Cyp6g2 is the major P450 epoxidase responsible for juvenile hormone biosynthesis in Drosophila melanogaster.

BACKGROUND: Juvenile hormones (JH) play crucial role in regulating development and reproduction in insects. The most common form of JH is JH III, derived from MF through epoxidation by CYP15 enzymes. However, in the higher dipterans, such as the fruitfly, Drosophila melanogaster, a bis-epoxide form of JHB3, accounted most of the JH detected. Moreover, these higher dipterans have lost the CYP15 gene from their genomes. As a result, the identity of the P450 epoxidase in the JH biosynthesis pathway in higher dipterans remains unknown. RESULTS: In this study, we show that Cyp6g2 serves as the major JH epoxidase responsible for the biosynthesis of JHB3 and JH III in D. melanogaster. The Cyp6g2 is predominantly expressed in the corpus allatum (CA), concurring with the expression pattern of jhamt, another well-studied gene that is crucial in the last steps of JH biosynthesis. Mutation in Cyp6g2 leads to severe disruptions in larval-pupal metamorphosis and exhibits reproductive deficiencies, exceeding those seen in jhamt mutants. Notably, Cyp6g2-/-::jhamt2 double mutants all died at the pupal stage but could be rescued through the topical application of JH analogs. JH titer analyses revealed that both Cyp6g2-/- mutant and jhamt2 mutant lacking JHB3 and JH III, while overexpression of Cyp6g2 or jhamt caused a significant increase in JHB3 and JH III titer. CONCLUSIONS: These findings collectively established that Cyp6g2 as the major JH epoxidase in the higher dipterans and laid the groundwork for the further understanding of JH biosynthesis. Moreover, these findings pave the way for developing specific Cyp6g2 inhibitors as insect growth regulators or insecticides.

Effect of Moxibustion on Inflammatory Cytokines for Low Back Pain: A Systematic Review, Meta-Analysis and Meta-Regression.

Background and Objective: Moxibustion is effective for low back pain (LBP), and inflammatory cytokines may play an important role in the mechanism of moxibustion treatment. The purpose of this meta-analysis was to explore the mechanism of moxibustion in LBP in terms of inflammatory cytokines. Methods: We searched China National Knowledge Infrastructure, Wanfang database, Cochrane Central Register of Controlled Trials, Ovid MEDLINE, Embase, PubMed, and Web of Science to identify eligible randomized controlled trials (RCTs). There was no restriction on the publication date. Results: Thirty RCTs measuring interleukin (IL-) 1, IL-1ß, IL-6, IL-12, IL-17, IL-23, and tumor necrosis factor (TNF-) α were included in this meta-analysis. Compared to controls: single moxibustion could effectively decrease levels IL-6 and IL-23 (SMD, -0.71, 95% CI: -1.25 to -0.17, p = 0.01; SMD, -1.61, 95% CI: -2.20 to -1.03, p < 0.01, respectively); combined moxibustion had significant effects on IL-1, IL-1ß, IL-6, IL-12, IL-17, and TNF-α (p < 0.05). Overall, for LBP, single or combined moxibustion could effectively down-regulate levels of pro-inflammatory cytokines (p = 0.007 and p < 0.00001, respectively). For safety of moxibustion, the incidence rate of side effects was similar to that of controls (RD, -0.01, 95% CI: -0.02 to 0.01, p = 0.59). Sensitivity analysis showed that the pooled estimates were robust, and publication bias analysis showed there was a significant small study effect (Egger's test p = 0.0000). High statistical heterogeneity existed between included RCTs, meta-regression showed there was no potential factor explaining the source of heterogeneity. Conclusion: For LBP, moxibustion can effectively decrease levels of IL-1, IL-1ß, IL-6, IL-12, IL-17, IL-23, and TNF-α to achieve analgesia. Because the side effects of moxibustion are transient, it is relatively safe for clinical use. However, based on high heterogeneity in this meta-analysis, rigorously designed RCTs are required to further confirm the results in this review.

Sterol Regulation of Development and 20-Hydroxyecdysone Biosynthetic and Signaling Genes in Drosophila melanogaster.

Ecdysteroids are crucial in regulating the growth and development of insects. In the fruit fly Drosophila melanogaster, both C27 and C28 ecdysteroids have been identified. While the biosynthetic pathway of the C27 ecdysteroid 20-hydroxyecdysone (20E) from cholesterol is relatively well understood, the biosynthetic pathway of C28 ecdysteroids from C28 or C29 dietary sterols remains unknown. In this study, we found that different dietary sterols (including the C27 sterols cholesterol and 7-dehydrocholesterol, the C28 sterols brassicasterol, campesterol, and ergosterol, and the C29 sterols ß-sitosterol, α-spinasterol, and stigmasterol) differentially affected the expression of 20E biosynthetic genes to varying degrees, but similarly activated 20E primary response gene expression in D. melanogaster Kc cells. We also found that a single dietary sterol was sufficient to support D. melanogaster growth and development. Furthermore, the expression levels of some 20E biosynthetic genes were significantly altered, whereas the expression of 20E signaling primary response genes remained unaffected when flies were reared on lipid-depleted diets supplemented with single sterol types. Overall, our study provided preliminary clues to suggest that the same enzymatic system responsible for the classical C27 ecdysteroid 20E biosynthetic pathway also participated in the conversion of C28 and C29 dietary sterols into C28 ecdysteroids.

The Expression of Plasmacytoid Dendritic Cells and TLR7/9-MyD88-IRAKs Pathway in Chronic Eczema Lesions.

Purpose: To investigate the expression of Plasmacytoid Dendritic Cells (pDCs) and TLR7/9-MyD88-IRAKs pathway in chronic eczema lesions. Patients and Methods: Lesional tissues and the surrounding healthy tissues were collected from 25 individuals with chronic eczema, and immunohistochemistry was used to detect and comparatively analyze the expression profile of CD123, CD2AP, toll-like receptor 7 (TLR7), toll-like receptor 9 (TLR9) along with interleukin-1 receptor-associated kinase 1 (IRAK1) and interferon regulatory factor 7 (IRF7) in signaling pathways. Results: The positive rates of CD2AP + pDC and CD123 + pDC in lesional tissues were significantly elevated compared to the surrounding healthy tissues (P < 0.05). They were distributed in both the epidermal and dermal layers of the lesional tissue, but the majority were in the dermal layer. The TLR7, TLR9, IRAK1 and IRF7 were more expressed in dermal layers of the lesional tissue with higher positive rates of expression compared to the surrounding healthy tissues (P < 0.05). IRAK1 and IRF7 were expressed in a small amount in the epidermal layer with higher positive rates of expression than in the surrounding healthy tissues (P < 0.05), while the positive rates of TLR7 and TLR9 expression in the epidermal layer were not statistically different from those in the surrounding healthy tissues (P > 0.05). Conclusion: PDC and TLR7/9-MyD88-IRAKs pathways are actively expressed in chronic eczema lesions and may be involved in pathogenesis and disease progression.

Study on the application of coal spontaneous combustion positive pressure beam tube classification monitoring and early warning.

To improve the accuracy of early warning on coal spontaneous combustion (CSC), this paper, on the basis of the principle of preferential selection of index gases in CSC process, carries out fitting analysis of the variation curve of index gas data with coal temperature by combining logistic fitting model, then establishes a CSC graded warning system based on positive pressure beam tube monitoring, and determines CO, O2, φ(CO)/φ(O2), C2H4, C2H6, φ(C2H4)/φ(C2H6) as the index gases for predicting and forecasting CSC, and accurately divides the CSC process into seven levels of early warning: safe, gray, blue, yellow, orange, red, and black. Applying the CSC positive pressure beam tube monitoring system to Dongtan coal mine and analyzing the error by manual sampling and sampling by positive pressure beam tube system, we find that the error is less than 0.1%. Monitoring of several working faces, we get that the CO and CH4 concentrations of 14,320 working face are higher than the normal level at the beginning of mining, and the 100 × CO/ΔO2 value is higher than the gray warning threshold of 0.1, and the warning level is gray warning. After taking timely preventive measures against coal oxidation and warming, the CO and CH4 concentrations return to the normal level and the warning level drops to the safe level. This paper improves the monitoring, identifying and early warning capabilities of underground CSC in its early stage.

Sacha Inchi Oil Press-Cake Protein Hydrolysates Exhibit Anti-Hyperuricemic Activity via Attenuating Renal Damage and Regulating Gut Microbiota.

The incidence of hyperuricemia has increased globally due to changes in dietary habits. The sacha inchi oil press-cake is generally discarded, resulting in the waste of resources and adverse environmental impact. For the purpose of developing sacha inchi oil press-cake and identifying natural components with anti-hyperuricemic activities, we systemically investigated the underlying mechanisms of sacha inchi oil press-cake protein hydrolysates (SISH) in the hyperuricemic rat model. SISH was obtained from sacha inchi oil press-cake proteins after trypsin treatment, and 24 peptides with small molecular weight (<1000 Da) were identified. The results of animal experiments showed that SISH significantly decreased the serum uric acid (UA) level by inhibiting the xanthine oxidase (XOD) activity and regulating the gene expression related to UA production and catabolism in hyperuricemia rats, such as Xdh and Hsh. In addition, SISH attenuated the renal damage and reduced the gene expression related to inflammation (Tlr4, Map3k8, Pik3cg, Pik3ap1, Ikbke, and Nlrp3), especially Tlr4, which has been considered a receptor of UA. Notably, SISH reversed high purine-induced gut microbiota dysbiosis, particularly by enhancing the relative abundance of butyric acid-producing bacteria (unidentified_Ruminococcaceae, Oscillibacter, Ruminiclostridium, Intestinimonas). This research provided new insights into the treatment of hyperuricemia.

Anti-diabetic and gut microbiota modulation effects of sacha inchi (Plukenetia volubilis L.) leaf extract in streptozotocin-induced type 1 diabetic mice.

BACKGROUND: Sacha inchi (Plukenetia volubilis L.) tea has been used as an adjuvant treatment for diabetes in Pu'er, in the Yunnan province of China. The effects of sacha inchi tea on diabetes and the underlying mechanisms remain unknown. This study was conducted to investigate the influence of a water extract of sacha inchi (P. volubilis L.) leaves (PWE) on hypoglycemic activity and gut microbiota composition in mice with streptozotocin (STZ)-induced type 1 diabetes mellitus (T1DM). During the 6 weeks of the study, T1DM mice were administered PWE intragastrically at 400 mg kg-1 body weight (BW) per day. RESULTS: Treatment with PWE reduced excessive loss of BW and excessive intake of food. It significantly decreased blood glucose levels and improved oral glucose tolerance. The treatment caused protective histopathological transformations in sections of the pancreas, leading to decreased insulin resistance and improved insulin sensitivity. Treatment with PWE also significantly ameliorated disorders of the gut microbiota structure and increased the richness and diversity of intestinal microbial species in T1DM mice. At the genus level, the populations of several crucial bacteria, such as Akkermansia, Parabacteroides, and Muribaculum increased in the PWE treatment group but the abundance of Ruminiclostridium and Oscillibacter decreased. CONCLUSIONS: Treatment with PWE can ameliorate hyperglycemic symptoms in STZ-induced T1DM mice, and the anti-diabetic effect of PWE was related to the amelioration of gut microbial structural disorder and the enrichment of functional bacteria. © 2022 Society of Chemical Industry.

Ambipolar Charge Storage in Type-I Core/Shell Semiconductor Quantum Dots toward Optoelectronic Transistor-Based Memories.

Efficient charge storage media play a pivotal role in transistor-based memories and thus are under intense research. In this work, the charge storage ability of type-I InP/ZnS core/shell quantum dots is well revealed through studying a pentacene-based organic transistor with the quantum dots (QDs) integrated. The quantum well-like energy band structure enables the QDs to directly confine either holes or electrons in the core, signifying a dielectric layer-free nonvolatile memory. Especially, the QDs in this device can be charged by electrons using light illumination as the exclusive method. The electron charging process is ascribed to the photoexcitation process in the InP-core and the hot holes induced. The QDs layer demonstrates an electron storage density of ≈5.0 × 1011  cm-2 and a hole storage density of ≈6.4 × 1011  cm-2 . Resultingly, the output device shows a fast response speed to gate voltage (10 µs), large memory window (42 V), good retention (>4.0 × 104 s), and reliable endurance. This work suggests that the core/shell quantum dot as a kind of charge storage medium is of great promise for optoelectronic memories.

Inhibition of LPA1 Signaling Impedes Conversion of Human Tenon's Fibroblasts into Myofibroblasts Via Suppressing TGF-ß/Smad2/3 Signaling.

Purpose: Lysophosphatidic acid (LPA) is a growth factor-like phospholipid that has been recognized as a profibrotic mediator in numerous tissues, yet, whether it plays a role in subconjunctival fibrosis remains to be investigated. Therefore, this study was designed to examine the effect of LPA1-3 signaling inhibitor, Ki16425 on the conversion of human Tenon's fibroblasts (HTFs) into myofibroblasts. Methods: Primary cultured HTFs were incubated with transforming growth factor-ß1 (TGF-ß1) alone or combined with Ki16425, the cell proliferation and migration were measured by Cell Counting Kit-8 and the scratch wound assay, respectively. HTFs contractility was evaluated with 3-dimensional (3D) Collagen Contraction assay. The mRNA and protein levels of α-smooth muscle actin (α-SMA), Snail and the phosphorylation levels of Smad2/3, p38MAPK, and ERK1/2 were determined by real-time quantitative polymerase chain reaction (RT-qPCR), western blot, and immunofluorescence staining. Results: Ki16425 significantly prevent the proliferation and migration of Tenon's fibroblasts (HTFs) in a dose-dependent manner. Furthermore, Ki16425 blocked HTFs myofibroblast differentiation via downregulation of mRNA and protein expression of α-SMA. 3D collagen gel contraction assay demonstrated that Ki16425 effectively inhibits myofibroblast contraction induced by TGF-ß1. Mechanistically, we revealed that Ki16425 reduces Smad2/3 but not p38MAPK or ERK1/2 phosphorylation by TGF-ß1. By using an LPA1-specific inhibitor, AM095, we confirmed that LPA1 signaling but not LPA2 or LPA3 is involved in TGF-ß1 induced HTFs activation. Conclusions: Our results show that inhibition of LPA1 signaling presents potent antifibrotic effect in HTFs, which may serve as a promising intervention strategy for preventing subconjunctival fibrosis caused by glaucoma filtration surgery.

A Tunneling Dielectric Layer Free Floating Gate Nonvolatile Memory Employing Type-I Core-Shell Quantum Dots as Discrete Charge-Trapping/Tunneling Centers.

A nonvolatile memory with a floating gate structure is fabricated using ZnSe@ZnS core-shell quantum dots as discrete charge-trapping/tunneling centers. Systematical investigation reveals that the spontaneous recovery of the trapped charges in the ZnSe core can be effectively avoided by the type-I energy band structure of the quantum dots. The surface oleic acid ligand surrounding the quantum dots can also play a role of energy barrier to prevent unintentional charge recovery. The device based on the quantum dots demonstrates a large memory window, stable retention, and good endurance. What is more, integrating charge-trapping and tunneling components into one quantum dot, which is solution synthesizable and processible, can largely simplify the processing of the floating gate nonvolatile memory. This research reveals the promising application potential of type-I core-shell nanoparticles as the discrete charge-trapping/tunneling centers in nonvolatile memory in terms of performance, cost, and flexibility.

MEK inhibition prevents TGF­ß1­induced myofibroblast transdifferentiation in human tenon fibroblasts.

Subconjunctival fibrosis represents the primary cause of postoperative failure of trabeculectomy, and at present there is a lack of effective intervention strategies. The present study aimed to investigate the effect of the mitogen­activated protein kinase kinase (MEK) inhibitor U0126 on human tenon fibroblast (HTF) myofibrosis transdifferentiation, and to illuminate the underlying molecular mechanisms involved. It was demonstrated that U0126 significantly inhibited the proliferation, migration and collagen contraction of HTFs stimulated with TGF­ß1. In addition, U0126 largely attenuated the TGF­ß1­induced conversion of HTFs into myofibroblasts, as indicated by a downregulation of the mRNA and protein expression of α­smooth muscle actin and zinc finger protein SNAI1, and by ameliorating the 3D­collagen contraction response. Mechanistically, U0126 suppressed the TGF­ß1­stimulated phosphorylation of mothers against decapentaplegic homolog 2/3, P38 mitogen­activated protein kinase and extracellular signal­regulated kinase 1/2, indicating that U0126 may inhibit HTF activation through the canonical and non­canonical signaling pathways of TGF­ß1. Therefore, U0126 exhibits a potent anti­fibrotic effect among HTFs, and the inhibition of MEK signaling may serve as an alternative intervention strategy for the treatment of trabeculectomy­associated fibrosis.

Structural, functional properties and immunomodulatory activity of isolated Inca peanut (Plukenetia volubilis L.) seed albumin fraction.

This work aimed to determine the structural, functional properties and immunomodulatory activity of an albumin fraction isolated from Inca peanut seed (AF-IPS). Structural characterization revealed that AF-IPS contains two polypeptides with molecular weight ranges of 25-45 and 10-15 kDa. AF-IPS is mainly composed of α-helix, ß-sheet, and ß-turn secondary structures. Differential scanning calorimetry analysis indicated that the denaturation temperature of AF-IPS at room temperature is 101.93 °C. AF-IPS possessed excellent protein solubility (63.0%), water holding capacity (1.59 g/g), foaming (350.2%) and emulsifying (13.0 mL/g) ability. Heat treatment improved protein solubility, oil holding capacity, and foaming and emulsifying ability. AF-IPS exhibited immunomodulatory activity by stimulating the proliferation and enhancing the TNF-α secretion of splenic lymphocytes, and through increasing the cellular lysosomal enzyme and pinocytic activities, and by moderately promoting the NO and H2O2 production of RAW264.7 cells. Our findings indicated that AF-IPS has potential applications in functional and pharmaceuticals foods.

Nintedanib inhibits TGF-ß-induced myofibroblast transdifferentiation in human Tenon's fibroblasts.

Purpose: This study aimed to investigate the effect of nintedanib on the conversion of human Tenon's fibroblasts (HTFs) into myofibroblasts and reveal the molecular mechanisms involved. Methods: Primary cultured HTFs were incubated with transforming growth factor ß1 (TGF-ß1) alone or combined with nintedanib, and cell proliferation and migration were measured by cell counting kit-8 (CCK8) and the scratch wound assay, respectively. HTF contractility was evaluated with a 3D collagen contraction assay. The mRNA and protein levels of α smooth muscle actin (α-SMA) and Snail and the phosphorylation levels of Smad2/3, p38 mitogen-activated protein kinase (p38MAPK), and extracellular signal-regulated kinase ½ (ERK1/2) were determined by quantitative reverse transcription polymerase chain reaction (RT-PCR), western blot, and immunofluorescence staining. Results: Nintedanib inhibited the proliferation and migration of HTFs in a dose-dependent manner. Furthermore, nintedanib prevented HTF myofibroblast differentiation via downregulation of mRNA and protein expression of α-SMA and Snail. A three-dimensional (3D) collagen gel contraction assay demonstrated that nintedanib effectively inhibits myofibroblast contraction induced by TGF-ß1. Mechanistically, we revealed that nintedanib reduces the TGF-ß1-induced phosphorylation of Smad2/3, p38MAPK, and ERK1/2, suggesting that nintedanib acts through both classic and nonclassic signaling pathways of TGF-ß1 to prevent HTF activation. Conclusions: Our study provides new evidence that nintedanib has potent antifibrotic effects in HTFs and suggests that it may be used as a potential therapeutic agent for subconjunctival fibrosis.

Sirt1 regulates acrosome biogenesis by modulating autophagic flux during spermiogenesis in mice.

Sirt1 is a member of the sirtuin family of proteins and has important roles in numerous biological processes. Sirt1-/- mice display an increased frequency of abnormal spermatozoa, but the mechanism of Sirt1 in spermiogenesis remains largely unknown. Here, we report that Sirt1 might be directly involved in spermiogenesis in germ cells but not in steroidogenic cells. Germ cell-specific Sirt1 knockout mice were almost completely infertile; the early mitotic and meiotic progression of germ cells in spermatogenesis were not obviously affected after Sirt1 depletion, but subsequent spermiogenesis was disrupted by a defect in acrosome biogenesis, which resulted in a phenotype similar to that observed in human globozoospermia. In addition, LC3 and Atg7 deacetylation was disrupted in spermatids after knocking out Sirt1, which affected the redistribution of LC3 from the nucleus to the cytoplasm and the activation of autophagy. Furthermore, Sirt1 depletion resulted in the failure of LC3 to be recruited to Golgi apparatus-derived vesicles and in the failure of GOPC and PICK1 to be recruited to nucleus-associated acrosomal vesicles. Taken together, these findings reveal that Sirt1 has a novel physiological function in acrosome biogenesis.

Fasudil inhibits LPS-induced migration of retinal microglial cells via regulating p38-MAPK signaling pathway.

PURPOSE: To investigate the effect and possible molecular mechanisms of fasudil on retinal microglial (RMG) cell migration. METHODS: Primary cultured RMG cells were incubated with lipopolysaccharide (LPS), fasudil, and/or SB203580 (a p38 inhibitor). RMG cell motility was determined with the scratch wound assay and the Transwell migration assay. The phosphorylation of p38 and levels of matrix metalloproteinase 2 (MMP-2) and MMP-9 were measured with western blot. RESULTS: In the scratch-induced migration assay, as well as in the Transwell migration assay, the results indicated that LPS stimulated the migratory potential of RMG cells and fasudil significantly reduced LPS-stimulated RMG cell migration in a concentration-dependent manner. However, fasudil had no effect on RMG cell migration in the absence of LPS stimulation. Moreover, fasudil reduced the level of phosphor-p38 mitogen-activated protein kinase (p-p38-MAPK) in a concentration-dependent manner, without effects on the levels of phospho-p44/42 (p-ERK1/2) and phospho-c-Jun N-terminal kinase (p-JNK). Cotreatment with SB203580 (a p38 inhibitor) and fasudil resulted in the synergistic reduction of MMP-2, MMP-9, and p-p38-MAPK, as well as a reduction in the LPS-stimulated migration capabilities of the RMG cells, suggesting fasudil suppresses the LPS-stimulated migration of RMG cells via directly downregulating the p38-MAPK signaling pathway. CONCLUSIONS: Our studies indicated that fasudil inhibited LPS-stimulated RMG cell migration via suppression of the p38-MAPK signaling pathway.