Preparation of porous polylactic acid nanofibers and application in non-electret high-efficiency filtration composites.

Air pollution caused by fine particulate matter (PM0.3) has drawn increasing attention as an overwhelming threat to public health. Electret treatment is commonly used to improve the filtration performance of commercial fibrous filter materials by enhancing the electrostatic adsorption effect, but it is greatly affected by environmental factors (especially humidity). Moreover, filter materials are generally non-degradable and non-recyclable, causing serious environmental pollution. Herein, a strategy to manufacture fully degradable polylactic acid (PLA) filtration composites based on porous PLA nanofibers prepared by electrospinning was investigated in this study. Porous, bead-on-string and conventional PLA nanofibers could be obtained by adjusting spinning condition parameters. The porous PLA nanofibers exhibited 9.8 times greater specific surface area (24.01 m2 g-1) and 18 times more cumulative pore volume (0.108 cm3 g-1) than conventional PLA nanofibers. More importantly, fibrous filtration composites based on porous PLA nanofibers possessed a high PM0.3 filtration efficiency (99.9989%), low pressure drop (90.35 Pa) and high air permeability (72.4 Pa-1) at an air flow rate of 32 L min-1 without electret treatment. The fibrous filtration composites based on conventional or bead-on-string PLA nanofibers also exhibited excellent filtration performance (>99.99%), but the associated high pressure drop and low air permeability limited their application.

A biodegradable oxidized starch/carboxymethyl chitosan film coated with pesticide-loaded ZIF-8 for tomato fusarium wilt control.

Film mulching is one of the most important methods to control soil-borne diseases. However, the traditional mulch may cause microplastic pollution and soil ecological damage. Herein, a biodegradable film was developed using oxidized starch and carboxymethyl chitosan and incorporated ZIF-8 carrying fludioxonil to sustainably control soil-borne disease. The microstructure, mechanical properties, optical properties, and water barrier properties of the composite films (Flu@ZIF-8-OS/CMCS) were investigated. The results show that Flu@ZIF-8-OS/CMCS had a smooth and uniform surface and excellent light transmittance. The excellent mechanical properties of the films were verified by tensile strength, elongation at break and Young's modulus. Higher contact angle and lower water vapor permeability indicate water retention capacity of the soil was improved through using Flu@ZIF-8-OS/CMCS. Furthermore, the release properties, biological activity, degradability and safety to soil organisms of Flu@ZIF-8-OS/CMCS was determined. The addition of ZIF-8 significantly improved the film's ability to retard the release of Flu, while the Flu@ZIF-8-OS/CMCS has good soil degradability. In vitro antifungal assays and pot experiments demonstrated excellent inhibitory activity against Fusarium oxysporum f. sp. Lycopersici. Flu@ZIF-8-OS/CMCS caused only 13.33 % mortality of earthworms within 7 d. This research provides a new approach to reducing microplastic pollution and effectively managing soil-borne diseases.

Construction and biological effects of a redox-enzyme dual-responsive lufenuron nano-controlled release formulation.

BACKGROUND: Pesticide formulations based on nanotechnology can effectively improve the efficiency of pesticide utilization and reduce pesticide residues in the environment. In this study, mesoporous silica nanoparticles containing disulfide bonds were synthesized by the sol-gel method, carboxylated and adsorbed with lufenuron, and grafted with cellulose to obtain a lufenuron-loaded nano-controlled release formulation (Luf@MSNs-ss-cellulose). RESULTS: The structure and properties of Luf@MSNs-ss-cellulose were characterized. The results showed that Luf@MSNs-ss-cellulose exhibits a regular spherical shape with 12.41% pesticide loading. The highest cumulative release rate (73.46%) of this pesticide-loaded nanoparticle was observed at 7 days in the environment of glutathione and cellulase, which shows redox-enzyme dual-responsive performance. As a result of cellulose grafting, Luf@MSNs-ss-cellulose had a small contact angle and high adhesion work on corn leaves, indicating good wetting and adhesion properties. After 14 days of spraying with 20 mg L-1 formulations in the long-term control efficacy experiment, the mortality of Luf@MSNs-ss-cellulose against Ostrinia furnacalis larvae (56.67%) was significantly higher than that of commercial Luf@EW (36.67%). Luf@MSNs-ss-cellulose is safer for earthworms and L02 cells. CONCLUSION: The nano-controlled release formulation obtained in this study achieved intelligent pesticide delivery in time and space under the environmental stimulation of glutathione and cellulase, providing an effective method for the development of novel pesticide delivery systems. © 2023 Society of Chemical Industry.

Sodium alginate-carboxymethyl chitosan hydrogels loaded with difenoconazole for pH-responsive release to control wheat crown rot.

Increasing concern about environmental pollution has driven the development of controlled release formulations for agrochemicals. Due to the advantages of degradability and responsiveness to environmental stimuli, polysaccharide-based hydrogel is an ideal carrier for agrochemicals controlled release. In this study, a method-easy polysaccharide hydrogel for controlled release of difenoconazole (DZ) was prepared with sodium alginate (SA) and carboxymethyl chitosan (CMCS). Due to its three-dimensional crosslinked mesh structure, the prepared hydrogels (CSDZ) showed an agrochemical load capacity of 9.03 % and an encapsulation efficiency of 68.64 %. The release rate is faster in alkaline solution, followed by neutral solution, and slowest in an acid environment, which is consistent with the swelling behavior. Furthermore, leaching studies showed that CSDZ hydrogels have excellent protective properties for encapsulated agrochemicals. Compared with technical DZ, the results of in vitro and pot antifungal testing showed that CSDZ had a better control effect against wheat crown rot (Fusarium pseudograminearum). Safety assessment studies indicated that CSDZ hydrogels exhibit good biocompatibility on nontargeted organisms (Daphnia magna, zebrafish and Eisenia fetida) and wheat. This study aims to provide a potentially promising approach for the preparation and application of biocompatible polysaccharide-based hydrogels for agrochemical-controlled release in sustainable disease management.

Preparation and Characterization of a Novel Artemisia Oil Packaging Film and Its Application in Mango Preservation.

In this work, a new food packaging film was synthesized via blending Artemisia oil (AO) into soybean protein isolate (SPI) and gelatin (Gel) for the postharvest storage of mango. The morphological architecture and mechanical properties of the films were characterized using scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FTIR), X-ray diffractometer (XRD), and other technologies. The results show that the prepared films had relatively flat surfaces with good mechanical properties. AO enhanced the light-blocking ability of the film, increased the hydrophobicity, and affected the moisture content and water solubility of the film to a certain extent. Furthermore, the antioxidant performance and antifungal (Colletotrichum gloeosporioides) capacity of the films increased with higher AO concentration due to the presence of the active components contained in AO. During mango storage applications, the films showed good freshness retention properties. The above results indicate that SPI-Gel films containing AO have excellent physicochemical and application properties and have great potential in the field of food packaging.

Construction of Fe3O4/xSiO2/ySiO2 Nanoparticles for Pesticide Removal from Water: Improved Dispersion Stability and Adsorption Capacity.

Highly efficient and reusable adsorbents for pesticide removal from wastewater have received increasing attention. In this study, Fe3O4 was synthesized using the solvothermal method. Fe3O4/xSiO2 and Fe3O4/xSiO2/ySiO2 were obtained through layer-by-layer silica (SiO2) coating on the surface of Fe3O4. SiO2 coating improved the dispersibility of the adsorbent, which can be separated from water rapidly under the action of the external magnetic field. The adsorption capacity of the adsorbent was investigated through removing pyraclostrobin from synthetic wastewater. The adsorbent showed the highest adsorption effect at the adsorbent concentration of 1 mg mL-1, at a pH of 7, and the adsorbent time of 110 min. The fitting model of the adsorption process conformed to the second-order kinetic model and the Langmuir model. The maximum adsorption capacity of Fe3O4/xSiO2/ySiO2 nanoparticles was 94.89 mg g-1, and the removal efficiency was about 96% at adsorption equilibrium. Acetone as the eluent can effectively desorb the adsorbent, and the desorbed adsorbent had high reusability. Particularly, the removal efficiency was still greater than 86% after 9 times of reuse. These results provide a reference for designing reusable nanoparticles to effectively absorb pesticides in wastewater.

Construction and formulation optimization of prothioconazole nanoemulsions for the control of Fusarium graminearum: Enhancing activity and reducing toxicity.

In this study, the optimal emulsifier for prothioconazole nanoemulsions was initially screened based on appearance, microscopic observation, mean droplet size and polydispersity index (PDI). In addition, the BoxBehnken design method is adopted, and the optimal formula is screened with an emulsification time, emulsifier content, and solvent content as a single factor. On this basis, the nanoemulsion meets FAO standards for various indicators. The contact angle of droplets on wheat leaves was significantly reduced. This nanoemulsion also showed good inhibitory activity against Fusarium graminearum (EC50 =1.94 mg L-1), low acute toxicity to zebrafish (LC50 =26.35 mg L-1) and good biosafety to BEAS-2B cells. The nanoemulsion reduced the adverse effects of pesticide on wheat seed germination and growth. This study can help promote the design and manufacture of stable, efficient and safe agricultural nanoemulsions, and is expected to benefit the sustainable development of green plant protection.

Morphology and Properties of Polylactic Acid Composites with Butenediol Vinyl Alcohol Copolymer Formed by Melt Blending.

Due to its poor toughness and hydrophilicity, the application of polylactic acid (PLA) in the field of absorbent sanitary materials is restricted. A butenediol vinyl alcohol copolymer (BVOH) was used to improve PLA via melt blending. The morphology, molecular structure, crystallization, thermal stability, tensile property, and hydrophilicity of PLA/BVOH composites with different mass ratios were investigated. The results show that the PLA/BVOH composites possessed a two-phase structure with good interfacial adhesion. The BVOH could effectively blend into PLA without a chemical reaction. The addition of the BVOH promoted the crystallization of PLA, improved the perfection of the crystalline region, and increased the glass transition temperature and melting temperature of PLA in the heating process. Moreover, the thermal stability of PLA was markedly improved by adding the BVOH. The addition of the BVOH also had a significant effect on the tensile property of the PLA/BVOH composites. When the content of the BVOH was 5 wt.%, the elongation at the break of the PLA/BVOH composites could reach 9.06% (increased by 76.3%). In addition, the hydrophilicity of PLA was also significantly improved, and the water contact angles decreased with the increase in the BVOH content and time. When the content of the BVOH was 10 wt.%, the water contact angle could reach 37.3° at 60 s, suggesting good hydrophilicity.

CRISPR-based targeted haplotype-resolved assembly of a megabase region.

Constructing high-quality haplotype-resolved genome assemblies has substantially improved the ability to detect and characterize genetic variants. A targeted approach providing readily access to the rich information from haplotype-resolved genome assemblies will be appealing to groups of basic researchers and medical scientists focused on specific genomic regions. Here, using the 4.5 megabase, notoriously difficult-to-assemble major histocompatibility complex (MHC) region as an example, we demonstrated an approach to construct haplotype-resolved assembly of the targeted genomic region with the CRISPR-based enrichment. Compared to the results from haplotype-resolved genome assembly, our targeted approach achieved comparable completeness and accuracy with reduced computing complexity, sequencing cost, as well as the amount of starting materials. Moreover, using the targeted assembled personal MHC haplotypes as the reference both improves the quantification accuracy for sequencing data and enables allele-specific functional genomics analyses of the MHC region. Given its highly efficient use of resources, our approach can greatly facilitate population genetic studies of targeted regions, and may pave a new way to elucidate the molecular mechanisms in disease etiology.

Identification of Scd5 as a functional regulator of visceral fat deposition and distribution.

Ectopic deposition of visceral adipose tissue (VAT) in abdomen is usually accompanied with systematic chaos of energy metabolism, a higher risk of cardiovascular diseases and type II diabetes. Here, we identified a previously unexplored gene Scd5 as a master regulator of fat distribution, which alone plays a significant role in determining the VAT accumulation. Firstly, zebrafish scd5 had the highest homology with human SCD5 compared to other SCDs in mouse and rat. We then observed that scd5-homozygous mutant zebrafish displayed a puffy, short and rounded apple-shaped figure. Whole-mount micro-CT scan showed that excessive VAT deposition and short spine are responsible for the abnormal body ratio. And the supplementation of ω3-polyunsaturated fatty acid (ω3-PUFA) in dietary significantly decreased VAT accumulation in scd5 -/- zebrafish. Lastly, transcriptional analyses revealed that the Wnt, PPAR, C/EBP, and fat synthesis signaling pathways are significantly affected in the VAT of scd5 -/- mutant and restored by ω3-PUFA.

Zonular defects in loxl1-deficient zebrafish.

BACKGROUND: To investigate the roles of the lysyl oxidase-like 1 (loxl1) gene in zebrafish eye development and the potency of loxl1 deficiency in mimicking the ocular manifestations of exfoliation syndrome (XFS). METHODS: CRISPR/Cas9 technology was used to generate a frameshift coding deletion in zebrafish loxl1. Expression profiles and ocular manifestations of the wildtype, heterozygous mutant (loxl1+/- ) and homozygous mutant (loxl1-/- ) zebrafish were analysed in a range of developmental stages from zebrafish larvae to dissected adult zebrafish eyes. RESULTS: The loxl1 deficiency caused zonular bundling disorders in juvenile zebrafish and accumulation of pearl-like particles adhering to the adult zebrafish zonule. The bundles appeared to lack form and were thinner in both loxl1+/- and loxl1-/- zebrafish compared with the wildtype (p < 0.01 for all Bonferroni post-hoc analyses). The zonule of loxl1-/- zebrafish appeared stretched, ragged and torn, with isolated fibres also detected. The particles in loxl1-/- zebrafish were more numerous (counts: 92.33 ± 10.02/100 µm2 vs. 58.33 ± 5.03/100 µm2 , p = 0.006), but smaller in size (diameter: 0.21 ± 0.03 µm vs. 0.43 ± 0.04 µm, p = 0.002) compared with those in loxl1+/- . Transmission electron microscopy revealed thinning or even loss of elastic lamina in loxl1+/- Bruch's membrane (BM) (thickness of elastic lamina: 92.94 ± 18.19 nm in the wildtype vs 35.65 ± 14.76 nm in loxl1+/- , p = 0.003). The breakage of BM was observed in loxl1-/- . CONCLUSIONS: The loxl1-/- zebrafish is a promising animal model of XFS zonular pathology.

Altered DNA methylation pattern reveals epigenetic regulation of Hox genes in thoracic aortic dissection and serves as a biomarker in disease diagnosis.

BACKGROUND: Thoracic aortic dissection (TAD) is a severe disease with limited understandings in its pathogenesis. Altered DNA methylation has been revealed to be involved in many diseases etiology. Few studies have examined the role of DNA methylation in the development of TAD. This study explored alterations of the DNA methylation landscape in TAD and examined the potential role of cell-free DNA (cfDNA) methylation as a biomarker in TAD diagnosis. RESULTS: Ascending aortic tissues from TAD patients (Stanford type A; n = 6) and healthy controls (n = 6) were first examined via whole-genome bisulfite sequencing (WGBS). While no obvious global methylation shift was observed, numerous differentially methylated regions (DMRs) were identified, with associated genes enriched in the areas of vasculature and heart development. We further confirmed the methylation and expression changes in homeobox (Hox) clusters with 10 independent samples using bisulfite pyrosequencing and quantitative real-time PCR (qPCR). Among these, HOXA5, HOXB6 and HOXC6 were significantly down-regulated in TAD samples relative to controls. To evaluate cfDNA methylation pattern as a biomarker in TAD diagnosis, cfDNA from TAD patients (Stanford type A; n = 7) and healthy controls (n = 4) were examined by WGBS. A prediction model was built using DMRs identified previously from aortic tissues on methylation data from cfDNA. Both high sensitivity (86%) and specificity (75%) were achieved in patient classification (AUC = 0.96). CONCLUSIONS: These findings showed an altered epigenetic regulation in TAD patients. This altered epigenetic regulation and subsequent altered expression of genes associated with vasculature and heart development, such as Hox family genes, may contribute to the loss of aortic integrity and TAD pathogenesis. Additionally, the cfDNA methylation in TAD was highly disease specific, which can be used as a non-invasive biomarker for disease prediction.

Acute exposure to N-Ethylpentylone induces developmental toxicity and dopaminergic receptor-regulated aberrances in zebrafish larvae.

N-Ethylpentylone (NEP) is one of the most recent novel stimulants, and there is limited understanding of its toxicity. Here we employed zebrafish model for analyzing the effects of NEP on early embryos and cardiovascular and nervous systems at late developmental stages. We first observed multi-malformations in early embryos and larvae after NEP administration, together with significant deregulations of brain and heart development-associated genes (neurog1, her6, elavl3, nkx2.5, nppa, nppb, tnnt2a) at transcriptional level. Low-dosed NEP treatment induced an anxiety-like phenotype in zebrafish larvae, while higher doses of NEP exerted an inhibitory effect on locomotion and heart rate. Besides, the expression of th (tyrosine hydroxylase) and th2 (tyrosine hydroxylase 2), identifying dopamine (DA) release, were significantly increased during one-hour free swimming after effective low-dosed NEP administration, along with the upregulation of gene fosab and fosb related to stress and anxiety response. D1R antagonist SCH23390 and D2R antagonist sulpiride partially alleviated the aberrances of locomotion and heart rate, indicating dopaminergic receptors were involved in the bidirectional dosage-dependent pattern of NEP-induced performance. Meanwhile, sulpiride offset the upregulated expression of th, th2 and fosab in the group of 1.5 µM NEP, which highlighted the significant role of D2R in NEP-induced locomotive effects. This study systematically described the developmental, neuronal and cardiac toxicity of NEP in zebrafish, and identified the dopaminergic receptors as one of the downstream effectors of NEP administration.

Manipulation of focal Wnt activity via synthetic cells in a double-humanized zebrafish model of tumorigenesis.

The canonical Wnt signaling pathway is activated in numerous contexts, including normal and cancerous tissues. Here, we describe a synthetic cell-based therapeutic strategy that inhibits aberrant Wnt activity in specific focuses without interfering with the normal tissues in vivo. As a proof of principle, we generated a triple transgenic zebrafish liver cancer model that conditionally expressed human MET and induced ectopic Wnt signaling in hepatocytes. Then, we generated a customized synthetic Notch receptor (synNotch) cascade to express Wnt inhibitor DKK1 in Jurkat T cells and human peripheral blood mononuclear cells (PBMCs) after recognizing MET as antigen. After that, the synNotch PBMCs were sorted and microinjected into different tissues of the zebrafish model. In MET-expressing cancerous liver tissues, the injected cells expressed DKK1 and inhibited the local proliferation and Wnt activity; while in the yolk sac without MET, the injected cells remained inactive. Overall, our studies revealed the use of synthetic cells with antigen receptors to improve the spatiotemporal accuracy of anti-Wnt therapy, and proposed that the genetically humanized zebrafish model may serve as a small-scale and highly optically accessible platform for the functional evaluation of human synthetic cells.

Combinatorial Normalization of Liver-Derived Cytokine Pathways Alleviates Hepatic Tumor-Associated Cachexia in Zebrafish.

The role and significance of liver-derived cytokines in cancer-associated cachexia syndrome remain elusive. Here we report that combinatorial counterbalances of the leptin and Igf1 signaling pathways in hepatocellular carcinoma (HCC) models significantly relieves cachexia. Double transgenic zebrafish models of HCC that stably displayed focal lesions, anorexia, and wasting of adipose and muscle tissues were first generated. Knockout of lepr or mc4r from these zebrafish partially restored appetite and exerted moderate or no effect on tissue wasting. However, genetic replenishment of Igf1 in a lepr-mutant background effectively relieved the cachexia-like phenotype without affecting tumor growth. Similarly, administration of napabucasin, a Stat3/Socs3 inhibitor, on the zebrafish HCC model, mammalian cell lines with exogenous IGF1, and two mouse xenograft models restored insulin sensitivity and rescued the wasting of nontumor tissues. Together, these results describe the synergistic impact of leptin and Igf1 normalization in treating certain HCC-associated cachexia as a practical strategy. SIGNIFICANCE: Disruption of leptin signaling with normalized Igf1 expression significantly rescues anorexia, muscle wasting, and adipose wasting in Ras- and Myc-driven zebrafish models of HCC.

Efficacy and Safety of Tirofiban in Clinical Patients With Acute Ischemic Stroke.

BACKGROUND: Intravenous thrombolysis and endovascular thrombectomy have been approved for acute ischemic stroke (AIS). However, only a minority of patients received these treatments in China. We aimed to evaluate the efficacy and safety of tirofiban in patients with AIS who were not undergoing early recanalization treatments. METHODS: Patients with mild-to-moderate stroke [National Institutes of Health Stroke Scale (NIHSS) score, 4-15] were enrolled in this study. Patients due to cardiogenic embolism were excluded. Eligible patients within 12 h from symptom onset were randomly assigned (1:1) to receive tirofiban (a loading dose of 0.4 µg/kg/min over 30 min and a maintenance dose of 0.1 µg/kg/min up to 48 h) followed by regular treatment or to receive regular treatment (aspirin at a dose of 100 mg per day for 90 days) (control). The primary outcome was the proportion of favorable functional outcomes at 90 days [defined as the modified Rankin Scale (mRS) score of 0-2]. The secondary outcomes included a shift in the distribution of the mRS scores at 90 days and the NIHSS score at 24 h and 7 days. The primary safety outcome was symptomatic intracranial hemorrhage (sICH) within 7 days after tirofiban treatment. RESULTS: A total of 380 eligible patients were randomly assigned to the tirofiban group (n = 190) or the control group (n = 190). The proportion of favorable functional outcomes was higher in the tirofiban group (79.1%) than that in the control group (67.8%) at 90 days [odds ratio (OR), 1.80; 95% CI, 1.12-2.90; p = 0.0155]. An improvement was also observed in the overall distribution of the 90-day mRS scores (adjusted common OR, 2.31; 95% CI, 1.58-3.39; p < 0.0001). Additionally, the median NIHSS score was lower in the tirofiban group than in the control group at 7 days (3 vs. 5, p < 0.0001). Next, we observed that the occurrence of sICH did not differ between the two groups. CONCLUSION: Our trial supports that tirofiban was safe and effective and might be a remedial treatment for patients with AIS who did not receive recanalization treatments. CLINICAL TRIAL REGISTRATION: http://www.chictr.org.cn/, identifier: ChiCTR2000031297.

Increased anxiety was found in serpini1 knockout zebrafish larval.

Serpini1, which encodes neuroserpin, has been implicated in the development and normal function of the nervous system. Mutations in serpini1 cause familial encephalopathy, a rare neurodegenerative disorder characterized with neuroserpin inclusion bodies. However, function of neuroserpin in the nervous system is not fully understood. In this study, we generated a novel serpini1 mutant zebrafish model to investigate the loss of function of neuroserpin. Serpini1- deficient mutation was created with the CRISPR/Cas9 technique. No severe morphological characteristics were found in serpini1- deficient zebrafish. Serpini1-/- zebrafish larvae did not cause locomotor defects but displayed anxiety-like behavior. Extension of motoneurons axon defect was observed in serpini1-/- zebrafish. Furthermore, RNA-sequencing analysis revealed that loss of serpini1 resulted in affected expression of neurodegeneration-related genes.

Dampened VEPH1 activates mTORC1 signaling by weakening the TSC1/TSC2 association in hepatocellular carcinoma.

BACKGROUND & AIMS: Abnormal activation of mTORC1 signaling occurs at high frequency in hepatocellular carcinoma (HCC). However, the underlying causes of this aberrant activation remain elusive. In this study, we identified ventricular zone expressed pleckstrin homology domain-containing 1 (VEPH1) as a novel tumor suppressor that acts via the mTORC1 axis. METHODS: We performed quantitative reverse-transcription PCR (92 pairs), western blot (30 pairs), and immunostaining (225 cases) assays in HCC tissue samples to evaluate VEPH1 expression. We explored the functional effects of VEPH1 on tumor growth and metastasis. Molecular and biochemical strategies were used to gain insight into mechanisms underlying the tumor-suppressive function of VEPH1. RESULTS: VEPH1 is frequently silenced in HCC tissues, primarily resulting from let-7d upregulation. Decreased VEPH1 expression is associated with poor prognosis and aggressive tumor phenotypes in patients with HCC. VEPH1 mediates its tumor-suppressing activity through regulation of cell proliferation, migration and invasion in vitro and in vivo. The VEPH1 fragments 580-625aa and 447-579 aa bind directly to TSC1 (719-1,164aa) and TSC2 (1-420 aa), respectively, enhancing TSC1/TCS2 binding and promoting translocation of TSC2 to the membrane, which leads to increased TSC2 Ser1387 phosphorylation. Subsequently, Rheb is inactivated by the GTPase activity of TSC2, inhibiting mTORC1 signaling and contributing to changes in HCC carcinogenesis and metastasis. Rapamycin, the mTOR inhibitor, can inhibit the pro-tumorigenic effect of VEPH1 knockdown. Loss of VEPH1 correlates with decreased TSC2 Ser1387 phosphorylation and increased mTOR activity in HCC specimens. CONCLUSIONS: The loss of VEPH1 leads to aberrantly activated mTORC1 signaling in HCC; rapamycin (or rapalogs) may serve as an effective treatment option for patients with HCC and dampened VEPH1 expression. LAY SUMMARY: Abnormally activated mammalian target of rapamycin (mTOR) signaling is associated with poor tumor differentiation, early tumor recurrence and worse overall survival in patients with hepatocellular carcinoma. Herein, we identify low VEPH1 expression as a potential cause of abnormally activated mTOR signaling in hepatocellular carcinoma tissues. mTOR inhibitors could thus be an effective treatment option for patients with HCC and low VEPH1 expression.

Combinatorial genetic replenishments in myocardial and outflow tract tissues restore heart function in tnnt2 mutant zebrafish.

Cardiac muscle troponin T (Tnnt2) mediates muscle contraction in response to calcium ion dynamics, and Tnnt2 mutations are associated with multiple types of cardiomyopathy. Here, we employed a zebrafish model to investigate the genetic replenishment strategies of using conditional and inducible promoters to rescue the deficiencies in the heart. tnnt2a mutations were induced in zebrafish via the CRISPR/Cas9 technique, and the mutants displayed heart arrest and dilated cardiomyopathy-like phenotypes. We first utilized the classic myocardial promoter of the myl7 and TetOn inducible system to restore tnnt2a expression in myocardial tissue in tnnt2a mutant zebrafish. However, this attempt failed to recover normal heart function and circulation, although heart pumping was partially restored. Further analyses via both RNA-seq and immunofluorescence indicated that Tnnt2a, which was also expressed in a novel group of myl7-negative smooth muscle cells on the outflow tract (OFT), was indispensably responsible for the normal mechanical dynamics of OFT. Lastly, tnnt2 expression induced by OFT cells in addition to the myocardial cells successfully rescued heart function and circulation in tnnt2a mutant zebrafish. Together, our results reveal the significance of OFT expression of Tnnt2 for cardiac function and demonstrate zebrafish larva as a powerful and convenient in vivo platform for studying cardiomyopathy and the relevant therapeutic strategies.