Progresses in the establishment, evaluation, and application of in vitro blood-brain barrier models.

The blood-brain barrier (BBB) is a barrier between the circulatory system and the central nervous system (CNS), contributing to CNS protection and maintaining the brain homeostasis. Establishment of in vitro BBB models that are closer to the microenvironment of the human brain is helpful for evaluating the potential and efficiency of a drug penetrating BBB and thus the clinical application value of the drug. The in vitro BBB models not only provide great convenience for screening new drugs that can access to CNS but also help people to have a deeper study on the mechanism of substances entering and leaving the brain, which makes people have greater opportunities in the treatment of CNS diseases.

Effects of latroeggtoxin-VI on dopamine and α-synuclein in PC12 cells and the implications for Parkinson's disease.

Background: Parkinson's disease (PD) is characterized by death of dopaminergic neurons leading to dopamine deficiency, excessive α-synuclein facilitating Lewy body formation, etc. Latroeggtoxin-VI (LETX-VI), a proteinaceous neurotoxin discovered from the eggs of spider L. tredecimguttatus, was previously found to promote the synthesis and release of PC12 cells, showing a great potential as a drug candidate for PD.

Insights into the mediation of Ca signaling in the promoting effects of LETX-VI on the synthesis and release of dopamine.

Latroeggtoxin-VI (LETX-VI) is an active protein and was previously demonstrated to have effects on the synthesis and release of dopamine. Hererin, the involvement of Ca signaling in the effects of LETX-VI on dopamine was systematically investigated, using PC12 cells as a neuron model. LETX-VI was shown to promote dopamine release from PC12 cells both in the presence and absence of extracellular Ca; however the presence of extracellular Ca was favorable for enhancing the promoting effects of LETX-VI on dopamine, because LETX-VI facilitated the influx of extracellular Ca through the L-type calcium channels in plasma membrane (PM) to increase cytosolic Ca concentration.

Analysis of differentially expressed genes discovers Latroeggtoxin VI-induced changes and SYNJ1 as a main target in PC12 cells.

Background: Previous preliminary work found that Latroeggtoxin-VI (LETX-VI), a proteinaceous neurotoxin from the eggs of spider Latrodectus tredecimguttatus, could promote the synthesis and release of dopamine in PC12 cells. However, the underlying mechanisms have not been fully clear. Here, the effects of LETX-VI on the gene expression profile and dopamine in PC12 cells were analyzed with the differential transcriptome-based strategies.

Latroeggtoxin-VI protects nerve cells and prevents depression by inhibiting NF-κB signaling pathway activation and excessive inflammation.

Depression has a high incidence and seriously endangers human health. Accumulated evidence indicates that targeting neuroinflammation is a potential avenue for neuroprotection and thus depression prevention. Herein, the effects of latroeggtoxin-VI (LETX-VI), a bioactive protein from the eggs of spider , on lipopolysaccharide (LPS)-induced inflammation and depression were systematically investigated using RAW264.

Rab3 and synaptotagmin proteins in the regulation of vesicle fusion and neurotransmitter release.

Ca-triggered neurotransmitter release involves complex regulatory mechanisms, including a series of protein-protein interactions. Three proteins, synaptobrevin (VAMP), synaptosomal-associated protein of 25kDa (SNAP-25) and syntaxin, constitute the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) core complex that plays key roles in controlling vesicle fusion and exocytosis. Many other proteins participate in the regulation of the processes via direct and/or indirect interaction with the SNARE complex.

Synaptotagmin 1-mediated cell membrane penetration and dopamine release enhancement by latroeggtoxin-VI.

Latroeggtoxin-VI (LETX-VI), a proteinaceous neurotoxin mined from the egg transcriptome of spider L. tredecimguttatus, was previously found to promote the release of dopamine from PC12 cells. However, the relevant molecular mechanism has not been fully clear.

Effects of JZTX-V on the wild type Kv4.3 Expressed in HEK293T and Molecular Determinants in the Voltage-sensing Domains of Kv4.3 Interacting with JZTX-V.

JZTX-V is a toxin isolated from the venom of the Chinese spider . Previous studies had shown that JZTX-V could inhibit the transient outward potassium current of Kv4.2 and Kv4.

Ultrasonographic assessment of preoperative gastric volume in patients with dyspepsia: a prospective observational study.

Background: Patients undergoing gastroenteroscopy during sedation are prone to aspiration, and most patients with dyspepsia have delayed gastric emptying. This study aimed to investigate the feasibility of measuring the gastric antrum cross-sectional area (CSA) to supply a novel clinical diagnostic reference value in patients with dyspepsia.

Methods: Patients with dyspepsia undergoing elective gastroscopy were included.

Clinical significance of high mobility group box 1/toll-like receptor 4 in obese diabetic patients.

High mobility group box 1 (HMGB1) is an alarmin that may link to obesity and type 2 diabetes mellitus (T2DM). The present study analyzed the correlation between HMGB1/ Toll-like receptor 4 (TLR4) and certain biochemical parameters in obese (OB) diabetic patients. 40 normal glucose tolerant subjects (NGT) and 40 patients with newly diagnosed T2DM were enrolled.

Biochemical and cytotoxic evaluation of latroeggtoxin-VI against PC12 cells.

Latroeggtoxin-VI (LETX-VI) is a peptide neurotoxin discovered from Latrodectus tredecimguttatus eggs. In the current study, the action features of the neurotoxin on PC12 cells were systematically investigated. LETX-VI could promote dopamine release from PC12 cells in the absence and presence of Ca2+, involving an even more complex action mechanism in the presence of Ca2+ and when the treatment time was longer.

Pull-Down Assay-Guided Insights into the Effects of Latroeggtoxin-VI on Nerve Cells.

Latroeggtoxin-VI (LETX-VI) is a peptide neurotoxin newly found from the eggs of spider To explore the mechanism of action of the LETX-VI on nerve cells, the effects of LETX-VI on PC12 cells, a commonly used neuron model, were analyzed using a pull-down assay-guided strategy. LETX-VI was shown to interact with 164 PC12 cell proteins that have diverse molecular functions such as binding, catalysis, regulation, structural activity, etc., thereby extensively affecting the biological processes in the PC12 cells, particularly protein metabolism, response to stimulus, substance transport, and nucleic acid metabolism, with 56.

[Gene cloning, heterologous expression and activity identification of latroeggtoxin-Ⅵ].

One of the distinct characters of Latrodectus tredecimguttatus is that its toxic components exist not only in the venomous glands, but also in the tissues outside the venomous glands and even in the eggs. Investigation on the toxins outside the venomous glands can deepen our understanding of spider toxins and discover new lead molecules with important application prospects. In order to explore the low-abundance proteinaceous toxins in the L.

Comparative Study on Bioactivities from Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Gives an Insight into the Fermentation Broth Showing Greater Antioxidative Activities.

Ganoderma lucidum is one of the most famous mushrooms in traditional Chinese medicine. At present, the fully utilized parts of G. lucidum are mainly fruiting body and spore powder.

Comparative proteomic analysis to probe into the differences in protein expression profiles and toxicity bases of Latrodectus tredecimguttatus spiderlings and adult spiders.

The early reports and our previous work confirmed the existence of the toxic proteinaceous components in the body of the L. tredecimguttatus newborn and adult spiders. For revealing the differences in the protein expression profiles and toxicity bases of the spiders at different developmental stages, the spiderling and adult spider proteins were comparatively analyzed using a proteomic strategy.

Newly Discovered Action of HpTx3 from Venom of Heteropoda on Na1.7 and Its Pharmacological Implications in Analgesia.

It has been reported that Heteropodatoxin3 (HpTx3), a peptidic neurotoxin purified from the venom of the spider species Heteropoda venatoria, could inhibit K4.2 channels. Our present study newly found that HpTx3 also has potent and selective inhibitory action on Na1.

Gel Absorption-Based Sample Preparation Method for Shotgun Analysis of Membrane Proteome.

Membrane proteins solubilized in a starting buffer containing high concentration of SDS are directly entrapped and immobilized into gel matrix when the membrane protein solution is absorbed by the vacuum-dried polyacrylamide gel. After the detergent and other salts are removed by washing, the proteins are subjected to in-gel digestion, and the tryptic peptides are extracted and analyzed by CapLC-MS/MS. The newly developed method not only avoids protein loss and the adverse protein modifications during gel-embedment but also improves the subsequent in-gel digestion and the recovery of tryptic peptides, particularly hydrophobic peptides.

Anti-Breast Cancer Activity of Latroeggtoxin-V Mined from the Transcriptome of Spider Eggs.

As a black widow spider, has poisonous components not only in venomous glands but also in eggs. Our previous work had carried out a transcriptome analysis of the spider eggs in an attempt to probe into the molecular basis of the egg toxicity. A proteinaceous toxin, named Latroeggtoxin-V, was mined from the identified transcriptome.

Molecular basis and mechanism underlying the insecticidal activity of venoms and toxins from Latrodectus spiders.

Latrodectus species are among the most venomous of spiders, with abundant toxic proteinaceous components in their venomous glands and other tissues, as well as their eggs. To date, several proteinaceous toxins with insecticidal potential, including α-insectotoxin and δ-insectotoxin, two of the most potent known insecticidal toxins, have been purified and characterized by comprehensively utilizing conventional biochemical techniques. This has greatly enhanced our knowledge of the molecular basis and mechanism of action of their toxicity.

Comparative characterization of rat hippocampal plasma membrane and mitochondrial membrane proteomes based on a sequential digestion-centered combinative strategy.

Plasma membrane (PM) and mitochondrial membrane (MM) proteins of rat hippocampal neurons were identified and comparatively characterized on the basis of a sequential digestion-centered combinative strategy for sample treatment. A total of 478 membrane proteins were identified, of which 240 had PM localization, 170 had MM localization, and 33 had both of the two subcellular localizations. Compared with the PM proteome, the MM proteome not only was smaller, more basic, and more hydrophobic, but also had a narrower protein molecular mass distribution range and a higher proportion of transmembrane proteins.

Cytotoxic and apoptotic activities of black widow spiderling extract against HeLa cells.

Black widow spiders contain toxic components not only in the venom glands but also in other parts of the spider body, including the legs and abdomen. Additionally, both the eggs and newborn spiderlings of the black widow spider contain venom. It is important to investigate their potential effects on cancer cells.

Rab3A Inhibition of Ca -Dependent Dopamine Release From PC12 Cells Involves Interaction With Synaptotagmin I.

Rab3 and synaptotagmin have been suggested to play important roles in the regulation of neurotransmitter release and, however, the molecular mechanism has not been completely clear. Here, we studied the effects of Rab3A and synaptotagmin I (Syt I) on dopamine release using PC12 cells as a model system. Rab3A was demonstrated to have effects on both Ca -independent and Ca -dependent dopamine releases from the PC12 cells.

Pull-down combined with proteomic strategy reveals functional diversity of synaptotagmin I.

Synaptotagmin I (Syt I) is most abundant in the brain and is involved in multiple cellular processes. Its two C2 domains, C2A and C2B, are the main functional regions. Our present study employed a pull-down combined with proteomic strategy to identify the C2 domain-interacting proteins to comprehensively understand the biological roles of the C2 domains and thus the functional diversity of Syt I.

Localization of Rab3A-binding site on C2A domain of synaptotagmin I to reveal its regulatory mechanism.

Synaptotagmin I (Syt I) functions in the regulation of neurotransmitter release and multiple other cellular processes through its C2 domain binding to other molecules. Our previous study demonstrated that Rab3A, a small GTP-binding protein, is a new interacting partner of Syt I and could bind to both of the C2 domains; the polylysine motif in C2B is a key site for Rab3A binding, but the binding site on C2A is not clear. In order to localize Rab3-binding site on C2A and reveal the relevant regulatory mechanism, in the present study we investigated the interaction between recombinant Rab3A and various C2A mutants.

Transcriptome Analysis to Understand the Toxicity of Latrodectus tredecimguttatus Eggs.

is a kind of highly venomous black widow spider, with toxicity coming from not only venomous glands but also other parts of its body as well as newborn spiderlings and eggs. Up to date, although eggs have been demonstrated to be rich in proteinaceous toxins, there is no systematic investigation on such active components at transcriptome level. In this study, we performed a high-throughput transcriptome sequencing of eggs with Illumina sequencing technology.