Primary perihilar bile duct neuroendocrine tumor: a case report and review of the literature.

Neuroendocrine tumors represent a rare neoplastic entity, with even rarer occurrences within the biliary tract system. The pathogenesis of such conditions remains enigmatic. Clinical manifestations and radiological evaluations exhibit limited specificity, rendering preoperative diagnoses challenging. As of now, definitive therapeutic modalities remain elusive. Surgical excision stands as the paramount approach for managing biliary neuroendocrine tumors. A thorough preoperative assessment should precede the formulation of a judicious surgical strategy. Postoperative targeted adjuvant therapies hold promise in enhancing therapeutic efficacy and retarding tumor recurrence. This article chronicles a case study detailing a neuroendocrine tumor's diagnostic and treatment course within the perihilar bile duct. Integrating pertinent literature, it encapsulates the clinical attributes and diagnostic and therapeutic advancements in biliary neuroendocrine tumors. The aspiration is to augment awareness of this category of ailments, mitigating the occurrence of both missed and erroneous diagnoses, and furnishing a reference for forthcoming clinical endeavors.

Impaired sensitivity to thyroid hormones is associated with albuminuria in the euthyroid population: results from NHANES.

PURPOSE: The relationship between thyroid hormone sensitivity and albuminuria remains unclear. We aimed to investigate the association between thyroid hormone sensitivity and the risk of albuminuria in a euthyroid population. METHODS: This cross-sectional study included 7634 euthyroid adults collected from the National Health and Nutrition Examination Survey (NHANES) between 2007 and 2012. Central sensitivity to thyroid hormones was evaluated using the thyroid-stimulating hormone index (TSHI), the thyrotrophic thyroxine resistance index (TT4RI), and the thyroid feedback quantile-based index (TFQI). Peripheral sensitivity to thyroid hormones was measured using the free triiodothyronine/free thyroxine (FT3/FT4) ratio. Furthermore, the independent relationship between sensitivity to thyroid hormones and albuminuria was assessed. RESULTS: The proportion of albuminuria increased with a higher interquartile range of TFQI levels (7.31% vs. 7.89% vs. 7.95% vs. 9.89%, P = 0.024). Furthermore, TFQI was found to be independently associated with the risk of albuminuria after adjusting for confounding factors (OR = 1.28, 95% CI 1.01-1.60, P = 0.037). Subgroup analysis revealed a significant relationship between TFQI and albuminuria, especially among individuals over 60. CONCLUSIONS: In euthyroid subjects, impaired central sensitivity to thyroid hormones is associated with albuminuria. TFQI holds significant potential as an epidemiological tool for quantifying the impact of impaired central sensitivity on the risk of albuminuria.

Associations Between Serum TNF-α, IL-6, hs-CRP and GLMD in Obese Children and Adolescents: A Cross-Sectional Study.

Purpose: To explore the relationships between serum tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), high-sensitivity C-reactive protein (hs-CRP) levels and glucolipid metabolism disorders (GLMD) in obese children and adolescents. Patients and Methods: In this cross-sectional study, 105 obese children and adolescents were selected for the detection of TNF-α, IL-6, hs-CRP, and glycolipid metabolism indicators. All participants were divided into elevated TNF-α group (≥8.1 pg/mL; n=49) and normal TNF-α group (<8.1 pg/mL; n=56), elevated IL-6 group (≥5.9 pg/mL; n=13) and normal IL-6 group (<5.9 pg/mL; n=92), elevated hs-CRP group (≥3.0 mg/L; n=44) and normal hs-CRP group (<3.0 mg/L; n=61), respectively. Results: Low-density lipoprotein cholesterol (LDL-C) in the elevated TNF-α group was higher than that in the normal TNF-α group (P=0.010). TNF-α was positively correlated with LDL-C (P=0.005). Fasting insulin (FINS) and homeostasis model assessment of insulin resistance (HOMA-IR) in the elevated IL-6 group were higher than those in the normal IL-6 group (all for P <0.05), while high-density lipoprotein cholesterol (HDL-C) in the elevated IL-6 group was lower than that in the normal IL-6 group (P<0.001). IL-6 was positively correlated with FINS, 2-hour postprandial insulin, HOMA-IR and triglyceride (all for P <0.01), while was negatively correlated with HDL-C (P=0.006). Moreover, hs-CRP was positively correlated with FINS and HOMA-IR (all for P <0.05). Conclusion: There may be correlations between serum TNF-α, IL-6, hs-CRP levels and GLMD in obese children and adolescents. Attention should be paid to monitoring serum inflammatory factors and preventing their elevation in obese children and adolescents, thus reducing the occurrence of GLMD.

Prim-O-glucosylcimifugin ameliorates aging-impaired endogenous tendon regeneration by rejuvenating senescent tendon stem/progenitor cells.

Adult tendon stem/progenitor cells (TSPCs) are essential for tendon maintenance, regeneration, and repair, yet they become susceptible to senescence with age, impairing the self-healing capacity of tendons. In this study, we employ a recently developed deep-learning-based efficacy prediction system to screen potential stemness-promoting and senescence-inhibiting drugs from natural products using the transcriptional signatures of stemness. The top-ranked candidate, prim-O-glucosylcimifugin (POG), a saposhnikovia root extract, could ameliorate TPSC senescent phenotypes caused by long-term passage and natural aging in rats and humans, as well as restore the self-renewal and proliferative capacities and tenogenic potential of aged TSPCs. In vivo, the systematic administration of POG or the local delivery of POG nanoparticles functionally rescued endogenous tendon regeneration and repair in aged rats to levels similar to those of normal animals. Mechanistically, POG protects TSPCs against functional impairment during both passage-induced and natural aging by simultaneously suppressing nuclear factor-κB and decreasing mTOR signaling with the induction of autophagy. Thus, the strategy of pharmacological intervention with the deep learning-predicted compound POG could rejuvenate aged TSPCs and improve the regenerative capacity of aged tendons.

Hip circumference has independent association with the risk of hyperuricemia in middle-aged but not in older male patients with type 2 diabetes mellitus.

BACKGROUND: Obesity and type 2 diabetes mellitus (T2DM) are risk factors for hyperuricemia. However, which anthropometric indices can better predict incident hyperuricemia in patients with T2DM remains inconsistent. This study aimed to examine the associations between hyperuricemia and different anthropometric indices in middle-aged and older male patients with T2DM. METHODS: In this retrospective study, a total of 1447 middle-aged (45-65 years, n = 791) and older (≥ 65 years, n = 656) male patients with T2DM were collected from December 2015 to January 2020 at Shanghai Xinhua Hospital. Hyperuricemia was defined as a serum uric acid level above 7.0 mg/dL. Weight, height, waist circumference (WC) and hip circumference (HC) were measured by trained nurses at visit. RESULTS: The median uric acid level of subjects was 5.6 (interquartile ranges: 4.7-6.7) mg/dl, and 279 (19.3%) were hyperuricemia, with 146 (18.5%) in the middle-aged group, and 133 (20.3%) in the older group. After adjusting for age, duration of T2DM, fasting plasma glucose and insulin, homeostasis model assessment-ß, aspartate aminotransferase, triglycerides, high-density lipoprotein cholesterol and estimated glomerular filtration rate, body mass index (BMI), WC, HC, and waist-to-height ratio (WHtR) were associated with a higher risk of hyperuricemia in both middle-aged and older group (P < 0.05). After further adjusting for BMI and WC, HC still showed a positive relationship with the risk of hyperuricemia (Odds Ratio = 1.51, 95% confidence intervals: 1.06-2.14) in the middle-aged group, but such relationship was not found in the older group. Moreover, according to receiver operating characteristic analysis, the optimal cutoff value was 101.3 cm of HC for hyperuricemia screening in the middle-aged male patients with T2DM. CONCLUSION: In middle-aged male patients with T2DM, more attention should be paid to HC with the cutoff value of 101.3 cm in clinical practice for early recognition of individuals with a high risk of hyperuricemia for targeted guidance on disease prevention, such as community screening.

Macrocycle Self-Assembly Hydrogel for High-Efficient Oil-Water Separation.

Supramolecular hydrogels involved macrocycles have been explored widely in recent years, but it remains challenging to develop hydrogel based on solitary macrocycle with super gelation capability. Here, the construction of lantern[33 ]arene-based hydrogel with low critical gelation concentration (0.05 wt%), which can be used for efficient oil-water separation, is reported. The lantern[33 ]arenes self-assemble into hydrogen-bonded organic nanoribbons, which intertwine into entangled fibers to form hydrogel. This hydrogel which exhibits reversible pH-responsiveness characteristics can be coated on stainless-steel mesh by in situ sol-gel transformation. The resultant mesh exhibits excellent oil-water separation efficiency (>99%) and flux (>6 × 104 L m-2 h-1 ). This lantern[33 ]arene-based hydrogel not only sheds additional light on the gelation mechanisms for supramolecular hydrogels, but also extends the application of macrocycle-based hydrogels as functional interfacial materials.

Malignant melanoma resection and reconstruction with the first manifestation of lumbar metastasis: A case report.

BACKGROUND: Malignant melanoma (MM) has shown an increasing incidence worldwide, and a potential to metastasize to almost any part of the body. Clinically, MM with bone metastasis as the initial manifestation is extremely rare. Spinal metastatic MM can cause spinal cord or nerve root compression, resulting in severe pain and paralysis. Currently, the primary clinical treatments for MM are surgical resection in conjunction with chemotherapy, radiotherapy, and immunotherapy. CASE SUMMARY: Here, we report the case of a 52-year-old male who presented to the clinic with progressive low back pain and limited nerve function. No primary lesion or spinal cord compression was detected from computed tomography and magnetic resonance imaging of the lumbar vertebrae and positron emission tomography scan. A lumbar puncture biopsy confirmed the diagnosis of lumbar spine metastatic MM. Following surgical resection, the patient's quality of life improved, symptoms were relieved, and comprehensive treatment was initiated, which prevented recurrence. CONCLUSION: Spinal metastatic MM is clinically rare, and may cause neurological symptoms, including paraplegia. Currently, the clinical treatment plan consists of surgical resection in combination with chemotherapy, radiotherapy, and immunotherapy.

Combination of IDO inhibitors and platinum(IV) prodrugs reverses low immune responses to enhance cancer chemotherapy and immunotherapy for osteosarcoma.

In recent years, immune checkpoint blockades (ICBs) have made great progress in the treatment of cancer. However, most ICBs have not yet been observed to be satisfactory in the treatment of osteosarcoma. Herein, we designed composite nanoparticles (NP-Pt-IDOi) from a reactive oxygen species (ROS) sensitive amphiphilic polymer (PHPM) with thiol-ketal bonds in the main chain to encapsulate a Pt(IV) prodrug (Pt(IV)-C12) and an indoleamine-(2/3)-dioxygenase (IDO) inhibitor (IDOi, NLG919). Once NP-Pt-IDOi enter the cancer cells, the polymeric nanoparticles could dissociate due to the intracellular ROS, and release Pt(IV)-C12 and NLG919. Pt(IV)-C12 induces DNA damage and activates the cGAS-STING pathway, increasing infiltration of CD8+ T cells in the tumor microenvironment. In addition, NLG919 inhibits tryptophan metabolism and enhances CD8+ T cell activity, ultimately activating anti-tumor immunity and enhancing the anti-tumor effects of platinum-based drugs. NP-Pt-IDOi were shown to have superior anti-cancer activity in vitro and in vivo in mouse models of osteosarcoma, providing a new clinical paradigm for combining chemotherapy with immunotherapy for osteosarcoma.

Phenotypic and functional characterization of two coelomocyte subsets in Apostichopus japonicus.

The hemocytes of invertebrates are composed of different cell subsets with different morphologies and structures. Different cell subsets have different immune functions, which play an important role in innate immune response against pathogens. However, the understanding of the classification of Apostichopus japonicus coelomocytes and the molecular basis of immune function of different cell subsets is very limited. In this study, two coelomocyte subpopulations of A. japonicus were isolated by Percoll density gradient centrifugation. They were identified from their morphological and structural characteristics, namely, spherical cells with a size of 10-12 µm spherical in shape and a large number of small granules inside; lymphocyte-like cells with a size of 4-5 µm spherical or oval in shape, and 1-3 filopodia. Functionally, the phagocytic capacity and lysosomal activity in spherical cells were significantly greater than those in lymphocyte-like cells. The results suggest that spherical cells may play a more critical role in the immune responses. Meanwhile, transcriptome sequencing analysis was performed to further clarify the functional differences between the two cell subsets. The data indicated significantly different gene expression patterns in them. Spherical cells tend to participate in immune defense, whereas lymphocyte-like cells tend to participate in energy metabolism. In addition, lymphocyte-like cells may convert oxidative phosphorylation to glycolysis by changing the manner of energy metabolism to quickly adapt to the energy demand of external stimuli. Spherical cells may respond to LPS stimulation through phagocytosis, and their response time is slower than that of lymphocyte-like cells. The expression of genes involved in endocytosis, phagocytosis, and lysosomal and humoral immunity in spherical cells was significantly higher than that in lymphocyte-like cells. These data provide valuable information for understanding the molecular basis of cellular and humoral immunity in A. japonicus.

Evolution of mitogen-activated protein kinase family and their immune function in Apostichopus japonicus.

The mitogen-activated protein kinase family plays an important role in cell differentiation, growth, proliferation, and survival. However, the current research on the mitogen-activated protein kinase (MAPK) family in invertebrates is limited to the individual gene, and the analysis has not been conducted at the family level. In the present study, echinoderm MAPK family was identified by genomic screening, and five members, including three ERK subfamily members, one c-Jun N-terminal kinase (JNK) subfamily, and one p38-MAPK member were detected. Phylogenetic analysis showed that three MAPK subfamilies were separated into three separated clusters, and ERK subfamily appeared earlier than the other two subfamilies. Synteny analysis revealed that the p38 subfamily might be derived from the continuous gene duplication events of MAPK14 subfamily in invertebrates, which displayed genome expansion via gene duplication in vertebrates. The role of MAPK family in echinoderm immune defense was determined by investigating the expression profiles of MAPKs in Vibrio splendidus-challenged Apostichopus japonicus and LPS-exposed coelomocytes. The result showed that five MAPK members displayed induced expression profiles both in vitro and in vivo, and the peak expression was detected at different time points. Our study provides new insights into the evolutionary history of the MAPK family and show the similar immune function among MAPK members.

Primary hepatic adenosquamous carcinoma: a case report and review of the literature.

Primary hepatic adenosquamous carcinoma is considered a rare subtype of intrahepatic cholangiocarcinoma, with fewer than 100 domestic and international cases reported. This malignancy exhibits a high degree of malignancy, strong invasiveness, and an unfavorable prognosis due to its propensity for early lymph node and intrahepatic metastasis. The etiology of this disease remains uncertain, and preoperative diagnosis is exceedingly challenging owing to the nonspecific clinical features and lack of specificity in imaging studies. Radical surgical resection is the most effective treatment for non-metastatic tumors, while targeted adjuvant therapy administered postoperatively can enhance therapeutic efficacy and delay tumor recurrence. This article documents the diagnostic and therapeutic course of a case of primary hepatic adenosquamous carcinoma treated at our medical institution, along with a comprehensive synthesis of the clinical characteristics and advances in the diagnosis and treatment of this disease, aiming to augment understanding and serve as a reference for future clinical endeavors.

Mn3+/Mn4+ ion-doped carbon dots as fenton-like catalysts for fluorescence dual-signal detection of dopamine.

Carbon dots (CDs), a new zero-dimensional material, have ignited a revolution in the fields of sensing, bioimaging, and biomedicine. However, the difficulty of preparing CDs with Fenton-like catalytic properties has seriously hindered their application in the diagnosis of oxidation/reduction biomolecules or metal ions. Here, an innovative method was successfully established to synthesize Mn3+/Mn4+ ion-doped blue-green fluorescent CDs with Fenton-like catalytic properties using manganese acetate as the manganese source. Specifically, the CDs prepared here were equipped with functional groups of -COOH, NH2, C=O, and Mn-O, offering the possibility to function as a fluorescence sensor. More importantly, the introduction of manganese acetate resulted in the preparation of CDs with Fenton-like catalytic properties, and the dual-signal fluorescence detection of dopamine (DA) was realized with linear ranges of 100-275 nM and 325-525 nM, and the detection limits were 3 and 12 nM, respectively. In addition, due to the Fenton-like catalytic activity of Mn3+/Mn4+ ion-doped CDs, the material has broad application prospects in the detection of oxidation/reduction biomolecules or metal ions related to disease diagnosis and prevention.

Sleep duration of lactating mothers and its relationship with feeding pattern, milk macronutrients and related serum factors: A combined longitudinal cohort and cross-sectional study.

Objective: Insufficient sleep is common in postpartum mothers. The main objectives of this study are to explore the sleep duration among Chinese lactating mothers and preliminarily investigate the relationship between sleep duration and feeding pattern. The secondary objectives are to investigate the relationships between sleep duration and milk macronutrients and between maternal-related indicators, including melatonin (MT), growth hormone (GH), ghrelin (GHRL), glucagon-like peptide-1 (GLP-1), prolactin (PRL), and cholecystokinin (CCK). Methods: The present study comprises a longitudinal and a cross-sectional cohort from December 2019 to December 2021. Postpartum lactating women living in Shanghai were recruited through online and offline recruitment. The subjects were included in the longitudinal cohort or cross-sectional study based on their lactation period at the time of recruitment. The longitudinal cohort included a total of 115 mothers. Human milk and feeding pattern were measured and collected at 2-4 months and 5-7 months postpartum. At four predetermined follow-up time points, data on sleep duration was collected (at the time of recruitment, 2-4 months postpartum, 5-7 months postpartum, and 12-17 months postpartum). The cross-sectional study included 35 lactating mothers (2-12 months postpartum) who reported their sleep duration and provided blood samples. Mid-infrared spectroscopy (MIRS) method was used to analyze the macronutrients of breast milk, while MT, GH, GHRL, GLP-1, PRL, and CCK in maternal blood were determined by ELISA. Results: The maternal sleep duration before pregnancy was 8.14 ± 1.18 h/d (n = 115), 7.27 ± 1.31 h/d (n = 113) for 2-4 months postpartum, 7.02 ± 1.05 h/d (n = 105) for 5-7 months postpartum, and 7.45 ± 1.05 h/d (n = 115) for 12-17 months postpartum. The incidence of insufficient sleep (<7 h/d) before pregnancy (12.17%) was significantly less than at any follow-up time after delivery (vs. 2-4 months postpartum, χ2 = 10.101, p = 0.001; vs. 5-7 months postpartum, χ2 = 15.281, p < 0.0001; vs. 12-17 months postpartum, χ2 = 6.426, p = 0.011). The percentage of insufficient maternal sleep was highest at 5-7 months postpartum (34.29%). No significant difference was found between the incidence of insufficient sleep at 5-7 months postpartum, 2-4 months postpartum (29.20%, χ2 = 0.650, p = 0.420), and 12-17 months postpartum (25.22%, χ2 = 2.168, p = 0.141). At 2-4 months postpartum, the frequency of formula feeding per day is related to reduced maternal sleep duration (Standardization coefficient ß = -0.265, p = 0.005, Adjusted R2 = 0.061). At 2-4 months and 5-7 months postpartum, the relationship between macronutrients in breast milk and the mother's sleep duration was insignificant (all p > 0.05). Other than the positive correlation found between maternal GHRL and sleep duration (r = 0.3661, p = 0.0305), no significant relationship was observed between sleep duration and other indexes (all p > 0.05). Conclusions: Postpartum mothers generally sleep less, but there is no correlation between insufficient sleep and the macronutrient content of breast milk. Formula feeding may be related to the mother's sleep loss, while breastfeeding (especially direct breastfeeding) may be related to increased maternal sleep duration. The findings suggest that sleep duration is related to maternal serum GHRL. More high-quality studies are needed to clarify the mechanism of these findings and provide a solid theoretical basis and support references for breastfeeding.

Genome-wide identification m6A modified circRNAs revealed their key roles in skin ulceration syndrome disease development in Apostichopus japonicus.

Circular RNAs (circRNAs) are novel endogenous non-coding RNAs (ncRNAs) and can be acted as competing endogenous RNAs (ceRNAs) to regulate microRNA (miRNA) and downstream gene expression. Recently, m6A modification has been found in circRNA, and m6A circRNAs also play important roles in various biological processes and a variety of diseases. Our previous study had been demonstrated that circRNAs were differentially expressed in skin ulceration syndrome (SUS) diseased sea cucumber Apostichopus japonicus. However, whether the function of circRNAs are dependent on m6A levels are largely unknown. Here, we firstly investigated the genome-wide map of m6A circRNAs in sea cucumbers with different stages of Vibrio splendidus challenge, that's Control group, SUS-diseased group, and SUS-resistant group. MeRIP-seq revealed that m6A abundances were enriched in circRNAs in all three groups, especially for SUS-resistant group. Among them, more than 62% of modified circRNAs harbor only a single m6A peak and about 55% of m6A sites in circRNAs were derived from sense overlapping in each group. After V. splendidus infection, we found that most of m6A peaks in circRNAs were upregulated and less were downregulated in both SUS-diseased and SUS-resistant groups when compared with Control. Furthermore, GO analysis indicated that the host genes of circRNAs with dysregulated m6A peaks in SUS-diseased and SUS-resistant groups were both mainly enriched in the adhesion pathway. More importantly, we discovered that more than 50% m6A circRNAs showed a positive correlation between the circRNAs expression and m6A methylation levels both in SUS-diseased and SUS-resistant groups. Therefore, a core circRNA-miRNA-mRNA (ceRNA) network whether influenced by m6A modification was constructed based on conjoint analysis. Our results indicated that several selected m6A circRNAs bind with miRNAs were mainly targeting to ubiquitylation system and adhesion pathway. What's more, three candidate m6A circRNAs and three target genes were validated by MeRIP-qPCR and qPCR, whose m6A levels in circRNA and mRNA expressions were consistent with disease occurrence or disease resistance. All of our current findings suggested that m6A circRNAs could play important roles during pathogen infection and might be served as a new molecular biomarker in SUS disease diagnose of A. japonicus.

Comparing human milk macronutrients measured using analyzers based on mid-infrared spectroscopy and ultrasound and the application of machine learning in data fitting.

OBJECTIVE: Fat, carbohydrates (mainly lactose) and protein in breast milk all provide indispensable benefits for the growth of newborns. The only source of nutrition in early infancy is breast milk, so the energy of breast milk is also crucial to the growth of infants. Some macronutrients composition in human breast milk varies greatly, which could affect its nutritional fulfillment to preterm infant needs. Therefore, rapid analysis of macronutrients (including lactose, fat and protein) and milk energy in breast milk is of clinical importance. This study compared the macronutrients results of a mid-infrared (MIR) analyzer and an ultrasound-based breast milk analyzer and unified the results by machine learning. METHODS: This cross-sectional study included breastfeeding mothers aged 22-40 enrolled between November 2019 and February 2021. Breast milk samples (n = 546) were collected from 244 mothers (from Day 1 to Day 1086 postpartum). A MIR milk analyzer (BETTERREN Co., HMIR-05, SH, CHINA) and an ultrasonic milk analyzer (HonÉ¡yanÉ¡ Co,. HMA 3000, Hebei, CHINA) were used to determine the human milk macronutrient composition. A total of 465 samples completed the tests in both analyzers. The results of the ultrasonic method were mathematically converted using machine learning, while the Bland-Altman method was used to determine the limits of agreement (LOA) between the adjusted results of the ultrasonic method and MIR results. RESULTS: The MIR and ultrasonic milk analyzer results were significantly different. The protein, fat, and energy determined using the MIR method were higher than those determined by the ultrasonic method, while lactose determined by the MIR method were lower (all p < 0.05). The consistency between the measured MIR and the adjusted ultrasound values was evaluated using the Bland-Altman analysis and the scatter diagram was generated to calculate the 95% LOA. After adjustments, 93.96% protein points (436 out of 465), 94.41% fat points (439 out of 465), 95.91% lactose points (446 out of 465) and 94.62% energy points (440 out of 465) were within the LOA range. The 95% LOA of protein, fat, lactose and energy were - 0.6 to 0.6 g/dl, -0.92 to 0.92 g/dl, -0.88 to 0.88 g/dl and - 40.2 to 40.4 kj/dl, respectively and clinically acceptable. The adjusted ultrasonic results were consistent with the MIR results, and LOA results were high (close to 95%). CONCLUSIONS: While the results of the breast milk rapid analyzers using the two methods varied significantly, they could still be considered comparable after data adjustments using linear regression algorithm in machine learning. Machine learning methods can play a role in data fitting using different analyzers.

Corrigendum: CircRNA75 and CircRNA72 Function as the Sponge of MicroRNA-200 to Suppress Coelomocyte Apoptosis Via Targeting Tollip in Apostichopus japonicus.

[This corrects the article DOI: 10.3389/fimmu.2021.770055.].

Global N6-methyladenosine methylation analysis reveals the positive correlation between m6A modification and mRNA abundance during Apostichopus japonicus disease development.

N6-methyladenosine (m6A), the most abundant epitranscriptomic modification in eukaryotic messenger RNA (mRNA), plays important roles in regulation of gene expression for fundamental biological processes and diverse physiological functions, including combating with pathogen infection. Here, we were first profile transcriptome-wide m6A sequencing in four stages of skin ulceration syndrome-diseased Apostichopus japonicus following Vibrio splendidus infection, including Control (healthy), Early (small ulcer), Later (extensive ulcer), and Resistant (no ulcer) groups. Our results revealed that three experimental groups were all extensively methylated by m6A and the proportion of the m6A modified genes were also significantly increased to 28.90% (Early), 27.97% (Later), and 29.98% (Resistant) when compared with Control group (15.15%), indicating m6A modification could be induced by V. splendidus infection. Intriguingly, we discovered a positive correlation between the m6A methylation level and mRNA abundance, indicating a positive regulatory role of m6A in sea cucumber gene expression during V. splendidus infection. Moreover, genes with specific and differentially expressed m6A methylation in Later group were both enriched in cell adhesion, while Early and Resistant groups were both mainly involved in DNA conformation change and chromosome organization when compared with Control, suggesting the higher-methylated m6A might serve as "conformational marker" and associated to the initiation of related anti-disease genes transcription in order to improve disease resistance of sea cucumber. Subsequently, we selected the pivotal genes enriched in cell adhesion pathway and found that the IggFc-binding protein (FcGBP) and Fibrocystin-L both had higher levels of m6A methylation and higher level of mRNA expressions in Later group. Conversely, Fibrinogen C domain-containing protein 1 (F1BCD1) gene presented as an antibacterial role in sea cucumber and showed higher mRNA expression and higher m6A methylation in Resistant group and lower mRNA level in Later group. The levels of m6A methylation and mRNA abundance of FcGBP and F1BCD1 genes indicates disease occurrence or disease resistant were also verified by MeRIP-qPCR. Overall, our study presents the first comprehensive characterize of dynamic m6A methylation modification in the different stages of disease in sea cucumber. These data provide an invaluable resource for future studies of function and biological significance of m6A in mRNA in marine invertebrates.

Src kinase mediates coelomocytes phagocytosis via interacting with focal adhesion kinase in Vibrio splendidus challenged Apostichopus japonicus.

Immune cells have many efficient ways to participate in the host immunity, including phagocytosis, which is an important pathway to eliminate pathogens. Only ß-integrin-mediated phagocytosis pathways have been confirmed in Apostichopus japonicus. The Src family kinases (SFKs), a class of non-receptor tyrosine kinases plays an important role in the regulation of phagocytic signals in invertebrates. However, the SFK-mediated phagocytic mechanism is largely unknown in A. japonicus. In this study, a novel SFK homologue (AjSrc) with a conservative SH3 domain, an SH2 domain, and a tyrosine kinase domain was identified from A. japonicus. Both gene and protein expression of AjSrc and phosphorylation levels increased under Vibrio splendidus challenged, reaching the highest level at 24 h. Knock-down of AjSrc could depress coelomocytes' phagocytosis by 25% compared to the control group. To better understand the mechanism of AjSrc-mediated phagocytosis, focal adhesion kinase (FAK) was identified by a Co-immunoprecipitation experiment to be verified as an interactive protein of AjSrc. The phagocytosis rates of coelomocytes were decreased by 33% and 37% in AjFAK and AjSrc + AjFAK interference groups compared with the control group, respectively. Furthermore, the phosphorylation level of AjFAK was increased and reached the maximum level at 24 h post V. splendidus infection, as the same as that of AjSrc. Our results suggested that AjSrc could mediate V. splendidus-induced coelomocytes' phagocytosis via interacting with AjFAK and co-phosphorylation. This study enriched the mechanism of phagocytosis in echinoderm and provided the new theoretical foundation for disease control of sea cucumber.

Effect of different anesthetic dose of pentobarbital on respiratory activity in rabbits.

Anesthetics inhibit the respiratory muscles and even cause upper airway to collapse. Diaphragm electromyography (EMGdi) and airflow signals are usually extracted to assess the degree of respiration inhibition by anesthetics. However, the ECG interference in EMGdi affects the accuracy of its time domain and frequency domain information extraction. We studied the changes in EMGdi (left EMGdi and right EMGdi) and airflow characteristics under two pentobarbital anesthetic doses. First, we filtered out the ECG in EMGdi based on the combination of stationary wavelet transform and the positioning of ECG to obtain EMGdi without ECG interference (EMGdip). The effectiveness of filtering algorithm was verified by calculating the power spectrum before and after noise reduction. Second, root mean square (RMS), average rectified value (ARV), and fixed sample entropy (fSampEn) were used to quantify EMGdi (left EMGdi, left EMGdip and right EMGdi). Median frequency (MF) and centroid frequency (fc) of EMGdi were calculated. Tidal volume, respiratory cycle duration and peak airflow were calculated from airflow. Finally, the average and standard deviation of these parameters for all rabbits (n = 10) were compared and analyzed under two anesthesia states. Our results indicate that anesthesia induced by an increase in pentobarbital dose leads to decrease in ventilation and EMGdi amplitude. There was no significant change in diaphragm power spectrum (MF and fc) with the increase of anesthesia dose.

Skeletal Muscle Mass Has Stronger Association With the Risk of Hyperuricemia Than Body Fat Mass in Obese Children and Adolescents.

Background: Hyperuricemia has been increasing among children with obesity in recent years. However, few studies in such a study group had explored the relationship between obesity-anthropometric indexes and hyperuricemia. This study aimed to examine the associations between hyperuricemia and different body components in children and adolescents with obesity, and further explore gender differences in these associations. Methods: In this cross-sectional study, a total of 271 obese children and adolescents (153 boys and 118 girls) aged 6-17 years were recruited from Shanghai Xinhua Hospital. Data about basic information, anthropometric assessments, body composition, and laboratory tests of participants were collected. Results: In this study, 73 boys (47.71%) and 57 girls (48.31%) were diagnosed to have hyperuricemia. The impacts of percentage of skeletal muscle (PSM) (OR = 1.221, P < 0.001) and skeletal muscle mass (SMM) (OR = 1.179, P < 0.001) on the risk of hyperuricemia was the largest, followed by hip circumference (HC) (OR = 1.109, P < 0.001), waist circumference (WC) (OR = 1.073, P < 0.001), and body fat mass (BFM) (OR = 1.056, P < 0.05) in whole sample, which was adjusted for age, gender and body mass index (BMI). After being stratified by gender, PSM (boys: OR = 1.309, P < 0.001) and SMM (boys: OR = 1.200, P < 0.001; girls: OR = 1.147, P < 0.05) were still the most predictors of hyperuricemia, followed by HC (boys: OR = 1.147, P < 0.001; girls: OR = 1.080, P < 0.05). WC showed a significant association with hyperuricemia only in boys (OR = 1.083, P < 0.05), while BFM showed no association with hyperuricemia in both gender groups after adjusting for age and BMI. Conclusion: Our findings suggested that SMM was a stronger predictor of hyperuricemia than BFM in children and adolescents with obesity, especially in boys.