NPInter v5.0: ncRNA interaction database in a new era.

Noncoding RNAs (ncRNAs) play key regulatory roles in biological processes by interacting with other biomolecules. With the development of high-throughput sequencing and experimental technologies, extensive ncRNA interactions have been accumulated. Therefore, we updated the NPInter database to a fifth version to document these interactions. ncRNA interaction entries were doubled from 1 100 618 to 2 596 695 by manual literature mining and high-throughput data processing. We integrated global RNA-DNA interactions from iMARGI, ChAR-seq and GRID-seq, greatly expanding the number of RNA-DNA interactions (from 888 915 to 8 329 382). In addition, we collected different types of RNA interaction between SARS-CoV-2 virus and its host from recently published studies. Long noncoding RNA (lncRNA) expression specificity in different cell types from tumor single cell RNA-seq (scRNA-seq) data were also integrated to provide a cell-type level view of interactions. A new module named RBP was built to display the interactions of RNA-binding proteins with annotations of localization, binding domains and functions. In conclusion, NPInter v5.0 (http://bigdata.ibp.ac.cn/npinter5/) provides informative and valuable ncRNA interactions for biological researchers.

Simple Framework for Simultaneous Analysis of Both Electrodes in Stoichiometric Lithium-Sulfur Batteries.

A battery is composed of two electrodes that depend on and interact with each other. However, galvanostatic charging-discharging measurement, the most widely used method for battery evaluation, cannot simultaneously reflect performance metrics [capacity, Coulombic efficiency (CE), and cycling stability] of both electrodes because the result is generally governed by the lower-capacity electrode of the cell, namely the limiting reagent of the battery reaction. In studying stoichiometric Li-S cells operating under application-relevant high-mass-loading and lean-electrolyte conditions, we take advantage of the two-stage discharging behavior of sulfur to construct a simple framework that allows us to analyze both electrodes simultaneously. The cell capacity and its decay are anode performance descriptors, whereas the first plateau capacity and cell CE are cathode performance descriptors. Our analysis within this frame identifies Li stripping/plating and polysulfide shuttling to be the limiting factors for the cycling performance of the stoichiometric Li-S cell. Using our newly developed framework, we examine various previously reported strategies to mitigate these bottleneck problems and find modifying the separator with a reduced graphene oxide layer to be an effective means, which improves the capacity retention rate of the cell to 99.7% per cycle.

Sodium Octahydridotriborate as a Solid Electrolyte with Excellent Stability Against Sodium-Metal Anode.

Solid-state sodium metal batteries have been extensively investigated because of their potential to improve safety, cost-effectiveness, and energy density. The development of such batteries urgently required a solid-state electrolyte with fast Na-ion conduction and favorable interfacial compatibility. Herein, the progress on developing the NaB3H8 solid-state electrolytes is reported, which show a liquid-like ionic conductivity of 0.05 S cm-1 at 56 °C with an activation energy of 0.35 eV after an order-disorder phase transformation, matching or surpassing the best single-anion hydridoborate conductors investigated up to now. The steady polarization voltage and significantly decreased resistance are achieved in the symmetric Na/NaB3H8/Na cell, indicating the great electrochemical stability and favorable interfacial contact with the Na metal of NaB3H8. Furthermore, a Na/NaB3H8/TiS2 battery, the first high-rate (up to 1 C) solid-state sodium metal battery using the single-anion hydridoborate electrolyte, is demonstrated, which exhibits superior rate capability (168.2 mAh g-1 at 0.1 C and 141.2 mAh g-1 at 1 C) and long-term cycling stability (70.9% capacity retention at 1 C after 300 cycles) at 30 °C. This work may present a new possibility to solve the interfacial limitations and find a new group of solid-state electrolytes for high-performance sodium metal batteries.

Brain potentials reveal how emotion filters native language access when bilinguals read words in their second language.

It is now well established that reading words in a second language (L2) automatically activates native language (L1) translations in bilinguals. Although there is evidence that access to such representations is inhibited when words have a negative emotional valence, the mechanism underlying such inhibition is elusive, and it is unknown whether inhibition arises online as L2 is being processed or whether negative valence affects subsequent L1 processing. Here, we recorded event-related brain potentials in Chinese-English bilinguals engaged in an implicit translation-priming paradigm involving L2 (English) word pairs. Participants performed a semantic relatedness task, unaware that word pairs could conceal a sound repetition if translated into Chinese. When emotional valence was manipulated in prime position (first word), we observed form repetition priming through L1 translations for positive but not for negative words. However, when emotional valence was manipulated in target position (second word), priming occurred for both positive and negative word valences. This result begins to elucidate the mechanism by which emotion regulates language processing in bilinguals: Negative words in L2 induce a refractory period during which cross-language lexical access is blocked. These findings show that despite being neuroanatomically distinct in the human brain, emotional (limbic) regulation systems can penetrate language processing.

A High-Rate and Long-Life Sodium Metal Battery Based on a NaB3H8 ⋠xNH3@NaB3H8 Composite Solid-State Electrolyte.

All-solid-state sodium metal batteries are promising for large-scale energy storage applications owing to their intrinsic safety and cost-effectiveness. However, they generally suffer from sodium dendrite growth or rapid capacity fading, especially at high rates, mainly due to poor wettability, sluggish ionic transport, or low interfacial stability of the solid electrolytes. Herein, we report a novel composite, NaB3H8 ⋅ xNH3@NaB3H8 (x<1), as a new class of solid electrolyte for high-rate batteries. NaB3H8 ⋅ xNH3@NaB3H8 is obtained from the sticky NaB3H8 ⋅ NH3 after removal of NH3 partially at room temperature. It delivers an ionic conductivity of 0.84 mS cm-1 at 25 °C and reaches 20.64 mS cm-1 at 45 °C after an order-disorder phase transformation. It also reveals a good capability of dendrite suppression and remarkable stability against sodium metal. These performances enable the all-solid-state Na//TiS2 battery with a high capacity of 232.4 mAh g-1 (97.2 % of theoretical capacity) and long-term cycling stability at 1 C. Notably, this battery shows superior long-life cycling stability even at 5 and 10 C, which has been rarely reported in all-solid-state sodium metal batteries. This work opens a new group of solid electrolytes, contributing to fast-charging or high-power-density sodium metal batteries.

Manipulating the Spin State of Spinel Octahedral Sites via a π-π Type Orbital Coupling to Boost Water Oxidation.

Spin state is often regarded as the crucial valve to release the reactivity of energy-related catalysts, yet it is also challenging to precisely manipulate, especially for the active center ions occupied at the specific geometric sites. Herein, a π-π type orbital coupling of 3d (Co)-2p (O)-4f (Ce) was employed to regulate the spin state of octahedral cobalt sites (CoOh) in the composite of Co3O4/CeO2. More specifically, the equivalent high-spin ratio of CoOh can reach to 54.7% via tuning the CeO2 content, thereby triggering the average eg filling (1.094) close to the theoretical optimum value. The corresponding catalyst exhibits a superior water oxidation performance with an overpotential of 251 mV at 10 mA cm-2, rivaling most cobalt-based oxides state-of-the-art. The π-π type coupling corroborated by the matched energy levels between Ce t1u/t2u-O and CoOh t2g-O π type bond in the calculated crystal orbital Hamilton population and partial density of states profiles, stimulates a π-donation between O 2p and π-symmetric Ce 4fyz2 orbital, consequently facilitating the electrons hopping from t2g to eg orbital of CoOh. This work offers an in-depth insight into understanding the 4f and 3d orbital coupling for spin state optimization in composite oxides.

Sandwich Structured Metal oxide/Reduced Graphene Oxide/Metal Oxide-Based Polymer Electrolyte Enables Continuous Inorganic-Organic Interphase for Fast Lithium-Ion Transportation.

Introducing inorganic fillers into organic poly(ethylene oxide)(PEO)-based electrolyte has attracted substantial attention to enhance its ionic conductivity and mechanical strength, but limited inorganic-organic interphases are always caused by isolated particles agglomeration. Herein, a variety of sandwich structured metal oxide/reduced graphene oxide(rGO)/metal oxide nanocomposites to optimize lithium-ion conduction by interconnected amorphous organic-inorganic interphases in lithium metal batteries, are proposed. With the support of high surface area rGO, the agglomeration of metal oxide particles is precluded, forming continuous amorphous organic-inorganic interphases with stacked layer-by-layer structure, thus creating 3D interconnected lithium-ion transportation channels vertically and laterally. Besides, metal oxide nanoparticles with hydroxyls possess high affinity toward bis(tri-fluoromethanesulfonyl)imide anions by hydrogen bindings between hydroxyls and fluorine and metal-oxygen bonds, releasing more free lithium ions. Consequently, PEO-ZnO/rGO/ZnO electrolyte delivers superior ionic conductivity of 1.02 × 10-4 S cm-1 at 25 °C and lithium-ion transference number of 0.38 at 60 °C. Furthermore, ZnO/rGO/ZnO insertion promotes the formation of LiF-rich stable solid electrolyte interface, endowing Li symmetric cells with long-term cycling stability over 900 hours. The corresponding LiFePO4 cathode possesses a high reversible specific capacity of 130 mAh g-1 at 0.5C after cycling 300 cycles with a poor capacity fading of 0.05% per cycle.

Exploration of Multi-Gene DNA Barcode Markers to Reveal the Broad Genetic Diversity of Field Ticks (Acari: Ixodidae) in a Tropical Environment of Hainan Island, China.

Ticks are hematophagous arthropods and obligate ectoparasites of humans and other animals. This study focused on the molecular discrimination of ticks in the tropical environment of Hainan according to multi-gene DNA barcode markers with the expectation of accurately distinguishing species. A total of 420 ticks, including 49 adult ticks, 203 nymphal ticks, and 168 larval ticks, were collected in the field, and the 49 adult ticks were identified as Rhipicephalus turanicus, Dermacentor marginatus, and Haemaphysalis longicornis. The mitochondrial 16S rRNA, ribosomal 28S rRNA D2, and ribosomal internal transcribed spacer 2 (ITS2) regions were used as DNA barcode markers to discriminate species. According to basic local alignment search tool analysis against the GenBank database, 16S rRNA positively identified ticks in the Rhipicephalus, Dermacentor, and Haemaphysalis genera; the 28S rRNA D2 region identified ticks in the Rhipicephalus and Dermacentor genera; and ITS2 identified ticks as D. marginatus. Pairwise sequence comparisons based on these three regions were visualized with a Sequence Demarcation Tool matrix. Substitution saturation tests using data analysis and molecular biology and evolution revealed little substitution saturation (Iss < Iss.c, p < 0.05) in the 16S rRNA region for the Haemaphysalis genus; 28S rRNA D2 region for the Rhipicephalus, Dermacentor, and Haemaphysalis genera; and ITS2 region for the Rhipicephalus and Dermacentor genera. Distinctive sequences for which it is difficult to obtain good matches with the sequences available in GenBank exist in the ticks of Hainan. Future studies should obtain complementary sequences to refine and update the database for the molecular characterization of ticks.

Prevalence of visual impairment in older people living with dementia and its impact: a scoping review.

BACKGROUND AND OBJECTIVES: Visual impairment (VI) and dementia both increase with age, and it is likely that many older people are living with both conditions. This scoping review aims to investigate the prevalence and types of VI among older people living with dementia, and the impact of VI on older people living with dementia and their caregivers. METHODS: This scoping review used Arksey and O'Malley's methodological framework. Studies in any setting involving people living with dementia and some assessment of either VI, eye diseases causing VI or the impact of VI were included. RESULTS: Thirty-six studies were included. Thirty-one studies reported the prevalence of VI in older people living with dementia, while ten studies reported on impacts of VI on people living with dementia. Only one study reported on impacts on caregivers. The prevalence of VI or specific eye diseases among older people living with dementia ranged from 0.2 to 74%. The impacts of VI on older people living with dementia included increased use of hospital services, increased disability and dependency, reduced social engagement, negative emotions, increased abnormal behaviours, loss of hobbies, difficulty in using visual aids or memory aids, and greater Neuropsychiatric Inventory symptoms. And the impacts on caregivers included increased conflict and physical exhaustion. CONCLUSION: VI is common in older people living with dementia and is associated with negative impacts on those with dementia and their caregivers. However, heterogeneity between studies in terms of setting and method for assessing and defining VI make it difficult to compare findings among studies. Further research is needed, particularly assessing the impact on caregivers.

Electroporation of SUMO-His-Cre protein triggers a specific recombinase-mediated cassette exchange in HEK 293T cells.

Cre recombinase is a widely-used genetic manipulation of genomic DNA. However, the conventional transfection of the DNA vectors expressing the Cre recombinase or viral transduction method yields low transfection efficiencies or insertion mutagenesis. The present paper evaluated whether the direct protein delivery of Cre recombinase through electroporation can induce the Cre-mediated recombination in the HEK 293T cells. Here, the small ubiquitin-related modifier (SUMO) -tagged His-Cre fusion protein was expressed in a soluble pattern in the Eschrichria coli (E.coli) cells, purified using affinity chromatography, and finally electroporated into the HEK 293T cells. These cells were previously transfected with three different Cre reporter vectors. The electroporation of the HEK 293T cells revealed either the activation of EGFP expression, or a decrease in RFP expression, and a concomitant increase in EGFP expression, indicating a desired recombinase-mediated cassette exchange (RMCE) event (conversion of RFP to EGFP), and a biological activity of the purified SUMO-His-Cre protein. The fusion protein is expected to serve in the Easi-CRISPR-LoxP-mediated genome editing to generate transgenic animal models.

Establishment of purification method for prokaryotic expression of Serpin gene for Dermatophagoides farinae.

This study aimed to develop an effective method for the expression and purification of the Dermatophagoides farinae serpin protein and to establish an experimental foundation for elucidating its role in the temperature stress response. The total RNA of D. farinae was extracted, and specific primers were designed for serpin amplification. Serpin was joined with pET32a vector and transformed into BL21 (DE3) cells. Expression of recombinant proteins was induced. Proteins were extracted by enzymatic lysis or enzymatic lysis combined with ultrasonication. Recombinant proteins were purified by Ni-NTA method. SDS-PAGE was conducted to evaluate protein expression, extraction, and purification efficiency. Agarose gel electrophoresis and sequencing analysis showed that the amplified serpin open reading frame was 1284 bp, encoding a hydrophilic and stable protein with a relative molecular weight of 48.30 kD. SDS-PAGE demonstrated that there was a specific band at 55-70 kD, which was consistent with the predicted size of the recombinant pET32a-Serpin protein. Enzymatic lysis combined with 30% ultrasonic power promoted the release of soluble protein more effectively than enzymatic lysis alone. 16 °C for 4 h was optimal for inducing expression. The optimal imidazole concentrations for washing non-His-tagged protein and eluting His-tagged protein were determined to be 20 mM and 200 mM, respectively. In this study, A prokaryotic expression and purification system for the D. farinae serpin protein was successfully established, providing a technical reference for functional gene research in mites at the protein level.

Primary ovarian carcinoid arising in associated mature cystic teratoma.

BACKGROUND: Primary ovarian carcinoid is a very rare ovarian low-grade neuroendocrine tumor, accounting for about 0.1% of all ovarian neoplasms. CASE PRESENTATION: We reported a case of primary ovarian carcinoid arising from a mature cystic teratoma in a 50-year-old woman. Intraoperative frozen section of left ovarian mass was assessed and a malignant epithelial tumor was considered. Morphologically, the main tumor was composed of cells forming trabeculae, and mature cystic teratoma was observed adjacent to the main part. Immunohistochemistry revealed that the trabecular cells were diffuse positive for pan Cytokeratin, CD56 and synaptophysin with low Ki-67 index (about 1%). CONCLUSIONS: Careful morphological observation combined with appropriate accessory examination are essential for the diagnosis of primary ovarian carcinoid arising from mature cystic teratoma. In addition, the classification criteria of the primary ovarian neuroendocrine tumor are discussed.

De novo transcriptome sequencing and functional annotation of Demodex canis.

Demodex canis is an important parasitic mite causing skin lesions in dogs. However, molecular studies on this species are limited because of the lack of transcriptomic data. To obtain functional genes of D. canis, mites were collected and RNA was extracted for transcriptome sequencing and functional annotation. Coding sequences (CDSs) of important functional genes were screened and identified using bioinformatics strategies. Six complete CDSs were amplified by specific primers, cloned and sequenced. Results demonstrated that the quantity of the RNA extracted from 100 mites was 12.8 ng and the RNA integrity number was 5.4, indicating that it could be used to construct a cDNA library. Transcriptome sequencing yielded 263,619 unigenes, of which 151,048 (57.3%) were functionally annotated. Using bioinformatics strategies, 58 total CDSs were identified as homologous to those of closely related mite species, including 16 types of allergen genes, 13 types of protease genes, and nine types of motion-related genes. The verified CDSs were almost identical to the CDSs of unigenes. Phylogenetic analyses of tropomyosin further revealed that the verified CDSs clustered successively with those of Demodex species and Tetranychus species in Raphignathae, which agreed with the morphological classification, demonstrating the reliability of the transcriptomic data. In conclusion, this study is the first to successfully sequence transcriptome of D. canis, perform functional annotation, and verify CDSs, which will provide ample data for further studies on the functional genes and molecular pathogenic mechanism of D. canis.

Molecular identification and DNA barcode screening of acaroid mites in ground flour dust.

Molecular identification of acaroid mites is difficult because of the scarcity of molecular data in GenBank. Here, acaroid mites collected from ground flour dust in Xi'an, China, were preliminarily morphologically classified/grouped. Universal primers were then designed to amplify and screen suitable DNA barcodes for identifying these mites. Sixty mite samples were morphologically classified into six groups. Groups 1-2 were identified to Dermatophagoides farinae and Tyrophagus putrescentiae, while Groups 3-6 were not identified to the species level. ITS2 exhibited higher efficiency in molecular identification in comparison with COI, 12S, and 16S. Groups 1-6 were identified as D. farinae, T. putrescentiae, Suidasia nesbitti, Chortoglyphus arcuatus, Lepidoglyphus destructor, and Gohieria sp., respectively. The phylogenetic results were consistent with the morphological classification. Group 6 was further identified as G. fusca according to the morphology of the reproductive foramen. We conclude that the use of ITS2 and the availability of universal primers provide an ideal DNA barcode for molecular identification of acaroid mites. The use of multiple target genetic markers in conjunction with morphological approaches will improve the accuracy of Acaridida identification.

The Xi'an longitudinal mother-child cohort study: design, study population and methods.

The large-scale Xi'an longitudinal mother-child cohort study has started to enroll pregnant women who attended Northwest Women's and Children's Hospital (NWCH) for antenatal care in early pregnancy (less than 20 weeks' gestation) from January 2013 and the enrollment will be ended in January 2023. We aimed to investigate the role of external factors (i.e., diet and environment) and internal (i.e., biological, genetic and epigenetic) on the short- and long-term outcomes of mothers and children up to at least 12 years. Mothers completed all routine prenatal care during pregnancy and four times of follow-up at 42 days, 3, 6 and 12 years after delivery, respectively. For children, birth information were obtained from routine medical records and the follow-up information were obtained from child health care clinics of NWCH at age 42 days, 6, 12 and 24 months, then by interviewing mothers every two years until 12 years old. A range of data (including biological, demographic, birth outcomes/birth defects and nutritional factors from both maternal and off-spring) were collected by both interviews and laboratory tests. By June 30th 2019, a total of 114,946 mothers and 124,454 live births had been recruited.

Platelet and Erythrocyte Extravasation across Inflamed Corneal Venules Depend on CD18, Neutrophils, and Mast Cell Degranulation.

Platelet extravasation during inflammation is under-appreciated. In wild-type (WT) mice, a central corneal epithelial abrasion initiates neutrophil (PMN) and platelet extravasation from peripheral limbal venules. The same injury in mice expressing low levels of the ß2-integrin, CD18 (CD18hypo mice) shows reduced platelet extravasation with PMN extravasation apparently unaffected. To better define the role of CD18 on platelet extravasation, we focused on two relevant cell types expressing CD18: PMNs and mast cells. Following corneal abrasion in WT mice, we observed not only extravasated PMNs and platelets but also extravasated erythrocytes (RBCs). Ultrastructural observations of engorged limbal venules showed platelets and RBCs passing through endothelial pores. In contrast, injured CD18hypo mice showed significantly less venule engorgement and markedly reduced platelet and RBC extravasation; mast cell degranulation was also reduced compared to WT mice. Corneal abrasion in mast cell-deficient (KitW-sh/W-sh) mice showed less venule engorgement, delayed PMN extravasation, reduced platelet and RBC extravasation and delayed wound healing compared to WT mice. Finally, antibody-induced depletion of circulating PMNs prior to corneal abrasion reduced mast cell degranulation, venule engorgement, and extravasation of PMNs, platelets, and RBCs. In summary, in the injured cornea, platelet and RBC extravasation depends on CD18, PMNs, and mast cell degranulation.

Temperature stress response: A novel important function of Dermatophagoides farinae allergens.

Dermatophagoides farinae, an important pathogen, has multiple allergens. However, their expression under physiological conditions are not understood. Our previous RNA-seq showed that allergens of D. farinae were up-regulated under temperature stress, implying that they may be involved in stress response. Here, we performed a comprehensive study. qRT-PCR detection indicated that 26 of the 34 allergens showed differential expression. Der f1 had the most abundant basic expression quantity. Der f 28.0201 (HSP70) and Der f3 had the same regulation pattern in 9 highly expressed transcripts, which only up-regulated at 41 °C and 43 °C, but Der f 28.0201 showed stronger regulation than Der f 3 (19.88-fold vs 6.02-fold). Whereas Der f 1, 2, 7, 21, 22, 27, and 30 were up-regulated under both heat and cold stress, and Der f 27 showed the strongest regulation ability among them. Der f 27 showed more significant up-regulation than Der f 28.0201 under heat stress (23.59-fold vs 19.88-fold), and Der f27 had more obvious up-regulation under cold than heat stress (30.70-fold vs 23.59-fold). The expression of Der f 27, 28.0201 and 1, and D. farinae survival rates significantly decreased following RNAi, indicating the upregulation of these allergens under temperature stress conferred thermo-tolerance or cold-tolerance to D. farinae. In this study, we described for the first time that these allergens have temperature-stress response functions. This new scientific discovery has important clinical value for revealing the more frequent and serious allergic diseases caused by D. farinae during the change of seasons.

Green synthesis of up- and down-conversion photoluminescent carbon dots from coffee beans for Fe3+ detection and cell imaging.

Carbon dots (CDs) are a developing stable, conductive, and environmentally friendly fluorescent nanomaterial with potential applications in biomedicine and sensing. In this study, CDs with up- and down-conversion photoluminescence characteristics were prepared via a one-pot hydrothermal method using coffee beans as the sole raw material. Fourier transform infrared spectra and X-ray photoelectron spectroscopy showed the presence of nitrogen and oxygen functional groups on the CDs surfaces. Transmission electron microscopy showed that the CDs were spherical with an average size of 4.6 nm. The CDs precursor and synthesis process are environmentally benign, and the resulting CDs exhibit excellent photostability, reversible fluorescence response to temperature, and good ionic-strength tolerance. Moreover, these CDs can be used as efficient fluorescence probes in selective and sensitive Fe3+ detection by a dual-readout assay. The down- and up-conversion fluorescence measurements from the CDs were in the linear range for 0-0.10 mM Fe3+ with the detection limits of 15.4 and 16.3 nM, respectively. Furthermore, the CDs were successfully applied to intracellular sensing and imaging Fe3+ ions, indicating their potential use in bioscience applications.

Screening of reference genes and quantitative real-time PCR detection and verification in Dermatophagoides farinae under temperature stress.

Dermatophagoides farinae is an important source of indoor allergens that shows strong tolerance to external temperatures. However, the regularity and mechanism of tolerance are still unclear. Based on our previous RNA-seq and annotation of D. farinae under temperature stress, it is planned to identify differentially expressed genes (DEGs) involved in the temperature stress response by quantitative real-time PCR (qRT-PCR). However, the lack of reference genes directly limited the detection and confirmation of DEGs. Accordingly, in this study, we have selected six candidates as reference genes in D. farinae: 60S RP L11, 60S RP L21, α tubulin, GAPDH, Der f Mal f 6, and calreticulin, and evaluated their expression stabilities as affected by heat and cold stresses, using geNorm, NormFinder, BestKeeper, comparative ΔCt and RefFinder methods. Then the expression level of 15 DEGs were detected and verified. geNorm analysis showed that α tubulin and calreticulin were the most stable reference genes under heat stress and cold stress of D. farinae. Similar evaluation results were obtained by NormFinder and BestKeeper, in which 60S RP L21 and α tubulin were the most stable reference genes. By comparative ΔCt method and a comprehensive evaluation of RefFinder, α tubulin was identified as the most ideal reference gene of D. farinae under heat and cold stresses. Furthermore, qRT-PCR detection results of 15 DEGs were almost identical to the RNA-seq results, indicating that α tubulin is stable as a reference gene. This study provided technical support for DEGs expression studies in D. farinae using qRT-PCR.

IL-20 promotes epithelial healing of the injured mouse cornea.

After corneal epithelial injury, the ensuing inflammatory response is necessary for efficient wound healing. While beneficial healing effects are attributed to recruited neutrophils and platelets, dysregulated inflammation (too little or too much) is associated with impaired wound healing. The purpose of this study was to use an established C57BL/6J mouse model of corneal injury to evaluate the potential modulatory role of interleukin-20 (IL-20) on the inflammatory and healing responses to epithelial wounding. In the uninjured cornea, immunofluorescence staining for IL-20 and its receptor, IL-20RA, was observed on basal epithelial cells at the limbus. After a 2 mm central epithelial abrasion, IL-20 staining was also observed in stromal keratocytes and ELISA studies showed a significant increase (nearly 3-fold) in IL-20 expression. Injured corneas healed more slowly when treated with a topical application of a neutralizing anti-IL-20 antibody. While corneal epithelial cell division and epithelial nerve recovery measured at 24 h post-injury were reduced compared to controls, neutrophil influx into the cornea was increased. In contrast, topical application of recombinant IL-20 (rIL-20) decreased corneal inflammation as evidenced by reductions in limbal vessel dilatation, platelet extravasation, neutrophil recruitment and CXCL1 expression. In wild type mice, topical rIL-20 had a limited effect on corneal wound healing and resulted in only a slight increase in epithelial cell division and epithelial nerve recovery; the rate of wound closure was unaffected. To clarify the effect of IL-20 on corneal wound healing, rIL-20 was topically applied to neutropenic wild type (WT) mice and mutant mice (ɣδ T cell deficient mice and CD11a deficient mice), all of which have well characterized reductions in neutrophil recruitment and delayed wound healing after corneal injury. In each case, rIL-20 restored corneal wound healing to baseline levels while neutrophil recruitment remained low. Thus, it appears that IL-20 plays a beneficial and direct role in corneal wound healing while negatively regulating neutrophil and platelet infiltration.