A Review of Sisal Fiber-Reinforced Geopolymers: Preparation, Microstructure, and Mechanical Properties.

The early strength of geopolymers (GPs) and their composites is higher, and the hardening speed is faster than that of ordinary cementitious materials. Due to their wide source of raw materials, low energy consumption in the production process, and lower emissions of pollutants, they are considered to have the most potential to replace ordinary Portland cement. However, similar to other inorganic materials, the GPs themselves have weak flexural and tensile strength and are sensitive to micro-cracks. Improving the toughness of GP materials can be achieved by adding an appropriate amount of fiber materials into the matrix. The use of discrete staple fibers shows great potential in improving the toughness of GPs. Sisal is a natural fiber that is reproducible and easy to obtain. Due to its good mechanical properties, low cost, and low carbon energy usage, sisal fiber (SF) is a GP composite reinforcement with potential development. In this paper, the research progress on the effect of SF on the properties of GP composites in recent decades is reviewed. It mainly includes the chemical composition and physical properties of SFs, the preparation technology of sisal-reinforced geopolymers (SFRGs), the microstructure analysis of the interface of SFs and the GP matrix, and the macroscopic mechanical properties of SFRGs. The properties of SFs make them have good bonding properties with the GP matrix. The addition of SFs can improve the flexural strength and tensile strength of GP composites, and SFRGs have good engineering application prospects.

Genetic mutation of TRPV2 induces anxiety by decreasing GABA-B R2 expression in hippocampus.

Transient receptor potential vanillic acid 2 (TRPV2) are well recognized for their contributions to neuronal development, cardiac function, immunity and cancer. However, the precise roles for this thermo TRPchannels in neurological disorder remain unknown. In this study, we employed the CRISPR/Cas9 system to generate genetic mutations of TRPV2. Genetic mutation of TRPV2 resulted in autistic-like phenotypes in mice accompanied with the disordered electrical signals recorded by multi-channels in vivo. To determine possible molecular mechanisms, western blotting was further used to assess the possible involvement of several autism-related proteins. The significantly decreased expression of the R2 subunit of the GABA-B receptor in the hippocampus was observed. Together, our findings suggest that genetic mutation of TRPV2 induces autism-like behavior, results in decreased expression of the R2 subunit of the GABA-B receptor.

miR-544 Regulates Dairy Goat Male Germline Stem Cell Self-Renewal via Targeting PLZF.

The balance between the self-renewal and differentiation of male germline stem cells (mGSCs) is critical for the initiation and maintenance of mammalian spermatogenesis. The promyelocytic leukemia zinc finger (PLZF), a zinc finger protein, is a critical factor for maintaining the self-renewal of mGSCs, so, evaluation of the PLZF pathway in mGSCs may provide a deeper insight into mammalian spermatogenesis. miRNA was also an important regulating factor for the self-renewal and differentiation of mGSCs; however, there is currently no data indicating that which miRNA regulate the self-renewal and differentiation of mGSCs via PLZF. Here, we predicted the prospective miRNA targeting to PLZF using the online Bioinformatics database-Targetscan, and performed an analysis of the dual-luciferase recombinant vector, psiCHCEKTM-2-PLZF-3'UTR. miR-544 mimics (miR-544m), miR-544 inhibitors (miR-544i), Control (NC, scrambled oligonucleotides transfection), pPLZF-IRES2-EGFP or PLZF siRNA were transfected into mGSCs; the cells proliferation was evaluated by BRDU incorporation assay and flow cytometry, and the mGSC marker, GFRa1, PLZF, KIT, DAZL, and VASA expression were analyzed by RT-qPCR, immunofluorescence and Western blot. The results showed that miR-544 regulates dairy goat male germline stem cell self-renewal via targeting PLZF. Our study identifies a new regulatory pathway for PLZF and expands upon the PLZF regulatory network in mGSCs.

Mg-Sr-Ca containing bioactive glass nanoparticles hydrogel modified mineralized collagen scaffold for bone repair.

The aim of this study is to explore the therapeutic effects of Mg-Sr-Ca containing bioactive glass nanoparticles sodium alginate hydrogel modified mineralized collagen scaffold (Mg-Sr-Ca-BGNs-SA-MC) on the repair of osteoporotic bone defect. During the study, Mg-Sr-Ca containing bioactive glass nanoparticles (Mg-Sr-Ca-BGNs) were synthesized using the sol-gel method, and the Mg-Sr-Ca-BGNs-SA-MC scaffold was synthesized by a simple method. The Mg-Sr-Ca-BGNs and the Mg-Sr-Ca-BGNs-SA-MC scaffold were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The elements of Mg, Sr, Ca and Si were effectively integrated into Mg-Sr-Ca-BGNs. SEM analysis revealed the presence of Mg-Sr-Ca-BGNs on the scaffold's surface. Furthermore, the cytotoxicity of the scaffolds were assessed using a live/dead assay. The result of the live/dead assay demonstrated that the scaffold materials were non-toxic to cell growth. More importantly, the in vivo study indicated that implanted scaffold promoted tissue regeneration and integration with newly formed bone. Overall, the Mg-Sr-Ca-BGNs-SA-MC scaffold is suitable for guided bone regeneration and beneficial to repair of osteoporotic bone defects.

A 10-m national-scale map of ground-mounted photovoltaic power stations in China of 2020.

We provide a remote sensing derived dataset for large-scale ground-mounted photovoltaic (PV) power stations in China of 2020, which has high spatial resolution of 10 meters. The dataset is based on the Google Earth Engine (GEE) cloud computing platform via random forest classifier and active learning strategy. Specifically, ground samples are carefully collected across China via both field survey and visual interpretation. Afterwards, spectral and texture features are calculated from publicly available Sentinel-2 imagery. Meanwhile, topographic features consisting of slope and aspect that are sensitive to PV locations are also included, aiming to construct a multi-dimensional and discriminative feature space. Finally, the trained random forest model is adopted to predict PV power stations of China parallelly on GEE. Technical validation has been carefully performed across China which achieved a satisfactory accuracy over 89%. Above all, as the first publicly released 10-m national-scale distribution dataset of China's ground-mounted PV power stations, it can provide data references for relevant researchers in fields such as energy, land, remote sensing and environmental sciences.

Genetic mutation of Tas2r104/Tas2r105/Tas2r114 cluster leads to a loss of taste perception to denatonium benzoate and cucurbitacin B.

BACKGROUND: Bitter taste receptors (Tas2rs) are generally considered to sense various bitter compounds to escape the intake of toxic substances. Bitter taste receptors have been found to widely express in extraoral tissues and have important physiological functions outside the gustatory system in vivo. METHODS: To investigate the physiological functions of the bitter taste receptor cluster Tas2r106/Tas2r104/Tas2r105/Tas2r114 in lingual and extraoral tissues, multiple Tas2rs mutant mice and Gnat3 were produced using CRISPR/Cas9 gene-editing technique. A mixture containing Cas9 and sgRNA mRNAs for Tas2rs and Gnat3 gene was microinjected into the cytoplasm of the zygotes. Then, T7EN1 assays and sequencing were used to screen genetic mutation at the target sites in founder mice. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunostaining were used to study the expression level of taste signaling cascade and bitter taste receptor in taste buds. Perception to taste substance was also studied using two-bottle preference tests. RESULTS: We successfully produced several Tas2rs and Gnat3 mutant mice using the CRISPR/Cas9 technique. Immunostaining results showed that the expression of GNAT3 and PLCB2 was not altered in Tas2rs mutant mice. But qRT-PCR results revealed the changed expression profile of mTas2rs gene in taste buds of these mutant mice. With two-bottle preference tests, these mutant mice eliminate responses to cycloheximide due to genetic mutation of Tas2r105. In addition, these mutant mice showed a loss of taste perception to quinine dihydrochloride, denatonium benzoate, and cucurbitacin B (CuB). Gnat3-mediated taste receptor and its signal pathway contribute to CuB perception. CONCLUSIONS: These findings implied that these mutant mice would be a valuable means to understand the biological functions of TAS2Rs in extraoral tissues and investigate bitter compound-induced responses mediated by these TAS2Rs in many extraoral tissues.

Characterization of genetic humanized mice with transgenic HLA DP401 or DRA but deficient in endogenous murine MHC class II genes upon Staphylococcus aureus pneumonia.

BACKGROUND: Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in "carrier" or "pathogenic" states. HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S. aureus infection. However, the contribution of HLA DP to S. aureus infection is unknown yet. METHODS: In this study, we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes. Neo-floxed IAß+/- mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice. After several rounds of traditional crossbreeding, we finally obtained HLA DP401-IAß-/- and HLA DRA-IAß-/- humanized mice, in which human DP401 or DRA0101 molecule was introduced into IAß-/- mice deficient in endogenous murine MHC class II molecules. A transnasal infection murine model of S. aureus pneumonia was induced in the humanized mice by administering 2 × 108 CFU of S. aureus Newman dropwise into the nasal cavity. The immune responses and histopathology changes were further assessed in lungs in these infected mice. RESULTS: We evaluated the local and systemic effects of S. aureus delivered intranasally in HLA DP401-IAß-/- and HLA DRA-IAß-/- transgenic mice. S. aureus Newman infection significantly increased the mRNA level of IL 12p40 in lungs in humanized mice. An increase in IFN-γ and IL-6 protein was observed in HLA DRA-IAß-/- mice. We observed a declining trend in the percentage of F4/80+ macrophages in lungs in HLA DP401-IAß-/- mice and a decreasing ratio of CD4+ to CD8+ T cells in lungs in IAß-/- mice and HLA DP401-IAß-/- mice. A decreasing ratio of Vß3+ to Vß8+ T cells was also found in the lymph node of IAß-/- mice and HLA DP401-IAß-/- mice. S. aureus Newman infection resulted in a weaker pathological injury in lungs in IAß-/- genetic background mice. CONCLUSION: These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S. aureus pneumonia and study what role DP molecule plays in S. aureus infection.

Dietary salt modulates the sodium chloride cotransporter expression likely through an aldosterone-mediated WNK4-ERK1/2 signaling pathway.

WNK is a serine/threonine kinase. Mutation in WNK1 or WNK4 kinase results in pseudohypoaldosteronism type II (PHA II) featuring hypertension, hyperkalemia and metabolic acidosis. Sodium chloride cotransporter (NCC) is known to be regulated by phosphorylation and trafficking. Dietary salt and hormonal stimulation, such as aldosterone, also affect the regulation of NCC. We have previously reported that WNK4 inhibits NCC protein expression. To determine whether dietary salt affects NCC abundance through WNK4-mediated mechanism, we investigated the effects of dietary salt change with or without aldosterone infusion (1 mg/kg/day) on NCC and WNK4 expression in rats. We found that high-salt (HS, 4% NaCl) diet significantly inhibits NCC mRNA expression and protein abundance while enhancing WNK4 mRNA and protein expression, whereas low-salt (LS, 0.07% NaCl) diet increases NCC mRNA expression and protein abundance while reducing WNK4 expression. We also found that aldosterone infusion in HS-fed rats increases NCC mRNA expression and protein abundance, but decreases WNK4 expression. Administration with spironolactone (0.1 g/kg/day) in LS-fed rats decreases NCC mRNA expression and protein abundance while increasing WNK4 expression. We further showed that ERK1/2 phosphorylation was increased in HS-fed rats, but decreased in LS-fed rats. In HEK293 cells, over-expressed WNK4 increases ERK1/2 phosphorylation, whereas knockdown of WNK4 expression decreases ERK1/2 phosphorylation. Aldosterone treatment for 3 h decreases ERK1/2 phosphorylation. These data suggest that dietary salt change affects NCC protein abundance in an aldosterone-dependent mechanism likely via the WNK4-ERK1/2-mediated pathway.

Preparation and Property Characterization of In2YSbO7/BiSnSbO6 Heterojunction Photocatalyst toward Photocatalytic Degradation of Indigo Carmine within Dye Wastewater under Visible-Light Irradiation.

In2YSbO7 and In2YSbO7/BiSnSbO6 heterojunction photocatalyst were prepared by a solvothermal method for the first time. The structural characteristics of In2YSbO7 had been represented. The outcomes showed that In2YSbO7 crystallized well and possessed pyrochlore constitution, a stable cubic crystal system and space group Fd3m. The lattice parameter of In2YSbO7 was discovered to be a = 11.102698 Å and the band gap energy of In2YSbO7 was discovered to be 2.68 eV, separately. After visible-light irradiation of 120 minutes (VLGI-120M), the removal rate (ROR) of indigo carmine (IC) reached 99.42% with In2YSbO7/BiSnSbO6 heterojunction (IBH) as a photocatalyst. The ROR of total organic carbon (TOC) reached 93.10% with IBH as a photocatalyst after VLGI-120M. Additionally, the dynamics constant k which was taken from the dynamic curve toward (DCT) IC density and VLGI time with IBH as a catalyst reached 0.02950 min-1. The dynamics constant k which came from the DCT TOC density and VLGI time with IBH as a photocatalyst reached 0.01783 min-1. The photocatalytic degradation of IC in dye wastewater (DW) with IBH as a photocatalyst under VLGI was in accordance with the first-order kinetic curves. IBH was used to degrade IC in DW for three cycles of experiments under VLGI, and the ROR of IC reached 98.74%, 96.89% and 94.88%, respectively, after VLGI-120M, indicating that IBH had high stability. Compared with superoxide anions or holes, hydroxyl radicals possessed the largest oxidative ability for removing IC in DW, as demonstrated by experiments with the addition of trapping agents. Lastly, the probable degradation mechanism and degradation pathway of IC were revealed in detail. The results showed that a visible-light-responsive heterojunction photocatalyst which possessed high catalytic activity and a photocatalytic reaction system which could effectively remove IC in DW were obtained. This work provided a fresh scientific research idea for improving the performance of a single catalyst.

Synthesis, Performance Measurement of Bi2SmSbO7/ZnBiYO4 Heterojunction Photocatalyst and Photocatalytic Degradation of Direct Orange within Dye Wastewater under Visible Light Irradiation.

Originally, the new catalyst Bi2SmSbO7 was synthesized by the hydrothermal synthesis method or by the solid-phase sintering method at a lofty temperature. A solvothermal method was utilized to prepare a Bi2SmSbO7/ZnBiYO4 heterojunction photocatalyst (BZHP). The crystal structure of Bi2SmSbO7 belonged to the pyrochlore structure and face-centered cubic crystal system by the space group of Fd3m. The cell parameter a was equivalent to 10.835(1) Å (Bi2SmSbO7). With Bi2SmSbO7/ZnBiYO4 heterojunction (BZH) as the photocatalyst, the removal rate (RR) of direct orange (DO) and the total organic carbon were 99.10% and 96.21% after visible light irradiation of 160 min (VLI-160M). The kinetic constant k toward DO concentration and visible light irradiation time (VLI) with BZH as photocatalyst reached 2.167 min−1. The kinetic constant k, which was concerned with total organic carbon, reached 0.047 min−1. The kinetic curve that came from DO degradation with BZH as a catalyst under VLI conformed to the second-order reaction kinetics. After VLI-160M, the photocatalytic degradation (PD) removal percentage of DO with BZH as the photocatalyst was 1.200 times, 1.268 times or 3.019 times that with Bi2SmSbO7 as the photocatalyst, ZnBiYO4 as the photocatalyst or with nitrogen-doped titanium dioxide as the photocatalyst. The photocatalytic activity (PA) was as following: BZH > Bi2SmSbO7 > ZnBiYO4 > nitrogen-doped titanium dioxide. After VLI-160M for three cycles of experiments with BZH as the photocatalyst, the RR of DO reached 98.03%, 96.73% and 95.43%, respectively, which meant that BZHP possessed high stability. By using the experiment of adding a trapping agent, the oxidative purifying capability for degradation of direct orange, which was in gradual depressed order, was as following: hydroxyl radical > superoxide anion > holes. Finally, the possible degradation pathway and degradation mechanism of DO were discussed systematically. A new high active heterojunction catalyst BZHP, which could efficiently remove toxic organic pollutants such as DO from dye wastewater after VLI, was obtained. Our research was meant to improve the photocatalytic property of the single photocatalyst.

Synthesis and Property Examination of Er2FeSbO7/BiTiSbO6 Heterojunction Composite Catalyst and Light-Catalyzed Retrogradation of Enrofloxacin in Pharmaceutical Waste Water under Visible Light Irradiation.

A new photocatalyst, Er2FeSbO7, was prepared by solid phase sintering using the high-temperature synthesis method for the first time in this paper. Er2FeSbO7/BiTiSbO6 heterojunction (EBH) catalyst was prepared by the solvent thermal method for the first time. Er2FeSbO7 compound crystallized in the pyrochlore-type architecture and cubelike crystal system; the interspace group of Er2FeSbO7 was Fd3m and the crystal cellular parameter a of Er2FeSbO7 was 10.179902 Å. The band gap (BDG) width of Er2FeSbO7 was 1.88 eV. After visible light irradiation of 150 minutes (VLGI-150min) with EBH as a photocatalyst, the removal rate (RR) of enrofloxacin (ENR) concentration was 99.16%, and the total organic carbon (TOC) concentration RR was 94.96%. The power mechanics invariable k toward ENR consistency and visible light irradiation (VLGI) time with EBH as a photocatalyzer attained 0.02296 min−1. The power mechanics invariable k which was involved with TOC attained 0.01535 min−1. The experimental results showed that the photocatalytic degradation (PCD) of ENR within pharmaceutical waste water with EBH as a photocatalyzer under VLGI was in keeping with the single-order reactivity power mechanics. The RR of ENR with EBH as a photocatalyzer was 1.151 times, 1.269 times or 2.524 times that with Er2FeSbO7 as a photocatalyst, BiTiSbO6 as a photocatalyst, or N-doping TiO2 (N-TO) as a photocatalyst after VLGI-150min. The photocatalytic activity, which ranged from high to low among above four photocatalysts, was as follows: EBHP > Er2FeSbO7 > BiTiSbO6 > N-TO. After VLGI-150min toward three periods of the project with EBH as a photocatalyst, the RR of ENR attained 98.00%, 96.76% and 95.60%. The results showed that the stability of EBH was very high. With appending trapping agent, it could be proved that the oxidative capability for degrading ENR, which ranged from strong to weak among three oxidic radicals, was as follows: superoxide anion > hydroxyl radicals (HRS) > holes. This work provides a scientific basis for the research and oriented leader development of efficient heterojunction catalysts.

Inter-Reader Variability of Volumetric Subsolid Pulmonary Nodule Radiomic Features.

OBJECTIVE: To evaluate the inter-observer consistency for subsolid pulmonary nodule radiomic features. MATERIALS AND METHODS: Subsolid nodules were selected by reviewing radiology reports of CT examinations performed December 1, 2015 to April 1, 2016. Patients with CTs at two time points were included in this study. There were 55 patients with subsolid nodules, of whom 14 had two nodules. Of 69 subsolid nodules, 66 were persistent at the second time point, yielding 135 lesions for segmentation. Two thoracic radiologists and an imaging fellow segmented the lesions using a semi-automated volumetry algorithm (Syngo.via Vb20, Siemens). Coefficient of variation (CV) was used to assess consistency of 91 quantitative measures extracted from the subsolid nodule segmentations, including first and higher order texture features. The accuracy of segmentation was visually graded by an experienced thoracic radiologist. Influencing factors on radiomic feature consistency and segmentation accuracy were assessed using generalized estimating equation analyses and the Exact Mann-Whitney test. RESULTS: Mean patient age was 71 (38-93 years), with 39 women and 16 men. Mean nodule volume was 1.39mL, range .03-48.2mL, for 135 nodules. Several radiomic features showed high inter-reader consistency (CV<5%), including entropy, uniformity, sphericity, and spherical disproportion. Descriptors such as surface area and energy had low consistency across inter-reader segmentations (CV>10%). Nodule percent solid component and attenuation influenced inter-reader variability of some radiomic features. The presence of contrast did not significantly affect the consistency of subsolid nodule radiomic features. Near perfect segmentation, within 5% of actual nodule size, was achieved in 68% of segmentations, and very good segmentation, within 25% of actual nodule size, in 94%. Morphologic features including nodule margin and shape (each p <0.01), and presence of air bronchograms (p = 0.004), bubble lucencies (p = 0.02) and broad pleural contact (p < 0.01) significantly affected the probability of near perfect segmentation. Stroke angle (p = 0.001) and length (p < 0.001) also significantly influenced probability of near perfect segmentation. CONCLUSIONS: The inter-observer consistency of radiomic features for subsolid pulmonary nodules varies, with high consistency for several features, including sphericity, spherical disproportion, and first and higher order entropy, and normalized non-uniformity. Nodule morphology influences the consistency of subsolid nodule radiomic features, and the accuracy of subsolid nodule segmentation.

Development of a humanized HLA-A30 transgenic mouse model.

BACKGROUND: There are remarkable genetic differences between animal major histocompatibility complex (MHC) systems and the human leukocyte antigen (HLA) system. HLA transgenic humanized mouse model systems offer a much better method to study the HLA-A-related principal mechanisms for vaccine development and HLA-A-restricted responses against infection in human. METHODS: A recombinant gene encoding the chimeric HLA-A30 monochain was constructed. This HHD molecule contains the following: α1-α2 domains of HLA-A30, α3 and cytoplasmic domains of H-2Db , linked at its N-terminus to the C-terminus of human ß2m by a 15-amino-acid peptide linker. The recombinant gene encoding the chimeric HLA-A30 monochain cassette was introduced into bacterial artificial chromosome (BAC) CH502-67J3 containing the HLA-A01 gene locus by Red-mediated homologous recombination. Modified BAC CH502-67J3 was microinjected into the pronuclei of wild-type mouse oocytes. This humanized mouse model was further used to assess the immune responses against influenza A virus (H1N1) pdm09 clinically isolated from human patients. Immune cell population, cytokine production, and histopathology in the lung were analyzed. RESULTS: We describe a novel human ß2m-HLA-A30 (α1α2)-H-2Db (α3 transmembrane cytoplasmic) (HHD) monochain transgenic mouse strain, which contains the intact HLA-A01 gene locus including 49 kb 5'-UTR and 74 kb 3'-UTR of HLA-A01*01. Five transgenic lines integrated into the large genomic region of HLA-A gene locus were obtained, and the robust expression of exogenous transgene was detected in various tissues from A30-18# and A30-19# lines encompassing the intact flanking sequences. Flow cytometry revealed that the introduction of a large genomic region in HLA-A gene locus can influence the immune cell constitution in humanized mice. Pdm09 infection caused a similar immune response among HLA-A30 Tg humanized mice and wild-type mice, and induced the rapid increase of cytokines, including IFN-γ, TNF-α, and IL-6, in both HLA-A30 humanized Tg mice and wild-type mice. The expression of HLA-A30 transgene was dramatically promoted in tissues from A30-9# line at 3 days post-infection (dpi). CONCLUSIONS: We established a promising preclinical research animal model of HLA-A30 Tg humanized mouse, which could accelerate the identification of novel HLA-A30-restricted epitopes and vaccine development, and support the study of HLA-A-restricted responses against infection in humans.

Generation and expression analysis of BAC humanized mice carrying HLA-DP401 haplotype.

Background: Human leukocyte antigen (HLA)-DP is much less studied than other HLA class II antigens, that is, HLA-DR and HLA-DQ, etc. However, the accumulating data have suggested the important roles of DP-restricted responses in the context of cancer, allergy, and infectious disease. Lack of animal models expressing these genes as authentic cis-haplotypes blocks our understanding for the role of HLA-DP haplotypes in immunity. Methods: To explore the potential cis-acting control elements involved in the transcriptional regulation of the HLA-DPA1/DPB1 gene, we performed the expression analysis using bacterial artificial chromosome (BAC)-based transgenic humanized mice in the C57BL/6 background, which carried the entire HLA-DP401 gene locus. We further developed a mouse model of Staphylococcus aureus pneumonia in HLA-DP401 humanized transgenic mice, and performed the analysis on the expression pattern of HLA-DP401 and immunological responses in the model. Results: In this study, we screened and identified a BAC clone spanning the entire HLA-DP gene locus. DNA from this clone was analyzed for integrity by pulsed-field gel electrophoresis and then microinjected into fertilized mouse oocytes to produce transgenic founder animals. Nine sets of PCR primers for regional markers with an average distance of 15 kb between each primer were used to confirm the integrity of the transgene in the five transgenic lines carrying the HLA-DPA1/DPB1 gene. Transgene copy numbers were determined by real-time PCR analysis. HLA-DP401 gene expression was analyzed at the mRNA and protein level. Although infection with S aureus Newman did not alter the percentage of immune cells in the spleen and thymus from the HLA-DP401-H2-Aß1 humanized mice. Increased expression of HLA-DP401 was observed in the thymus of the humanized mice infected by S aureus. Conclusions: We generated several BAC transgenic mice, and analyzed the expression of HLA-DPA1/DPB1 in those mice. A model of Saureus-induced pneumonia in the HLA-DP401-H2-Aß1-/- humanized mice was further developed, and S aureus infection upregulated the HLA-DP401 expression in thymus of those humanized mice. These findings demonstrate the potential of those HLA-DPA1/DPB1 transgenic humanized mice for developing animal models of infectious diseases and MHC-associated immunological diseases.

Epigenetic Landscapes of Single-Cell Chromatin Accessibility and Transcriptomic Immune Profiles of T Cells in COVID-19 Patients.

T cells play a critical role in coronavirus diseases. How they do so in COVID-19 may be revealed by analyzing the epigenetic chromatin accessibility of cis- and trans-regulatory elements and creating transcriptomic immune profiles. We performed single-cell assay for transposase-accessible chromatin (scATAC) and single-cell RNA (scRNA) sequencing (seq) on the peripheral blood mononuclear cells (PBMCs) of severely ill/critical patients (SCPs) infected with COVID-19, moderate patients (MPs), and healthy volunteer controls (HCs). About 76,570 and 107,862 single cells were used, respectively, for analyzing the characteristics of chromatin accessibility and transcriptomic immune profiles by the application of scATAC-seq (nine cases) and scRNA-seq (15 cases). The scATAC-seq detected 28,535 different peaks in the three groups; among these peaks, 41.6 and 10.7% were located in the promoter and enhancer regions, respectively. Compared to HCs, among the peak-located genes in the total T cells and its subsets, CD4+ T and CD8+ T cells, from SCPs and MPs were enriched with inflammatory pathways, such as mitogen-activated protein kinase (MAPK) signaling pathway and tumor necrosis factor (TNF) signaling pathway. The motifs of TBX21 were less accessible in the CD4+ T cells of SCPs compared with those in MPs. Furthermore, the scRNA-seq showed that the proportion of T cells, especially the CD4+ T cells, was decreased in SCPs and MPs compared with those in HCs. Transcriptomic results revealed that histone-related genes, and inflammatory genes, such as NFKBIA, S100A9, and PIK3R1, were highly expressed in the total T cells, CD4+ T and CD8+ T cells, both in the cases of SCPs and MPs. In the CD4+ T cells, decreased T helper-1 (Th1) cells were observed in SCPs and MPs. In the CD8+T cells, activation markers, such as CD69 and HLA class II genes (HLA-DRA, HLA-DRB1, and HLA-DRB5), were significantly upregulated in SCPs. An integrated analysis of the data from scATAC-seq and scRNA-seq showed some consistency between the approaches. Cumulatively, we have generated a landscape of chromatin epigenetic status and transcriptomic immune profiles of T cells in patients with COVID-19. This has provided a deeper dissection of the characteristics of the T cells involved at a higher resolution than from previously obtained data merely by the scRNA-seq analysis. Our data led us to suggest that the T-cell inflammatory states accompanied with defective functions in the CD4+ T cells of SCPs may be the key factors for determining the pathogenesis of and recovery from COVID-19.

The construction and application of lentiviral overexpression vector of goat miR-204 in testis.

The miRNA gene in DNA is first transcribed to Pri-miRNA, and then processed to Pre-miRNA, a stem-loop RNA segment (precursor) and further to miRNA which binds to mRNA by Dicer protein complex. It was confirmed that goat miR-204 could regulate the expressions of Sirt1 and the SSCs' (Spermatogonial Stem Cells) important genes Oct4 and Plzf, and inhibit the proliferation of dairy goat SSCs in vitro in our previous work. So, the research in vivo was needed next. In this study, the recombinant lentivirus vector pCDH-CMV-mir204-EF1-GreenPuro containing a goat chi-pri-mir-204 gene DNA segment was structured, and transfected into 293 T cells for packaged lentivirus, which then were injected into mouse seminiferous tubules. After 7 days, the goat miR-204 and the related genes such as Sirt1 and Plzf were detected in the mouse testis. This work laid a good foundation for further study of miR-204 biological function in vivo.

Reconstitution of male germline cell specification from mouse embryonic stem cells using defined factors in vitro.

In vitro induction of functional haploid cells from embryonic stem cells (ESCs) has been reported by several groups. However, these reports either involve complex induction process with undefined induction factors or show low-induction efficiency. Here, we report complete meiosis in vitro from ESCs with defined induction factors. ESCs were first induced into primordial germ cell-like cells, which were further induced into male germline cells, including spermatogonial stem cell-like cells (SSCLCs) and spermatid-like cells. Importantly, the obtained SSCLCs were functional as infertile male mice sired healthy offspring via SSCLC transplantation. Further, we found that eukaryotic translation initiation factor 2 subunit 3 and structural gene Y-linked (Eif2s3y) was essential for spermatogenesis. Eif2s3y-overexpressing ESCs showed enhanced spermatogenesis in vitro, as demonstrated by higher expression levels of SSC-specific markers during SSCLC induction process, improved reproductive ability recovery of infertile male mice, and increased efficiency of haploid cell induction. Our work provides a convenient and efficient approach to obtain functional male germline cells.

Ablation of NTPDase2+ cells inhibits the formation of filiform papillae in tongue tip.

BACKGROUND: Lingual epithelia in the tongue tip are among the most rapidly regenerating tissues, but the mechanism of cell genesis in this tissue is still unknown. Previous study has suggested the existence of multiple stem cell pools in lingual epithelia and papillae. Like K14+ and Sox2+ cells, NTPDase2+ cells have characteristics of stem cells. METHODS: We employed a system using doxycycline to conditionally ablate NTPDase2+ cells in lingual epithelia and papillae by regulated expression of the diphtheria toxin A (DTA) gene. Transgenic lines, which expressed the rtTA gene in NTPDase2+ cells, were produced by pronuclear injection of zygotes from C57BL/6 mice using the BAC clone RP23-47P18. The NTPDase2-rtTA transgenic mice were crossed with the TetO-DTA transgenic animals. The double transgenic mice were treated with doxycycline. Doxycycline (Dox) was diluted in 5% sucrose in water to a final concentration of 0.3-0.5 mg/mL and supplied as drinking water. RESULTS: After 15 days of Dox induction, the expression of NTPDase2, Sox2 and K14 was ablated from lingual epithelia. DTA expression in NTPDase2+ cells did not inhibit the turnover of GNAT3+ or PLCß2+ cells in taste buds, nor the expression of S100ß beneath lingual epithelia and papillae. After 35 days ablation of NTPDase2+ cells, the basic structure of lingual epithelia and papillae remained intact. However, the ratio of cell to total tissue area was decreased in lingual epithelia and circumvallate (CV) papillae. DTA expression also inhibited the regeneration of filiform papillae on the dorsal surface of the tongue tip. CONCLUSIONS: These studies provide important insights into the understanding of dynamic equilibrium among the multiple stem cell populations present in the lingual epithelia and papillae.

Melatonin Ameliorates Busulfan-Induced Spermatogonial Stem Cell Oxidative Apoptosis in Mouse Testes.

AIMS: Many men endure immunosuppressive or anticancer treatments that contain alkylating agents before the age of sexual maturity, especially the increasing number of preadolescent males who undergo busulfan treatment for myeloablative conditioning before hematopoietic stem cell transplantation. Before sperm production, there are no sperm available for cryopreservation. Thus, it is necessary to identify a solution to ameliorate the busulfan-induced damage of spermatogonial stem cells (SSCs). RESULTS: In this study, we demonstrated that melatonin relieved the previously described SSC loss and apoptosis in mouse testes. Melatonin increased the expression of manganese superoxide dismutase (MnSOD), which regulated the production of busulfan-induced reactive oxygen species (ROS). Moreover, melatonin promoted sirtuin type 1 (SIRT1) expression. SIRT1 participated in the deacetylation of p53, which promotes p53 ubiquitin degradation. Decreased concentrations of deacetylated p53 resulted in spermatogonial cell resistance to apoptosis. Acute T cell leukemia cell assay demonstrated that melatonin does not affect busulfan-induced cancer cell apoptosis and ROS. INNOVATION: The current evidence suggests that melatonin may alleviate the side effects of alkylating drugs, such as busulfan. CONCLUSION: Melatonin promoted MnSOD and SIRT1 expression, which successfully ameliorated busulfan-induced SSC apoptosis caused by high concentrations of ROS and p53. Antioxid. Redox Signal. 28, 385-400.

Image Quality on Dual-energy CTPA Virtual Monoenergetic Images: Quantitative and Qualitative Assessment.

RATIONALE AND OBJECTIVES: This study aims to determine the optimal photon energy for image quality of the pulmonary arteries (PAs) on dual-energy computed tomography (CT) pulmonary angiography (CTPA) utilizing low volumes of iodinated contrast. MATERIALS AND METHODS: The study received institutional review board exemption and was Health Insurance Portability and Accountability Act compliant. Adults (n = 56) who underwent dual-energy CTPA with 50-60 cc of iodinated contrast on a third-generation dual-source multidetector CT were retrospectively and consecutively identified. Twelve virtual monoenergetic kiloelectron volt (keV) image data sets (40-150 keV, 10-keV increments) were generated with a second-generation noise-reducing algorithm. Standard regions of interest were placed on main, right, left, and right interlobar pulmonary arteries; pectoralis muscle; and extrathoracic air. Attenuation [mean CT number (Hounsfield unit, HU)], noise [standard deviation (HU)], signal to noise (SNR), and contrast to noise ratio were evaluated. Three blinded chest radiologists rated (from 1 to 5, with 5 being the best) randomized monoenergetic and weighted-average images for attenuation and noise. P <.05 was considered significant. RESULTS: Region of interest mean CT number increased as keV decreased, with 40 keV having the highest value (P < .001). Mean SNR was highest for 40-60 keV (P <.05) (14.5-14.7) and was higher (P <.05) than all remaining energies (90-150 keV) for all vessel regions combined. Contrast to noise ratio was highest for 40 keV (P <.001) and decreased as keV increased. SNR was highest at 60 and 70 keV, only slightly higher than 40-50 keV (P <.05). Reader scores for 40-50 keV were greater than other energies and weighted-average images (P <.05). CONCLUSIONS: Kiloelectron volt images of 40-50 keV from the second-generation algorithm optimize attenuation on dual-energy CTPA and can potentially aid in interpretation and avoiding nondiagnostic examinations.