The ubiquitin ligase Stub1 negatively modulates regulatory T cell suppressive activity by promoting degradation of the transcription factor Foxp3.

Regulatory T (Treg) cells suppress inflammatory immune responses and autoimmunity caused by self-reactive T cells. The key Treg cell transcription factor Foxp3 is downregulated during inflammation to allow for the acquisition of effector T cell-like functions. Here, we demonstrate that stress signals elicited by proinflammatory cytokines and lipopolysaccharides lead to the degradation of Foxp3 through the action of the E3 ubiquitin ligase Stub1. Stub1 interacted with Foxp3 to promote its K48-linked polyubiquitination in an Hsp70-dependent manner. Knockdown of endogenous Stub1 or Hsp70 prevented Foxp3 degradation. Furthermore, the overexpression of Stub1 in Treg cells abrogated their ability to suppress inflammatory immune responses in vitro and in vivo and conferred a T-helper-1-cell-like phenotype. Our results demonstrate the critical role of the stress-activated Stub1-Hsp70 complex in promoting Treg cell inactivation, thus providing a potential therapeutic target for the intervention against autoimmune disease, infection, and cancer.

Inflammation negatively regulates FOXP3 and regulatory T-cell function via DBC1.

Forkhead box P3 (FOXP3)-positive Treg cells are crucial for maintaining immune homeostasis. FOXP3 cooperates with its binding partners to elicit Treg cells' signature and function, but the molecular mechanisms underlying the modulation of the FOXP3 complex remain unclear. Here we report that Deleted in breast cancer 1 (DBC1) is a key subunit of the FOXP3 complex. We found that DBC1 interacts physically with FOXP3, and depletion of DBC1 attenuates FOXP3 degradation in inflammatory conditions. Treg cells from Dbc1-deficient mice were more resistant to inflammation-mediated abrogation of Foxp3 expression and function and delayed the onset and severity of experimental autoimmune encephalomyelitis and colitis in mice. These findings establish a previously unidentified mechanism regulating FOXP3 stability during inflammation and reveal a pathway for potential therapeutic modulation and intervention in inflammatory diseases.

TEX15: A DNA repair gene associated with prostate cancer risk in Han Chinese.

BACKGROUND: Both common and rare genetic variants may contribute to risk of developing prostate cancer. Genome-wide association studies (GWASs) have identified ∼100 independent, common variants associated with prostate cancer risk. However, little is known about the association of rare variants (minor allele frequency [MAF] <1%) in the genome with prostate cancer risk. METHODS: A two-stage study was used to test the association of rare, deleterious coding variants, annotated using predictive algorithms, with prostate cancer risk in Chinese men. Predicted rare, deleterious coding variants in the Illumina HumanExome-12 v1.1 beadchip were first evaluated in 1343 prostate cancer patients and 1008 controls. Significant variants were then validated in an additional 1816 prostate cancer patients and 1549 controls. RESULTS: In the discovery stage, 14 predicted rare, deleterious coding variants were significantly associated with prostate cancer risk (P < 0.01). In the confirmation stage, Q1631H in TEX15 (rs142485241), a DNA repair gene, was significantly associated with prostate cancer risk (P = 0.0069). The estimated odds ratio (OR) of the variant in the combined analysis was 3.24 (95% Confidence Interval 1.85-6.06), P = 8.81 × 10-5 . Additionally, rs28756990 (V741F) at MLH3 (P = 0.06) and rs2961144 (I126V) at OR2A5 (P = 0.065) were marginally associated with prostate cancer risk in the replication stage. CONCLUSIONS: Our study provided preliminary evidence that the rare variant Q1631H in DNA repair gene TEX15 is associated with prostate cancer risk. This finding complements known common prostate cancer risk-associated variants and suggests the possible role of DNA repair genes in prostate cancer development.

Fn14 hepatic progenitor cells are associated with liver fibrosis in biliary atresia.

PURPOSE: The liver in biliary atresia (BA) is characterized by progressing fibrosis which is promoted by unclear reasons. We aimed to understand the factors influencing liver fibrosis. This study hypothesized that HPCs (hepatic progenitor cells) are activated and associated with liver fibrosis in biliary atresia. METHODS: Liver samples from biliary atresia patients are as BA group, and the normal liver derived from hepatoblastoma infants during operation are control group. The extent of fibrosis in liver samples was blindly evaluated by two experienced pathologists depending on Ishak system. The BA liver samples were divided into mild liver fibrosis group (grade I-IV, BAa) and severe liver fibrosis group (grade V-VI, BAb) to detect Fn14 protein expression. RESULTS: In mRNA level, Fn14 expression was 21.23 ± 8.3 vs. 1.00 ± 0.17, p = 0.023 < 0.05 and CD133 expression was 6.02 ± 2.16 vs. 1.14 ± 0.75, p = 0.008 < 0.01 between BA group and control group. Fn14 cells co-expressed the progenitor marker CD133 in liver, and activated in BA. Fn14 andα-SMA were co-location in fibrous area in liver. Compared to the control group, Fn14, CD133, and α-SMA protein expression were 2.10 ± 0.53 vs. 0.97 ± 0.2, p = 0.001, 2.23 ± 0.57 vs. 1.00 ± 0.03, p = 0.000, 4.96 ± 2.4 vs. 1.00 ± 0.22, p = 0.001. The Fn14 protein expression was 2.60 ± 0.35 vs. 1.86 ± 0.42, p = 0.012, between BAb and BAa group. CONCLUSION: Fn14 cells, which co-express the progenitor marker CD133 in liver, are HPCs and activated in BA. Fn14 + HPCs are associated with liver fibrosis in BA.

[Evaluation on Ability to Detect the Intracranial Hematoma with Different Density Using C-Arm Cone-beam Computed Tomography Based on Animal Model].

This study aims to evaluate the ability of C-arm cone-beam CT to detect intracranial hematomas in canine models. Twenty one healthy canines were divided into seven groups and each group had three animals. Autologous blood and contrast agent (3 mL) were slowly injected into the left/right frontal lobes of each animal. Canines in the first group, the control group, were only injected with autologous blood without contrast agent. Each animal in all the 7 groups was scanned with C-arm cone-beam CT and multislice computed tomography (MSCT) after 5 minutes. The attenuation values and their standard deviations of the hematoma and uniformed brain tissues were measured to calculate the image noise, signal to noise ratio (SNR) and contrast to noise ratio (CNR). A scale with scores 1-3 was used to rate the quality of the reconstructed image of different hematoma as a subjective evaluation, and all the experimental data were processed with statistical treatment. The results revealed that when the density of hematoma was less than 65 HU, hematomata were not very clear on C-arm CT images, and when the density of hematoma was more than 65 HU, hematomata showed clearly on both C-arm CT and MSCT images and the scores of them were close. The coherence between the two physicians was very reliable. The same results were obtained with C-arm cone-beam CT and MSCT grades in measuring SD value, SNR, and CNR. The reasonable choice of density detection range of intracranial hematoma with C-arm cone-beam CT could be effectively applied to monitoring the intracranial hemorrhage during interventional diagnosis and treatment.

PIM1 kinase phosphorylates the human transcription factor FOXP3 at serine 422 to negatively regulate its activity under inflammation.

Previous reports have suggested that human CD4(+) CD25(hi)FOXP3(+) T regulatory cells (Tregs) have functional plasticity and may differentiate into effector T cells under inflammation. The molecular mechanisms underlying these findings remain unclear. Here we identified the residue serine 422 of human FOXP3 as a phosphorylation site that regulates its function, which is not present in murine Foxp3. PIM1 kinase, which is highly expressed in human Tregs, was found to be able to interact with and to phosphorylate human FOXP3 at serine 422. T cell receptor (TCR) signaling inhibits PIM1 induction, whereas IL-6 promotes PIM1 expression in in vitro expanded human Tregs. PIM1 negatively regulates FOXP3 chromatin binding activity by specifically phosphorylating FOXP3 at Ser(422). Our data also suggest that phosphorylation of FOXP3 at the Ser(418) site could prevent FOXP3 phosphorylation at Ser(422) mediated by PIM1. Knockdown of PIM1 in in vitro expanded human Tregs promoted FOXP3-induced target gene expression, including CD25, CTLA4, and glucocorticoid-induced tumor necrosis factor receptor (GITR), or weakened FOXP3-suppressed IL-2 gene expression and enhanced the immunosuppressive activity of Tregs. Furthermore, PIM1-specific inhibitor boosted FOXP3 DNA binding activity in in vitro expanded primary Tregs and also enhanced their suppressive activity toward the proliferation of T effector cells. Taken together, our findings suggest that PIM1 could be a new potential therapeutic target in the prevention and treatment of human-specific autoimmune diseases because of its ability to modulate the immunosuppressive activity of human Tregs.

60-kDa Tat-interactive protein (TIP60) positively regulates Th-inducing POK (ThPOK)-mediated repression of eomesodermin in human CD4+ T cells.

The abundant expression of IFNγ in Th-inducing POK (ThPOK)-deficient CD4(+) T cells requires the activation of Eomesodermin (Eomes); however, the underlying mechanism of this phenomenon remains unclear. Here we report that ThPOK binds directly to the promoter region of the Eomes gene to repress its expression in CD4(+) T cells. We identified the histone acetyltransferase TIP60 as a co-repressor of ThPOK-target genes, where ectopically expressed TIP60 increased ThPOK protein stability by promoting its acetylation at its Lys(360) residue to then augment the transcriptional repression of Eomes. Moreover, knockdown of endogenous TIP60 abolished the stabilization of ThPOK in CD4(+) T cells, which led to the transcriptional activation of Eomes and increased production of IFNγ. Our results reveal a novel pathway by which TIP60 and ThPOK synergistically suppresses Eomes function and IFNγ production, which could contribute to the regulation of inflammation.

Identification of the E3 deubiquitinase ubiquitin-specific peptidase 21 (USP21) as a positive regulator of the transcription factor GATA3.

The expression of the transcription factor GATA3 in FOXP3(+) regulatory T (Treg) cells is crucial for their physiological function in limiting inflammatory responses. Although other studies have shown how T cell receptor (TcR) signals induce the up-regulation of GATA3 expression in Treg cells, the underlying mechanism that maintains GATA3 expression in Treg cells remains unclear. Here, we show how USP21 interacts with and stabilizes GATA3 by mediating its deubiquitination. In a T cell line model, we found that TcR stimulation promoted USP21 expression, which was further up-regulated in the presence of FOXP3. The USP21 mutant C221A reduced its capacity to stabilize GATA3 expression, and its knockdown led to the down-regulation of GATA3 protein expression in Treg cells. Furthermore, we found that FOXP3 could directly bind to the USP21 gene promoter and activated its transcription upon TcR stimulation. Finally, USP21, GATA3, and FOXP3 were found up-regulated in Treg cells that were isolated from asthmatic subjects. In summary, we have identified a USP21-mediated pathway that promotes GATA3 stabilization and expression at the post-translational level. We propose that this pathway forms an important signaling loop that stabilizes the expression of GATA3 in Treg cells.

CT Quantification of Interstitial Lung Abnormalities and Changes of Agerelated Pathomorphology.

BACKGROUND: Interstitial lung abnormalities (ILA) are associated with further disease progression, increased mortality risk, and decline in lung function in the elderly, which deserves enough attention. OBJECTIVE: The objective of this study was to quantify the extent of interstitial lung abnormalities (ILA) in a non-smoking asymptomatic urban cohort in China using low-dose CT (LDCT) and to analyze the age-related pathological changes. METHODS: We retrospectively analyzed clinical data and chest LDCT images from a cohort of 733 subjects who were categorized into 3 groups: 18-39, 40-59, and ≥60 years old according to age. Furthermore, we selected 40 cases of wax-embedded lung tissue blocks archived after pulmonary bullectomy and the same age groups were categorized. Four representative CT signs of ILA, including interlobular septal thickening (ILST), intralobular interstitial thickening (ILIT), ground-glass opacity (GGO), and reticular shadow (RS), were semi-quantified based on the percentage of the affected area. The scores and distribution of four CT signs of ILA were compared between different sex and age groups. The age-related pathological changes were analyzed. RESULTS: The ILA findings were found predominantly in the lower lobes and the subpleural region. The semi-quantitative scores of four CT signs in all subjects under 40 were 0. However, in subjects over 40 years old, the scores gradually increased with age, although most of them remained low. The size of the alveoli increased, the number of alveoli decreased, the alveolar septum became thinner, and the number of ATII cells increased with age. A statistically significant difference was observed among the different age groups (χ2=50.624, P=0.033; χ2=80.000, P=0.043; χ2=33.833, P=0.000; χ2=13.525, P=0.031). The macrophage population and the percentage of collagen fibers in the alveolar septum increased, while the percentage of elastic fibers decreased with age. There was no significant difference among the different age groups (χ2=19.817, P=0.506; χ2=52.419, P=0. 682; χ2=54.868, P=0.518). CONCLUSION: When the four CT signs mentioned above are in the upper central area, and the score has a medium or high score, it is crucial to determine the underlying pathological causes. ILA may be the result of chronic lung injury.

The Phylogeography of Deciduous Tree Ulmus macrocarpa (Ulmaceae) in Northern China.

Disentangling how climate oscillations and geographical events significantly influence plants' genetic architecture and demographic history is a central topic in phytogeography. The deciduous ancient tree species Ulmus macrocarpa is primarily distributed throughout Northern China and has timber and horticultural value. In the current study, we studied the phylogenic architecture and demographical history of U. macrocarpa using chloroplast DNA with ecological niche modeling. The results indicated that the populations' genetic differentiation coefficient (NST) value was significantly greater than the haplotype frequency (GST) (p < 0.05), suggesting that U. macrocarpa had a clear phylogeographical structure. Phylogenetic inference showed that the putative chloroplast haplotypes could be divided into three groups, in which the group Ⅰ was considered to be ancestral. Despite significant genetic differentiation among these groups, gene flow was detected. The common ancestor of all haplotypes was inferred to originate in the middle-late Miocene, followed by the haplotype overwhelming diversification that occurred in the Quaternary. Combined with demography pattern and ecological niche modeling, we speculated that the surrounding areas of Shanxi and Inner Mongolia were potential refugia for U. macrocarpa during the glacial period in Northern China. Our results illuminated the demography pattern of U. macrocarpa and provided clues and references for further population genetics investigations of precious tree species distributed in Northern China.

Peripheral interstitial lung abnormalities on LDCT in an asymptomatic, nonsmoking Chinese urban cohort.

To retrospectively investigate the imaging features and the related influencing factors of peripheral interstitial lung abnormalities (PILA) that caused "normal aging" by low-dose computed tomography (LDCT) in an nonsmoking, asymptomatic Chinese urban cohort. The clinical data of 733 subjects who underwent chest LDCT were retrospectively collected. The computed tomography (CT) signs of PILA (interlobular septal thickening [ILST], intralobular interstitial thickening [ILIT], ground-glass opacity [GGO], reticular shadow [RS], subpleural line [SL]) were evaluated at 6 levels and statistically analyzed. The effects of age, sex, body mass index (BMI), blood pressure (BP), and blood biochemistry parameters on ILST, ILIT, and RS were analyzed by Binary Logistic regression analysis. Significant age differences in PILA were found. None of the 5 PILA CT signs (GGO, ILST, ILIT, RS, and SL) was observed in subjects under 40 years old, while in subjects over 40 years old, the incidence of PILA increased with age. All 5 CT signs of PILA were significantly different among the subjects aged 18 to 49, 50 to 69, and 70 to 79 (P < .05). There was no significant sex difference in PILA. Among age, sex, BMI, BP, and laboratory biochemistry parameters, only age had a significant effect on ILST, ILIT, and RS. LDCT can be used as a noninvasive method to evaluate the PILA. PILA were mainly affected by age, while sex, BMI, BP, and laboratory biochemistry parameters had little effect on PILA. PILA observed before the age of 40 years should be considered an abnormal finding, whereas it is common in individuals over 70.

Variations in the nutrient concentration and composition in Liaodong Bay under long-term human activities.

This paper analyzed the long-term variations in nutrients in Liaodong Bay and their potential influencing factors based on historical data from 1978 to 2019. Under the influence of both human activities and natural changes, the concentration of DIN increased approximately 4-fold from the end of the 1990s to the mid-2010s, while DIP and DSi concentrations decreased from the beginning to the end of the 1980s and have since increased again. Asynchronous changes in nutrient levels have led to changes in the nutrient composition, which has caused a series of ecological effects. The total phytoplankton abundance decreased from the 1980s to the end of the 1990s and then increased again. Additionally, the phytoplankton composition shifted from a diatom-dominated to a dinoflagellate-dominated system, and the dominant species of zooplankton changed. Harmful algal blooms (HABs) rarely occurred before the 1980s but have frequently occurred since the end of the 1990s.

Radiomics Based on Lumbar Spine Magnetic Resonance Imaging to Detect Osteoporosis.

RATIONALE AND OBJECTIVES: Signal intensity of the lumbar spine in magnetic resonance imaging (MRI) correlates to bone mineral density (BMD). This study aims to explore a lumbar spine magnetic resonance imaging based on the radiomics model for detecting osteoporosis. MATERIALS AND METHODS: A total of 109 patients, who underwent both dual-energy X-ray absorptiometry (DEXA) and MRI of the lumbar spine, were recruited. Among these patients, 38 patients were normal, 32 patients had osteopenia, and 39 patients had osteoporosis, according to the DEXA results. A total of 396 × 2 radiomic features were extracted from the T1WI and T2WI images of the segmentation images in the lumbar magnetic resonance imaging. The correlated radiomic features were selected to establish the radiomic classification model. Then, the classification models (based on T1WI, T2WI, and T1WI+T2WI) of normal vs. osteopenia, normal vs. osteoporosis, and osteopenia vs. osteoporosis were established. The performance of the classification models was evaluated through the estimated area under the receiver operating characteristic curve. RESULTS: The area under the receiver operating characteristic curves based on T1WI, T2WI, and T1WI+T2WI were 0.772, 0.772, and 0.810, respectively, for the models of normal vs. osteopenia, 0.724, 0.682, and 0.797, respectively, for the models of normal vs. osteoporosis, and 0.730, 0.734, and 0.769, respectively, for the models of osteopenia vs. osteoporosis. CONCLUSION: Radiomic models established based on lumbar spine MRI can be used to detect osteoporosis.

Hydrogen Promotes the M1 Macrophage Conversion During the Polarization of Macrophages in Necrotizing Enterocolitis.

Background: Hydrogen is protective against intestinal injury in necrotizing enterocolitis (NEC), mainly through to alleviate inflammation response. The M1 macrophages can promote inflammation. We hypothesized that hydrogen would promote the M1 macrophages conversion during the polarization and reduce the inflammatory factors in NEC. Methods: We used M1 and M2 macrophages induced from RAW264.7 cells and bone marrow-derived macrophages, models of NEC and macrophages derived from spleens, abdominal lymph nodes and lamina propria in model mice. Cytokines, CD16/32 and CD206 were measured by quantitative PCR, flow cytometry. Nuclear factor-κB (NF-κB) p65 were determined by western blot. Histology staining were used to assess the severity of NEC. Results: Macrophages were successfully polarized to M1 or M2 by assessing the expression of inflammatory factors. Pro-inflammatory factors and CD16/32 in M1 macrophages were decreased, and the expression of CD16/32 in lamina propria were inhibited after treatment with hydrogen, but the changes has no effects in other tissues. Hydrogen inhibited the NF-κB p65 in M1 macrophages nucleus and distal ileum of NEC. HE staining showed hydrogen could attenuate the severity of NEC. Conclusion: Hydrogen could attenuate the severity of NEC through promoting M1 macrophages conversion by inhibited the expression of NF-κB p65 in the nucleus.

Stage-resolved Hi-C analyses reveal meiotic chromosome organizational features influencing homolog alignment.

During meiosis, chromosomes exhibit dramatic changes in morphology and intranuclear positioning. How these changes influence homolog pairing, alignment, and recombination remain elusive. Using Hi-C, we systematically mapped 3D genome architecture throughout all meiotic prophase substages during mouse spermatogenesis. Our data uncover two major chromosome organizational features varying along the chromosome axis during early meiotic prophase, when homolog alignment occurs. First, transcriptionally active and inactive genomic regions form alternating domains consisting of shorter and longer chromatin loops, respectively. Second, the force-transmitting LINC complex promotes the alignment of ends of different chromosomes over a range of up to 20% of chromosome length. Both features correlate with the pattern of homolog interactions and the distribution of recombination events. Collectively, our data reveal the influences of transcription and force on meiotic chromosome structure and suggest chromosome organization may provide an infrastructure for the modulation of meiotic recombination in higher eukaryotes.

Roles of LmCDA1 and LmCDA2 in cuticle formation in the foregut and hindgut of Locusta migratoria.

Chitin deacetylases (CDAs, including CDA1 and CDA2) are considered key enzymes for body cuticle formation and tracheal morphogenesis in various insect species. However, their functions in the formation of the cuticular intima of the foregut and hindgut are unclear. Here, we investigated the roles of their respective genes LmCDA1 and LmCDA2 in this process, in the hemimetabolous insect Locusta migratoria. Transcripts of LmCDA1 and LmCDA2 were highly expressed both before and after molting in the foregut. In the hindgut, their expression was high only before molting. In both the foregut and hindgut, LmCDA1 protein was localized in the basal half of the chitin matrix (procuticle), whereas LmCDA2 was detected in the upper half of the procuticle. Knockdown of LmCDA1 by RNA interference (RNAi) in 5th-instar nymphs caused no visible defects of the hindgut cuticle. By contrast, the chitinous lamellae of the cuticular intima in the foregut of knockdown animals were less compact than in control animals. RNAi against LmCDA2 led to thickening of both the foregut and hindgut cuticles, with a greater number of thinner laminae than in the respective control cuticles. Taken together, our results show that LmCDA1 and LmCDA2 have distinct, but overlapping, functions in chitin organization in the foregut cuticle. However, in the hindgut, this process seems independent of LmCDA1 activity but requires LmCDA2 function. Thus, the CDAs reflect tissue-specific differences in cuticular organization and function, which need further detailed molecular and histological analyses for full comprehension.

Prognostic value of miR-892a in gastric cancer and its regulatory effect on tumor progression.

BACKGROUND: Gastric cancer is a prevalent malignant around the world. Aberrantly expression of microRNAs (miRNAs) contributes to the progression of tumors. The aim of this study was to investigate the expression and role of miR-892a in gastric cancer. METHODS: A total of 119 gastric cancer patients were enrolled in this study. And the expression of miR-892a in gastric cancer tissues and cells was measured using RT-qPCR analysis. Kaplan-Meier plotter and multivariate Cox regression analysis were used to explore the prognostic value of miR-892a in gastric cancer. The biological function of miR-892a in gastric cancer cells was evaluated using CCK-8 assays and Transwell assays. RESULTS: The expression of miR-892a was high-expressed in gastric cancer tissues and cells. The miR-892a expression was associated with tumor size, differentiation, lymph node metastasis, and TNM stages. Gastric cancer patients with high miR-892a expression showed a short overall survival rate. Overexpression of miR-892a promoted cell proliferation, migration, and invasion of gastric cancer cells. CONCLUSION: miR-892a was upregulated and predictor of poor prognosis in gastric cancer patients. The miR-892a in gastric cancer cells significantly promoted cell proliferative, migratory, and invasive properties. Furthermore, miR-892a may be served as a prognostic marker as well as a therapeutic target for gastric cancer.

Intra- and inter-seasonal variations in the hydrological characteristics and nutrient conditions in the southwestern Yellow Sea during spring to summer.

Based on field survey in the southwestern Yellow Sea (SWYS) during April-September 2017, the spatiotemporal variations in the hydrological characteristics and nutrient conditions were coupled and analyzed; the intra-seasonal variations in the upwelling in the front of the Yellow Sea Cold Water Mass (YSCWM) and impacts on nutrient transport were explored. The coastal area was controlled by the low-salinity high-nutrient Lubei Coastal Current, Subei Coastal Current, and Yangtze River Diluted Water from north to south; at bottom, the northeastern SWYS was controlled by the low-temperature high-salinity high-nutrient YSCWM. Temperature, salinity and nutrient fronts formed around YSCWM. The upwelling velocity in the front increased during April to late June and decreased in early September; the upwelled fluxes of dissolved inorganic nitrogen (0.29×103-7.77×103 µmol·m-2d-1), phosphate (0.02×103-0.27×103 µmol·m-2d-1) and silicate (0.98×103-8.75×103 µmol·m-2d-1) showed similar variations during April-September. The upwelled nutrients could potentially contribute to local green tide development and phytoplankton growth during spring-summer.

Human ß-defensin-3 reduces excessive autophagy in intestinal epithelial cells and in experimental necrotizing enterocolitis.

Necrotizing enterocolitis (NEC) is a leading cause of mortality in preterm newborns. Intestinal barrier dysfunction is one key event in NEC pathogenesis. Human ß-defensin-3 (hBD3), one member of cationic host defence peptides, was reported to reduce the development of necrotizing enterocolitis in a neonatal rat model. And autophagy was induced in the intestine of human and animals with NEC. We hypothesized that regulation of autophagy might play a critical role in hBD3-mediated protection against NEC injury. Autophagy activity was evaluated both in intestinal epithelial cells and in NEC models. Newborn Sprague-Dawley rats were divided randomly into four groups: Control + NS, Control + rapamycin, NEC + NS, and NEC + hBD3. Body weight, histological score, survival time, enterocyte migration and mucosal barrier were recorded. Our results showed that hBD3 pretreatment could effectively inhibit autophagy activity in cultured IEC-6 and Caco2 enterocytes, and CXCR4 might be involved in hBD3-mediated autophagy suppression. Moreover, hBD3-induced inhibition of autophagy significantly promoted the intestinal epithelial cell migration by wound healing assay and transwell migration assay. In the rat model of NEC, hBD3 could noticeably reduce the expression of autophagy-activated proteins, down-regulate the expression of inflammatory mediators, and promote the mucosal integrity. Our data suggest an additional role of hBD3-mediated protection against intestinal mucosal injury: inhibition of over-activated autophagy in enterocytes.

Complete mitochondrial genome of two Thitarodes species (Lepidoptera, Hepialidae), the host moths of Ophiocordyceps sinensis and phylogenetic implications.

Thitarodes (Lepidoptera, Hepialidae) is the only genus that hosts to the Ophiocordyceps sinensis, a traditional Chinese medicine considered as a powerful medicinal supplement. In this study, the complete mitochondrial genomes (mitogenomes) of two species, T. damxungensis and T. pui, have been sequenced, which are 15,928 bp and 15,362 bp in size respectively, and both contain 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs and an AT-rich region. Like other hepialoids, the gene arrangement of the mitogenomes of T. damxungensis and T. pui is identical to the ancestral arrangement but differs from those of other lepidopteran species on account of the different arrangements of trnM, trnI, and trnQ. The size of AT-rich region is 545 bp in T. damxungensis and 1030 bp in T. pui. Tandem repetition in the AT-rich region is responsible for the length difference of the A + T-rich region in both species. In Hepialidae, the phylogenetic study based on the dataset of the sequences that combined the protein-coding genes and RNA genes suggested that the species T. yunnanensis should still belong to the genus Thitarodes rather than Ahamns, which is different from the results based on the traditional phylogeny.