Inhibitory Effect Research Articles

The Inhibitory Effects of the Herbals Secondary Metabolites (7α-acetoxyroyleanone, Curzerene, Incensole, Harmaline, and Cannabidiol) on COVID-19: A Molecular Docking Study.

Since the COVID-19 outbreak in early 2020, researchers and studies are continuing to find drugs and/or vaccines against the disease. As shown before, medicinal plants can be very good sources against viruses because of their secondary compounds which may cure diseases and help in survival of patients. There is a growing trend in the filed patents in this field. In the present study, we test and suggest the inhibitory potential of five herbal based extracts including 7α-acetoxyroyleanone, Curzerene, Incensole, Harmaline, and Cannabidiol with antivirus activity on the models of the significant antiviral targets for COVID-19 like spike glycoprotein, Papain-like protease (PLpro), non-structural protein 15 (NSP15), RNA-dependent RNA polymerase and core protease by molecular docking study. The Salvia rythida root was extracted, dried, and pulverized by a milling machine. The aqueous phase and the dichloromethane phase of the root extractive were separated by two-phase extraction using a separatory funnel. The separation was performed using the column chromatography method. The model of the important antivirus drug target of COVID-19 was obtained from the Protein Data Bank (PDB) and modified. TO study the binding difference between the studied molecules, the docking study was performed. These herbal compounds are extracted from Salvia rhytidea, Curcuma zeodaria, Frankincense, Peganum harmala, and Cannabis herbs, respectively. The binding energies of all compounds on COVID-19 main targets are located in the limited area of 2.22-5.30 kcal/mol. This range of binding energies can support our hypothesis for the presence of the inhibitory effects of the secondary metabolites of mentioned structures on COVID-19. Generally, among the investigated herbal structures, Cannabidiol and 7α- acetoxyroyleanone compounds with the highest binding energy have the most inhibitory potential. The least inhibitory effects are related to the Curzerene and Incensole structures by the lowest binding affinity. The general arrangement of the basis of the potential barrier of binding energies is in the order below: Cannabidiol > 7α-acetoxyroyleanone > Harmaline> Incensole > Curzerene. Finally, the range of docking scores for investigated herbal compounds on the mentioned targets indicates that the probably inhibitory effects on these targets obey the following order: main protease> RNA-dependent RNA polymerase> PLpro> NSP15> spike glycoprotein.

Dual actions of chloroinconazide on pepper blight in Capsicum annuum: disruption of Phytophthora capsici mycelium and activation of CaCNGC9-mediated SA signaling.

Pepper blight, caused by Phytophthora capsici, is a devastating disease that seriously threatens pepper production worldwide. With the emergence of resistance in P. capsici against conventional fungicides, there is an urgent need to explore novel alternatives for pepper blight management. This study aims to assess the inhibitory effect of chloroinconazide (CHI), a compound synthesized from tryptophan, against pepper blight, and to explore its potential mechanisms of action. The results demonstrated that CHI effectively targeted P. capsici, disrupting its growth and mycelial structure, which resulted in the release of dissolved intracellular substances. Additionally, CHI significantly inhibited the sporangium formation, zoospores release, and zoospores germination, thereby reducing the re-infection of P. capsici. In contrast, the commercial pesticide methylaxyl only inhibited mycelial growth and had limited effect on re-infection, while azoxystrobin inhibited re-infection but had a weak inhibitory effect on mycelial growth. Furthermore, CHI activated the salicylic acid (SA) signaling pathway-mediated immune response to inhibit P. capsici infection in pepper, with this activation being contingent upon cyclic nucleotide-gated ion channel CaCNGC9. CHI exhibited potent dual inhibitory effects on P. capsici by disrupting mycelial structure and activating the CaCNGC9-mediated SA signaling pathway. These dual mechanisms of action suggested that CHI could serve as a promising alternative chemical fungicide for the effective management of pepper blight, offering a new approach to control this devastating disease. Our findings highlighted the potential of CHI as a sustainable and efficient solution to combat the increasing resistance of P. capsici to conventional fungicides, ensuring better crop protection and yield. © 2024 Society of Chemical Industry.

Inhibitory effects of phenylpropionamides from the hemp seed husk on tyrosinase

Tyrosinase is an enzyme involved in melanin production. To identify inhibitors of this enzyme in natural products, extracts and fractions of hemp seed husk were screened for tyrosinase inhibition. Phenylpropionamides 1–9, which were isolated from the ethyl acetate fraction, had inhibitory effects; compounds 3 and 4 had the greatest inhibitory effects. Cannabisin A (3) was a non-competitive and cannabisin B (4) was a competitive inhibitor. The binding site and interaction on tyrosinase of each inhibitor were calculated by computational chemistry. Potential inhibitor 3 decreased the melanin content and tyrosinase activity in B16F10 melanoma cells. Therefore, cannabisin A (3) has potential as a tyrosinase inhibitor.

Effect of olive leaf extract on the quality of Cantal cheese

This study aimed to evaluate the impact of olive leaf extract (OLE) on the quality of Cantal cheese. The cheeses were fortified with different concentrations (Control, 1, 2, and 3 %) of OLE and analysed for quality during storage. The cheeses were evaluated on days 1, 28, and 63 for physicochemical, compositional, textural, rheological parameters, colour, microbiological analysis, and antioxidant activity. The MIR and fluorescence spectroscopies were used to study the changes in the cheese structure. A positive impact of OLE was observed on the antioxidant capacity. A mild inhibitory effect was observed against some beneficial bacteria (such as Lactococcus and Lactobacillus), while a strong inhibitory effect was found against pathogen microorganisms. No negative impact was found on the texture and chemical composition. Significant colour and structural changes were observed. The results indicate the potential of OLE to be used as bioactive ingredient to improve the quality of cheese.

Harnessing potential COX-2 engagement for boosting anticancer activity of substituted 2-mercapto-4(3H)-quinazolinones with promising EGFR/VEGFR-2 inhibitory activities

We designed and synthesized new quinazolinone-tethered phenyl thiourea/thiadiazole derivatives 4–26. Based on their structural characteristics, these compounds were proposed to have a multi-target mode of action for their anticancer activities. Using the MTT assay method, antiproliferative effects were assessed against three human cancer cell lines (HEPG-2, MCF-7, and HCT-116). In vitro assessment for enzymatic inhibitory activity of the most active compounds 4, 9 and 20 was done for EGFR, VEGFR-2 and COX-2 as potential targets. The screened compounds showed low micromolar IC50 inhibitory effects against the three targets. Compound 9 demonstrated similar EGFR/VEGFR-2 inhibitory effect to the control drugs and potential inhibitory activity for COX-2 enzyme. In MCF-7 cells, the most active analog 9 caused 41.02% total apoptosis, and arrested the cell cycle at the G2/M phase. Taken as a whole, the findings of this study provide significant new understandings into the relationship between COX inhibition and cancer therapy. Furthermore, the outcomes showcased the encouraging efficacy of these compounds with a multi-target mechanism, making them excellent choices for additional research and development into possible anticancer drug.

Antimalarial Effects of Nano Chloroquine Loaded Curcumin In vivo.

Malaria is still the deadliest parasitic disease caused by Plasmodium spp. Due to drug resistance and their unpleasant side effects, of conventional researchers are enormously seeking to achieve antimalarial drugs with more curative effective, less toxic and cost-affordable drugs using more advanced technology such as nanodrugs. The present study aimed to examine the antimalarial effects of a novel synthesized nonochloroquine-loaded curcumin relying on dendrimer G2 in susceptible mice. Antimalarial activity and toxicity of the nanocomposite were examined on BALB/C mice with microscopy, checking RBCs morphology and related enzymatic activity rate. The maximum inhibitory effect of the nanocomposite was seen at 10 mg/kg, killing 98% of P. berghei compared to sole chloroquine, whereas ED50 was reported at 5.5 mg/kg. The safety of the synthesized nanocomposite was confirmed with biochemical tests with no detrimental effects on mice. The sustainability and longevity of the nanodrug increased significantly with the NDC-CQ assay compared to the control groups. The study showed that nonochloroquine-loaded curcumin had a promising inhibitory effect on P. berghei growth in infected mice compared to standard drugs. However, further studies and clinical trials with large samples are recommended to study different aspects of using nanodrug.

Heterocycle-functional steroidal derivatives: Design, synthesis, bioevaluation and SARs of steroidal pyrazolo[1,5-a]pyrimidines as potential ALK inhibitors

Two series of heterocyclic steroidal pyrazolo[1,5-a]pyrimidines derived from dehydroepiandrosterone (DHEA) and epiandrosterone (EPIA) were designed and synthesized, and these compounds were screened for their potential antiproliferation activities. The preliminary bioassay indicated that some of target compounds exhibited significantly good antiproliferation activities against human melanoma cell line (A875) and human hepatocellular carcinoma (Huh-7) cell lines compared with 5-fluorouracil (5-FU), and some of which present good antiproliferative activities as potential ALK inhibitors. The detailed analysis of structure–activity relationships (SARs) based on the inhibition activities, kinase assay, and molecular docking demonstrated that the antiproliferation activities of these steroidal pyrazolo[1,5-a]pyrimidine might be affected by the β-hydroxyl group of steroidal scaffold and the N atom of pyridine heterocycle. Especially, compound 4c has certain inhibitory effects on the tyrosine protein kinases ALK, CDK2/CyclinE1, FAK, CDK5/P35, CDK9/CyclinT1, CDK5/P25, PIM2, CDK2/CyclinA2, CDK1/CyclinB1, et c., and which displayed highest inhibitory effect on the kinases of ALK with inhibition rate 40.63 % at the concentration of 10 μM, which induced cell death in A875 cells at least partly (initially), by apoptosis.

Indene as an alternative hydrogen carrier: Performance of supported noble metal catalysts for indene and indane hydrogenation

We propose in this article the use of indene, obtained from the light fraction of coal tar rectification, as a Liquid Organic Hydrogen Carrier (LOHC), with a thorough study of its hydrogenation. The hydrogenation process, conducted on noble metal catalysts (Pd, Pt, Rh, and Ru) supported by activated carbon or alumina, involves the rapid conversion of indene to indane, followed by the formation of cis- and trans-hydrindane. Carbon-supported catalysts exhibited faster reactions rates, with catalytic activities varying among metal phases, Rh and Ru being the most active. Indene was found to have an inhibitory effect on the hydrogenation reactions. While Ru catalysts enhance cracking, Rh catalysts are the most selective for total hydrogenation. The study highlights that Ru catalysts favour cracking reactions more than Rh. The proposed reaction mechanisms involve successive steps and follow a Langmuir-Hinshelwood type kinetics, confirming the inhibitory effect of indene.

Syringin as a novel therapeutic agent for renal cell carcinoma by targeting EGFR/PI3K/Akt pathway and enhancing sunitinib efficacy

Syringin, a natural bioactive compound extracted from Acanthopanax senticosus, has demonstrated potential therapeutic value in cancer treatment. However, its efficacy in treating renal cell carcinoma (RCC) remains unexplored. This study aims to investigate the therapeutic effects and underlying mechanisms of Syringin in RCC. In this study, network pharmacology, molecular docking validation, and bioinformatics were employed to predict the mechanisms by which Syringin affects RCC. In vitro experiments showed that Syringin inhibited RCC cell viability and reduced the IC50 of Sunitinib, enhancing its therapeutic effect. Syringin also inhibited RCC cell proliferation and migration and promoted apoptosis. The combination of Syringin and Sunitinib demonstrated an enhanced inhibitory effect. Western blot analysis confirmed that Syringin’s anti-RCC effects are mediated through the EGFR/PI3K/Akt pathway. In conclusion, our findings indicate that Syringin exhibits inhibitory effects on RCC cells and enhances their sensitivity to Sunitinib, offering a novel approach to exploring treatment strategies for Sunitinib-resistant RCC

TPCA-1 compound, inhibiting testis-specific serine/threonine protein kinase 3 for potential male sterile in Bombyx mori.

Protein kinases are a type of transferase enzyme that catalyze the phosphorylation of protein substrates, including receptor proteins. Testis-specific serine/threonine kinases (TSSKs) are a highly conserved group of protein kinases found in various organisms. They play an essential role in male reproduction by influencing sperm development and function. In this study, we report on the characterization of BmTSSK3, a TSSK from the silkworm, Bombyx mori. We found that BmTSSK3 is specifically expressed in the testis and localized to the sperm flagella, particularly in the sperm tail cyst. Furthermore, we developed BmTSSK3 inhibitors through molecular docking and binding assays. Small molecules 5-(4-Fluorophenyl)-2-ureidothiophene-3-carboxamide (TPCA-1)and Imidurea were identified to bind to BmTSSK3. Using site-specific mutation technology, we identified amino acid residues R134 and S184 as crucial binding sites for small molecules. RNA interference assay and Western blot analysis showed that knockdown of BmTSSK3 significantly decreased histone 3 phosphorylation. To confirm the inhibitory effect of these small molecules, we treated silkworm testes with TPCA-1 and observed a strong inhibitory effect. TPCA-1 is an inhibitor of BmTSSK3, which raises its potential as a future candidate for male sterility of the silkworm. Thus, this study may offer a novel strategy for sterile silkworms as well as insects. © 2024 Society of Chemical Industry.

Influence of coexisting anions on the one-step electrochemical reduction and precipitation removal of Cr(VI): Implications for advanced wastewater treatment

Electroreduction of Cr(VI) coupled with in-situ precipitation of Cr(III) on the cathode is a promising method for removing Cr(VI) from wastewaters. However, the influence of coexisting anions in wastewaters on the electrochemical removal process remains unclear. This study investigated the impact of common inorganic anions, including nitrate (NO3−), chloride (Cl−), phosphate (PO43−) and sulfate (SO42−), on the electrochemical removal processes of Cr(VI). The results indicated that HCrO4− was directly electrochemically reduced to Cr3+, and the OH− generated through electro-mediated water reduction could complex with Cr3+, thereby transforming Cr3+ into chromium hydroxide (Cr(OH)3) coated at cathode. Coexisting anions would partially penetrate the alkaline Cr(III) complexes, inhibiting the formation of Cr(OH)3 passivation layer and promoting the electroreduction of Cr(VI), whose penetration ability followed the order of SO42− > PO43− > Cl− > NO3−. Both the inhibitory effect on Cr(III) precipitation and promoting effect on Cr(VI) reduction were intensified with increasing concentrations of these anions in the range of 1−100 mmol L−1. Accordingly, after electrolysis of 10 mg L−1 Cr(VI) at an initial pH of 3.0 and −0.2 V (vs. Ag/AgCl), the highest electrochemical reduction ratio of Cr(VI) (99.9%) was achieved in the presence of 100 mmol L−1 SO42−, while the total Cr removal ratio was minimal (3.3%). In contrast, the presence of NO3− at 1 mmol L−1 resulted in a nearly lowest reduction ratio of Cr(VI) (92.9%), with the maximum total Cr removal ratio (92.8%). These findings provide new insights into the electrochemical removal mechanisms of Cr(VI) in complex solution environments.

Hydrolysis rate and mechanism of water-dispersed polyesters with different sulfonate group contents in the cutinase-catalyzed system.

Water-dispersed polyester (WPET) has irreplaceable and wide application prospects in the textile field, but it is easy to form printing and dyeing wastewater during traditional fabric treatment. In this paper, cutinase-catalyzed green hydrolysis of WPET was achieved. The main purpose of this paper is to study the difference between two PEG-free WPETs with different -SO3- content in the cutinase-catalyzed hydrolysis process. Differences in solution properties, enzyme activity, SSIPA, and TPA release as well as functional groups and thermal properties of the hydrolysis products of the systems during cutinase hydrolysis were analyzed in detail for 14.1 % S-0 % PEG and 9.5 % S-0 % PEG. Results show that the structure of WPET has an influential effect on its hydrolysis rate in cutinase-catalyzed systems, which mainly originates from the -SO3- content. SSIPA released by WPET during hydrolysis has an inhibitory effect on cutinase enzyme activity. The hydrolysis process of WPET with higher -SO3- content releases more SSIPA, which in turn inhibits the enzyme activity of cutinase in the reaction system, ultimately leading to a slower hydrolysis rate of WPET. These findings provide new ideas for cutinase to target WPETs with different structures and are important for the green development of the textile industry.

Construction of a N-CDs/AuNCs@ZIF-8-assisted ratiometric fluorescent nanosensor for glyphosate detection in edible and medicinal malt

Glyphosate (Gly) is a widely-used herbicide in food production, while, the residue of which due to the long-term and excessive overspray poses serious threats to public health. The development of reliable methods for its sensitive detection is of great urgency. In this study, a novel ratiometric fluorescent nanosensor by encapsulating N-doped carbon dots (N-CDs) and gold nanoclusters (AuNCs) in zeolitic imidazole framework-8 (ZIF-8) as the dual-emissive fluorescence probes at 410 and 650 nm, respectively, was constructed for Gly detection. Due to the adsorption property of ZIF-8, the N-CDs/AuNCs@ZIF-8 nanoprobes accumulated Cu2+ to quench the red fluorescence of AuNCs, and the blue fluorescence of N-CDs was stable. While thiocholine, a product of acetylthiocholine, hydrolyzed by acetylcholinesterase could coordinate with Cu2+, resulting in significant fluorescence recovery of AuNCs. This phenomenon was utilized for the quantitation of Gly, due to its inhibitory effect on acetylcholinesterase activity. By calculating the fluorescence intensity ratio (I650/I410), Gly in real sample could be accurately determined in a concentration range of 2–100 ng/mL with a limit of detection of 1.92 ng/mL because of the anti-interference and the self-correction ability of the two fluorescence signals. The N-CDs/AuNCs@ZIF-8 nanoprobes-assisted ratiometric fluorescent nanosensor exhibited unique merits of rapid construction, simple operation, high specificity, and good accuracy for Gly in edible and medicinal malt samples with recoveries of 93.7–108.2 %. This study presents a multiple tool for the versatile sensing of trace pesticides in more food matrices, which can be extended to a full range of environmental and food safety applications.

Diosmin reduces the stability of Snail and Cyclin D1 by targeting FAK to inhibit NSCLC progression

BackgroundIn different tumours, focal adhesion kinase (FAK), a nonreceptor tyrosine kinase, is upregulated and hence, it represents a promising target for cancer therapy. However, the development of FAK kinase inhibitors has faced a number of challenges. It is therefore imperative that new, effective FAK kinase inhibitors be identified promptly. MethodsSmall molecules that target FAK were identified through molecular docking and validated through surface plasmon resonance (SPR) and cell thermal shift analysis. We investigated the pharmacological effects of FAK kinase inhibitors using CCK-8, colony formation, EdU, and Transwell assays and cell cycle analysis. The molecular mechanism was determined via methods such as coimmunoprecipitation, RNA pull-down and RNA immunoprecipitation. ResultsHere, we confirmed that diosmin (Dio) is an inhibitor of FAK and demonstrated its anti-proliferative and anti-metastatic effects in lung adenocarcinoma. Mechanistically, Dio inhibited tumour proliferation and metastasis by impeding the catalytic activity of FAK. Dio activated the ubiquitin proteasome pathway to induce Cyclin D1 degradation, while inhibiting tumour proliferation and reversing the epithelial mesenchymal transition (EMT) process by reducing the mRNA stability of Snail, thereby inhibiting cancer metastasis. In addition, the inhibitory effect of Dio on lung adenocarcinoma was validated in a mouse xenograft model. ConclusionThese results support the tumour-promoting role of FAK in lung adenocarcinoma by stabilizing Cyclin D1 and Snail and suggest that Dio is a promising candidate for FAK inhibition.

One-pot chemobiological cascade strategy for synthesizing furyl hydroxymethyl ketone from corncob

Furyl hydroxymethyl ketone (FHK) is essential in synthesizing pharmaceuticals and biologically active molecules. However, green and sustainable methods for synthesizing FHK from renewable feedstocks remain challenging. This research proposed a simple one-pot chemobiocatalytic cascade strategy for effectually transforming renewable biomass into FHK via solid acid catalysis and whole-cells hydroxymethylation. Using whole-cells (SMPDC cells) containing pyruvate decarboxylase and small ubiquitin-like modifier (SUMO) fusion tag as biocatalysts, non-natural substrates furfural and formaldehyde were introduced into a C-C ligation bioreaction, establishing a biocatalytic pathway for FHK synthesis. SMPDC cells exhibited excellent biocatalytic activity and high tolerance to high furfural concentrations (up to 200mM), a well-known effective microbial inhibitor. Optimal reaction conditions were identified, enabling the manufacturing of FHK from corncob- and D-xylose-derived furfural with productivities of 0.12g FHK/(1g corncob + 0.09g HCHO) and 0.39g FHK/(1g D-xylose + 0.28g HCHO), respectively. This strategy demonstrated the potential for synthesizing valuable chemicals from low-cost biomass, providing a sustainable alternative to traditional chemical synthesis.

Liang-Ge-San protects against viral infection-induced acute lung injury through inhibiting α7nAChR-mediated mitophagy.

Acute lung injury (ALI) is the main cause of death in clinical respiratory virus infection. Liang-Ge-San (LGS), a famous traditional Chinese formula, has been proved to be effective in treating ALI caused by lipopolysaccharide. However, the effects of LGS on ALI induced by viral infections remain uncertain. To investigate the effect and mechanism of action of LGS on viral infection-induced ALI. The inhibitory effects of LGS on virus-induced inflammation in vitro were evaluated by qRT-PCR and ELISA. The protein expression of α7nAChR was examined by Western blotting. α7nAChR was inhibited by the transfection of siRNA or methyllycaconitine citrate (MLA, an α7nAChR inhibitor) to investigate the role of α7nAChR in the anti-inflammatory effect of LGS. Adoptive culture and co-culture systems of macrophages RAW264.7 and alveolar epithelial cells MLE-12 were established to mimic their interaction. Western blotting, immunofluorescence, flow cytometry and transmission electron microscopy were used to examine the effects of LGS on mitophagy inhibition. In vivo, ALI mouse models induced by SARS-CoV-2, H1N1 or Poly(I:C) infection were established to explore the therapeutic effect and mechanism of LGS. LGS reduced the release of IL-6, TNF-α and IL-1β and increased the expression of α7nAChR in virus-infected RAW264.7 cells. The blockage of α7nAChR counteracted the anti-inflammatory effect of LGS. Moreover, LGS significantly inhibited autophagy in MLE-12 cells induced by Poly(I:C) in adoptive culture and co-culture systems of RAW264.7 and MLE-12 cells, which could be attenuated after the inhibition of α7nAChR in RAW264.7 cells by decreasing the secretion of IL-6, TNF-α and IL-1β. Further study showed that LGS suppressed TNF-α-induced mitochondrial damage and mitophagy by inhibiting the generation of ROS in MLE-12 cells. In vivo, LGS significantly prolonged the survival time, alleviated pathological injury and acute inflammation of ALI mice induced by SARS-CoV-2, H1N1 or Poly(I:C) infection which were associated with the inhibition of α7nAChR-mediated mitophagy. This study first demonstrates that LGS inhibits virus infection-induced inflammation in RAW246.7 cells by increasing the expression of α7nAChR, thereby inhibiting mitophagy induction in MLE-12 cells to alleviate ALI. This work indicates that LGS may serve as a candidate drug for treating ALI/ARDS caused by viral infection.

C1q/tumor necrosis factor-related protein-6 suppresses the angiotensin II-induced differentiation of cardiac fibroblasts to myofibroblasts via activation of the AMPK pathway.

C1q/tumor necrosis factor-related protein-6 (CTRP6) has multiple protective effects against cardiovascular diseases. Myofibroblast differentiation plays a critical role in cardiac fibrosis under various cardiac pathological conditions. The aim of the present study was to determine the effects of CTRP6 on cardiac fibrosis, and to identify the possible mechanisms of action. Toward this end, we measured the expression of fibrotic markers, including collagen I, collagen III, CTGF, and TGFβ1, and assessed the effects of CTRP6 on cardiac fibroblast differentiation into myofibroblasts. CTRP6 inhibited the expression of the angiotensin II (Ang II)-induced myofibroblast markers α-SMA and SM22, and of profibrotic molecules, including collagen I, collagen III, CTGF, TGFβ1, MMP2, MMP9, and TIMP1. Furthermore, CTRP6 significantly attenuated the proliferation and migration of cardiac fibroblasts incubated with Ang II and activated the phosphorylation of AMP-activated protein kinase (AMPK). Incubation with an AMPK inhibitor reversed the subsequent inhibitory effects of CTRP6 on Ang II-induced myofibroblast differentiation. Therefore, CTRP6 suppresses cardiac fibrosis by inhibition of myofibroblast differentiation via AMPK pathway activation, suggesting CTRP6 as a target for the treatment of cardiac fibrosis.

Long-term therapeutic efficacy and safety profiles of hpCas13d RNA editing in treating early-onset hypertrophic cardiomyopathy

AimsThrough evolving a precise RNA nuclease, hpCas13d, we have successfully inhibited hypertrophic cardiomyopathy in a compound heterozygous model. However, further investigation is needed to assess the long-term therapeutic effects and safety profiles of hpCas13d treatment. Materials and methodsAAV-hpCas13d[RQ] was subcutaneously injected into neonatal Myh6RH/RQ mice. Sequential echocardiography analyses were conducted at 4 months and 12 months to evaluate the sustained therapeutic effects of hpCas13d. Electrocardiography was employed to assess cardiac arrhythmias, and mice were euthanized at 12 months. Quantification of Myh6RQ degradation induced by hpCas13d[RQ] was performed using digital droplet PCR and cDNA sequencing. Histological analysis, RNA sequencing, and proteomic analyses were utilized to examine the inhibitory effects on pathological phenotypes and downstream signaling pathways. Biodistribution, tissue damage, and host immune response to AAV-hpCas13d[RQ] were assessed to evaluate long-term safety profiles. Key findingsThe allele-specific RNA degradation persisted for 12 months in AAV-hpCas13d[RQ]-treated Myh6RH/RQ mice. Partial degradation of pathogenic Myh6RQ transcripts proved adequate for the long-term inhibition of cardiac hypertrophy, arrhythmias, fibrosis, and cellular apoptosis in Myh6RH/RQ mice. RNA sequencing and proteomic analyses revealed that hpCas13d[RQ] treatment impeded hypertrophy and fibrosis, mitochondrial dysfunction, and abnormalities in ion channels downstream of mutant Myh6. Prolonged treatment with AAV-hpCas13d from the neonatal stage did not induce significant tissue damage, liver toxicity, humoral responses, or cellular immune reactions against the AAV9 capsid and bacterial hpCas13d. SignificanceThese results underscore the promising translational potential of AAV-hpCas13d in treating cardiovascular diseases and advancing in vivo gene therapy.

Impact of controlling shareholders’ equity pledges on firms’ labour investment efficiency

Controlling shareholders in the Chinese capital market are increasingly obtaining pledged loans from financial institutions through equity pledges. This study analyses the relationship between controlling shareholders’ equity pledges and the labour investment efficiency of listed companies using a sample of listed companies from 2009 to 2022. Controlling shareholders’ equity pledges reduce labour investment efficiency by exacerbating capital pressure and refocusing business investment. Furthermore, the inhibitory effect of equity pledges on labour investment efficiency is more pronounced in companies with lower audit quality, lower equity checks and balances and higher degrees of equity pledges.

Lithospermic acid inhibits dengue virus infection through binding with envelope proteins

The dengue virus has emerged as a global pandemic, highlighting the urgent need for the immediate development of antiviral therapeutics. Lithospermum erythrorhizon, a medicinal plant commonly used in China for various ailments including viral infections, inflammation, rheumatism, and cancer, showed promising antiviral properties in our research. Specifically, both the ethanol extract of Lithospermum erythrorhizon and its active component, lithospermic acid, demonstrated significant inhibitory effects on Dengue virus (DENV) replication in Vero cells, with EC50 values of 6.50 μg/mL(95% CI: 2.25 to 18.98)and 15.00 μM(95% CI: 12.13 to 18.07), respectively. Notably, lithospermic acid exhibited potent antiviral activity across multiple cell lines against DENV, impeding virus replication and specifically impeding the expression of viral E and NS3 proteins during the early stages of DENV infection. Experimental assays involving RNase digestion and sucrose density gradient analysis confirmed that lithospermic acid did not directly inactivate DENV but rather interfered with viral processes. Furthermore, the compound was found to bind to the E protein of DENV, effectively inhibiting viral infection and mitigating the cytopathic effects induced by DENV. Collectively, these findings underscore the potential of lithospermic acid as a promising candidate for the development of therapeutic strategies targeting DENV infection.