Combined transcriptome and widely targeted metabolome analysis reveals the potential mechanism of HupA biosynthesis and antioxidant activity in Huperzia serrata.

Introduction: Huperzia serrata is a traditional Chinese herb that has gained much attention for its production of Huperzine A (HupA). HupA has shown promise on treating Alzheimer's disease (AD). However, the biosynthetic pathway and molecular mechanism of HupA in H. serrata are still not well understood. Methods: Integrated transcriptome and metabolome analysis was performed to reveal the molecular mechanisms related to HupA biosynthesis and antioxidant activity in Huperzia serrata. Results: HT (in vitro H. serrata thallus) exhibits higher antioxidant activity and lower cytotoxicity than WH (wild H. serrata). Through hierarchical clustering analysis and qRT-PCR verification, 7 important enzyme genes and 13 transcription factors (TFs) related to HupA biosynthesis were detected. Among them, the average |log2FC| value of CYP (Cytochrome P450) and CAO (Copper amine oxidase) was the largest. Metabolomic analysis identified 12 metabolites involved in the HupA biosynthesis and 29 metabolites related to antioxidant activity. KEGG co-enrichment analysis revealed that tropane, piperidine and pyridine alkaloid biosynthesis were involved in the HupA biosynthesis pathway. Furthermore, the phenylpropanoid, phenylalanine, and flavonoid biosynthesis pathway were found to regulate the antioxidant activity of H. serrata. The study also identified seven important genes related to the regulation of antioxidant activity, including PrAO (primary-amine oxidase). Based on the above joint analysis, the biosynthetic pathway of HupA and potential mechanisms of antioxidant in H. serrata was constructed. Discussion: Through differential transcriptome and metabolome analysis, DEGs and DAMs involved in HupA biosynthesis and antioxidant-related were identified, and the potential metabolic pathway related to HupA biosynthesis and antioxidant in Huperzia serrata were constructed. This study would provide valuable insights into the HupA biosynthesis mechanism and the H. serrata thallus medicinal value.

A Unique Approach: Biomimetic Graphdiyne-Based Nanoplatform to Treat Prostate Cancer by Combining Cuproptosis and Enhanced Chemodynamic Therapy.

Purpose: Current treatment approaches for Prostate cancer (PCa) often come with debilitating side effects and limited therapeutic outcomes. There is urgent need for an alternative effective and safe treatment for PCa. Methods: We developed a nanoplatform to target prostate cancer cells based on graphdiyne (GDY) and a copper-based metal-organic framework (GDY-CuMOF), that carries the chemotherapy drug doxorubicin (DOX) for cancer treatment. Moreover, to provide GDY-CuMOF@DOX with homotypic targeting capability, we coated the PCa cell membrane (DU145 cell membrane, DCM) onto the surface of GDY-CuMOF@DOX, thus obtaining a biomimetic nanoplatform (DCM@GDY-CuMOF@DOX). The nanoplatform was characterized by using transmission electron microscope, atomic force microscope, X-ray diffraction, etc. Drug release behavior, antitumor effects in vivo and in vitro, and biosafety of the nanoplatform were evaluated. Results: We found that GDY-CuMOF exhibited a remarkable capability to load DOX mainly through π-conjugation and pore adsorption, and it responsively released DOX and generated Cu+ in the presence of glutathione (GSH). In vivo experiments demonstrated that this nanoplatform exhibits remarkable cell-killing efficiency by generating lethal reactive oxygen species (ROS) and mediating cuproptosis. In addition, DCM@GDY-CuMOF@DOX effectively suppresses tumor growth in vivo without causing any apparent side effects. Conclusion: The constructed DCM@GDY-CuMOF@DOX nanoplatform integrates tumor targeting, drug-responsive release and combination with cuproptosis and chemodynamic therapy, offering insights for further biomedical research on efficient PCa treatment.

Amplifying Frequency Up-Converted Infrared Signals with a Molecular Optomechanical Cavity.

Frequency up-conversion, enabled by molecular optomechanical coupling, has recently emerged as a promising approach for converting infrared signals into the visible range through quantum coherent conversion of signals. However, detecting these converted signals poses a significant challenge due to their inherently weak signal intensity. In this work, we propose an amplification mechanism capable of enhancing the signal intensity by a factor of 1000 or more for the frequency up-converted infrared signal in a molecular optomechanical system. The mechanism takes advantage of the strong coupling enhancement with molecular collective mode and the Stokes sideband pump. This work demonstrates a feasible approach for up-converting infrared signals to the visible range.

Multiple-photon bundle emission in the n-photon Jaynes-Cummings model.

We study the multiple-photon bundle emission in the n-photon Jaynes-Cummings model composed of a two-level system coupled to a single-mode optical field via the n-photon exciting process. Here, the two-level system is strongly driven by a near-resonant monochromatic field, and hence the system can work in the Mollow regime, in which a super-Rabi oscillation between the zero-photon state and the n-photon state can take place under proper resonant conditions. We calculate the photon number populations and the standard equal-time high-order correlation functions, and find that the multiple-photon bundle emission can occur in this system. The multiple-photon bundle emission is also confirmed by investigating the quantum trajectories of the state populations and both the standard and generalized time-delay second-order correlation functions for multiple-photon bundle. Our work paves the way towards the study of multiple-photon quantum coherent devices, with potential application in quantum information sciences and technologies.

First Report of Stemphylium Leaf Blight Caused by Stemphylium eturmiunum on Welsh Onion in China.

Welsh onion (Allium fistulosum L. ) is native to Southwest China (Tomoo and Shinji 2012), with a cultivated area of about 550,000 ha and a production of 20 million tons in 2020, and widely cultivated throughout the world as a condiment. During April 2020, a Stemphylium-like disease occurred on a three-hectare field in Nanchang County (28°33'N, 115°57'E) was first observed and then monitored in Ningdu (26°23'N, 115°59'E) and Leping (28°50'N, 117°18'E), Jiangxi Province, China. The disease incidence was estimated to be nearly 30% in a survey of 300 plants. Disease symptoms were oval or irregular lesions at the center of outer leaves. The lesions were light brown at the edges and black-violet in the middle. Previous studies reported that the disease was mainly caused by S. vesicarium (Misawa et al. 2012). Besides, S. botryosum (Misawa et al. 2011) and S. solani (Dumin et al. 2021) can also cause this disease. To investigate the causal agent of the disease in Jiangxi, pieces of tissue from the margins of the symptomatic leaves were surface sterilized with 75% ethanol for 15 s, rinsed in sterile water three times, and placed on Potato Dextrose Agar (PDA) followed by incubation at 25°C in the dark for 5 days. Colonies were initially yellowish-brown and gradually turned into reddish-brown after 2 weeks. Conidia were solitary, oblong or ellipsoid, mid to deep brown (22-45×14-25 µm) with transverse and longitudinal septa (Fig. S1). Morphological characteristics were consistent with S. eturmiunum reported by Simmons (Simmons 2001). For molecular identification, genomic DNA was extracted from two representative isolates. ITS and gapdh genes of the isolates were amplified with primers ITS1/ITS4 (White et al. 1990) and gdp1/gdp2 (Berbee et al. 1999). BLASTn analysis showed that ITS sequences (OM846570, OM846571) of the two isolates both > 99% identity with S. eturmiunum strain CBS 109845 (MH862841) (Vu et al. 2018); and also had 100% identity for gapdh gene sequences (OM867337, OM867338) with S. eturmiunum strain CBS 109845 (KU850689) (Woudenberg et al. 2017). Phylogenetic tree based on ITS and gapdh gene sequences were constructed using a maximum-likelihood method with 1000 bootstraps indicated that the two isolates were clustered with S. eturmiunum (Fig. S2). Based on morphological characteristics and molecular analysis, the fungus was identified as S. eturmiunum. A pathogenicity test was performed on healthy welsh onion leaves to fulfill Koch's postulates. Twenty leaves were inoculated with a spore suspension of 1×104 conidia/mL followed by incubation at 25℃ (relative humidity > 90%) in the greenhouse. Negative controls were inoculated with sterile water. The experiment was repeated three times. Purple brown lesions were observed on all inoculated leaves at 7 days post-inoculation that similar to those observed in the field, and control remained symptomless. S. eturmiunum was re-isolated from the diseased leaves. It has reported that S. eturmiunum can infect tomato, garlic, sweet cherry and many other crops and cause different diseases. To the best of our knowledge, this is the first report of S. eturmiunum can cause Stemphylium leaf blight on welsh onion in China. Appropriate strategies should be developed to manage this disease.

Ferrocene-Based Polymeric Nanoparticles Carrying Doxorubicin for Oncotherapeutic Combination of Chemotherapy and Ferroptosis.

Mono-chemotherapy has significant side effects and unsatisfactory efficacy, limiting its clinical application. Therefore, a combination of multiple treatments is becoming more common in oncotherapy. Chemotherapy combined with the induction of ferroptosis is a potential new oncotherapy. Furthermore, polymeric nanoparticles (NPs) can improve the antitumor efficacy and decrease the toxicity of drugs. Herein, a polymeric NP, mPEG-b-PPLGFc@Dox, is synthesized to decrease the toxicity of doxorubicin (Dox) and enhance the efficacy of chemotherapy by combining it with the induction of ferroptosis. First, mPEG-b-PPLGFc@Dox is oxidized by endogenous H2 O2 and releases Dox, which leads to an increase of H2 O2 by breaking the redox balance. The Fe(II) group of ferrocene converts H2 O2 into ·OH, inducing subsequent ferroptosis. Furthermore, glutathione peroxidase 4, a biomarker of ferroptosis, is suppressed and the lipid peroxidation level is elevated in cells incubated with mPEG-b-PPLGFc@Dox compared to those treated with Dox alone, indicating ferroptosis induction by mPEG-b-PPLGFc@Dox. In vivo, the antitumor efficacy of mPEG-b-PPLGFc@Dox is higher than that of free Dox. Moreover, the loss of body weight in mice treated mPEG-b-PPLGFc@Dox is lower than in those treated with free Dox, indicating that mPEG-b-PPLGFc@Dox is less toxic than free Dox. In conclusion, mPEG-b-PPLGFc@Dox not only has higher antitumor efficacy but it reduces the damage to normal tissue.

Enantiodiscrimination of chiral molecules via quantum correlation function.

We propose a method to realize enantiodiscrimination of chiral molecules based on quantum correlation function in a driven cavity-molecule system, where the chiral molecule is coupled with a quantized cavity field and two classical light fields to form a cyclic three-level model. According to the inherent properties of electric-dipole transition moments of chiral molecules, there is a π-phase difference in the overall phase of the cyclic three-level model for the left- and right-handed chiral molecules. Thus, the correlation function depends on this overall phase and is chirality-dependent. The analytical and numerical results indicate that the left- and right-handed chiral molecules can be discriminated by detecting quantum correlation function. Our work opens up a promising route to discriminate molecular chirality, which is an extremely important task in pharmacology and biochemistry.

Untargeted metabolomics reveals alterations in the metabolic reprogramming of prostate cancer cells by double-stranded DNA-modified gold nanoparticles.

Metabolic reprogramming plays an important role in the development of prostate cancer (PCa). However, there are few reports on the effects of nanomaterials as vectors on cancer metabolic reprogramming. Herein, a type of nanoparticle with good biocompatibility was synthesized by modifying the double-stranded of DNA containing a sulfhydryl group on the surface of gold nanoparticles (AuNPs-dsDNA) through salt-aging conjugation methods. The resultant AuNPs-dsDNA complexes possessed low toxicity to PC3 and DU145 cells in vitro. There was also no obvious hepatorenal toxicity after intravenous injection of AuNPs-dsDNA complexes in vivo, which indicated that these nanoparticles had good biological compatibilities. We investigated their biological functions using prostate cancer cells. Seahorse assay showed that AuNPs-dsDNA complexes could increase glycolysis and glycolysis capacity both in PC3 and DU145 cells. We further detected the expression of glycolysis-related genes by qPCR assay, and found that PKM2, PDHA, and LDHA were significantly upregulated. Furthermore, untargeted metabolomics revealed that PC (18:2(9Z,12Z)/18:2(9Z,12Z)) and PC (18:0/18:2 (9Z,12Z)) levels were decreased and inosinic acid level was increased in PC3 cells. Whereas (3S,6E,10E)-1,6,10,14-Phytatetraen-3-ol, Plasmenyl-PE 36:5 and Cer (d18:2/18:2) were decreased, PE 21:3 and 1-pyrrolidinecarboxaldehyde were increased in DU145 cells after co-culturing with AuNPs-dsDNA. In summary, we found that AuNPs and AuNPs-dsDNA complexes possibly regulate the metabolic reprogramming of cancer cells mainly through the lipid metabolic pathways, which could compensate for the previously mentioned phenomenon of enhanced glycolysis and glycolysis capacity. This will provide an important theoretical basis for our future research on the characteristic targeted design of nanomaterials for cancer metabolism.

Manipulating Mesenchymal Stem Cell Differentiation on Nanopattern Constructed through Cell-Mediated Mineralization.

The extracellular matrix microenvironment, including chemical constituents and topological structure, plays key role in regulating the cell behavior, such as adhesion, proliferation, differentiation, apoptosis, etc. Until now, to investigate the relationship between surface texture and cell response, various ordered patterns have been prepared on the surface of different matrixes, whereas almost all these strategies depend on advanced instruments or severe synthesis conditions. Herein, cell-mediated mineralization method has been applied to construct nanopattern on the surface of ß-TCP scaffold. The formation process, morphology, and composition of the final pattern were characterized, and a possible mineralization mechanism has been proposed. Moreover, the cell behavior on the nanopattern has been investigated, and the results showed that the mouse bone marrow mesenchyme stem cells (mBMSCs) display good affinity with the nanopattern, which was manifested by the good proliferation and osteogenic differentiation status of cells. The synthetic strategy may shed light to construct advanced topological structures on other matrixes for bone repair.

Carbon Nanospheres Exert Antitumor Effects Associated with Downregulation of 4E-BP1 Expression on Prostate Cancer.

INTRODUCTION: Although carbon nanospheres (CNPs) are promising nanomaterials in cancer treatment, how they affect prostate cancer (PCa) remains unclear. METHODS: In this study, scanning electron microscopy (SEM), X-ray diffraction (XRD), and Raman spectroscopy were used to confirm the successful synthesis of CNPs. CCK-8, flow cytometry, Transwell, wound healing, Western blot and immunohistochemistry (IHC) assays were performed to evaluate the antitumor effect of CNPs toward the two kinds of prostate cancer cell lines PC3 and DU145. RESULTS: Our results showed that CNPs inhibited cell growth, invasion, and migration and induced apoptosis and autophagy in PCa cells. Multifactor detection of a single Akt phosphorylation pathway and Western blot results suggested the suppression of 4E-BP1 in PCa cells after incubation with CNPs. The results from animal experiments also suggested the antitumor effect of CNPs and reduced 4E-BP1 expression in PCa tissue samples from BALB/c nude mice administered a local subcutaneous injection of CNPs.

Study on the Diagnosis of Gastric Cancer by Magnetic Beads Extraction and Mass Spectrometry.

OBJECTIVE: This study constructed a model for the early diagnosis of gastric cancer by comparing the serum peptides profiles of patients with advanced gastric cancer and healthy people. And that model may be the potential to be applied for the efficacy evaluation and recurrence monitoring in gastric cancer. METHODS: Serums of 30 healthy people and 30 advanced gastric cancer patients were matched by age and gender were collected. The serum peptide spectrum was obtained by MB-WCX concentration and MALDI-TOF MS analysis. Based on the analysis of the efficiency of differential peptides in the diagnosis of gastric cancer, we first established a model for the diagnosis of gastric cancer based on differential peptides and then carried out external verification. The diagnostic reliability of this model was further tested by compared with carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9). RESULTS: In this present study, we found the expression of two peptide peaks with a molecular weight of 2863 Da and 2953 Da were significantly increased in gastric cancer serum, while the expression of two peptide peaks with a molecular weight of 1945 Da and 2082 Da were significantly decreased. Depending on the characteristics of peptide expression, we constructed a diagnostic model, we compared the sensitivity and specificity of the model established by 2953 Da/1945 Da, and found this model is significantly higher than CEA and CA19-9. CONCLUSION: There were some differences in serum peptides profiles between patients with advanced gastric cancer and healthy people. The serum peptide diagnostic models based on 2953 Da and 1945 Da have high diagnostic efficiency for advanced gastric cancer. Our result indicated that this model was well worth further validation for clinical diagnosis.

Enhancement of photon blockade effect via quantum interference.

We study the photon blockade effect in a coupled cavity system, which is formed by a linear cavity coupled to a Kerr-type nonlinear cavity via a photon-hopping interaction. We explain the physical phenomenon from the viewpoint of the conventional and unconventional photon blockade effects. The corresponding physical mechanisms of the two kinds of photon blockade effects are based on the anharmonicity in the eigenenergy spectrum and the destructive quantum interference between two different transition paths, respectively. In particular, we find that the photon blockade via destructive quantum interference also exists in the conventional photon blockade regime and that the unconventional photon blockade occurs in both the weak- and strong-Kerr nonlinearity cases. The photon blockade effect can be observed by calculating the second-order correlation function of the cavity field. This model is general and hence it can be implemented in various experimental setups such as coupled optical-cavity systems, coupled photon-magnon systems, and coupled superconducting-resonator systems. We present some discussions on the experimental feasibility.

Whole Genome Re-sequencing Reveals Natural Variation and Adaptive Evolution of Phytophthora sojae.

Due to the monocultural basis of agricultural crops, mutated plant microbes with increased pathogenicity can easily spread in the field and lead to serious yield losses. As a major threat to a wide range of crop plants, oomycete pathogens continuously undergo adaptive evolution to overcome plant defense barriers. However, the genetic basis of their evolution at the molecular level remains largely unknown. Here, we investigated the nature variation and the population genomics of the soybean pathogen Phytophthora sojae by high-throughput genome re-sequencing. Genomic variation analysis revealed uneven "two-speed" evolutionary pattern with genes in gene-sparse regions (GSRs) showing higher rates of structural polymorphisms and positive selection. GSRs are enriched in effector genes and transposase-related genes. Our results also suggested that the NADH oxidase and MIP transporter gene families undergo rapid and diversifying selection. Furthermore, we demonstrated that P. sojae isolates possess varying numbers of RxLR effectors with diverse sequences, totaling 471 members. Among them, 42 core RxLR effectors are assumed to be important for infection. Finally, we observed that Avr genes exhibit abundant sequence variation in P. sojae isolates. Several novel variants lead to the evading of host resistance, including a complete deletion in Avr3c and amino acid mutations in Avr1a. Taken together, our results provide an adaptive landscape of P. sojae at single-nucleotide resolution, as well as resources for further resistance breeding and disease prevention against this important plant pathogen.

A Phytophthora capsici Effector Targets ACD11 Binding Partners that Regulate ROS-Mediated Defense Response in Arabidopsis.

Reactive oxygen species (ROS) play a vital role in plant immune response, but the genes involved in the regulation of ROS are scantily reported. Phytophthora pathogens produce a large number of effectors to promote infection, but the modes of action adopted are largely unknown. Here, we report that RxLR207 could activate ROS-mediated cell death in Nicotiana benthamiana and was essential for virulence of P. capsici. We found that this effector targeted BPA1 (binding partner of ACD11) and four members of BPLs (BPA1-Like proteins) in Arabidopsis, and the bpa1 and bpl mutants had enhanced ROS accumulation and cell death under biotic or abiotic stresses. Furthermore, we showed that BPA1 and several BPLs functioned redundantly in plant immunity to P. capsici. We discovered that BPA1 and all six BPLs interacted with ACD11, and stabilization of ACD11 was impaired in the bpa1, bpl2, bpl3, and bpl4 mutants. RxLR207 could promote the degradation of BPA1, BPL1, BPL2, and BPL4 to disrupt ACD11 stabilization in a 26S proteasome-dependent manner. Taken together, these findings indicate the important roles of Arabidopsis BPA1 and its homologs in ROS homeostasis and defense response, highlighting the usefulness of a pathogen effector-directed approach as a promising strategy for the discovery of novel plant immune regulators.

Folate-decorated PEG-PLGA nanoparticles with silica shells for capecitabine controlled and targeted delivery.

Di-block polymer of poly (lactic-co-glycolic acid)-poly (ethylene glycol) (PLGA-PEG) end-capped with capecitabine (CAP) at the hydrophobic domain and folate (FA) at hydrophilic domain were synthesized respectively. The products were extensively studied by nuclear magnetic resonance ((1)H NMR) and gel permeation chromatography (GPC) measurement. By using emulsion-solvent evaporation method, the two conjugates, drug CAP and tetramethoxysilane (TMOS) were mixed to form the CAP entrapped nanoparticles (NPs) with the FA moieties exposed on NPs surface, while simultaneously forming a cross-linked silica shell out of hydrophobic PLGA core domain. The testing results showed the CAP-loaded NPs presented suitable physical stability, favorable size around 200 nm, negative zeta potential charge (-28.43 ± 2.55 mV) and high encapsulation efficiency (69%). Both silica shell cross-linked drug-loaded NPs (SSCL NPs) and none silica shell cross-linked NPs (NSSCL NPs) provided an initial burst release and followed by a sustained two-stage release of the CAP. Straight lines approximate the steady-state for the two-stage release, and the K/K' of the two stages are 1.96304 and 1.78697 respectively suggesting the silica shell influenced the release of first stage significantly.

Temozolomide loaded PLGA-based superparamagnetic nanoparticles for magnetic resonance imaging and treatment of malignant glioma.

Polysorbate 80 coated temozolomide-loaded PLGA-based superparamagnetic nanoparticles (P80-TMZ/SPIO-NPs) were successfully synthesized and characterized as drug carriers and diagnosis agent for malignant brain glioma. The mean size of P80-TMZ/SPIO-NPs was 220 nm with narrow hydrodynamic particle size distribution. The superparamagnetic characteristic of P80-TMZ/SPIO-NPs was proved by vibration simple magnetometer. P80-TMZ/SPIO-NPs exhibited high drug loading and encapsulation efficiency as well as good sustained drug release performance for 15 days. MTT assay demonstrated the antiproliferative effect of P80-TMZ/SPIO-NPs for C6 glioma cells. Significant cellular uptake of P80-TMZ/SPIO-NPs was evaluated in C6 glioma cells by fluorescence microscopy, Prussian blue staining, and atomic absorption spectrophotometer (AAS) for qualitative and quantitative study, respectively. MRI scanning analyses in vitro indicated that P80-TMZ/SPIO-NPs could be used as a good MRI contrast agent. Polysorbate 80 coated temozolomide-loaded PLGA-based superparamagnetic nanoparticles could be able to promise a multifunctional theragnostic carrier of brain cancer.