Activation of the Nrf2/ARE signaling pathway ameliorates hyperlipidemia-induced renal tubular epithelial cell injury by inhibiting mtROS-mediated NLRP3 inflammasome activation.

Dyslipidemia is the most prevalent independent risk factor for patients with chronic kidney disease (CKD). Lipid-induced NLRP3 inflammasome activation in kidney-resident cells exacerbates renal injury by causing sterile inflammation. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that modulates the cellular redox balance; however, the exact role of Nrf2 signaling and its regulation of the NLRP3 inflammasome in hyperlipidemia-induced kidney injury are poorly understood. In this study, we demonstrated that activation of the mtROS-NLRP3 inflammasome pathway is a critical contributor to renal tubular epithelial cell (RTEC) apoptosis under hyperlipidemia. In addition, the Nrf2/ARE signaling pathway is activated in renal tubular epithelial cells under hyperlipidemia conditions both in vivo and in vitro, and Nrf2 silencing accelerated palmitic acid (PA)-induced mtROS production, mitochondrial injury, and NLRP3 inflammasome activation. However, the activation of Nrf2 with tBHQ ameliorated mtROS production, mitochondrial injury, NLRP3 inflammasome activation, and cell apoptosis in PA-induced HK-2 cells and in the kidneys of HFD-induced obese rats. Furthermore, mechanistic studies showed that the potential mechanism of Nrf2-induced NLRP3 inflammasome inhibition involved reducing mtROS generation. Taken together, our results demonstrate that the Nrf2/ARE signaling pathway attenuates hyperlipidemia-induced renal injury through its antioxidative and anti-inflammatory effects through the downregulation of mtROS-mediated NLRP3 inflammasome activation.

Association between perioperative red blood cell transfusions and postoperative venous thromboembolism: A systematic review and meta-analysis.

BACKGROUND: Whether perioperative red blood cell transfusions increases the risk of postoperative venous thromboembolism is controversial and uncertain.We aims to explore the relationship between perioperative red blood cell transfusions and the risk of postoperative venous thromboembolism by conducting a meta-analysis. OBJECTIVE: To conduct a meta-analysis to systematically evaluate the relationship between perioperative red blood cell transfusions and the risk of postoperative venous thromboembolism. METHODS: PubMed, Embase, Cochrane, and Web of Science databases were searched to identify studies examining the relationship between perioperative red blood cell transfusions and the risk of postoperative venous thromboembolism. The databases were searched from establishment to August 2023.Two researchers independently screened literature and extracted data according to inclusion and exclusion criteria. Newcastle-ottawa Scale was used for quality assessment. Meta-analysis of data was performed using RevMan 5.4 software. RESULTS: A total of 15 studies involving 1,880,990 patients were included in this study.Meta-analysis showed that perioperative red blood cell transfusions increased the risk of postoperative venous thromboembolism [OR = 1.61, 95%CI (1.37, 1.89), P < 0.001]. Subgroup analyses showed that the transfusion dose,transfusion timing,study population and follow-up time were closely related to the risk of postoperative venous thromboembolism. CONCLUSIONS: In summary, this meta-analysis demonstrated a significant positive association between perioperative red blood cell transfusions and postoperative venous thromboembolism.Healthcare professionals should pay attention to the influence of blood transfusions on postoperative venous thromboembolism, strengthen management and prevention.

CD36-mediated podocyte lipotoxicity promotes foot process effacement.

Background: Lipid metabolism disorders lead to lipotoxicity. The hyperlipidemia-induced early stage of renal injury mainly manifests as podocyte damage. CD36 mediates fatty acid uptake and the subsequent accumulation of toxic lipid metabolites, resulting in podocyte lipotoxicity. Methods: Male Sprague-Dawley rats were divided into two groups: the normal control group and the high-fat diet group (HFD). Podocytes were cultured and treated with palmitic acid (PA) and sulfo-N-succinimidyl oleate (SSO). Protein expression was measured by immunofluorescence and western blot analysis. Boron-dipyrromethene staining and Oil Red O staining was used to analyze fatty acid accumulation. Results: Podocyte foot process (FP) effacement and marked proteinuria occurred in the HFD group. CD36 protein expression was upregulated in the HFD group and in PA-treated podocytes. PA-treated podocytes showed increased fatty acid accumulation, reactive oxygen species (ROS) production, and actin cytoskeleton rearrangement. However, pretreatment with the CD36 inhibitor SSO decreased lipid accumulation and ROS production and alleviated actin cytoskeleton rearrangement in podocytes. The antioxidant N-acetylcysteine suppressed PA-induced podocyte FP effacement and ROS generation. Conclusions: CD36 participated in fatty acid-induced FP effacement in podocytes via oxidative stress, and CD36 inhibitors may be helpful for early treatment of kidney injury.

Analysis of Traditional Chinese Medicine Symptoms in Children with Spastic Cerebral Palsy: A Data Mining Study.

Background: Cerebral palsy (CP) ranks as a major cause of motor disabilities in children, with spastic CP making up roughly 70-80% of all CP cases. The primary objective of our study is to identify characteristics of Traditional Chinese Medicine(TCM) symptom of spastic CP, thereby establishing correlations between the TCM symptom and the disease, providing a more scientific theoretical foundation for TCM treatments on spastic CP, enabling a deeper comprehension of clinical interventions, ultimately, improving rehabilitation outcomes in TCM treatment for spastic CP. Methods: We conducted a data mining study on TCM symptom of spastic CP children aged 4-14 years old treated at Xi'an Encephalopathy Hospital Affiliated to Shaanxi University of Chinese Medicine, from October 2021 to March 2023. The medical records of all eligible and complete spastic CP patients were extracted, processed for data cleansing, transformed, and subsequently analyzed to discern distinctive TCM symptom. K-Means Clustering Analysis and Association Rule Analysis were used for data mining. Results: Core symptoms identified for spastic CP encompassed "Motor Dysfunction", "Impaired Speech", "Delayed Development", "Limb Stiffness", "Rigidity in the limbs", "Intellectual Impairment", "Timidity and susceptibility to startle responses", "Muscle Wasting", and "Pale or Dull Complexion". Among the top-ranking associations of symptom, patterns emerge wherein "Motor dysfunction" intertwine with "Impaired speech", "Motor dysfunction" coexist with "Delayed development", and "Impaired speech" are accompanied by "Delayed development". Conclusion: This study identified the core symptom of spastic CP and tentatively suggests that the clinical manifestations of spastic CP are essentially consistent with the TCM pattern "liver exuberance and spleen weakness". This finding has facilitated the preliminary establishment of correlations between TCM pattern differentiation and the disease in medicine. It is anticipated that this correlation will bring tangible benefits to a larger number of children with spastic CP.

Association of Systemic Immune-Inflammation Index and Systemic Inflammation Response Index with Diabetic Kidney Disease in Patients with Type 2 Diabetes Mellitus.

Purpose: To evaluate the association of the systemic immune-inflammatory index (SII) and systemic inflammatory response index (SIRI) with the clinical and pathological features and progression of diabetic kidney disease (DKD). Patients and Methods: We analyzed 303 patients with type 2 diabetes mellitus (T2DM), classifying them into distinct groups: T2DM, early DKD (EDKD), and clinical DKD (Cli-DKD). Variations in SII and SIRI levels across these groups and their association with renal function were assessed. Logistic regression analysis was used to identify independent risk factors for DKD. Additionally, in 43 patients with biopsy-confirmed DKD, we analyzed the relationship between SII, SIRI, and pathological changes. Kaplan-Meier survival analysis and the Cox proportional hazards model were used to assess the influence of SII and SIRI levels on outcomes in patients with DKD. Results: SII and SIRI were significantly higher in the Cli-DKD group than in the T2DM and EDKD groups, and were positively correlated with the urinary albumin-creatinine ratio and negatively correlated with estimated glomerular filtration rate. Notably, SIRI was identified as an independent risk factor for DKD development. Additionally, a lower SII score was associated with a higher cumulative survival rate. Conclusion: This study demonstrates an association between SII, SIRI, and renal function in patients with T2DM. A high SIRI was an independent risk factor for DKD, while an elevated SII was associated with an increased risk of kidney disease progression in biopsy-confirmed DKD cases. Our findings underscore the potential implications of utilizing SII and SIRI as cost-effective and readily available inflammatory indicators for monitoring DKD in primary care settings.

Metabolomics study reveals blood biomarkers for early diagnosis of chronic kidney disease and IgA nephropathy: A retrospective cross-sectional study.

BACKGROUND AND AIMS: Chronic kidney disease (CKD) causes low quality of life and alarming morbidity and mortality. The crucial to retard CKD progression is to diagnose early for timely treatment. IgA nephropathy (IgAN) is a typical CKD and the most common glomerulonephritis. Both CKD and IgAN lack accurate and sensitive blood biomarkers for early diagnosis. Here we report the potential of plasma biomarkers for early diagnosis of CKD and IgAN. MATERIALS AND METHODS: Plasma levels of metabolites derived from tryptophan were quantified with an LC-MS/MS-based metabolomics for two cohorts. Based on the predictive probability of each metabolite, multivariate models including logistic regression and random forest were used to establish the early diagnostic biomarkers for CKD and IgAN. RESULTS: The plasma melatonin diagnosed early CKD (stages Ⅰ-Ⅱ) with an accuracy exceeding 95%, and a panel of melatonin and tryptophan achieved a remarkable 100% accuracy in diagnosing early CKD. Furthermore, indole-3-lactic acid had an excellent ability to distinguish IgAN among CKD patients. Based on the CKD screening and IgAN diagnosis primarily contributed by melatonin and indole-3-lactic acid, early IgAN could be diagnosed with an accuracy of over 85%. CONCLUSIONS: This study provides promising plasma biomarkers for early diagnosis of CKD and IgAN.

Role of TFEB-autophagy lysosomal pathway in palmitic acid induced renal tubular epithelial cell injury.

Lysosomal dysfunction and impaired autophagic flux are involved in the pathogenesis of lipotoxicity in the kidney. Here, we investigated the role of transcription factor EB (TFEB), a master regulator of autophagy-lysosomal pathway, in palmitic acid induced renal tubular epithelial cells injury. We examined lipid accumulation, autophagic flux, expression of Ps211-TFEB, and nuclear translocation of TFEB in HK-2 cells overloaded with palmitic acid (PA). By utilizing immunohistochemistry, we detected TFEB expression in renal biopsy tissues from patients with diabetic nephropathy and normal renal tissue adjacent to surgically removed renal carcinoma (controls), as well as kidney tissues from rat fed with high-fat diet (HFD) and low-fat diet (LFD). We found significant lipid accumulation, increased apoptosis, accompanied with elevated Ps211-TFEB, decreased nuclear TFEB, reduced lysosome biogenesis and insufficient autophagy in HK-2 cells treated with PA. Kidney tissues from patients with diabetic nephropathy had lower nuclear and total levels of TFEB than that in control kidney tissues. Level of renal nuclear TFEB in HFD rats was also lower than that in LFD rats. Exogenous overexpression of TFEB increased the nuclear TFEB level in HK-2 cells treated with PA, promoted lysosomal biogenesis, improved autophagic flux, reduced lipid accumulation and apoptosis. Our results collectively indicate that PA is a strong inducer for TFEB phosphorylation modification at ser211 accompanied with lower nuclear translocation of TFEB. Impairment of TFEB-mediated lysosomal biogenesis and function by palmitic acid may lead to insufficient autophagy and promote HK-2 cells injury.

TAC3 regulates GnRH/gonadotropin synthesis in female chickens.

Neurokinin B (NKB), a peptide encoded by the tachykinin 3 (TAC3), is critical for reproduction in all studied species. However, its potential roles in birds are less clear. Using the female chicken (c-) as a model, we showed that cTAC3 is composed of five exons with a full-length cDNA of 787 bp, which was predicted to generate the mature NKB peptide containing 10 amino acids. Using cell-based luciferase reporter assays, we demonstrated that cNKB could effectively and specifically activate tachykinin receptor 3 (TACR3) in HEK293 cells, suggesting its physiological function is likely achieved via activating cTACR3 signaling. Notably, cTAC3 and cTACR3 were predominantly and abundantly expressed in the hypothalamus of hens and meanwhile the mRNA expression of cTAC3 was continuously increased during development, suggesting that NKB-TACR3 may emerge as important components of the neuroendocrine reproductive axis. In support, intraperitoneal injection of cNKB could significantly promote hypothalamic cGnRH-Ι, and pituitary cFSHß and cLHß expression in female chickens. Surprisingly, cTAC3 and cTACR3 were also expressed in the pituitary gland, and cNKB treatment significantly increased cLHß and cFSHß expression in cultured primary pituitary cells, suggesting cNKB can also act directly at the pituitary level to stimulate gonadotropin synthesis. Collectively, our results reveal that cNKB functionally regulate GnRH/gonadotropin synthesis in female chickens.

[Blood Lipid Indicators and Different Clinical Classifications of Dyslipidemia and Diabetic Kidney Disease: Correlation and Predictive Value].

Objective: To explore the relationship between blood lipid indicators and different clinical classifications of dyslipidemia and diabetic kidney disease (DKD) and to compare the value of different clinical classifications of dyslipidemia for predicting DKD. Methods: Continuously enrollment of subjects was conducted at the First Affiliated Hospital of Chongqing Medical University and the Yongchuan Hospital of Chongqing Medical University between October 2020 and October 2021. A total of 356 type 2 diabetes mellitus (T2DM) patients admitted to the two hospitals were enrolled. They were divided into DKD group ( n=126) and simple T2DM group ( n=230) according to whether their T2DM was combined with DKD. In addition, 250 healthy individuals undergoing physical examination during the same period were enrolled for the control group. The blood pressure, blood lipid, blood glucose, and the kidney function indicators of the three groups were measured. The effects of different classifications of dyslipidemia on DKD were analyzed with unconditional logistic regression models, the receiver operating characteristic (ROC) curve was constructed, the area under the curve ( AUC) of ROC was calculated, and the value of different classifications of dyslipidemia for predicting DKD was analyzed. Results: The diastolic blood pressure (DBP), systolic blood pressure (SBP), total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), serum creatinine (Scr), uric acid (UA), and glycosylated hemoglobin A1c (HbA1c) of the DKD group and the simple T2DM group were significantly higher than those of the control group, while the high-density lipoprotein cholesterol (HDL-C) levels of the DKD group and the simple T2DM group were lower than that of the control group (all P<0.05). The disease course of T2DM, DBP, SBP, TC, TG, Scr, UA and HbA1c of the DKD group were significantly higher than those of the T2DM group (all P<0.05). After adjusting for the effects of T2DM disease course, DBP, SBP, Scr, UA and HbA1c, the results showed that TC ( OR=1.426, 95% CI: 1.088-1.868) and TG ( OR=1.404, 95% CI: 1.075-1.833) were independent risk factors for DKD, and that hypercholesterolemia ( OR=1.817, 95% CI: 1.040-3.177) and mixed hyperlipidemia ( OR=2.148, 95% CI: 1.110-4.159) were independent risk factors for DKD (all P<0.05). The AUC (95% CI) of hypercholesterolemia was 0.789 (0.729-0.871). The AUC (95% CI) of mixed hyperlipidemia was 0.671 (0.579-0.760). Hypercholesterolemia showed better predictive value for the diagnosis and prediction of DKD. Conclusion: Among the blood lipid indicators, TC and TG are independent risk factors of DKD. In the clinical classifications of dyslipidemia, hypercholesterolemia and mixed hyperlipidemia are independent risk factors of DKD. Hypercholesterolemia can be used as a predictor to screen for DKD among T2DM patients and is well suited for extensive application in outpatient screening.

Analysis of Traditional Chinese Medicine Symptoms in Children with Spastic Cerebral Palsy, a Protocol for Data Mining.

Background: Cerebral palsy (CP) is characterized by abnormal pronunciation, posture, and movement. Clinically, CP can be categorized into various motor syndromes, including spastic hemiplegia, diplegia, quadriplegia, involuntary movement, ataxia, and mixed types. Among these, spastic CP represents over 70-80% of all CP cases. The primary objective of our study is to identify the top and core Traditional Chinese Medicine (TCM) symptoms and analysis their association rules in children with spastic cerebral palsy, thereby enhancing the theoretical foundations of TCM treatment on spastic CP. Methods: The study will be conducted on children aged 4 to 14 years with spastic CP who are undergoing treatment at Xi'an Encephalopathy Hospital Affiliated to Shaanxi University of Chinese Medicine. Basic information about the patients and their TCM symptoms will be collected on the first day of admission. This information will include age, gender, birth history, family history, disease classification, and TCM symptoms (including symptoms, tongue, and pulse). Once the data is collected, it will be exported from the electronic medical record system for further analysis. Descriptive statistics will be performed using Excel 2019, while exploratory factor analysis and cluster analysis will be conducted using SPSS Statistics 22. Additionally, association rule analysis will be carried out using SPSS Modeler 18. Results: This study will investigate the most top TCM symptoms in children with spastic CP and explore the association rules between these symptoms, mapping the presentation of spastic CP onto symptoms identified within TCM. Conclusion: Our findings will provide the distinctive characteristics of TCM symptoms in children with spastic CP, furnishing evidence-based support to clinicians and patients in making well-informed decisions collaboratively.

Acupuncture and Tuina Treatment for Gross Motor Function in Children with Spastic Cerebral Palsy: A Monocentric Clinical Study.

Objective: Cerebral palsy (CP) is a condition characterized by abnormal pronunciation, posture, and movement, particularly spastic CP, which involves Gross motor dysfunction due to increased muscle tone and stiffness. This monocentric clinical study aims to evaluate the effectiveness of acupuncture and tuina (AT) in improving gross motor function and alleviating associated symptoms in children diagnosed with spastic CP. Methods: A total of 83 eligible patients received AT treatment, while 85 patients received conventional rehabilitation treatment. Both groups underwent a 12-week treatment period following the research protocol. Pre- and post-treatment assessments included the Modified Ashworth Muscle Tension Scale (MAS), Gross Motor Function Measure (GMFM-D and GMFM-E), 6-min walking distance measurement (6MWD), and Modified Children's Functional Independence Rating Scale (WeeFIM). Results: After 12 weeks of treatment, when compared with baseline, the scores of MAS in both AT group and control group are decreased (p<0.01, p<0.01), the scores of GMFM-D, GMFME, 6MWD, WeeFIM in both group are increased (p<0.01 in all indicators). When compared with control group, AT group had significantly lower MAS scores compared to the control group (p<0.01), indicating reduced muscle tension. Moreover, AT group showed significantly higher scores in GMFM-D, GMFM-E, 6MWD, and WeeFIM compared to the control group (p<0.01 in all indicators), indicating improved gross motor function and functional independence. The study also revealed an inverse correlation between the children's age and treatment efficacy (r= -0.496, p<0.01 in AT group, r=-0.540, p<0.01 in control group), highlighting the importance of early intervention in the management of CP in children. Conclusion: These findings suggest that AT may effectively enhance gross motor function and alleviate associated symptoms in children diagnosed with spastic CP. Moreover, early initiation of treatment is crucial to maximize therapeutic efficacy in children with spastic CP. Trial Registration: Chinese Clinical Trial Registry, ChiCTR2200059823. Registered on 12 May 2022.

Active immunization against gonadotropin-releasing hormone affects thymic T cell production, migration, and colonization in male rat lymphoid tissue.

Active immunization against gonadotropin-releasing hormone (GnRH) inhibits animal reproduction and has become a friendly alternative to surgical castration, which has been reported to affect the proportion of thymic T cell subpopulations. The effects of active immunization against GnRH on T cell migration from the thymus to the periphery and T cell distribution in lymphoid tissues remain unclear. Here, we showed that active immunization against GnRH increased thymic size and weight, enlarged the number of thymocytes, and enhanced CD4+ recent thymic emigrants (RTEs) and CD8+ RTEs migration to the blood and spleen. Active immunization against GnRH had no significant effect on naïve CD4+, naïve CD8+, CD4+ memory/activated, or CD8+ memory/activated T cells. In addition, active immunization against GnRH increased the proportion of CD3+ T cells in the spleen and lymph nodes. The percentages of CD3+CD4+ and CD3+CD8+ T cells in the blood, spleen, and lymph nodes were not significantly affected by GnRH immunization. Overall, these results enhance our understanding of thymic T cell production, migration, and colonization in rat lymphoid tissues affected by GnRH immunization.

Effect of Purslane (Portulaca oleracea L.) on Intestinal Morphology, Digestion Activity and Microbiome of Chinese Pond Turtle (Mauremys reevesii) during Aeromonas hydrophila Infection.

Large-scale mortality due to Aeromonas hydrophila (A. hydrophila) infection has considerably decreased the yield of the Chinese pond turtle (Mauremys reevesii). Purslane is a naturally active substance with a wide range of pharmacological functions, but its antibacterial effect on Chinese pond turtles infected by A. hydrophila infection is still unknown. In this study, we investigated the effect of purslane on intestinal morphology, digestion activity, and microbiome of Chinese pond turtles during A. hydrophila infection. The results showed that purslane promoted epidermal neogenesis of the limbs and increased the survival and feeding rates of Chinese pond turtles during A. hydrophila infection. Histopathological observation and enzyme activity assay indicated that purslane improved the intestinal morphology and digestive enzyme (α-amylase, lipase and pepsin) activities of Chinese pond turtle during A. hydrophila infection. Microbiome analysis revealed that purslane increased the diversity of intestinal microbiota with a significant decrease in the proportion of potentially pathogenic bacteria (such as Citrobacter freundii, Eimeria praecox, and Salmonella enterica) and an increase in the abundance of probiotics (such as uncultured Lactobacillus). In conclusion, our study uncovers that purslane improves intestinal health to protect Chinese pond turtles against A. hydrophila infection.

SGU-Net: Shape-Guided Ultralight Network for Abdominal Image Segmentation.

Convolutional neural networks (CNNs) have achieved significant success in medical image segmentation. However, they also suffer from the requirement of a large number of parameters, leading to a difficulty of deploying CNNs to low-source hardwares, e.g., embedded systems and mobile devices. Although some compacted or small memory-hungry models have been reported, most of them may cause degradation in segmentation accuracy. To address this issue, we propose a shape-guided ultralight network (SGU-Net) with extremely low computational costs. The proposed SGU-Net includes two main contributions: it first presents an ultralight convolution that is able to implement double separable convolutions simultaneously, i.e., asymmetric convolution and depthwise separable convolution. The proposed ultralight convolution not only effectively reduces the number of parameters but also enhances the robustness of SGU-Net. Secondly, our SGUNet employs an additional adversarial shape-constraint to let the network learn shape representation of targets, which can significantly improve the segmentation accuracy for abdomen medical images using self-supervision. The SGU-Net is extensively tested on four public benchmark datasets, LiTS, CHAOS, NIH-TCIA and 3Dircbdb. Experimental results show that SGU-Net achieves higher segmentation accuracy using lower memory costs, and outperforms state-of-the-art networks. Moreover, we apply our ultralight convolution into a 3D volume segmentation network, which obtains a comparable performance with fewer parameters and memory usage. The available code of SGUNet is released at https://github.com/SUST-reynole/SGUNet.

Efficacy of Immunization against a Novel Synthetic 13-Amino Acid Betaglycan-Binding Peptide Sequence of Inhibin α Subunit on Promoting Fertility in Female Rats.

Inhibins suppress the FSH production in pituitary gonadotrope cells by robustly antagonizing activin signaling by competitively binding to activin type II receptors (ACTR II). The binding of inhibin A to ACTR II requires the presence of its co-receptor, namely, betaglycan. In humans, the critical binding site for betaglycan to inhibin A was identified on the inhibin α subunit. Through conservation analysis, we found that a core 13-amino-acid peptide sequence within the betaglycan-binding epitope on human inhibin α subunit is highly conserved across species. Based on the tandem sequence of such a conserved 13-amino-acid betaglycan-binding epitope (INHα13AA-T), we developed a novel inhibin vaccine and tested its efficacy in promoting female fertility using the female rat as a model. Compared with placebo-immunized controls, INHα13AA-T immunization induced a marked (p < 0.05) antibody generation, enhanced (p < 0.05) ovarian follicle development, and increased ovulation rate and litter sizes. Mechanistically, INHα13AA-T immunization promoted (p < 0.05) pituitary Fshb transcription and increased (p < 0.05) serum FSH and 17ß-estradiol concentrations. In summary, active immunization against INHα13AA-T potently increased FSH levels, ovarian follicle development, ovulation rate and litter sizes, thus causing super-fertility in females. Therefore, immunization against INHα13AA is a promising alternative to the conventional approach of multiple ovulation and super-fertility in mammals.

Spexin2 Is a Novel Food Regulator in Gallus gallus.

Spexin2 (SPX2), a paralog of SPX1, is a newly identified gene in non-mammalian vertebrates. Limited studies in fish have evidenced its important role in food intake and energy balance modulation. However, little is known about its biological functions in birds. Using the chicken (c-) as a model, we cloned the full-length cDNA of SPX2 by using RACE-PCR. It is 1189 base pair (bp) in length and predicted to generate a protein of 75 amino acids that contains a 14 amino acids mature peptide. Tissue distribution analysis showed that cSPX2 transcripts were detected in a wide array of tissues, with abundant expression in the pituitary, testis, and adrenal gland. cSPX2 was also observed to be ubiquitously expressed in chicken brain regions, with the highest expression in the hypothalamus. Its expression was significantly upregulated in the hypothalamus after 24 or 36 h of food deprivation, and the feeding behavior of chicks was obviously suppressed after peripheral injection with cSPX2. Mechanistically, further studies evidenced that cSPX2 acts as a satiety factor via upregulating cocaine and amphetamine regulated transcript (CART) and downregulating agouti-related neuropeptide (AGRP) in hypothalamus. Using a pGL4-SRE-luciferase reporter system, cSPX2 was demonstrated to effectively activate a chicken galanin II type receptor (cGALR2), a cGALR2-like receptor (cGALR2L), and a galanin III type receptor (cGALR3), with the highest binding affinity for cGALR2L. Collectively, we firstly identified that cSPX2 serves as a novel appetite monitor in chicken. Our findings will help clarify the physiological functions of SPX2 in birds as well as its functional evolution in vertebrates.

Self-assembled micropillar arrays via near-field electrospinning.

Self-assembly in near-field electrospinning is reported for the first time in this paper, which realized the conversion from two-dimensional planar printing to three-dimensional (3D) structures. Repeatedly stacked fibres formed a micropillar array structure (MPAS) with intervals on the deposition paths by adding carbonyl iron powder particles to a polyethylene oxide (PEO) solution. The growth process of the self-assembled MPAS is documented, and the mechanism of the self-assembled MPAS is proposed. In addition, the effects of substrate speed and injection speed on self-assembly were investigated. Electric field distribution simulations show that the electric field strength around the MPAS is enhanced by nearly ten times so that the micropillar can attract the jet for further deposition. Self-assembly can obtain MPASs with arbitrary paths on different substrates, and the interval of the MPAS can be controlled by using bulging substrates. Furthermore, a self-assembled MPAS has been successfully used to prepare mold cavities, which can be used to prepare MPASs of other materials. Due to their small feature size, large surface area and structural periodicity, micropillar arrays will have promising applications, such as hydrophobicity of surfaces and electrochemical detection. Self-assembly in near-field electrospinning can significantly reduce the preparation cost of an MPAS and provide new processes and ideas.

PNX14 but not PNX20 as a novel regulator of preadipocyte differentiation via activating Epac-ERK signaling pathway in Gallus gallus.

Small integral membrane protein 20 (SMIM20) could generate two main peptides, PNX14 and PNX20, which participate in multiple biological roles such as reproduction, inflammation and energy metabolism in mammals. However, little is known about their physiological functions in non-mammalian vertebrates. Using chicken (c-) as an animal model, we found cSMIM20 was moderately expressed in adipose tissues, and its expression was gradually increased during the differentiation of chicken preadipocytes, suggesting that it may play an important role in chicken adipogenesis. Further research showed cPNX14 could facilitate the differentiation of chicken preadipocytes into mature adipocytes by enhancing expression of adipogenic genes including PPARγ, CEBPα and FABP4, and promoting the formation of lipid droplets. This pro-adipogenic effect of cPNX14 was completely attenuated by Epac-specific and ERK inhibitor. Interestingly, cPNX20 failed to regulate the adipogenic genes and lipid droplet content. Collectively, our findings reveal that cPNX14 but not cPNX20 can serve as a novel adipogenesis mediator by activating the Epac-ERK signaling pathway in chickens.

Mechanic Insight into the Distinct and Common Roles of Ovariectomy Versus Adrenalectomy on Adipose Tissue Remodeling in Female Mice.

Dysfunctions of the ovaries and adrenal glands are both evidenced to cause aberrant adipose tissue (AT) remodeling and resultant metabolic disorders, but their distinct and common roles are poorly understood. In this study, through biochemical, histological and RNA-seq analyses, we comprehensively explored the mechanisms underpinning subcutaneous (SAT) and visceral adipose tissue (VAT) remodeling, in response to ovariectomy (OVX) versus adrenalectomy (ADX) in female mice. OVX promoted adipocyte differentiation and fat accumulation in both SAT and VAT, by potentiating the Pparg signaling, while ADX universally prevented the cell proliferation and extracellular matrix organization in both SAT and VAT, likely by inactivating the Nr3c1 signaling, thus causing lipoatrophy in females. ADX, but not OVX, exerted great effects on the intrinsic difference between SAT and VAT. Specifically, ADX reversed a large cluster of genes differentially expressed between SAT and VAT, by activating 12 key transcription factors, and thereby caused senescent cell accumulation, massive B cell infiltration and the development of selective inflammatory response in SAT. Commonly, both OVX and ADX enhance circadian rhythmicity in VAT, and impair cell proliferation, neurogenesis, tissue morphogenesis, as well as extracellular matrix organization in SAT, thus causing dysfunction of adipose tissues and concomitant metabolic disorders.

UNC93B1 facilitates TLR18-mediated NF-κB signal activation in Schizothorax prenanti.

Toll-like receptor 18 (TLR18), a non-mammalian TLR, has been believed to play an important role in anti-bacterial immunity of teleost fishes. UNC93B1 is a classical molecular chaperone that mediates TLRs transport from endoplasmic reticulum to the located membrane. However, TLR18-mediated signal transduction mechanism and the regulatory effect of UNC93B1 to TLR18 are still unclear in teleost fishes. In this study, the coding sequences of TLR18 and UNC93B1 were cloned from Schizothorax prenanti, named spTLR18 and spUNC93B1, respectively. The spTLR18 and spUNC93B1 are 2583 bp and 1878 bp in length, encode 860 and 625 amino acids, respectively. The spTLR18 widely expressed in various tissues with the highest expression level in liver. After stimulation of Aeromonas hydrophila, lipopolysaccharide (LPS) and Poly(I:C), the expression levels of spTLR18 were significantly increased in spleen and head kidney. The spTLR18 located in the cell membrane, while spUNC93B1 located in the cytoplasm. Luciferase and overexpression analysis showed that spTLR18 activated NF-κB and type I IFN signal pathways, and spTLR18-mediated NF-κB activation might depend on the adaptor molecule MyD88. Besides, spUNC93B1 positively regulates spTLR18-mediated NF-κB signal. Our study first uncovers TLR18-UNC93B1-mediated signal transduction mechanism, which contributes to the understanding of TLR signaling pathway in teleost fishes.