A dual-response fluorescent probe for norepinephrine and viscosity and its application in depression research.

Depression is a serious mental disease that causes grievous harm to human health and quality of life. The vesicular exocytosis of noradrenaline (NE), rather than its intrinsic intracellular concentration, is more associated with depression. Based on the reports on exocytosis of NE, it is reasonable to assume that the viscosity of cells has an important effect on the release of NE. Herein, a dual-response fluorescent probe (RHO-DCO-NE) for detecting NE and viscosity was designed and synthesized. The probe can simultaneously detect NE concentration and viscosity level with negligible crosstalk between the two channels. We utilized the probe to study the effect of viscosity changes on the NE release of PC12 and the corticosterone-induced PC12 cells. The experiment data revealed that the decrease in viscosity level can accelerate the release of NE of depression cell models. The finding provides new insight into the study of the pathological mechanisms of depression.

Accurate detection depression cell model with a dual-locked fluorescence probe in response to noradrenaline and HClO.

Due to the serious harm of depression to human health and quality of life, an accurate diagnosis of depression is warranted. For the complex etiology of depression, a single biomarker diagnostic method often leads to misdiagnosis. As noradrenaline and HClO are closely related to depression, a "dual-locked" fluorescence probe R-NE-HClO for diagnosing of depression through the simultaneous detection of noradrenaline and HClO was designed and synthesized. Fluorescence of R-NE-HClO can only be restored in the presence of both noradrenaline and HClO. The probe demonstrates excellent selectivity for noradrenaline and HClO and low cytotoxicity in cell imaging experiments. It is to be observed that we successfully applied the probe to accurately detect depressed cells which provides a possible tool for diagnosing depression.

Identification of a broad sarbecovirus neutralizing antibody targeting a conserved epitope on the receptor-binding domain.

Omicron, as the emerging variant with enhanced vaccine tolerance, has sharply disrupted most therapeutic antibodies. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) belongs to the subgenus Sarbecovirus, members of which share high sequence similarity. Herein, we report one sarbecovirus antibody, 5817, which has broad-spectrum neutralization capacity against SARS-CoV-2 variants of concern (VOCs) and SARS-CoV, as well as related bat and pangolin viruses. 5817 can hardly compete with six classes of receptor-binding-domain-targeted antibodies grouped by structural classifications. No obvious impairment in the potency is detected against SARS-CoV-2 Omicron and subvariants. The cryoelectron microscopy (cryo-EM) structure of neutralizing antibody 5817 in complex with Omicron spike reveals a highly conserved epitope, only existing at the receptor-binding domain (RBD) open state. Prophylactic and therapeutic administration of 5817 potently protects mice from SARS-CoV-2 Beta, Delta, Omicron, and SARS-CoV infection. This study reveals a highly conserved cryptic epitope targeted by a broad sarbecovirus neutralizing antibody, which would be beneficial to meet the potential threat of pre-emergent SARS-CoV-2 VOCs.

The highly selective rhodol-based putrescine probe and visual sensors for on-site detection of putrescine in food spoilage.

Putrescine (Butane-1,4-diamine) has been regarded as a vital marker of spoiling protein-rich foods, especially meat and seafood. The detection of putrescine in food is considered a convenient and powerful method for evaluating the degree of spoilage of protein-rich foods. Herein, a novel rhodol-based fluorescent probe RSMA (formyl-rhodol Schiff base with methoxyaniline) was developed to detect putrescine. RSMA exhibited excellent linearity (R2 = 0.9912) in the concentration range of 0-45 µM of putrescine with a detection limit as low as 0.45 µM. Although RSMA had moderate responses to some aliphatic diamines, the selectivity of RSMA for putrescine was one of the best reported in the literature so far. Moreover, RSMA was successfully fabricated to solid-state sensors for on-site detection of putrescine in shrimp, that demonstrated its application in monitoring food spoilage.

A potent and broad-spectrum neutralizing nanobody for SARS-CoV-2 viruses, including all major Omicron strains.

SARS-CoV-2 viruses are highly transmissible and immune evasive. It is critical to develop broad-spectrum prophylactic and therapeutic antibodies for potential future pandemics. Here, we used the phage display method to discover nanobodies (Nbs) for neutralizing SARS-CoV-2 viruses especially Omicron strains. The leading nanobody (Nb), namely, Nb4, with excellent physicochemical properties, can neutralize Delta and Omicron subtypes, including BA.1, BA.1.1 (BA.1 + R346K), BA.2, BA.5, BQ.1, and XBB.1. The crystal structure of Nb4 in complex with the receptor-binding domain (RBD) of BA.1 Spike protein reveals that Nb4 interacts with an epitope on the RBD overlapping with the receptor-binding motif, and thus competes with angiotensin-converting enzyme 2 (ACE2) binding. Nb4 is expected to be effective for neutralizing most recent Omicron variants, since the epitopes are evolutionarily conserved among them. Indeed, trivalent Nb4 interacts with the XBB1.5 Spike protein with low nM affinity and competes for ACE2 binding. Prophylactic and therapeutic experiments in mice indicated that Nb4 could reduce the Omicron virus loads in the lung. In particular, in prophylactic experiments, intranasal administration of multivalent Nb4 completely protected mice from Omicron infection. Taken together, these results demonstrated that Nb4 could serve as a potent and broad-spectrum prophylactic and therapeutic Nb for COVID-19.

Preparation and Magnetic Manipulation of Fe3O4/Acrylic Resin Core-Shell Microspheres.

Core-shell microspheres refer to duo-layer or multilayer microspheres, which are widely used in drug delivery, microreactors, etc. Accurate manipulation of microspheres is a research hot spot, while traditional manipulation methods including ultrasonic manipulation and laser manipulation still face some limitations. In this study, magnetic core-shell microspheres were adopted to realize the accurate manipulation of microspheres. Combined with microfluidic technology, polystyrene sulfonic acid (PSSA)/Fe3O4 magnetic fluid was utilized as the core material and photosensitive acrylic resin became the shell material. After UV curing, a magnetic core-shell microsphere with an average size of 55 µm could be achieved, and the diameter was uniform and controllable. By adjusting the flow rate of the dispersed phase, the dual-core microspheres with different core particle sizes that ranged from 9.3 to 28.4 µm could be prepared. Experimental results showed that the prepared Fe3O4/acrylic resin core-shell microspheres can be used as functionalized microspheres that have good magnetic response properties and self-assembly ability. In addition, the magnetic manipulation and self-assembly of the prepared core-shell microspheres were presented with different external magnetic fields. The magnetic core-shell microspheres have shown great potential in the fields of biomedical engineering and targeted delivery of drugs.

Identification of GnRH-like peptide and its potential signaling pathway involved in the oocyte meiotic maturation in the Chinese mitten crab, Eriocheir sinensis.

Gonadotropin-releasing hormone (GnRH) plays a pivotal role in reproductive regulation in vertebrates. However, GnRH was rarely isolated and its function remains poorly characterized in invertebrates. The existence of GnRH in ecdysozoa has been controversial for a long. Here, we isolated and identified two GnRH-like peptides from brain tissues in Eriocheir sinensis. Immunolocalization showed that the presence of EsGnRH-like peptide in brain, ovary and hepatopancreas. Synthetic EsGnRH-like peptides can induce germinal vesicle breakdown (GVBD) of oocyte. Similar to vertebrates, ovarian transcriptomic analysis revealed a GnRH signaling pathway in the crab, in which most genes exhibited dramatically high expression at GVBD. RNAi knockdown of EsGnRHR suppressed the expression of most genes in the pathway. Co-transfection of the expression plasmid for EsGnRHR with reporter plasmid bearing CRE-luc or SRE-luc response element into 293T cells showed that EsGnRHR transduces its signal via cAMP and Ca2+ signaling transduction pathways. In vitro incubation of the crab oocyte with EsGnRH-like peptide confirmed the cAMP-PKA cascade and Ca2+ mobilization signaling cascade but lack of a PKC cascade. Our data present the first direct evidence of the existence of GnRH-like peptides in the crab and demonstrated its conserved role in the oocyte meiotic maturation as a primitive neurohormone.

Dialdehyde Cellulose Solution as Reducing Agent: Preparation of Uniform Silver Nanoparticles and In Situ Synthesis of Antibacterial Composite Films with High Barrier Properties.

The demand for antimicrobial materials is gradually increasing due to the threat of infections and diseases caused by microorganisms. Silver nanoparticles (AgNPs) are widely used because of their broad-spectrum antimicrobial properties, but their synthesis methods are often environmentally harmful and AgNPs difficult to isolate, which limits their application in several fields. In this study, an aqueous solution of dialdehyde cellulose (DAC) was prepared and used as a reducing agent to synthesize AgNPs in an efficient and environmentally friendly process. The synthesized AgNPs can be easily separated from the reducing agent to expand their applications. In addition, the AgNPs were immobilized in situ on dialdehyde cellulose to form antibacterial composite films. The results showed that the prepared silver nanoparticles were mainly spherical and uniformly dispersed, with an average size of about 25 nm under optimal conditions. Moreover, the dialdehyde cellulose-nanosilver (DAC@Ag) composite films had excellent mechanical properties, positive transparency, ultraviolet-blocking properties, and effective antibacterial activity against E. coli and S. aureus. Notably, the composite films exhibited excellent oxygen and water vapor barrier properties, with WVT and ORT of 136.41 g/m2·24 h (30 °C, 75% RH) and <0.02 cm3/m2·24 h·0.1 MPa (30 °C, 75% RH), respectively, better than commercial PE films. Hence, this study not only provides an environmentally friendly method for the preparation of silver nanoparticles, but also offers a simple and novel strategy for the in situ synthesis of silver-loaded antibacterial composite films.

A near infrared two-channel fluorescent probe for the detection of hydrogen sulfide and viscosity with a negligible crosstalk influence.

Both imbalance of H2S production and the change of viscosity in cells are associated with many diseases such as inflammation, Alzheimer's disease, and Parkinson's disease. Thus, the development of two-channel fluorescent probes for the detection of H2S and viscosity is of great significance for the study of pathogenic mechanisms. Herein, we design a two-channel NIR fluorescent probe RHO-DCO-DNP, which was able to selectively respond to H2S in one channel (λex = 580 nm, λem = 760 nm) and to viscosity in another channel (λex = 400 nm, λem = 585 nm). It should be emphasized that there is a negligible impact from the crosstalk between the two optical channels and the two targets. In addition, with the low cytotoxicity and unique dual lysosome/mitochondria targeting capability, the probe was successfully applied to the sensing of H2S and viscosity in normal cells and inflammation cells through fluorescent imaging. The probe could be a promising molecular tool for exploring the pathological role of H2S, viscosity, and both of them.

The enterprise's external knowledge acquisition capability and technological diversification: From the perspective of intellectual property strategy.

Enterprises need intellectual property rights to protect their core knowledge, and technological diversification is an important strategic measure for enterprises to improve innovation performance. From the perspective of external resource acquisition, this study explores the mechanism of external knowledge acquisition capability (internal absorptive capability and external relational learning) on firm's technological diversification. It considers the impact of firm's innovation capability and external environmental uncertainty. The survey data of 258 Chinese pharmaceutical companies were obtained through questionnaire surveys, and various theoretical hypotheses were validated using regression analysis methods. The results show that internal absorptive capacity, external relational learning, and their interaction have a significant positive impact on technological diversification; the innovation capacity and the uncertainty of the external environment also affect enterprises' technological diversification.

Transcriptional changes revealed water acidification leads to the immune response and ovary maturation delay in the Chinese mitten crab Eriocheir sinensis.

Nowadays, due to increasing carbon dioxide released, water acidification poses a series of serious impacts on aquatic organisms. To evaluate the effects of water acidification on crustaceans, we focused on the Chinese mitten crab Eriocheir sinensis, which is a spawning migration and farmed species in China. Based on histological and oocyte transparent liquid observation, we found that the acidified environment significantly delayed the ovarian maturation of E. sinensis. Moreover, RNA-seq was applied to obtain gene expression profile from the crab's gills and ovaries in response to acidified environment. Compared with control groups, a total of 5471 differentially expressed genes (DEGs) were identified in acidified gills and 485 DEGs were identified in acidified ovaries. Enrichment analysis indicated that some pathways also responded to the acidified environment, such as PI3K-Akt signaling pathway, Chemokine signaling pathway, apoptosis, and toll-like receptor signaling pathway. Subsequently, some DEGs involved in immune response (ALF, Cathepsin A, HSP70, HSP90, and catalase) and ovarian maturation (Cyclin B, Fem-1a, Fem-1b, and Fem-1c) were selected to further validate the influence of water acidification on gene expression by qRT-PCR. The results showed that the expression level of immune-related genes was significantly increased to response to the water acidification, while the ovarian maturation-related genes were significantly decreased. Overall, our data suggested that E. sinensis was sensitive to the reduced pH. This comparative transcriptome also provides valuable molecular information on the mechanisms of the crustaceans responding to acidified environment.

Identification and characterization of sex-biased and differentially expressed miRNAs in gonadal developments of the Chinese mitten crab, Eriocheir sinensis.

MicroRNA (miRNA) is a posttranscriptional downregulator that plays a vital role in a wide variety of biological processes. In this study, we constructed five ovarian and testicular small RNA libraries using two somatic libraries as reference controls for the identification of sex-biased miRNAs and gonadal differentially expressed miRNAs (DEMs) of the Chinese mitten crab, Eriocheir sinensis. A total of 535 known and 243 novel miRNAs were identified, including 312 sex-biased miRNAs and 402 gonadal DEMs. KEGG pathway analysis showed that DEM target genes were statistically enriched in MAPK, Wnt, and GnRH signaling pathway, and so on. A number of the sex-biased miRNAs target genes associated with sex determination/differentiation, such as IAG, Dsx, Dmrt1, and Fem1, while others target the genes related to gonadal development, such as P450s, Wnt, Ef1, and Tra-2c. Dual-luciferase reporter assay in vitro further confirmed that miR-34 and let-7b can downregulate IAG expression, miR-9-5p, let-7d, let-7b, and miR-8915 can downregulate Dsx. Taken together, these data strongly suggest a potential role for the sex-biased miRNAs in sex determination/differentiation and gonadal development in the crab.