Nobiletin derivative, 5-acetoxy-6,7,8,3',4'-pentamethoxyflavone, inhibits neuroinflammation through the inhibition of TLR4/MyD88/MAPK signaling pathways and STAT3 in microglia.

OBJECTIVE: Microglia in the central nervous system regulate neuroinflammation that leads to a wide range of neuropathological alterations. The present study investigated the anti-neuroinflammatory properties of nobiletin (Nob) derivative, 5-acetoxy-6,7,8,3',4'-pentamethoxyflavone (5-Ac-Nob), in lipopolysaccharide (LPS)-activated BV2 microglia. MATERIALS AND METHODS: By using the MTT assay, Griess method, flow cytometry, and enzyme-linked immunosorbent assay (ELISA), we determined the cell viability, the levels of nitric oxide (NO), reactive oxygen species (ROS), and pro-inflammatory factors (interleukin 1 beta; IL-1ß, interleukin 6; IL-6, tumor necrosis factor alpha; TNF-α and prostaglandin E2; PGE2) in LPS-stimulated BV2 microglia. Toll-like receptor 4 (TLR4)-mediated myeloid differentiation primary response gene 88 (MyD88)/nuclear factor-kappa B (NF-κB), mitogen-activated protein kinase (MAPK) signaling pathway and signal transducer and activator of transcription 3 (STAT3) were measured by western blotting. Analysis of NO generation and mRNA of pro-inflammatory cytokines was confirmed in the zebrafish model. RESULTS: 5-Ac-Nob reduced cell death, the levels of NO, ROS, inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), and pro-inflammatory factors in LPS-activated BV-2 microglial cells. TLR4-mediated MyD88/NF-κB and MAPK pathway (p38, ERK and JNK) after exposure to 5-Ac-Nob was also suppressed. Moreover, 5-Ac-Nob inhibited phosphorylated STAT3 proteins expression in LPS-induced BV-2 microglial cells. Furthermore, we confirmed that 5-Ac-Nob decreased LPS-induced NO generation and mRNA of pro-inflammatory cytokines in the zebrafish model. CONCLUSIONS: Our findings suggest that 5-Ac-Nob represses neuroinflammatory responses by inhibiting TLR4-mediated signaling pathway and STAT3. As a result of these findings, 5-Ac-Nob has potential as an anti-inflammatory agent against microglia-mediated neuroinflammatory disorders.

Complete mitochondrial genome of Distenia punctulatoides (Coleoptera: Chrysomeloidea: Disteniinae) and its phylogenetic implications.

The family Disteniidae is a moderately large and widely distributed lineage. Distenia punctulatoides belongs to the family Disteniidae from the cerambycoid assemblage. Here, we report the complete mitogenome of D. punctulatoides, which is 15,675 bp in length. It contains 37 genes and a noncoding control region, which are arranged in the same order as that of the putative ancestor of beetles. The total base composition of the new mitogenome is 40.2% for A, 17.1% for C, 10.0% for G, and 32.7% for T. The new mitogenomic organization, nucleotide composition, and codon usage do not differ significantly from other beetles. Using available complete mitogenomes, the high-level phylogeny of the family Disteniidae was explored. The phylogenetic analyses showed that Disteniidae were monophyletic, and the genus Distenia grouped with the genus Clytomelegena as sister groups. Combining the morphological and molecular data, Typodryas Thomson, 1864 is suggested to be a junior synonym of Distenia Lepeletier and Audinet-Serville, 1828.

A Heparinase Sensor Based on a Ternary System of Hg2+-Heparin-Osmium Nanoparticles.

A visual assay for the detection of heparinase was developed on the basis of a ternary system of Hg2+-heparin-osmium nanoparticles (OsNPs). First, heparin-capped OsNPs (heparin-OsNPs) were synthesized by a facile reduction method using heparin as the protecting/stabilizing agent. The oxidase-like activity of heparin-OsNPs, however, turned out to be low, which somewhat limits their application. We discovered that Hg2+ can significantly/specifically boost the oxidase-like activity of heparin-OsNPs via electrostatic interaction. The oxidase-like activity of heparin-OsNPs toward the oxidation of the substrate, 3,3',5,5'-tetramethylbenzidine, by dissolved O2 was found to increase by 76-fold in the presence of Hg2+. More significantly, heparin in heparin-OsNPs could be specifically hydrolyzed into small fragments in the presence of heparinase, which resulted in the weakening of the oxidase-like activity of Hg2+/heparin-OsNPs. On the basis of these findings, a linear response of the sensor for heparinase was obtained in the range 20-1000 µg/L with a low detection limit (15 µg/L), which is comparable to those of other reported sensors. Further, the colorimetric sensor was employed for the detection of heparinase in human serum samples with satisfactory results. We speculate that combining such surface modification of the osmium nanozyme with a sensing element could be an interesting direction for promoting nanozyme research in medical diagnosis.

Sandensolide Induces Oxidative Stress-Mediated Apoptosis in Oral Cancer Cells and in Zebrafish Xenograft Model.

Presently, natural sources and herbs are being sought for the treatment of human oral squamous cell carcinoma (OSCC) in order to alleviate the side effects of chemotherapy. This study investigates the effect of sandensolide, a cembrane isolated from Sinularia flexibilis, to inhibit human OSCC cell growth with the aim of developing a new drug for the treatment of oral cancer. In vitro cultured human OSCC models (Ca9.22, SCC9 and HSC-3 cell lines) and oral normal cells (HGF-1), as well as a zebrafish xenograft model, were used to test the cytotoxicity of sandensolide (MTT assay), as well as to perform cell cycle analysis and Western blotting. Both the in vitro bioassay and the zebrafish xenograft model demonstrated the anti-oral cancer effect of sandensolide. Moreover, sandensolide was able to significantly suppress colony formation and induce apoptosis, as well as cell cycle arrest, in OSCC by regulating multiple key proteins. Induction of reactive oxygen species (ROS) was observed in sandensolide-treated oral cancer cells. However, these apoptotic changes were rescued by NAC pretreatment. These findings contribute to the knowledge of the model of action of sandensolide, which may induce oxidative stress-mediated cell death pathways as a potential agent in oral cancer therapeutics.

Non-basic amino acids in the ROMK1 channels via an appropriate distance modulate PIP2 regulated pHi-gating.

The ROMK1 (Kir1.1) channel activity is predominantly regulated by intracellular pH (pHi) and phosphatidylinositol 4,5-bisphosphate (PIP2). Although several residues were reported to be involved in the regulation of pHi associated with PIP2 interaction, the detailed molecular mechanism remains unclear. We perform experiments in ROMK1 pHi-gating with electrophysiology combined with mutational and structural analysis. In the present study, non basic residues of C-terminal region (S219, N215, I192, L216 and L220) in ROMK1 channels have been found to mediate channel-PIP2 interaction and pHi gating. Further, our structural results show these residues with an appropriate distance to interact with membrane PIP2. Meanwhile, a cluster of basic residues (R188, R217 and K218), which was previously discovered regarding the interaction with PIP2, exists in this appropriate distance to discriminate the regulation of channel-PIP2 interaction and pHi-gating. This appropriate distance can be observed with high conservation in the Kir channel family. Our results provide insight that an appropriate distance cooperates with the electrostatics interaction of channel-PIP2 to regulate pHi-gating.

The Aggregation Pheromone of Phyllotreta striolata (Coleoptera: Chrysomelidae) Revisited.

Aggregations of the striped flea beetle Phyllotreta striolata on their crucifer host plants are mediated by volatiles emitted from feeding males. The male-specific sesquiterpene, (6R,7S)-himachala-9,11-diene (compound A), was shown previously to be physiologically and behaviorally active, but compound A was attractive only when combined with unnaturally high doses of the host plant volatile allyl isothiocyanate (AITC) in field trapping experiments. This indicated that our understanding of the chemical communication in this species is incomplete. Another male-specific sesquiterpenoid, (3S,9R,9aS)-3-hydroxy-3,5,5,9-tetramethyl-5,6,7,8,9,9a-hexahydro-1H-benzo[7]annulen-2(3H)-one (compound G), has been reported from an American P. striolata population. We confirmed the presence of compound G, and investigated its interaction with compound A and AITC in a P. striolata population in Taiwan. Compound G was attractive to Taiwanese P. striolata in laboratory bioassays, but significantly more beetles were attracted to a blend of compounds A and G. Under the same conditions, P. striolata showed no preference for the blend of A and G combined with a range of doses of AITC over the sesquiterpenoid blend alone. The sesquiterpenoid blend was tested further in field trapping experiments and attracted significantly more beetles than traps baited with compound A and ecologically relevant amounts of AITC. We conclude that A and G are components of the male-specific aggregation pheromone of P. striolata in Taiwan, and that the attractiveness of the pheromone is not reliant on the presence of AITC. Our results further indicate that the male-specific sesquiterpenoid blends differ qualitatively between the Taiwanese and American populations of P. striolata.

Protective Effects of Costunolide against Hydrogen Peroxide-Induced Injury in PC12 Cells.

Oxidative stress-mediated cellular injury has been considered as a major cause of neurodegenerative diseases including Alzheimer's and Parkinson's diseases. The scavenging of reactive oxygen species (ROS) mediated by antioxidants may be a potential strategy for retarding the diseases' progression. Costunolide (CS) is a well-known sesquiterpene lactone, used as a popular herbal remedy, which possesses anti-inflammatory and antioxidant activity. This study aimed to investigate the protective role of CS against the cytotoxicity induced by hydrogen peroxide (H2O2) and to elucidate potential protective mechanisms in PC12 cells. The results showed that the treatment of PC12 cells with CS prior to H2O2 exposure effectively increased the cell viability. Furthermore, it decreased the intracellular ROS, stabilized the mitochondria membrane potential (MMP), and reduced apoptosis-related protein such as caspase 3. In addition, CS treatment attenuated the cell injury by H2O2 through the inhibition of phosphorylation of p38 and the extracellular signal-regulated kinase (ERK). These results demonstrated that CS is promising as a potential therapeutic candidate for neurodegenerative diseases resulting from oxidative damage and further research on this topic should be encouraged.

Taxonomic studies on the genera Echinovelleda Breuning, 1936 and Propedicellus Huang, Huang & Liu, 2020 (Coleoptera, Cerambycidae, Lamiinae, Lamiini).

The genera Echinovelleda Breuning, 1936 and Propedicellus Huang, Huang & Liu, 2020 are revised. The latterisconsideredto be ajunior synonym of theformer based on a comprehensive morphological investigation, especially on the characteristics of male endophallus. Two new species are described from China, viz. Echinovelleda mumuae Bi & Mu sp. nov. from Yunnan and Guangxi, and E. protochinensis Bi & Lin sp. nov. from Yunnan and Sichuan. New records are reported for previously described taxa including one new country record of a morphologically similar genus, Hechinoschema Thomson, 1857 from China. Illustrations of habitus, endophallic structure, major diagnostic features for all studied taxa, as well as a distributional map are provided.

The Antimicrobial Peptide Tilapia Piscidin 4 Induced the Apoptosis of Bladder Cancer Through ERK/SIRT1/PGC-1α Signaling Pathway.

Marine antimicrobial peptides have been demonstrated in numerous studies to possess anti-cancer properties. This research investigation aimed to explore the fundamental molecular mechanisms underlying the antitumor activity of Tilapia piscidin 4 (TP4), an antimicrobial peptide, in human bladder cancer. TP4 exhibited a remarkable inhibitory effect on the proliferation of bladder cancer cells through cell cycle arrest at the G2/M phase. Additionally, TP4 upregulated the expression of cleaved caspase-3, caspase-9, and PARP, leading to the activation of apoptotic pathways in bladder cancer cells. TP4 exhibit a marked rise in mitochondria reactive oxygen species, leading to the subsequent loss of potential for the mitochondrial membrane. Furthermore, the inhibition of mitochondrial oxidative phosphorylation resulted in a decrease in downstream ATP production. Meanwhile, TP4-treated bladder cancer cells showed an increase in Bax and ERK but a decrease in SIRT1, PGC-1α, and Bcl2. ERK activation, SIRT1/PGC-1α-axis, and TP4-induced apoptosis were all significantly reversed by the ERK inhibitor SCH772984. Finally, the inhibitory effect of TP4 on tumor growth has been confirmed in a zebrafish bladder cancer xenotransplantation model. These findings suggest that TP4 may be a potential agents for human bladder cancer through apoptosis induction, ERK activation, and the promotion of SIRT1-mediated signaling pathways.

Blamada rubripronota, a new genus and species of the tribe Saperdini (Coleoptera: Cerambycidae: Lamiinae) from Southeast Asia.

A new saperdine species belonging to a new genus, Blamada rubripronota gen. et sp. nov., is described from Laos, Vietnam and China. The genus differs from other genera of the tribe Saperdini in having the antennal scape bearing an expanded and ridged ring at apex, and second antennomere relatively longer (more than 1/4 of scape in length) than that of other saperdine taxa.

A hitherto overlooked article by Gressitt in 1941 (Insecta, Coleoptera, Cerambycidae, Lamiinae).

An article published by Gressitt (1941) has been ignored by all longicornists, including Gressitt himself. However, according to ICZN, it meets all criteria as an official publication, and this status affects three taxa that were formally described therein: Bacchisa (Bacchisa) rigida (Gressitt, 1941) = Chreonomarigida Gressitt, 1941 = Chreonomarigida Gressitt, 1942 homonym and syn. nov.; Tetraophthalmussikang (Gressitt, 1941) = Chreonomasikang Gressitt, 1941 = Chreonomasikanga Gressitt, 1942, syn. nov.; Anastathesparvahainana Gressitt, 1941 = Anastathesparvahainana Gressitt, 1942, homonym and syn. nov.

A new species of the genus Clyzomedus Pascoe, 1865 from China (Coleoptera: Cerambycidae: Lamiinae: Mesosini).

A new species Clyzomedus pani sp. nov. is described from Beijing, Shaanxi, Henan and Anhui, China. Quercus acutissima Carruth (Fagaceae) is the possible host plant. Images of habitus, external morphology and genitalia are provided. It is one of the currently discovered new species from Capital of China, since for the Biodiversity Survey and Assessment Project of Beijing.

Plumbagin induces the apoptosis of drug-resistant oral cancer in vitro and in vivo through ROS-mediated endoplasmic reticulum stress and mitochondrial dysfunction.

BACKGROUND: Oral cancer is one of the leading causes of cancer-related deaths worldwide. Chemotherapy is widely used in the treatment of oral cancer, but its clinical efficacy is limited by drug resistance. Hence, novel compounds capable of overcoming drug-resistance are urgently needed. PURPOSE: Plumbagin (PG), a natural compound isolated from Plumbago zeylanica L, has been used to treat various cancers. In this study, we investigated the anticancer effects of PG on drug-resistant oral cancer (CR-SAS) cells, as well as the underlying mechanism. METHODS: MTT assays were used to evaluate the effect of PG on the viability of CR-SAS cells. Apoptosis and reactive oxygen species (ROS) production by the cells were determined using flow cytometry. Protein expression levels were detected by western blotting. RESULTS: The results show that PG reduces the viability and causes the apoptosis of CR-SAS cells. PG is able to induce intracellular and mitochondrial ROS generation that leads to mitochondrial dysfunction. Furthermore, endoplasmic reticulum (ER) stress was triggered in PG-treated CR-SAS cells. The inhibition of ROS using N-acetylcysteine (NAC) abrogated the PG-induced ER stress and apoptosis, as well as the reduction in cell viability. Meanwhile, similar results were observed both in zebrafish and in murine models of drug-resistant oral cancer. CONCLUSION: Our results indicate that PG induces the apoptosis of CR-SAS cells via the ROS-mediated ER stress pathway and mitochondrial dysfunction. It will be interesting to develop the natural compound PG for the treatment of drug-resistant oral cancer.

Blockade of acid-sensing ion channels protects articular chondrocytes from acid-induced apoptotic injury.

OBJECTIVE: Acid-sensing ion channels (ASICs) are members of the degenerin/epithelial sodium channel (DEG/ENaC) protein superfamily and play a critical role in acid-induced cell injury. In this study, we examined whether drugs such as amiloride that block ASICs could attenuate acid-induced apoptotic injury to articular chondrocytes. METHODS: Articular chondrocytes were isolated from Sprague-Dawley rats, and their phenotype was determined by toluidine blue and immunocytochemical staining. Articular chondrocyte viability assay was performed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). Apoptosis of chondrocytes was observed by the terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling method as well as propidium iodide labeling methods. Intracellular calcium ([Ca(2+)](i)) was analyzed by a Ca(2+)-imaging method. In addition, the expression levels of calpain and calcineurin in articular chondrocytes were examined by real-time PCR and immunocytochemical staining. The activity of caspase-3 was evaluated by spectrophotometric assays. RESULTS: Positive staining for glycosaminoglycan and collagen II was seen in articular chondrocytes. Blocking acid-sensing ion channels significantly decreased the cell death percentage and increased cell viability following acid exposure. After pretreated with amiloride, acid-induced [Ca(2+)](i) rises were reduced. Amiloride also inhibited calpain and calcineurin expression levels in acid-induced chondrocytes, and inhibited caspase-3 activity. CONCLUSION: The data presented in this study provided some experimental evidence that blocking ASICs could protect acid-induced apoptotic injury to chondrocytes.

A study on the genus Anaches Pascoe, 1865 (Coleoptera, Cerambycidae, Lamiinae, Pteropliini), with a new species and two new synonyms.

A new species, Anaches m-signatus sp. nov. is described from Zhejiang, Hunan and Guangxi Provinces, China. Sthenias semicylindricus Hayashi, 1974 and Sthenias murzini Lazarev, 2020 are synonymized with Anaches medioalbus (Breuning, 1956). Three similar species are compared with additional pictures and descriptions.

Taxonomic studies on the genera Meges Pascoe, 1866 and Pseudomeges Breuning, 1944 from China (Coleoptera, Cerambycidae, Lamiinae, Lamiini).

The genus Meges Pascoe, 1866 stat. res. is resurrected from synonyms of Monochamus Dejean, 1821 and is made a senior synonym of Magninia Clermont, 1932 syn. nov. Meges currently contains two species, i.e. Meges gravidus (Pascoe, 1858) and Meges tonkineus (Clermont, 1932) comb. nov. The male of the latter species is described for the first time. A related genus Pseudomeges Breuning, 1944 is investigated for comparison and Pseudomeges aureus Bi, Chen Lin sp. nov. (, Jn bn wi m tin ni) is described from Yunnan, China. New localities are reported. Illustrations of habitus, endophallic structures and major diagnostic features for all involved taxa are provided.

A new species, Cylindroeme yunnanensis sp. nov. from China (Coleoptera, Cerambycidae, Cerambycinae).

A new species, Cylindroeme yunnanensis sp. nov., is described and illustrated from Yunnan Province of China. Cylindroeme vietnamica Vives, 2019 is recorded from China for the first time. A key to the species of Cylindroeme is provided.

Trichodermin inhibits the growth of oral cancer through apoptosis-induced mitochondrial dysfunction and HDAC-2-mediated signaling.

Trichodermin (TCD), a trichothecene first isolated from marine Trichoderma viride, is an inhibitor of eukaryotic protein synthesis. However, the potential effects of TCD on human oral squamous cell carcinoma (OSCC) cells and the underlying molecular mechanisms remain unknown. In this study, the exposure of OSCC cells (Ca922 and HSC-3 cells) to TCD suppressed cell proliferation assessed using MTT assays and colony formation assays. TCD inhibited the migration and invasion of OSCC cells (Ca922 and HSC-3 cells) through the downregulation of matrix metalloproteinase 9. After treatment of OSCC cells with TCD, the G2/M phase was arrested, caspase-related apoptosis (cleaved caspase-3 and PARP expression) was induced, and the protein level of x-linked inhibitor of apoptosis was reduced. Meanwhile, the TCD-induced cell death was reversed by the pan-caspase inhibitor Z-VAD-FMK. Furthermore, TCD diminished mitochondrial membrane potential, mitochondrial oxidative phosphorylation and glycolytic function in OSCC cells. In addition, TCD decreased the levels of histone deacetylase 2 (HDAC-2) and downstream signaling proteins, including phosphorylated STAT3 and NF-κB. Finally, TCD significantly suppressed tumor growth in a zebrafish OSCC xenotransplantation model. Overall, this evidence demonstrates that TCD is a novel promising strategy for the treatment of OSCCs.

6-methoxyflavone suppresses neuroinflammation in lipopolysaccharide- stimulated microglia through the inhibition of TLR4/MyD88/p38 MAPK/NF-κB dependent pathways and the activation of HO-1/NQO-1 signaling.

BACKGROUND: Microglia-related neuroinflammation is associated with a variety of neurodegenerative diseases. Flavonoids have demonstrated different pharmacological effects, such as antioxidation, neuroprotection and anti-inflammation However, the effect of flavonoid 6-methoxyflavone (6-MeOF) on microglia-mediated neuroinflammation remain unknown. PURPOSE: The current study aim to study the antineuroinflammatory effects of 6-MeOF in lipopolysaccharide- (LPS-) induced microglia in vitro and in vivo. METHODS: Pretreatment of BV2 microglia cells with 6-MeOF for 1 h then stimulated with LPS (100 ng/ml) for 24 h. The expression levels of pro-inflammatory factors, NO and reactive oxygen species (ROS) were performed by the enzyme-linked immunosorbent assay (ELISA), Griess assay and flow cytometry. Western blotting was used to assess MAPK, NF-κB signal transducer and antioxidant enzymes-related proteins. Analysis of ROS and microglial morphology was confirmed in the zebrafish and mice brain, respectively. RESULTS: Our results demonstrated that 6-MeOF dose-dependently prevent cell death and decreased the levels of pro-inflammatory mediators in LPS-stimulated BV2 microglia cells. Phosphorylated NF-κB/IκB and TLR4/MyD88/p38 MAPK/JNK proteins after exposure to 6-MeOF was suppressed in LPS-activated BV-2 microglial cells. 6-MeOF also presented antioxidant activity by reduction of NO, ROS, iNOS and COX-2 and the induction of the level of HO-1 and NQO1 expressions in LPS-activated BV2 microglial cells. Furthermore, we demonstrated that 6-MeOF inhibited LPS-induced NO generation in an experimental zebrafish model and prevent the LPS-induced microgliosis in the prefrontal cortex and substantia nigra of mice. CONCLUSION: These results explored that 6-MeOF possesses potential as anti-inflammatory and anti-oxidant agents against microglia-associated neuroinflammatory disorders.

Tsounkranaglenea hefferni gen. et sp. nov. from Sabah, Malaysia (Coleoptera, Cerambycidae, Lamiinae: Saperdini).

A new saperdine species belonging to a new genus, Tsounkranaglenea hefferni gen. et sp. nov., is described from Sabah, Malaysia. The new genus differs from other genera of the tribe Saperdini by the special male sternite VII modified into a rake-shape, with the apex of the ventral plate of the median lobe unusually emarginated.