clues about the fundamental causes of neurological and neurodevelopmental disorders.

Root hemiparasitism in Malania oleifera (Olacaceae), a neglected aspect in research of the highly valued tree species.

Malania oleifera (Olacaceae) is a valued tree species, mostly because its seeds have high precious fatty acid content (particularly nervonic acid). However, seedling mortality rates are often high and regeneration of this tree has been problematic, which greatly hinders its utilization at a large scale. Cultivation difficulties of some tree species in the family Olacaceae have been attributed to their root hemiparasitic habit. Prompted by field observations and the taxonomic proximity of M. oleifera to root hemiparasites in Olacaceae, we hypothesized that tuberous structures observed on the roots of M. oleifera are parasitic organs known as haustoria. To test this hypothesis, we collected root samples from M. oleifera plants of various ages and growth conditions, investigated the morphological and anatomical features of tuberous structures and their connections to neighboring roots. Our analyses confirmed that M. oleifera are root hemiparasites. To the best of our knowledge, this is the first empirical report on root hemiparasitism in M. oleifera. Because life strategies of root hemiparasitic plants differ greatly from autotrophic plants, the root hemiparasitic habit needs to be taken into account for successful seedling regeneration of M. oleifera. This study establishes the foundation for investigations into a long-neglected but essential aspect in research of these highly valued tree species.

A neglected alliance in battles against parasitic plants: arbuscular mycorrhizal and rhizobial symbioses alleviate damage to a legume host by root hemiparasitic Pedicularis species.

Despite their ubiquitous distribution and significant ecological roles, soil microorganisms have long been neglected in investigations addressing parasitic plant-host interactions. Because nutrient deprivation is a primary cause of host damage by parasitic plants, we hypothesized that beneficial soil microorganisms conferring nutrient benefits to parasitized hosts may play important roles in alleviating damage. We conducted a pot cultivation experiment to test the inoculation effect of an arbuscular mycorrhizal fungus (Glomus mosseae), a rhizobium (Rhizobium leguminosarum) and their interactive effects, on alleviation of damage to a legume host (Trifolium repens) by two root hemiparasitic plants with different nutrient requirements (N-demanding Pedicularis rex and P-demanding P. tricolor). Strong interactive effects between inoculation regimes and hemiparasite identity were observed. The relative benefits of microbial inoculation were related to hemiparasite nutrient requirements. Dual inoculation with the rhizobium strongly enhanced promotional arbuscular mycorrhizal effects on hosts parasitized by P. rex, but reduced the arbuscular mycorrhizal promotion on hosts parasitized by P. tricolor. Our results demonstrate substantial contribution of arbuscular mycorrhizal and rhizobial symbioses to alleviating damage to the legume host by root hemiparasites, and suggest that soil microorganisms are critical factors regulating host-parasite interactions and should be taken into account in future studies.

Analysis of hidradenitis suppurativa-linked mutations in four genes and the effects of PSEN1-P242LfsX11 on cytokine and chemokine expression in macrophages.

Hidradenitis suppurativa (HS), or acne inversa, is a chronic inflammatory skin disorder characterized clinically with acne-like lesions in apocrine gland-bearing skin, follicular occlusion and recurrent inflammation. Thirty-four unique mutations in patients with HS have been found in three genes encoding the γ-secretase complex: nicastrin (NCSTN), presenilin 1 (PSEN1), presenilin enhancer 2 (PSENEN) and in POGLUT1, an endoplasmic reticulum O-glucosyltransferase involved in Notch signaling. We have carried out a system review and have performed a functional analysis of the 34 unique reported HS-linked mutations in NCSTN, PSEN1, PSENEN and POGLUT1. We have also examined the effects of the HS-linked PSEN1-P242LfsX11 mutation on cytokine and chemokine expression in macrophages. Mutations in NCSTN are predicted to cause loss of function, to result in loss of transmembrane (TM) domain, to affect NCSTN substrate recruitment sites, to cause loss or creation of new ligand binging sites and to alter post-translational modifications and disulfide bonds. PSEN1-P242LfsX11 occurs at the opposite side of TM5 from Alzheimer's disease-linked PSEN1 mutations. All of the PSENEN mutations occur on TM regions that are predicted to disrupt membrane function. POGLUT1 mutations lead to an early termination of protein synthesis and are predicted to affect ligand binding function. In addition, PSEN1-P242LfsX11 mediates cytokine and chemokine expression and prolongs tumor necrosis factor α production on the inflammatory processes in THP-1 cells and phorbol-12-myristate-13-acetate-differentiated macrophages in response to lipopolysaccharide stimulation. These in silico analyses are instructive for functional studies of the HS-linked mutations. The PSEN1-P242LfsX11 mutation mediates cytokine and chemokine expression in macrophages.

N-P fertilization did not reduce AMF abundance or diversity but alter AMF composition in an alpine grassland infested by a root hemiparasitic plant.

Fertilization has been shown to have suppressive effects on arbuscular mycorrhizal fungi (AMF) and root hemiparasites separately in numerous investigations, but its effects on AMF in the presence of root hemiparasites remain untested. In view of the contrasting nutritional effects of AMF and root hemiparasites on host plants, we tested the hypothesis that fertilization may not show strong suppressive effects on AMF when a plant community was infested by abundant hemiparasitic plants. Plants and soil samples were collected from experimental field plots in Bayanbulak Grassland, where N and P fertilizers had been applied for three continuous years for control against a spreading root hemiparasite, Pedicularis kansuensis. Shoot and root biomass of each plant functional group were determined. Root AMF colonization levels, soil spore abundance, and extraradical hyphae length density were measured for three soil depths (0-10 cm, 10-20 cm, 20-30 cm). Partial 18S rRNA gene sequencing was used to detect AMF diversity and community composition. In addition, we analyzed the relationship between relative abundance of different AMF genera and environmental factors using Spearman's correlation method. In contrast to suppressive effects reported by many previous studies, fertilization showed no significant effects on AMF root colonization or AMF species diversity in the soil. Instead, a marked increase in soil spore abundance and extraradical hyphae length density were observed. However, fertilization altered relative abundance and AMF composition in the soil. Our results support the hypothesis that fertilization does not significantly influence the abundance and diversity of AMF in a plant community infested by P. kansuensis.

Segmentation of Drosophila heart in optical coherence microscopy images using convolutional neural networks.

Convolutional neural networks (CNNs) are powerful tools for image segmentation and classification. Here, we use this method to identify and mark the heart region of Drosophila at different developmental stages in the cross-sectional images acquired by a custom optical coherence microscopy (OCM) system. With our well-trained CNN model, the heart regions through multiple heartbeat cycles can be marked with an intersection over union of ~86%. Various morphological and dynamical cardiac parameters can be quantified accurately with automatically segmented heart regions. This study demonstrates an efficient heart segmentation method to analyze OCM images of the beating heart in Drosophila.

CD90+ cardiac fibroblasts reduce fibrosis of acute myocardial injury in rats.

OBJECTIVE: To explore the differentiation tendency of CD90+ cardiac fibroblast (CFs) into cardiomyogenic cells in vitro and repair functions in acute myocardial infarction rats. METHODS: CD90+ subpopulation was sorted from rat CFs by flow cytometry. 10 µmoL/L of 5-Azacytosine (5-aza) was used to induce differentiation of CFs into cardiomyogenic cells. An acute myocardial infarction model was prepared by ligation of the rat left anterior descending coronary artery. After nuclei were labeled by DAPI, induced CD90+ CFs were injected into the infarction marginal zone. Before coronary ligation, 40 min after ligation, and at 7 and 14 days after cell transplantation, cardiac function changes were detected by ultrasound imaging system respectively. cTnT and endothelial cell marker VIII factor were detected by immunofluorescence staining. Infarct size was examined by TTC staining. Fibrosis was evaluated with masson's trichrome staining, vimentin, type I and type III collagen staining. RESULTS: CD90+ CFs sorted by flow cytometry was 34.9%. On day 28 after induction, the cTnT positive rate was 61.17 ±â€¯9.75%. Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) at days 7 and 14 after transplantation were significantly increased compared with those before transplantation (P < 0.05). LVEF and LVFS at the 14th day after transplantation were also significantly increased compared with those at the 7th day (LVEF: 61.40 ±â€¯2.45% vs. 56.25 ±â€¯2.9%, LVFS: 33.21 ±â€¯0.68% vs. 30.26 ±â€¯2.06%, P < 0.01). Additionally, small numbers of CD90+ CFs differentiated into cardiomyocytes and became involved in neovascularization. CD90+ CFs and CFs reduced myocardial infarct size at days 14. It was significantly smaller in rats with CFs transplantation group than those in MI group(24.78 ±â€¯2.28% vs. 31.28 ±â€¯2.83%, P < 0.05), and it was also significantly smaller in rats with CD90+CFs transplantation group than those in CFs transplantation group (17.47 ±â€¯4.15% vs. 24.78 ±â€¯2.28%, P < 0.05). Meanwhile, The percentage of fibrotic area and vimentin, type I and type III collagen in the infarct border zone and infarct area were both significantly reduced in CD90 + CFs. CONCLUSION: CD90+ CFs is the preponderant subpopulation of cardiomyogenic differentiation, with potential use as seed cells in basic and clinical research of heart regeneration and repair.