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During the pupal‐adult eclosion process of holometabolous insects, the old cuticle is shed and replaced by a completely different new cuticle that requires tanning and expansion, along with extensive extracellular matrix (ECM) remodeling. In vertebrates, matrix metalloproteinases (MMPs), a class of zinc‐dependent endopeptidases, play key roles in regulating the ECM that surrounds cells. However, little is known about these extracellular proteinases available in insects. The small hive beetle (SHB), Aethina tumida, is a widespread invasive parasite of honey bees. In this study, 6 MMP homologs were identified in the SHB genome. RNA interference experiments showed that all 6 AtMmps are not required for the larval‐pupal transition, only AtMmp2 was essential for pupal‐adult eclosion in SHB. Knockdown of AtMmp2 resulted in eclosion defects and wing expansion failure, as well as mortality within 3 d of adult eclosion. Transcriptomic analysis revealed that knockdown of AtMmp2 significantly increased expression of the Toll and Imd pathways, chitin metabolism, and cross‐linking (such as the pro‐phenoloxidase activating cascade pathway and the tyrosine‐mediated cuticle sclerotization and pigmentation pathway). These data revealed evolutionarily conserved functions of Mmp2 in controlling adult eclosion and wing expansion, also provided a preliminary exploration of the novel function of regulating Toll and Imd pathways, as well as new insights into how MMPs regulate insect development and defense barriers. [ABSTRACT FROM AUTHOR]

Brain transcriptional regulatory network for behavior demonstrates that brain gene expression in the honey bee can be accurately predicted from the expression transcription factors (TFs), but roles for specific TFs are less understood. Mushroom bodies (MBs) are important for learning, memory and sensory integration in the honey bee brain. A TFs, Mblk‐1, expressed preferentially in the large‐type Kenyon cells of the honeybee MBs is predicted to be involved in brain function by regulating transcription of its target genes in honey bee. However, its function and the mechanism of regulation in behavior of honey bee is still obscure. Here we show that Mblk‐1 had significantly higher expression in the brains of forager bees relative to nurse bees. Mblk‐1 was significantly inhibited in bees fed small interfering RNA. In addition, inhibition of Mblk‐1 decreased sucrose responsiveness in foragers. Finally, we determined that Mblk‐1 regulated the messenger RNA of AmGR1. These findings suggest that Mblk‐1 may target AmGR1 to regulate the sucrose responsiveness of foragers. [ABSTRACT FROM AUTHOR]

A new distance metric between interval-valued fuzzy sets is proposed. The four special logic metric spaces based on the distance are structured. we analysis and compare the structures of the four logic metric spaces. It is shown that the Łukasiewicz logic metric space and Goguen logic metric space are more suitable for interval-valued fuzzy reasoning. Moreover, the robustness of interval-valued fuzzy reasoning triple I methods are studied in the two logic metric spaces. We prove that fuzzy reasoning triple I methods based on the interval-valued Łukasiewicz residuated implication and interval-valued Goguen residuated implication are robust. [ABSTRACT FROM AUTHOR]

In this paper, a new tangent similarity between single valued neutrosophic sets is given, which contain tangent similarity [23] as a special case. A best-worst multi-criteria decision making method based on the single valued neutrosophic sets is proposed. To achieve this goal, we design an algorithm to identify the best and worst criteria through computing the out-degrees and in-degrees of the collective single valued neutrosophic preference relation directed network, and then calculate the optimal weight vector of attributes. Moreover, a mathematical model corresponding to the definitions of consistent single valued neutrosophic preference relation is contracted. Finaly, the evaluation values of all alternatives are calculated by the proposed new tangent similarity. This approach can make the process of decision making more vivid, intuitive and dynamic in the decision-making process. To illustrate the effectiveness of the proposed method, we used it to solve appointment registration systems problems. [ABSTRACT FROM AUTHOR]

Melioidosis is an infectious disease caused by Burkholderia pseudomallei , mainly found in Southeast Asia and northern Australia. In Hainan, sporadic cases were first described in 1990; since then, more cases have been identified. No systematic study has yet been done to detect the environmental source of the organism and its true extent in Hainan. This study is aimed to confirm the prevalence of B. pseudomallei in soil samples in Hainan. 1080 soil samples from 18 different counties were collected from 3 sampling points of 360 sites. They were screened for the presence of B. pseudomallei by Ashdown selective media. Suspected colonies of B. pseudomallei were confirmed by biochemical test and a specific PCR assay. 48 of 360 sites (13.3%) were positive for B. pseudomallei , including all coastal counties in Hainan Island. This study revealed the prevalence and distribution of B. pseudomallei in the soil environment in Hainan Island of southern China and may be helpful to understand the distribution of B. pseudomallei and to access its epidemiological importance. [ABSTRACT FROM AUTHOR]

XN4 might induce DNA damage and apoptotic cell death through reactive oxygen species (ROS). The inhibition of proliferation of K562 and K562/G01 cells was measured by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide). The mRNA levels of NADPH oxidase 1-5 (Nox1-5) genes were evaluated by qRT-PCR. The levels of extracellular reactive oxygen species (ROS), DNA damage, apoptosis, and cell cycle progression were examined by flow cytometry (FCM). Protein levels were analyzed by immunoblotting. XN4 significantly inhibited the proliferation of K562 and K562/G01 cells, with IC50 values of 3.75±0.07 µM and 2.63±0.43 µM, respectively. XN4 significantly increased the levels of Nox4 and Nox5 mRNA, stimulating the generation of intracellular ROS, inducing DNA damage and activating ATM-γ-H2AX signaling, which increased the number of cells in the S and G2/M phase of the cell cycle. Subsequently, XN4 induced apoptotic cell death by activating caspase-3 and PARP. Moreover, the above effects were all reversed by the ROS scavenger N-acetylcysteine (NAC). Additionally, XN4 can induce apoptosis in progenitor/stem cells isolated from CML patients’ bone marrow. In conclusion, XN4-induced DNA damage and cell apoptosis in CML cells is mediated by the generation of ROS. [ABSTRACT FROM AUTHOR]

Among the most classic measures in single valued neutrosophic sets (SVNS) theory, the distance is an important tool to compare and calculate degree of difference between SVNS. Although there exist some types of distance for single valued neutrosophic sets, most of them lack of strictly axiomatic definition and exist counter-intuitive cases. In this paper, a novel distance between single valued neutrosophic sets based on matrix norm is given. Then we proved that the new distance satisfies the axiomatic definition of the metric. Finally, the distance is applicated to pattern recognition and medical diagnoses. [ABSTRACT FROM AUTHOR]

Abstract: We here report the sequence and functional analysis of cstB of Azospirillum brasilense Sp7. The predicted cstB contains C-terminal two PAS domains and N-terminal part which has similarity with CheB–CheR fusion protein. cstB mutants had reduced swarming ability compared to that of A. brasilense wild-type strain, implying that cstB was involved in chemotaxis in A. brasilense. A microscopic analysis revealed that cstB mutants developed mature cyst cells more quickly than wild type, indicating that cstB is involved in cyst formation. cstB mutants were affected in colony morphology and the production of exopolysaccharides (EPS) which are essential for A. brasilense cells to differentiate into cyst-like forms. These observations suggested that cstB was a multi-effector involved in cyst development and chemotaxis in A. brasilense. [Copyright &y& Elsevier]

Simple Summary: The small hive beetle is a destructive pest of honeybees, causing severe economic damage to the apiculture industry. Chemosensory genes play key roles in insect behavior, such as foraging and mating partners. However, the chemosensory genes are lacking in the small hive beetle. In order to better understand its chemosensory process at the molecular level, a total of 130 chemosensory genes, including 38 odorant receptors, 24 ionotropic receptors, 14 gustatory receptors, 3 sensory neuron membrane proteins, 29 odorant binding proteins, and 22 chemosensory proteins were identified from the transcriptomic data of antennae and forelegs. Reverse-transcription PCR showed that 3 OBPs (AtumOBP3, 26 and 28) and 3 CSPs (AtumCSP7, 8 and 21) were highly expressed in antennae. Overall, this study could provide a basis for elucidating functions of these chemosensory genes at the molecular level. Aethina tumida is a parasite and predator of honeybee causing severe loss to the bee industry. No effective and environmentally friendly methods are available to control this pest at present. Chemosensory genes play key roles in insect behavior which can potentially be used as targets for developing environmentally friendly pest control agents. In this study, the putative chemosensory genes in antennae and forelegs of A. tumida involved in olfaction or contact chemical communication of adults were investigated using RNA transcriptome sequencing and PCR methods. Based on transcriptomic data, unigenes encoding 38 odorant receptors (ORs), 24 ionotropic receptors (IRs), 14 gustatory receptors (GRs), 3 sensory neuron membrane proteins (SNMPs), 29 odorant binding proteins (OBPs), and 22 chemosensory proteins (CSPs) were identified. The analyses of tissue expression profiles revealed that genes encoding 38 ORs, 13 antennal IRs, 11 GRs, 1 SNMP, 24 OBPs, and 12 CSPs were predominately expressed in antennae. No significant differences in expression levels of these genes were found between males and females. Genes encoding 5 non-NMDA iGluRs, 3 GRs, 2 SNMPs, 5 OBPs, and 12 CSPs were predominately expressed in forelegs. RT-PCR assays for SNMPs, OBPs and CSPs further revealed that 3 OBPs (AtumOBP3, 26 and 28) and 3 CSPs (AtumCSP7, 8 and 21) were highly expressed in antennae. Our results enrich the gene inventory of A. tumida and facilitate the discovery of potential novel targets for developing new pest control measures. [ABSTRACT FROM AUTHOR]

Cytochrome P450 monooxygenases (CYPs) serve many functions in insects, from the regulation of development to xenobiotic detoxification. Several conserved CYPs have been shown to play a role in insect growth and development. CYP303A1 is a highly conserved CYP with a single ortholog in most insects, but its underlying molecular characteristics and specific physiological functions remain poorly understood. In Drosophila melanogaster and Locusta migratoria , CYP303A1 is indispensable for eclosion to adult. Here, we report additional functions of the locust gene LmCYP303A1 in nymphal molts, cuticular lipid deposition and insecticide penetration. RT-qPCR revealed that LmCYP303A1 had a high expression level before ecdysis and was highly expressed in integument, wing pads, foregut and hindgut. Suppression of LmCYP303A1 expression by RNA interference (RNAi) caused a lethal phenotype with molting defect from nymph to nymph. In addition, LmCYP303A1 RNAi resulted in locusts being more susceptible to desiccation and to insecticide toxicity. Furthermore, knockdown of LmCYP303A1 efficiently suppressed the transcript level of key genes (ELO7 , FAR15 and CYP4G102) responsible for cuticular hydrocarbon (CHC) synthesis, which led to a decrease in some CHC levels. Taken together, our results suggest that one of the functions of LmCYP303A1 is to regulate the biosynthesis of CHC, which plays critical roles in protecting locusts from water loss and insecticide penetration. Unlabelled Image • LmCYP303A1 is highly expressed before ecdysis and mainly distributed in the tissues of ectoderm development. • Suppression of LmCYP303A1 caused a lethal phenotype with molting defect from nymphs to nymphs. • LmCYP303A1 RNAi resulted in locusts being more susceptible to desiccation and insecticide exposure. • RNAi suppressed the expression of key genes responsible for CHC synthesis and led to a decrease of CHC levels. [ABSTRACT FROM AUTHOR]

Insect cytochrome P450 monooxygenases (CYPs) play essential roles in both xenobiotic metabolism and developmental processes. However, the exact physiological function of many CYP genes remains largely unknown. Screening the expression of the CYP genes from the CYP2 and mitochondrial CYP clans of Drosophila melanogaster revealed that Cyp303a1 is highly expressed in the pupal stage. Knockdown of CYP303A1 transcripts by RNAi using the Gal4/UAS system with a ubiquitous driver (tubulin-Gal4) in Drosophila or by dsRNA injection in the last nymph stage of Locusta migratoria resulted in severe defects in eclosion and lethality during and after adult emergence. In Drosophila , tissue-specific RNAi of Cyp303a1 with a wing-specific driver (MS1096-Gal4) revealed that Cyp303a1 was essential for wing extension. Stage-specific RNAi of Cyp303a1 using Gal80ts for thermal-dependent-suppression found that the expression of Cyp303a1 at the middle pupal stage was absolutely required. Meanwhile, Cyp303a1 mutants exhibited more than 80% lethality at the late embryonic development stages. Embryonic lethality of the Cyp303a1 mutants was fully rescued by the ubiquitous overexpression of exogenous Cyp303a1. Taken together, we conclude that Cyp303a1 is indispensable for embryonic development and adult eclosion in D. melanogaster , the latter role being conserved over 400 million years of insect evolution. Image 1 • Cyp303a1 is highly expressed in the pupal stage with the highest level in the wings at 48h APF. • Knockdown of CYP303A1 in Drosophila and Locusta resulted in lethality during and after adult emergence. • Cyp303a1 mutants are arrested in the late embryonic stage, which can be rescued by the ubiquitous expression of Cyp303a1. [ABSTRACT FROM AUTHOR]

Oral cancer is a disease with high morbidity and mortality worldwide and greatly impacts the quality of life, especially in patients with advanced stages. Photodynamic therapy (PDT) is one of the most effective clinical treatments for oral cancers. However, most clinically applied photosensitizers have several deficiencies, including oxygen dependence, poor aqueous solubility, and a lack of tumor-targeting ability. Herein, the carrier-free multifunctional Sorafenib (Sor), chlorin e6 (Ce6), and Fe3+ self-assembly co-delivery nanoparticles (Sor-Ce6 NPs) were constructed via combining a ferroptosis inducer Sor and a photosensitizer Ce6 for synergetic therapy. The as-synthesized Sor-Ce6 NPs presented excellent colloidal stability and water dispersity with good in vivo tumor-targeting ability. More significantly, the low dose of Sor-Ce6 NPs had little dark toxicity but produced significantly enhanced ROS and supplied O 2 sustainably to increase phototoxicity through ferroptosis pathway. Notably, the Sor-Ce6 NPs showed significantly higher in vitro and in vivo anti-tumor efficacy than the Sor/Ce6 mixture due to the improvement of cellular uptake and the incorporation of foreign Fe ions in the system, which also confer the T 1 magnetic resonance-guided imaging ability to the formed Sor-Ce6 NPs. Our study demonstrates a promising self-assembled strategy for overcoming hypoxia-related PDT resistance for oral cancer treatment. • Carrier-free Sorafenib-Ce6 self-assemblies are designed by molecular simulation • The effect of oxygen-dependent PDT is boosted by inducing ferroptosis in oral cancer • Exogenous Fe3+ enhanced Sorafenib's ferroptosis inducing effect in oral cancer • Sor-Ce6 NPs is suitable for fluorescence and T 1 MR dual-modal imaging-guided PDT [Display omitted] [ABSTRACT FROM AUTHOR]

Background: As an important catecholamine neurotransmitter in invertebrates and vertebrates, dopamine plays multiple roles in the life of the honey bee. Dopamine receptors (DA), which specifically bind to dopamine to activate downstream cascades, have been reported to be involved in honey bee reproduction, division of labour, as well as learning and motor behaviour. However, how dopamine receptors regulate honey bee behavior remains uninvestigated. Results: The expression level of Amdop2 in the brain increased with the age of worker bees, which was just the opposite trend of ame-let-7. Inhibition of ame-let-7 through feeding an inhibitor upregulated Amdop2 expression; conversely, overexpression of ame-let-7 through a mimic downregulated Amdop2. Moreover, knockdown of Amdop2 in forager brain led to significantly higher sucrose responsiveness, which is similar to the phenotype of overexpression of ame-let-7. Finally, we confirmed that ame-let-7 directly targets Amdop2 in vitro by a luciferase reporter assay. Conclusions: ame-let-7 is involved in the dopamine receptor signaling pathway to modulate the sucrose sensitivity in honey bees. Specifically, it down-regulates Amdop2, which then induces higher responses to sucrose. These results further unraveled the diverse mechanisms of the dopamine pathway in the regulation of insect behavior. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Three Mmps genes have been identified in the silkworm. • Inhibition of Mmps activity inhibited fat body cell dissociation and ovary development. • Overexpression of each Mmp promoted these developmental processes. • Mmp3 and Mmp1 have the strongest and weakest stimulatory effects, respectively. Abstract Matrix metalloproteinases (Mmps) are pivotal extracellular proteinases participating in tissue remodeling. Three Mmps genes have been identified from the silkworm, Bombyx mori , and their expression levels and enzyme activity are consistent with progressive fat body cell dissociation during the early pupal stages. Using both loss-of-function and gain-of-function experiments, we have demonstrated that Mmps are functionally required for fat body cell dissociation and ovary development in female pupae. Moderate inhibition of Mmps activity via inhibitor treatments delayed fat body cell dissociation and ovary development, while severe inhibition blocked these developmental processes and eventually led to pupal lethality. Individual RNAi knockdown of each Mmp delayed fat body cell dissociation, with the strongest and weakest phenotypes occurring for Mmp3 and Mmp1 , respectively. By contrast, overexpression of each Mmp promoted fat body cell dissociation and ovary development, with the strongest stimulatory effects for Mmp3 overexpression and the weakest effects for Mmp1 overexpression. This is the first time to show that Mmps induce fat body cell dissociation in Lepidoptera, and we also hypothesize that Mmps-induced fat body cell dissociation is required for ovary development in this insect species. [ABSTRACT FROM AUTHOR]

Proteolysis-targeting chimaera (PROTAC) has received extensive attention in industry. However, there are still some limitations that hinder its further development. In a previous study, our group first demonstrated that the HSP90 degrader BP3 synthesised by the principle of PROTACs showed therapeutic potential for cancer. However, its application was hindered by its high molecular weight and water insolubility. Herein, we aimed to improve these properties of HSP90-PROTAC BP3 by encapsulating it into human serum albumin nanoparticles (BP3@HSA NPs). The results demonstrated that BP3@HSA NPs showed a uniform spherical shape with a size of 141.01 ± 1.07 nm and polydispersity index < 0.2; moreover, BP3@HSA NPs were more readily taken up by breast cancer cells and had a stronger inhibitory effect in vitro than free BP3. BP3@HSA NPs also demonstrated the ability to degrade HSP90. Mechanistically, the improved inhibitory effect of BP3@HSA NPs on breast cancer cells was related to its stronger ability to induce cell cycle arrest and apoptosis. Furthermore, BP3@HSA NPs improved PK properties and showed stronger tumour suppression in mice. Taken together, this study demonstrated that hydrophobic HSP90-PROTAC BP3 nanoparticles encapsulated by human serum albumin could improve the safety and antitumour efficacy of BP3. [ABSTRACT FROM AUTHOR]

Lipids are either taken up from food sources or produced internally in specialized tissues such as the liver. Among others, both routes of lipid metabolism involve cytochrome P450 monooxygenases (CYPs). We sought to analyze the function of Cyp311a1 that has been shown to be expressed in the midgut of the fruit fly Drosophila melanogaster. Using a GFP-tagged version of CYP311A1 that is expressed under the control of its endogenous promoter, we show that Cyp311a1 localizes to the endoplasmic reticulum in epithelial cells of the anterior midgut. In larvae with reduced Cyp311a1 expression in the anterior midgut, compared to control larvae, the apical plasma membrane of the respective epithelial cells contains less and shorter microvilli. In addition, we observed reduction of neutral lipids in the fat body, the insect liver, and decreased phosphatidylethanolamine (PE) and triacylglycerols (TAG) amounts in the whole body of these larvae. Probably as a consequence, they cease to grow and eventually die. The microvillus defects in larvae with reduced Cyp311a1 expression are restored by supplying PE, a major phospholipid of plasma membranes, to the food. Moreover, the growth arrest phenotype of these larvae is partially rescued. Together, these results suggest that the anterior midgut is an import hub in lipid distribution and that the midgut-specific CYP311A1 contributes to this function by participating in shaping microvilli in a PE-dependent manner. [ABSTRACT FROM AUTHOR]

• Try to use exosomes in anti-tuberculosis immunotherapy to provide new ideas for the major infectious diseases. • The immunological mechanism of NK exosome anti-Mtb from apoptosis to pyroptosis was completely studied. • Provide some immunotherapy for the treatment of MDR-TB. Murine Natural Killer cells were cultivated in vitro to isolate NK-derived exosomes. Subsequent quantification via qPCR confirmed enrichment of miR-1249-3p. Ana-1 murine macrophages were cultured in vitro and subsequently inoculated with Mycobacterium tuberculosis (MTB) strain H37Rv. NK-exo and NK-exo miR-1249-3p were separately applied to the infection model, followed by immunological assays conducted post-48-hour co-culture. Western blot analyses corroborated that NK-exo exhibited exosomal marker proteins Granzyme A (GzmA), Granzyme B (GzmB), and Perforin (PFN), alongside a notable enrichment of miR-1249-3p. Functionally, NK-exo augmented the expression levels of Caspase-9,-8, and -3, as well as PARP, while attenuating the expression of NLRP3, ASC, and Cleaved-Caspase-1. Furthermore, qPCR demonstrated an up-regulation of Caspase-9, -8, and -3, along with pro-apoptotic factors Bax and Bid, and a concomitant down-regulation of the anti-apoptotic factor Bcl-2. The expression levels of inflammatory markers ASC, NLRP3, Cleaved-Caspase-1, and IL-1β were concomitantly decreased. ELISA findings indicated diminished levels of TNF-α and ROS secretion. NK-exo miR-1249-3p specifically targeted and attenuated the expression of SKOR-1, engendering up-regulation of apoptosis-associated proteins and down-regulation of inflammation-related proteins, consequently affecting cellular fate.Our empirical evidence substantiates that NK-exo induces macrophage apoptosis, thereby mitigating MTB survival. Furthermore, NK-exo miR-1249-3p directly targets and inhibits SKOR-1 expression, leading to macrophage apoptosis and consequently hampering the proliferation of MTB. The data implicate the potential therapeutic relevance of NK-exo and miR-1249-3p in managing drug-resistant tuberculosis. [ABSTRACT FROM AUTHOR]

[Display omitted] Juvenile hormone (JH) and 20-hydroxyecdysone (20E) coordinately regulate development and metamorphosis in insects. Two JH intracellular receptors, methoprene-tolerant (Met) and germ-cell expressed (Gce), have been identified in the fruit fly Drosophila melanogaster. To investigate JH membrane signaling pathway without the interference from JH intracellular signaling, we characterized phosphoproteome profiles of the Met gce double mutant in the absence or presence of JH in both chronic and acute phases. Functioning through a potential receptor tyrosine kinase and phospholipase C pathway, JH membrane signaling activated protein kinase C (PKC) which phosphorylated ultraspiracle (USP) at Ser35, the PKC phosphorylation site required for the maximal action of 20E through its nuclear receptor complex EcR-USP. The usp S35A mutant, in which Ser was replaced with Ala at position 35 by genome editing, showed decreased expression of Halloween genes that are responsible for ecdysone biosynthesis and thus attenuated 20E signaling that delayed developmental timing. The usp S35A mutant also showed lower Yorkie activity that reduced body size. Altogether, JH membrane signaling phosphorylates USP at Ser35 and thus potentiates 20E action that regulates the normal fly development. This study helps better understand the complex JH signaling network. [ABSTRACT FROM AUTHOR]

Purpose. This research was designed to investigate the protective effect of rifampicin (RIF) loaded by N-(2-hydroxypropyl) methylacrylamide- (HPMA-) polylactic acid (PLA) nanopolymer on macrophages infected with Mycobacterium tuberculosis (MTB). Methods. We first induced H37Rv to infect macrophages to build a cell model. Then, the HPMA-PLA nanopolymer loaded with RIF was prepared to treat MTB-infected macrophages. The macrophage activity was tested by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the nitric oxide (NO) in cells was measured through Griess reagent, and the bacterial activity of MTB was observed via the colony-forming unit (CFU) assay. The inflammation-related factors in cells were detected via the enzyme-linked immunosorbent assay (ELISA), the apoptosis of macrophages was examined via flow cytometry, and the expression of apoptosis-related proteins was determined by western blot (WB). Results. HPMA-PLA had no obvious toxicity to macrophages. The expression of NO and inflammatory factors in macrophages infected with MTB increased significantly, but the apoptosis rate was not significantly different from that of uninfected cells. However, after treatment with HPMA-PLA-RIF or free RIF, the inflammatory reaction of infected cells was inhibited, the expression of NO was decreased, the apoptosis rate was increased, and the bacterial activity in cells was decreased, with statistically significant differences; moreover, HPMA-PLA-RIF was more effective than free RIF. Conclusions. HPMA-PLA-RIF has a high protective effect on macrophages infected with MTB, with high safety. Its protective mechanism is at least partly through inhibiting the production of NO and inflammatory response, which can inhibit bacterial activity and induce cell apoptosis. [ABSTRACT FROM AUTHOR]

Abstract: Paenibacillus sabinae T27 (CCBAU 10202=DSM 17841) is a gram-positive, spore-forming diazotroph with high nitrogenase activities. Three nifH clusters were cloned from P. sabinae T27. Phylogenetic analysis revealed that NifH1, NifH2 and NifH3 cluster with Cyanobacterium. Each of the coding regions of nifH1, nifH2 and nifH3 from P. sabinae T27 under the control of the nifH promoter of Klebsiella pneumoniae could partially restore nitrogenase activity of K. pneumoniae nifH − mutant strain 1795, which has no nitrogenase activity. This suggests that the three nifH genes from P. sabinae T27 have some function in nitrogen fixation. RT-PCR showed that all three nifH genes were expressed under nitrogen-fixing growth conditions. Using promoter vectors which have promoterless lacZ gene, three putative promoter regions of nifH genes were identified. [Copyright &y& Elsevier]

Summary: Our previous study indicated org35 was involved in chemotaxis and interacted with nitrogen fixation transcriptional activator NifA via PAS domain. In order to reveal the role of org35 in nitrogen regulation, the downstream target genes of org35 were identified. We here report differentially expressed genes in org35 mutants comparing with wild type Sp7 by means of cDNA-AFLP. Four up-regulated transcript-derived fragments (TDFs) homologues of chemotaxis transduction proteins were found, including CheW, methyl-accepting chemotaxis protein and response regulator CheY-like receiver. Three distinct TDFs (AB46, AB58 and AB63) were similar to PHB de-polymerase C-terminus, cell shape-determining protein and flagellin domain protein. And 11 TDFs showed similarities with signal transduction proteins, including homologous protein of the nitrogen regulation protein NtrY and nitrate/nitrite response regulator protein NarL. These data suggested that the Azospirillum brasilense org35 was a multi-effecter and involved in chemotaxis, cyst development and regulation of nitrogen fixation. [ABSTRACT FROM AUTHOR]

Abstract: We here report the sequence and functional analysis of org35 of Azospirillum brasilense Sp7, which was originally identified to be able to interact with NifA in yeast-two-hybrid system. The org35 encodes a hybrid two-component system protein, including N-terminal PAS domains, a histidine kinase (HPK) domain and a response regulator (RR) domain in C-terminal. To determine the function of the Org35, a deletion–insertion mutant in PAS domain [named Sp7353] and a complemental strain Sp7353C were constructed. The mutant had reduced chemotaxis ability compared to that of wild-type, and the complemental strain was similar to the wild-type strain. These data suggested that the A. brasilense org35 played a key role in chemotaxis. Variants containing different domains of the org35 were expressed, and the functions of these domains were studied in vitro. Phosphorylation assays in vitro demonstrated that the HPK domain of Org35 possessed the autokinase activity and that the phosphorylated HPK was able to transfer phosphate groups to the RR domain. The result indicated Org35 was a phosphorylation-communicating protein. [Copyright &y& Elsevier]

Reports that the interaction between P&subu; an Nifa in A, brasilense Sp7 was investigated by using the yeast two-hybrid system. Results showed that P&subu; directly interacted with the entire Nifa protein and its N-terminal domain, but did not interact with the central domain and the C-terminal domain of NifA.

• The relationship between TLR2 polymorphism and TB susceptibility is not as close as we thought before. • Screening more gene mutations to provide new ideas for prevention and treatment of TB. • TLR2 rs5743708 polymorphism exhibits a significant association with a higher tuberculosis risk. • The polymorphism of TLR2 gene was systematically reviewed for the first time. Toll-like receptor 2 (TLR2) is a crucial factor in the development of tuberculosis. However, no studies have explored the association between TLR2 polymorphisms and tuberculosis susceptibility. This study aimed to explore the correlation between tuberculosis susceptibility and TLR2 polymorphisms (rs3804099, rs3804100, rs1898830, rs5743708, rs121917864, and (-196-174) del). All relevant online databases including PubMed, CNKI, WANFANG DATA, and METSTR-FMRS were systematically searched. STATA17.0 (Stata Corp LP, College Station, Texas, USA) was used. A total of 37 studies, covering six polymorphisms and comprising 9,474 cases and 10,295 controls, were included in this analysis. rs3804099(C vs T: OR = 1.00, 95 % CI: 0.93–1.08, CC + TC vs TT: OR = 1.04, 95 % CI: 0.98–1.10), rs3804100 (C vs T: OR = 1.19, 95 % CI: 0.93–1.07, CC + TC vs TT: OR = 0.97, 95 % CI: 0.89–1.06), rs1898830(G vs A: OR = 0.90, 95 % CI: 0.81–1.00, GG + AG vs AA: OR = 0.87, 95 % CI: 0.67–1.12), (-196 ∼174) del polymorphism (Del vs Ins: OR = 0.93,95 % CI: 0.76–1.14, DD + DI vs II: OR = 0.92,95 % CI: 0.72–1.17). This study indicated that only the TLR2 rs5743708 polymorphism exhibited a significant association with a higher tuberculosis risk, while TLR2 rs3804099, rs3804100, rs1898830, rs121917864, and (-196-174) del polymorphisms were not associated with tuberculosis susceptibility. [ABSTRACT FROM AUTHOR]

Heat shock protein 90 (Hsp90), a highly conserved and widely expressed molecular chaperone, is mainly responsible for maintaining the correct folding of client proteins and is closely related to the stability and activation of tumour-related proteins. Hsp90α, the major isoform of Hsp90, can promote tumour cell migration and metastasis, and is abundantly secreted in highly invasive tumours. To date, most pan-Hsp90 inhibitors have been limited in their applications due to high toxicity. Herein, we described the candidate compound X10g based on a proteolysis-targeting chimaera (PROTAC) strategy that potently and selectively degraded Hsp90α. The results showed that X10g inhibited tumours better with lower toxicity in vivo. These findings demonstrate that synthesized selective Hsp90α degrader X10g provides a new strategy for breast cancer therapy. [Display omitted] • Designed and synthesized a series of PROTACs that selectively targeted Hsp90α. • X10g was found to selectively degrade Hsp90α without affecting Hsp90β. • X10g showed antitumour activity in vivo and had no significant effect on the body weights of mice. [ABSTRACT FROM AUTHOR]

Objective: To analyze the mechanism of the halo vest-assisted Z-shape elevating-pulling reduction technique for cervical unilateral locked facets, and confirm the safety of the spinal cord under the epidural pressure that occurs during the reduction process. Methods: Eleven osteoligamentous whole coronal and cervical spine specimens were established as skull-neck-thorax models of cervical unilateral locked facets at the C5/6 level. The halo vest-assisted Z-shape elevating-pulling reduction technique was then applied to reduce the locked facets. The changes in the epidural pressure in five cervical positions (cervical physiological curvature, cervical lateral bending, cervical unilateral locked facets, cervical unilateral perched facets, and reduction) were measured by a pressure sensor during the reduction procedure. The models simultaneously underwent multi-angle radiographic examination and CT scanning. Results: Successful closed reduction was achieved via the halo vest-assisted Z-shape elevating-pulling reduction technique in all 11 models. The epidural pressure in the cervical unilateral locked facets position was significantly higher than that in the other four cervical positions (P < 0.005). There was no significant difference in the epidural pressures measured during cervical lateral bending, cervical unilateral perched facets, and reduction. Conclusions: Maximum epidural pressures were measured in the position of cervical unilateral locked facets. The halo vest-assisted Z-shape elevating-pulling reduction technique achieved spinal decompression without causing secondary spinal cord injury. The halo vest-assisted Z-shape elevating-pulling reduction technique is safe and effective, and has a high success rate of reduction. [ABSTRACT FROM AUTHOR]

Recent studies showed that the distribution of hepatitis B virus (HBV) genotype exhibited geographical and ethnic characteristics. Haikou city is the largest city on Hainan Island that geographically isolated from mainland of China, and is the home of multiple ethnic groups. The aim of the study was to investigate the characteristics of the HBV genotype/subgenotype distribution in Haikou city. HBV DNA was isolated from180 serum samples derived from the Han and Li groups. The HBV genotype was detected by polymerase chain reaction using genotype-specific primers and was further determined by full-length genome sequences. The results revealed that the genotype B (37.2%) and C (62.8%) were the predominant HBV genotypes in Haikou, regardless of ethnic background., Additionally, the genotype distribution was not significantly different regarding ethnicity, sex or level of serum HBV DNA. Moreover, there were multiple subgenotypes circulating in the region. In conclusion, our study revealed the diverse HBV genotypes/subgenotypes in Haikou. These findings provide a preliminary study of the distribution of HBV genotypes circulating on Hainan Island. • There are no significant geographical and ethnic distribution of HBV genotypes in Hainan Island • The genotype B and C are the predominant HBV genotype in Haikou city • There were diverse circulation of HBV subgenotypes found in Haikou [ABSTRACT FROM AUTHOR]

[Display omitted] Poly ADP-ribose polymerase 1 (PARP1) plays an essential role in DNA repair signaling, rendering it an attractive target for cancer treatment. Despite the success of PARP1 inhibitors (PARPis), only a few patients can currently benefit from PARPis. Moreover, drug resistance to PARPis occurs during clinical treatment. Natural and acquired resistance to PARPis has forced us to seek new therapeutic approaches that target PARP1. Here, we synthesized a series of compounds by proteolysis-targeting chimera (PROTAC) technology to directly degrade the PARP1 protein. We found that CN0 (compound 3) with no polyethylene glycol (PEG) linker can degrade the PARP1 protein through the proteasome pathway. More importantly, CN0 could inhibit DNA damage repair, resulting in highly efficient accumulation of cytosolic DNA fragments due to unresolved unrepaired DNA lesions when combined with daunorubicin (DNR). Therefore, CN0 can activate the cyclic GMP-AMP synthase/stimulator of the interferon gene (cGAS/STING) pathway of innate immunity and then spread the resulting inflammatory signals, thereby reshaping the tumor microenvironment, which may eventually enhance T cell killing of tumor cells. [ABSTRACT FROM AUTHOR]

[Display omitted] The therapeutic strategy of poly (ADP-ribose) polymerase (PARP) inhibition of BRCA1/2 mutant cancers has been overwhelmingly successful, however, the highly aggressive triple negative breast cancers (TNBC) that receptor protein tyrosine kinase (RTKs) is known to be overexpressed are not sensitive to PARP inhibitors. Our research focused on exploring PARP inhibitors incorporating a bicyclic tetrahydropyridine pyrimidine. All synthesized compounds were more potent than Olaparib (ola) in killing tumor cells, especially in TNBC. Furthermore, compound 7 exhibited strong inhibitory effects on PARP enzymatic activity, moreover, the expression of EGFR and phosphorylated EGFR was inhibited by compound 7. Therefore, compound 7 can effectively inhibit TNBC cells with high expression of EGFR. In addition, significant synergistic effect of anti-tumor effect of new PARP inhibitors and adriamycin was also observed. [ABSTRACT FROM AUTHOR]

Heat shock protein 90 (HSP90) is involved in the stabilization and activation of oncoproteins, rendering it essential for oncogenic transformation. However, the HSP90 inhibitors evaluated to date have not led to the expected effects in cancer therapy. Herein, we systematically described the design, synthesis, and evaluation of HSP90 degraders based upon the proteolysis-targeting chimera (PROTAC) strategy. The results showed that the candidate compound 16b (BP3) potently degraded HSP90 and effectively inhibited the growth of human breast cancer cells. When used as a single agent, BP3 led to effective tumor suppression in mice. These findings demonstrate that our HSP90-targeting PROTAC strategy has potential novel applications in breast cancer therapy. [Display omitted] • A series of PROTACs targeting HSP90 were designed for the first time and subsequently synthesized. • BP3 had a certain degradation effect on HSP90 protein in MCF-7 cells. • BP3 had a significant inhibition of breast cancer cells in vitro or in vivo compared with control. [ABSTRACT FROM AUTHOR]

Trichosanthin (TCS), an active component extracted from the root tubers of traditional Chinese medical herb Tian-Hua-Fen of the Cucurbitaceae family, has long been used for medical purpose in China; there is increasing interest in developing TCS as cancer therapeutic agents. The present study was to investigate the growth arrest of K562 cells and its molecular mechanisms, which the drugs induced by TCS and the possible functional interaction of TCS with imatinib (STI571) to K562 cells. Trypan blue exclusive staining was used to access the cell growth inhibition; western blot was used to evaluate the p210Bcr-Abl, phosphorylated tyrosine kinase (PTK), and some signaling molecules involving in cell proliferation and apoptosis in K562 cells. TCS and imatinib inhibited K562 cells at a time- and dose-dependent manners, respectively; TCS down-regulated p210Bcr-Abl at a time- and dose-dependent manners; TCS synergistically enhanced imatinib-induced K562 cell growth arrest and down-regulation of p210Bcr-Abl, PTK activities, procaspase-3, Hsp90,NF-κB and PKC. The results suggest that TCS not only by itself involves but also synergizes activities of imatinib to induce K562 cell growth arrest, down-regulation of p210Bcr-Abl and its downstream signals and to stimulate the effect of the tyrosine kinase inhibition. [ABSTRACT FROM AUTHOR]

The roles and therapeutic potential of long noncoding RNAs (lncRNAs) in acute myeloid leukemia (AML) have attracted increased attention. However, many lncRNAs have not been annotated in AML, and their predictive value for AML therapy remains unclear. In this study, we identified a novel large intergenic noncoding RNA uc002jit.1 (D43770) from a lncRNA microarray. We first proved uc002jit.1 is a target gene of nuclear factor kappa B/RELA, RELA regulated uc002jit.1 transcription by binding to its promoter. Additionally, uc002jit.1 knockdown impaired the stability of poly (ADP-ribose) polymerase 1 (PARP1) mRNA, and then reduced PARP1 protein content and PARylation level upon DNA damage, thus inhibiting DNA damage repair in AML cells. Moreover, uc002jit.1 knockdown significantly inhibited AML cells proliferation and increased the sensitivity to chemotherapeutic drugs in vitro as well as in a mouse model in vivo. Overall, our study indicated that uc002jit.1 may be associated with the occurrence and prognosis of AML and could be a new diagnostic/prognostic biomarker and therapeutic target for AML. Image 1 • The novel lincRNA uc002jit.1(D43770) is a target gene of RELA. • uc002jit.1 affects the stability of PARP1 mRNA, thus regulating DNA damage repair in AML cells. • uc002jit.1 has significant biological functions in cell proliferation and sensitivity to chemotherapeutic drugs. [ABSTRACT FROM AUTHOR]

Poly (ADP-ribose) polymerase (PARP) inhibitors have achieved great success in clinical application, especially for the prolonged survival of cisplatin-sensitive ovarian cancer patients. However, there are still many patients who do not respond to PARP inhibitors. Novel PARP inhibitors with higher activity are urgently needed. Herein we report a series of compounds by molecular hybridization PARP-1 inhibitor Olaparib (Ola) with HSP90 inhibitor C0817 (one curcumin derivative). All synthesized compounds were evaluated for their antiproliferative activity in vitro, and some were further assessed for their inhibitory activities of the PARP enzyme and HSP90 affinity. Our results indicated that compound 4 could bind to HSP90 and cause static quenching, indicating that compound 4 was able to bind to HSP90, moreover, downstream molecular breast cancer 1 (BRAC-1) was reduced. In conclusion, dual target inhibitors of PARP and HSP90 exhibited stronger selective cytotoxicities against cancer. [ABSTRACT FROM AUTHOR]

3D-QSAR, molecular docking and molecular dynamics (MD) simulations were performed to investigate the mechanism of ACE-inhibitory (ACEi) peptides with tryptophan C-terminal in aqueous solutions. In this work, COMFA (Q2 = 0.85, R2 = 0.993) and COMSIA (Q2 = 0.706, R2 = 0.997) models of di- and tri-peptides were established. Novel tripeptides were predicted by the 3D-QSAR models. Different conformations and interactions between VKW with high activity and GTW with low activity were compared by MD simulation. The results revealed VKW had stronger electrostatic interaction and binding affinity with key residues of ACE. ACE-VKW complex performed more stable than ACE-GTW. Thus, VKW showed a higher activity. Moreover, ACEi tripeptides with Trp C-terminal potentiate the combination with Zn of ACE active pockets. ACE inhibitory tripeptides of different bioactivity were used to investigate the mechanism between ACE and peptides in aqueous solutions by 3D-QSAR and molecular simulation. The ligands could interact with the key residues and ions of ACE active sites. Electrostatic binding, van der Waal interactions and hydrophobic effects had impact on the combination. The electrostatic binding energy of high-activity molecular was stronger than low-activity molecular. It was in accordance with result of 3D-QSAR models. ACE combined with VKW was more stable. Amino acid Trp was beneficial for the combination with zinc ion. Zn was important to the stabilization of ligands and receptor. Unlabelled Image • Different active inhibitors peptides revealed action mechanism using MD simulation and 3D-QSAR. • Complex with highly reactive ACEi peptide exhibited stronger stability and better electrostatic interactions. • Zinc ion was important for the stabilization of ACE complex. • Amino acid Trp was beneficial for the combination of the ligand with Zn. [ABSTRACT FROM AUTHOR]