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We report compound heterozygous variants in TOE1 in siblings of Chinese origin who presented with dyskinesia and intellectual disabilities. Our report provides further information regarding the etiology and pathogenesis of pontocerebellar hypoplasia type 7 syndrome (PCH7). Clinical manifestations were obtained, and genomic DNA was collected from family members. Whole-exome and Sanger sequencing were performed to identify associated genetic variants. Bioinformatics analysis was conducted to identify and characterize the pathogenicity of the heterozygous variants. Following long-term rehabilitation, both siblings showed minimal improvement, and their condition tended to progress. Whole-exome sequencing revealed two unreported heterozygous variants, NM_025077: c.C553T (p.R185W) and NM_025077: c.G562T (p.V188L), in the TOE1 gene mapped to 1p34.1. Sanger sequencing confirmed that the two variants in the proband and her brother were inherited from their parents. The NM_025077: c.C553T (p.R185W) variant was inherited from the father, and the NM_025077: c.G562T (p.V188L) variant was inherited from the mother. Although the two variants in the TOE1 gene have not been reported previously, they were associated with PCH7 based on integrated analysis. Thus, our report contributes to our knowledge regarding the etiology and phenotype of PCH 7., (© 2023 Wiley Periodicals LLC.)

Background & Aims: Breast regression protein 39 (BRP39) (Chi3L1) and its human homolog YKL-40, is an established biomarker of liver fibrosis in nonalcoholic steatohepatitis (NASH) patients, but its role in NASH pathogenesis remains unclear. We recently identified Chi3L1 as one of the top up-regulated genes in mice with inducible gain-of-function NOD-like receptor protein 3 (NLRP3) activation that mimics several liver features of NASH. This study aimed to investigate the effects of BRP39 deficiency on NLRP3-induced liver inflammation using tamoxifen-inducible Nlrp3 knockin mice sufficient (Nlrp3 A350V CRT) and deficient for BRP39 (Nlrp3 A350V /BRP -/- CRT)., Methods: Using Nlrp3 A350V CRT mice and Nlrp3 A350V BRP -/- CRT, we investigated the consequences of BRP39 deficiency influencing NLRP3-induced liver inflammation., Results: Our results showed that BRP39 deficiency in NLRP3-induced inflammation improved body weight and liver weight. Moreover, liver inflammation, fibrosis, and hepatic stellate cell activation were reduced significantly, corresponding to significantly decreased Ly6C+ infiltrating macrophages, CD68+ osteopontin-positive hepatic lipid-associated macrophages, and activated Lymphocyte antigen 6 complex locus G6D positive (Ly6G+) and citrullinated histone H3 postivie (H3Cit+) neutrophil accumulation in the liver. Further investigation showed that circulatory neutrophils from NLRP3-induced BRP39-deficient mice have impaired chemotaxis and migration ability, and this was confirmed by RNA bulk sequencing showing reduced immune activation, migration, and signaling responses in neutrophils., Conclusions: These data showcase the importance of BRP39 in regulating the NLRP3 inflammasome during liver inflammation and fibrotic NASH by altering cellular activation, recruitment, and infiltration during disease progression, and revealing BRP39 to be a potential therapeutic target for future treatment of inflammatory NASH and its associated diseases., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

In recent years, there has been a rapid expansion in our understanding of regulated cell death, leading to the discovery of novel mechanisms that govern diverse cell death pathways. One recently discovered type of cell death is pyroptosis, initially identified in the 1990s as a caspase-1-dependent lytic cell death. However, further investigations have redefined pyroptosis as a regulated cell death that relies on the activation of pore-forming proteins, particularly the gasdermin family. Among the key regulators of pyroptosis is the inflammasome sensor NOD-like receptor 3 (NLRP3), a critical innate immune sensor responsible for regulating the activation of caspase-1 and gasdermin D. A deeper understanding of pyroptosis and its interplay with other forms of regulated cell death is emerging, shedding light on a complex regulatory network controlling pore-forming proteins and cell fate. Cell death processes play a central role in diseases such as metabolic dysfunction-associated steatotic liver disease, metabolic dysfunction-associated steatohepatitis, autoinflammatory disorders, and cancer. Cell death often acts as a starting point in these diseases, making it an appealing target for drug development. Yet, the complete molecular mechanisms are not fully understood, and new discoveries reveal promising novel avenues for therapeutic interventions. In this review, we summarize recent evidence on pathways and proteins controlling pyroptosis and gasdermins. Furthermore, we will address the role of pyroptosis and the gasdermin family in metabolic dysfunction-associated steatotic liver disease and steatohepatitis. Additionally, we highlight new potential therapeutic targets for treating metabolic dysfunction-associated steatohepatitis and other inflammatory-associated diseases., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Stoess, Leszczynska, Kui and Feldstein.)

Chitinase-3-like 1 protein (CHI3L1) is a secreted glycoprotein, strongly correlated with fibrosis severity in chronic liver diseases including non-alcoholic steatohepatitis (NASH). However, the mechanisms by which CHI3L1 contributes to fibrogenesis remain undefined. Here, we showed that infiltrating monocyte-derived liver macrophages represent the main source of CHI3L1 in murine NASH. We developed a floxed CHI3L1 knock-out (KO) mouse to further study the cell-specific role of CHI3L1 ablation. Wildtype (WT) and myeloid cell-specific CHI3L1 KO mice (Cre Lyz ) were challenged with a highly inflammatory and fibrotic dietary model of NASH by administering choline-deficient high-fat diet for 10 weeks. Macrophage accumulation and inflammatory cell recruitment were significantly ameliorated in the Cre Lyz group compared to WT (F4/80 IHC p < 0.0001, CD11b IHC p < 0.0001). Additionally, hepatic stellate cell (HSC) activation and fibrosis were strongly decreased in this group (α-SMA IHC p < 0.0001, picrosirius red staining p < 0.0001). In vitro studies were performed stimulating bone marrow derived macrophages, THP-1 (human monocytes) and LX2 (human HSCs) cells with recombinant CHI3L1 to dissect its relationship with fibrosis development. Results showed an important role of CHI3L1 regulating fibrosis-promoting factors by macrophages (TGFB1 p < 0.05, CTGF p < 0.01) while directly activating HSCs (ACTA2 p < 0.01, COL1A1 p < 0.01), involving IL13Rα2 as the potential mediator. Our findings uncovered a novel role of CHI3L1 derived from liver macrophages in NASH progression and identifies this protein as a potential anti-fibrotic therapeutic target. KEY MESSAGES: We showed that CHI3L1 expression is increased in murine CDAA-HFAT diet NASH model, and that infiltrating macrophages are a key source of CHI3L1 production. Myeloid cell-specific CreLyz CHI3L1 knock-out in mice fed with CDAA-HFAT diet improved the NASH phenotype, with significantly reduced accumulation of pro-inflammatory macrophages and neutrophils compared with WT group. DEG and qPCR analysis of genes in CreLyz CHI3L1 knock-out mouse liver showed the mechanistic role of CHI3L1 in cellular chemotaxis. HSC is directly activated by CHI3L1 via receptor IL13Rα2, leading to upregulation of collagen deposition and pro-fibrotic gene, TIMP-1 and TIMP-2 release in whole liver. Direct stimulation of macrophages with CHI3L1 leads to upregulated expression of HSC-activation factors, suggesting its role in modulating macrophage-HSC crosstalk., (© 2023. The Author(s).)

STAT6 (signal transducer and activator of transcription 6) is a transcription factor that plays a central role in the pathophysiology of allergic inflammation. We have identified 16 patients from 10 families spanning three continents with a profound phenotype of early-life onset allergic immune dysregulation, widespread treatment-resistant atopic dermatitis, hypereosinophilia with esosinophilic gastrointestinal disease, asthma, elevated serum IgE, IgE-mediated food allergies, and anaphylaxis. The cases were either sporadic (seven kindreds) or followed an autosomal dominant inheritance pattern (three kindreds). All patients carried monoallelic rare variants in STAT6 and functional studies established their gain-of-function (GOF) phenotype with sustained STAT6 phosphorylation, increased STAT6 target gene expression, and TH2 skewing. Precision treatment with the anti-IL-4Rα antibody, dupilumab, was highly effective improving both clinical manifestations and immunological biomarkers. This study identifies heterozygous GOF variants in STAT6 as a novel autosomal dominant allergic disorder. We anticipate that our discovery of multiple kindreds with germline STAT6 GOF variants will facilitate the recognition of more affected individuals and the full definition of this new primary atopic disorder., (© 2023 Sharma et al.)

Due to the higher birth rate of preterm infants and improvements in their management, metabolic bone disease of prematurity (MBDP) has a high incidence and is attracting attention. However, clear indicators for the early diagnosis of MBDP are lacking. We aimed to explore simple and feasible early warning indicators for diagnosing MBDP. Our study collected case data of premature infants from two medical centers in Chongqing from January 2020 to February 2022. According to the inclusion and exclusion criteria, data from 136 cases were collected. The correlation between 14 variables in each case and the occurrence of MBDP was analyzed. According to area under the receiver operating characteristic curve (AUROC) analysis, the best cutoff value for each variable was determined. Potential predictors were selected, and Least Absolute Shrinkage and Selection Operator (LASSO) regression analysis was used to establish the association of two models with MBDP, whose results were used to develop a diagnostic nomogram. Furthermore, a model decision curve was analyzed. Four predictors were selected from 14 clinical variables by LASSO regression, and Model I was established, including the following characteristics: height (>36 cm), head circumference (≤29.49 cm), total serum calcium (Ca) (>2.13 mmol/L), and alkaline phosphatase (ALP) (>344 U/L) levels. A single predictor, the ALP level (>344 U/L), was used to establish Model II. The AUROC values of the two models were 0.959 for Model I and 0.929 for Model II. In conclusion, in this study, two diagnostic models of MBDP were developed using four combinations of predictors and ALP as a single predictor. Both models showed good sensitivity and specificity for the early diagnosis of metabolic bone disease (MBD), and an ALP level of 344 U/L was defined as a simple and effective diagnostic threshold. In future studies, using larger samples, diagnostic threshold values of ALP for premature infants of different ages should be established, and internal and external validations are needed to improve the adaptability of the current model., Competing Interests: Declaration of competing interest There are no conflicts of interest to declare., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Heparanase has been identified as a universal tumor-associated antigen, but heparanase epitope peptides are difficult to recognize. Therefore, it is necessary to explore novel strategies to ensure efficient delivery to antigen-presenting cells. Here, we established a novel immunotherapy model targeting antigens to dendritic cell (DC) receptors using a combination of heparanase CD4 + and CD8 + T-cell epitope peptides to achieve an efficient cytotoxic T-cell response, which was associated with strong activation of DCs. First, pegylated poly(lactic-coglycolic acid) (PLGA) nanoparticles (NPs) were used to encapsulate a combined heparanase CD4 + and CD8 + T-cell epitope alone or in combination with Toll-like receptor 3 and 7 ligands as a model antigen to enhance immunogenicity. The ligands were then targeted to DC cell-surface molecules using a DEC-205 antibody. The binding and internalization of these PLGA NPs and the activation of DCs, the T-cell response and the tumor-killing effect were assessed. The results showed that PLGA NPs encapsulating epitope peptides (mHpa399 + mHpa519) could be targeted to and internalized by DCs more efficiently, stimulating higher levels of IL-12 production, T-cell proliferation and IFN-γ production by T cells in vitro. Moreover, vaccination with DEC-205-targeted PLGA NPs encapsulating combined epitope peptides exhibited higher tumor-killing efficacy both in vitro and in vivo. In conclusion, delivery of PLGA NP vaccines targeting DEC-205 based on heparanase CD4 + and CD8 + T-cell epitopes are suitable immunogens for antitumor immunotherapy and have promising potential for clinical applications., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Serological tumor markers are useful for the detection of malignancies and evaluation of disease progression. These markers are not checked as part of a routine examination for patients with benign diseases and without any clinical suspicion of malignancy. However, some markers appear to be elevated in patients with benign diseases and without malignancies. We present a case of pyogenic liver abscesses with an elevated serum carcinoembryonic antigen (CEA) level associated with neither evidence of malignancy nor elevation of other tumor markers such as carbohydrate antigen (CA 19-9) and alpha-fetoprotein (AFP) levels. The serological level of CEA decreased and subsequently became within normal limits with treatment. This case also demonstrates that diabetic patients with a liver abscess may present with no infectious symptoms and that fine-needle aspiration is as effective as catheter drainage in the treatment of pyogenic liver abscess., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Medical Ultrasound.)

Objective: Alkaline phosphatase (ALP) is a ubiquitous enzyme in humans that can be used for diagnosing childhood diseases. Infants have the highest rapid growth rate and are susceptible to metabolic bone diseases. In infants, ALP activities exhibit significant month-wise variations, and authoritative standards are lacking. The present study aimed to provide a reference for the diagnosis of diseases related to abnormal ALP activities in infants., Methods: This study included 24,618 samples collected from infants aged 0-12 months from three medical centers in Chongqing, China. Samples of infants diagnosed with diseases that may affect ALP activity have been exclude. ALP activity was analyzed using an automatic biochemical analyzer. A percentile curve for ALP activity in male and female infants was constructed using MATLAB, and the skewness-median-coefficient of variation method was employed for curve fitting., Results: ALP activity in male and female infants peaked at 0-4 months; the peak appeared at 1-2 months and declined gradually thereafter. After 4-5 months of age, the ALP activities declined further, with the lowest values observed at 11-12 months of age. A comparison between the data from this study and a those from a published German study indicates that Chinese infants exhibited peak ALP activity later and subsequent decline greater than German infants., Conclusions: A percentile curve was constructed for month-wise ALP activity in male and female infants, which could provide a reference for diagnosing diseases related to abnormal ALP activity in infants., (Copyright © 2021. Published by Elsevier Inc.)

Background & Aims: Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease worldwide. The NLRP3 inflammasome, a platform for caspase-1 activation and release of interleukin 1β, is increasingly recognized in the induction of inflammation and liver fibrosis during NAFLD. However, the cell-specific contribution of NLRP3 inflammasome activation in NAFLD remains unknown., Methods: To investigate the role of NLRP3 inflammasome activation in hepatocytes, hepatic stellate cells (HSCs) and myeloid cells, a conditional Nlrp3 knock-out mouse was generated and bred to cell-specific Cre mice. Both acute and chronic liver injury models were used: lipopolysaccharide/adenosine-triphosphate to induce in vivo NLRP3 activation, choline-deficient, L-amino acid-defined high-fat diet, and Western-type diet to induce fibrotic nonalcoholic steatohepatitis (NASH). In vitro co-culture studies were performed to dissect the crosstalk between myeloid cells and HSCs., Results: Myeloid-specific deletion of Nlrp3 blunted the systemic and hepatic increase in interleukin 1β induced by lipopolysaccharide/adenosine-triphosphate injection. In the choline-deficient, L-amino acid-defined high-fat diet model of fibrotic NASH, myeloid-specific Nlrp3 knock-out but not hepatocyte- or HSC-specific knock-out mice showed significant reduction in inflammation independent of steatosis development. Moreover, myeloid-specific Nlrp3 knock-out mice showed ameliorated liver fibrosis and decreased HSC activation. These results were validated in the Western-type diet model. In vitro co-cultured studies with human cell lines demonstrated that HSC can be activated by inflammasome stimulation in monocytes, and this effect was significantly reduced if NLRP3 was downregulated in monocytes., Conclusions: The study provides new insights in the cell-specific role of NLRP3 in liver inflammation and fibrosis. NLRP3 inflammasome activation in myeloid cells was identified as crucial for the progression of NAFLD to fibrotic NASH. These results may have implications for the development of cell-specific strategies for modulation of NLRP3 activation for treatment of fibrotic NASH., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Talaromyce marneffei is an important thermally dimorphic pathogen causing disseminated mycoses in immunocompromised individuals in southeast Asia. Previous studies have suggested that NLRP3 inflammasome plays a critical role in antifungal immunity. However, the mechanism underlying the role of NLRP3 inflammasome activation in host defense against T. marneffei remains unclear. We show that T. marneffei yeasts but not conidia induce potent IL-1β production. The IL-1β response to T. marneffei yeasts is differently regulated in different cell types; T. marneffei yeasts alone are able to induce IL-1β production in human PBMCs and monocytes, whereas LPS priming is essential for IL-1β response to yeasts. We also find that Dectin-1/Syk signaling pathway mediates pro-IL-1β production, and NLRP3-ASC-caspase-1 inflammasome is assembled to trigger the processing of pro-IL-1β into IL-1β. In vivo , mice deficient in NLRP3 or caspase-1 exhibit higher mortality rate and fungal load compared to wild-type mice after systemic T. marneffei infection, which correlates with the diminished recruitment of CD4 T cells into granulomas in knockout mice. Thus, our study first demonstrates that NLRP3 inflammasome contributes to host defense against T. marneffei infection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ma, Chan, Tan, Kui, Tsang, Pei, Lau, Woo and Lee.)

Background: This study aimed to evaluate the correlation and consistency between traditional head measurement and structured light three-dimensional (3D) scanning parameters when measuring infant skull shape., Methods: A total of 76 infants aged 3 months to 2.5 years old were included in the study. Head circumference (HC) was measured with a tape measure. The transverse, anteroposterior, and oblique diameters were measured using a spreading caliper, and the cranial vault asymmetry index (CVAI) and a cranial index (CI) of symmetry were calculated; 76 cases were measured successfully. The above indexes were measured using a structured light 3D scanning system (71 cases were measured with success). Thus, in the end, the valid data of 71 cases were analyzed, and the measurements of the two approaches were compared., Results: The 95% confidence interval of traditional head measurement and structured light 3D scanning was between 0.633 and 0.988. Pearson's correlation coefficient indicated a high correlation between the two methods (r=0.793-0.980). The correlation coefficients of the transverse diameter, anteroposterior diameter, and HC, and the CI of symmetry were higher than 0.9. The lowest correlation coefficient for the CVAI was 0.793. The P values of the above measurement data were all <0.001, which indicated that they were closely related. A Bland-Altman plot indicated reasonable consistency between the two methods., Conclusions: Both traditional head measurement and structured light 3D scanning are suitable for the measurement of infant head shape. However, while traditional head measurement using a spreading caliper is economical and simple, making it suitable for general screening at a basic level, structured light 3D scanning can deliver additional parameters, which is useful for infants with an abnormal head shape. The latter is also convenient for designing a customized helmet for skull correction when needed., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://dx.doi.org/10.21037/tp-21-186). The authors have no conflicts of interest to declare., (2021 Translational Pediatrics. All rights reserved.)

PDB-1 is a new C-27-carboxylated-lupane-triterpenoid derivative isolated from Potentilla discolor Bunge. In our previous research, PDB-1 was suggested to have an obvious selectivity for tumor cells. This study focused on clarifying PDB-1's anticancer mechanism in the inhibition of proliferation and in the induction of apoptosis and autophagy in A549 cells. In general, A549 cells were treated with PDB-1 for different times, and cell survival was assessed by a CCK8 assay. The assessment of intracellular reactive oxygen species, a mitochondrial membrane potential assay, a cell cycle assay, an annexin V-FITC/PI assay, and MDC staining were performed in A549 cells treated with PDB-1. Moreover, the mRNA and protein expression of cell cycle-, apoptosis- and autophagy-related factors were detected by RT-qPCR and western blotting. The results showed that PDB-1 inhibited A549 cell proliferation and colony formation in a dose- and time-dependent manner. The decrease in the viability of A549 cells was due to a G2/M cell cycle arrest. Moreover, PDB-1 induced cell apoptosis, accompanied by an increase in the Bax/Bcl-2 ratio and an increase in the expression levels of cleaved caspase-3/caspase-9. We also found that PDB-1 induced autophagy by increasing the conversion of LC3-I to LC3-II and elevating Beclin-1. In addition, further studies indicated that pretreatment with a specific PI3K inhibitor (LY294002) enhanced the effects of PDB-1 on the expression of proteins associated with apoptosis and autophagy, demonstrating that the PI3K/Akt/mTOR pathway was related to PDB-1-induced apoptosis and autophagy. These results indicated that PDB-1 may be considered a potential candidate for the future treatment of lung adenocarcinoma. These findings should benefit the development of the C 14 -COOH type of pentacyclic triterpenoids., (Copyright © 2020. Published by Elsevier Masson SAS.)

Aims: Preeclampsia (PE) accounts for the foremost cause of maternal and fetal mortality worldwide, whereas, there are no effective treatments for the disease yet. Long non-coding RNAs (lncRNAs) play critical roles in various human disorders, including PE. Here, we identified an up-regulated lncRNA HOTAIR, and explored its underlying mechanisms in PE., Main Methods: qRT-PCR analysis was used to examine HOTAIR expression in PE tissues and cell lines. Trophoblast proliferation was examined by colony formation and 5-Ethynyl-2'-deoxyuridine (EdU) incorporation assays. Trophoblast migration and invasion was determined by transwell and wound healing assays. Bioinformatics analysis was performed to verify the regulation HOTAIR on miRNAs. The interaction between HOTAIR and EZH2 was detected using RNA immunoprecipitation assay (RIP). Chromatin immunoprecipitation (CHIP) assay was also performed to verify that the negative regulation of HOTAIR on miR-106a was dependent on the epigenetic repressor EZH2., Key Findings: HOTAIR was up-regulated in PE placenta tissues, which repressed the proliferation, migration and invasion of trophoblast cells. HOTAIR significantly repressed miR-106a expression and the reduced miR-106a level was also observed in placentas from PE patients. Additionally, miR-106a mimic enhanced the migration and invasion of trophoblast cells. Further mechanistic analyses implied that the action of HOTAIR is moderately attributable to its repression of miR-106a via association with EZH2., Significance: High level of HOTAIR repressed the proliferation, migration and invasion of trophoblast cells through targeting miR-106 in an EZH2-dependent manner, which may provide new insights into the roles of HOTAIR and miR-106a as potential regulators in PE., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Objective: Alkaline phosphatase (ALP) serves as a biomarker for diagnosing several types of diseases in adults; nonetheless, its use is restricted in children because of changes in ALP activity during different physiological phases. The present study aimed to investigate ALP activity and its dynamics in children of different ages to establish the reference values for ALP activity in children., Methods: Total 167,625 samples of children aged 0-18 years were enrolled in this study. ALP activity was measured using the 4-nitrophenyl-1-phosphate disodium salt (4-NPP)-2-amino-2-methyl-1-propanol (AMP) method with an automatic biochemical analyzer. Patients with known diagnoses that may affect ALP activity were excluded. A percentile curve was plotted using MATLAB software, and the curve was fitted using the skewness-median-coefficient of variation (LMS) method., Results: ALP activity reached the highest peak at 12-13 years of age and then gradually decreased to the lowest peak at 18-19 years of age in boys, whereas it reached the highest at 10-11 years and then gradually reduced to the lowest at 17-18 years in girls. Furthermore, the highest peak of ALP activity appeared substantially earlier in children of either sex in China than in those in Germany., Conclusions: We showed the dynamics of ALP activity in both boys and girls between the ages of 0 and 18 years in China and compared the difference in ALP activity between children in China and Germany. Our findings provide a reference for clinicians., (Copyright © 2020 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

MicroRNA‑21 (miR‑21) is reported to exhibit cancer‑promoting activity in various types of cancer. It has been previously demonstrated that miR‑21 is overexpressed in bladder tumor tissue compared with normal mucosa. However, the functional mechanism of miR‑21 in bladder cancer remains largely unknown. Thus, the current study aimed to determine the roles of miR‑21 in autophagy and the malignant development of bladder cancer in T24 cells. Upregulation or downregulation of miR‑21 was achieved following the transfection of miR‑21 mimic or miR‑21 inhibitor. An MTT assay was additionally performed to measure cell growth. Wound healing and transwell invasion assays were used to detect cell migration and invasion. The apoptotic potential and cell cycle were examined via flow cytometry and reverse transcription‑quantitative PCR was performed to evaluate the expression of phosphatase and tensin homolog (PTEN), beclin 1, microtubule‑associated protein l light chain 3B (LC3‑II), cyclin D1, caspase‑3, E‑cadherin, matrix metallopeptidase‑9 (MMP‑9) and vimentin. The results revealed that the proliferation, migration and invasion of T24 cells was greatly increased in the miR‑21 mimic group, while apoptosis was greatly inhibited. Additionally, T24 cells treated with miR‑21 mimic exhibited G1‑phase arrest. In the miR‑21 mimic group, the expression of PTEN, beclin 1, LC3‑II, caspase‑3 and E‑cadherin were decreased, while the expression of cyclin D1, MMP‑9 and vimentin were increased. Opposite effects were observed in the miR‑21 inhibitor group. The data of the current study may indicate that miR‑21 overexpression inhibited autophagy and promoted the proliferation, migration, invasion and epithelial to mesenchymal transition of bladder cancer T24 cells. The results may further elucidate the molecular mechanism of miR‑21 in the development of bladder cancer.

Talaromyces (Penicillium) marneffei is an AIDS-defining infection in Southeast Asia and is associated with high mortality. It is rare in non-immunosuppressed individuals, especially children. Little is known about host immune response and genetic susceptibility to this endemic fungus. Genetic defects in the interferon-gamma (IFN-γ)/STAT1 signaling pathway, CD40/CD40 ligand- and IL12/IL12-receptor-mediated crosstalk between phagocytes and T-cells, and STAT3-mediated Th17 differentiation have been reported in HIV-negative children with talaromycosis and other endemic mycoses such as histoplasmosis, coccidioidomycosis, and paracoccidioidomycosis. There is a need to design a diagnostic algorithm to evaluate such patients. In this article, we review a cohort of pediatric patients with disseminated talaromycosis referred to the Asian Primary Immunodeficiency Network for genetic diagnosis of PID. Using these illustrative cases, we propose a diagnostics pipeline that begins with immunoglobulin pattern (IgG, IgA, IgM, and IgE) and enumeration of lymphocyte subpopulations (T-, B-, and NK-cells). The former could provide clues for hyper-IgM syndrome and hyper-IgE syndrome. Flow cytometric evaluation of CD40L expression should be performed for patients suspected to have X-linked hyper-IgM syndrome. Defects in interferon-mediated JAK-STAT signaling are evaluated by STAT1 phosphorylation studies by flow cytometry. STAT1 hyperphosphorylation in response to IFN-α or IFN-γ and delayed dephosphorylation is diagnostic for gain-of-function STAT1 disorder, while absent STAT1 phosphorylation in response to IFN-γ but normal response to IFN-α is suggestive of IFN-γ receptor deficiency. This simple and rapid diagnostic algorithm will be useful in guiding genetic studies for patients with disseminated talaromycosis requiring immunological investigations., (Copyright © 2019 Lee, Lao-araya, Yang, Chan, Ma, Pei, Kui, Mao, Yang, Zhao, Trakultivakorn and Lau.)

We developed a facile synthesis for a series of multifunctionalized polyesters by Passerini three-component polymerization (Passerini-3CP) in a "one-pot" method at room temperature using serial dicarboxylic acids, dialdehyde, and tert -butyl isocyanide as monomers. First, the effects of monomer feed ratio, monomer concentration, and different dicarboxylic acids involved in the polymerization were systematically investigated. The in situ FTIR and GPC measurements have suggested a step-growth mechanism for Passerini-3CP. Second, five succinic acid end-capped polyethylene glycols (S-PEGs) with different molecular weights of 400, 800, 1000, 2000, and 4000 g/mol were prepared and selected as dicarboxylic acids for the subsequent Passerini-3CP to fabricate the thermosensitive and biocompatible polyesters. Among the five resulting polyesters, four polyesters from S-PEG-400, S-PEG-800, S-PEG-1000, and S-PEG-2000 show reversible response to the external temperature, and the lower critical solution temperature (LCST) in water is in the range of 28.5-84.2 °C. Through the copolymerization of S-PEG-400 and S-PEG-800, the LCSTs for functional polyesters can be conveniently controlled to be 38.7, 42.3, and 58.0 °C, respectively. After 24-72 h of incubation in polyester solution, the viability rate of HeLa cells reached up to 80-107%, showing its excellent biocompatibility. The cleavable polyesters were also prepared by integrating S-S bonds onto their backbones in Passerini-3CP of 3,3'-dithiodipropionic acid as one comonomer for the biomedical applications. With the aid of the hydrophobicity of doxorubicin (DOX) and thermosensitivity of polyesters, the doxorubicin-loaded carriers with the size of 200-400 nm and core-shell structure were easily obtained by dialysis below LCST and subsequent heating to LCST. The effective release of DOX from the carriers can be triggered by the characteristic reaction of l-glutathione (GSH) with S-S bonds in the functionalized polyester backbones.

Background: Implantation of nonabsorbable polypropylene (PP) mesh in the vagina is the main surgical treatment for pelvic organ prolapse (POP); however, clinical outcomes remain controversial and far from satisfactory. In particular, reducing the exposure or erosion of vaginal implants to obtain improved functional reconstruction is challenging. There is an urgent need for the development of new materials and/or products for POP treatment. A nanofibrous biomimetic mesh was recently developed to address this issue., Objective: In this study, the basic properties of the newly developed mesh, including structural characteristics, mechanical properties, biological response of human umbilical cord mesenchymal stem cells in vitro, and tissue regeneration and biocompatibility in vivo, were evaluated and compared with those of Gynemesh™PS., Methods: Scanning electron microscopy and uniaxial tensile methods were used to evaluate microstructure and mechanical properties, respectively. Mesenchymal stem cell growth on the meshes was observed by fluorescence microscopy to visualize the expression of enhanced red fluorescent protein. Twenty-four mature female Sprague Dawley rats were randomly assigned to two groups: group 1 (nanofibrous biomimetic mesh, Medprin, Germany, n=12) and group 2 (Gynemesh(TM)PS, Ethicon, USA; n=12). The posterior vaginal wall was incised from the introitus, and the mesh was then implanted. Three implants of each type were tested for 1, 4, 8 and 12 weeks. Connective tissue organization, inflammation, vascularization, and regenerated tissue were histologically assessed., Results: The nanofibrous biomimetic mesh is a relatively heavy material and exhibited lower porosity than Gynemesh(TM)PS. The new mesh was stiffer than Gynemesh(TM)PS (p<0.001) but supported human umbilical cord mesenchymal stem cell attachment. Erosion of the grafts did not occur in any animal. The nanofibrous biomimetic mesh was encapsulated by a thicker layer of connective tissue and was associated with significantly greater inflammatory scores compared with Gynemesh(TM)PS. At 12 weeks, the vascularization of the new mesh was greater than that of Gynemesh(TM)PS (p<0.05). No significant difference in the thickness of the smooth muscle layer following implantation was observed between the two groups (p>0.05)., Conclusions: The nanofibrous biomimetic mesh is a candidate for reinforcing pelvic reconstruction. The mesh could be improved by decreasing its weight and stiffness and increasing its porosity. This mesh could serve as a carrier for stem cells in future regenerative medicine and tissue engineering research., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The innate immune system senses cytosolic dsDNA and bacterial cyclic dinucleotides and initiates signaling via the adaptor STING to induce type 1 interferon (IFN) response. We demonstrate here that BTK-deficient cells have impaired IFN-β production and TBK1/IRF3 activation when stimulated with agonists or infected with pathogens that activate STING signaling. BTK interacts with STING and DDX41 helicase. The kinase and SH3/SH2 interaction domains of BTK bind, respectively, the DEAD-box domain of DDX41 and transmembrane region of STING. BTK phosphorylates DDX41, and its kinase activities are critical for STING-mediated IFN-β production. We show that Tyr364 and Tyr414 of DDX41 are critical for its recognition of AT-rich DNA and binding to STING, and tandem mass spectrometry identifies Tyr414 as the BTK phosphorylation site. Modeling studies further indicate that phospho-Tyr414 strengthens DDX41's interaction with STING. Hence, BTK plays a critical role in the activation of DDX41 helicase and STING signaling., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: We report here a rare case of anterior superior iliac spine avulsion fracture that presented initially as meralgia paraesthetica., Case Report: A 14-year-old male sprinter presented with anterior superior iliac spine avulsion fracture, which was not observed on initial plain radiograph of the hip, but was diagnosed by ultrasound. Both clinical presentations and electrophysiological studies indicated meralgia paraesthetica. The lateral femoral cutaneous nerve of the thigh was probably compressed by an inguinal haematoma resulting from sartorius muscle strain, which was detected on musculoskeletal ultrasound. Computed tomography of the pelvis confirmed anterior superior iliac spine avulsion fracture., Conclusion: Meralgia paraesthetica in adolescents can be due to anterior superior iliac spine avulsion fracture. Sonography is a valuable tool for screening for muscular haematoma and occult fractures, which may allow clinicians to diagnose the nature of the muscle injury, and thus guide the most appropriate therapeutic strategy.

Despite the improved survival rate among systemic lupus erythematosus (SLE) patients, there are many factors associated with the mortality of SLE. In the current study, death-related factors of patients associated with course of disease were surveyed. Retrospective study was used. Mortalities among these three groups (group A, B and C, the course of disease was ≤ 5 years, 5-10 years and > 10 years, respectively) were calculated and compared. Various factors related to mortality were analyzed. Male SLE patients died relatively more than female patients. The total mortality was 8.5 %. The mortalities were significant difference in group A, B and C which were 9.4, 4.8 and 8.9 %, respectively. The mortalities of group A and group C were significantly higher than that of group B, but there was no significant difference between mortalities of group A and group C. The most common death-related factor was infection, followed by involved disorders in renal, brain, multisystem, heart, etc. The mortalities resulted from neuropsychiatric systemic lupus erythematosus (NPSLE), pulmonary infection, involved digestive system and hematological system were significantly different between three groups. There was no difference between mortalities of group A and group C associated with pulmonary infection and NPSLE. Patients in group C died more than in group A from involved renal, heart, multisystem, etc, while group A had more patients than group C who died of pulmonary infection, involved hematological system. In conclusion, Male SLE patients have worse outcome than female patients. Infection and active SLE are not only contributors to the death of early stage patients, but also to that of later stage patients.

Background: Chinese primary care settings have a heavy patient load, shortage of physicians, limited medical resources and low medical literacy, making it difficult to screen for developmental disorders in infants. The Infant Neurological International Battery (INFANIB) for the assessment of neuromotor developmental disorders in infants aged 0 ~ 18 months is widely applied in community health service centers because of its simplicity, time-saving advantages and short learning curve. We aimed to develop and assess a Chinese version of the INFANIB., Methods: A Chinese version of the INFANIB was developed. Fifty-five preterm and 49 full-term infants with high risk of neurodevelopmental delays were assessed using the Chinese version of the INFANIB at 3, 7 and 10 months after birth. The Peabody Developmental Motor Scale (PDMS) was simultaneously used to assess the children with abnormalities and diagnose cerebral palsy. The sensitivity, specificity, positive predictive value and negative predictive value of the scale were calculated., Results: At birth, a higher proportion of full-term infants had asphyxia (p < 0.001), brain damage ( p = 0.003) and hyperbilirubinemia ( p = 0.022). The interclass correlation coefficient and intraclass correlation coefficient values for the INFANIB at 3, 7 and 10 months were >0.8, indicating excellent reliability with regard to inter- and intraobserver differences. The specificity, sensitivity, positive predictive value and negative predictive value were high for both high-risk premature infants and full-term infants at the age of 10 months. For premature infants at the age of 7 months or below, INFANIB had low validity for detecting abnormalities., Conclusions: The Chinese version of the INFANIB can be useful for screening infants with high-risk for neuromotor abnormality in Chinese primary care settings.

The neuromedin B receptor (Nmbr) is an important physiological regulator of spontaneous activities and stress responses through different cascades as well as its autocrine and paracrine effects. Previous studies have revealed that neuromedin B (Nmb) and its receptor signal via the Rela (also known as p65)/Il6 pathway in a mouse model of pregnancy. This study investigated the mechanism of Nmbr signaling via the Rela/p65-Il6 pathway and regulation of the concentration of intracellular free calcium ([Ca(2+)](i)) during the onset of labor in primary mouse myometrial cell cultures isolated from mice in term labor. Data demonstrated Nmbr agonist-mediated upregulation of the DNA binding activity of Rela/p65, Il6 expression, and [Ca(2+)](i) in a concentration-dependent manner. Furthermore, a significant correlation was observed between DNA binding activity of Rela/p65 and Il6 expression. Moreover, this up-regulation was blocked by Nmbr and Rela/p65 knockdown, achieved by RNA interference (RNAi) technology. No significant differences were identified in the inhibition of Il6 expression as a result of Nmbr or Rela/p65 knockdown. However, significant differences were observed between the [Ca(2+)](i) in Rela/p65-specific group and that in the Nmbr-specific small interfering RNA (siRNA)-treated groups. These data demonstrated that the Nmb/Nmbr interaction in pregnant myometrial primary cells in vitro predominantly influenced uterine activity through regulation of Il6 expression via the Rela/p65 pathway, although the effects of Nmbr on [Ca(2+)](i) involved several pathways that remain to be elucidated.

The biomedical applications of poly(l-lactide) (PLLA) were limited by its high crystallinity. In this paper, the copolymerization of trimethylene carbonate (TMC) and l-lactide (LLA) was carried out to improve the flexibility of PLLA. The effects of feeding dose, reaction temperature and polymerization time were investigated, and the copolymers were characterized with (1)H nuclear magnetic resonance, Fourier transform infrared reflection, gel permeation chromatography differential scanning calorimetry, thermogravimetric analysis and x-ray diffraction. The copolymers were electrospun to form porous films to study their cell compatibility. The results showed that the composition of the copolymer was nearly the same as that in the feeding dose, and the molecular weight of the copolymer decreased with increasing TMC content. The decrease in the reaction temperature and polymerization time would increase the molecular weight, but the composition deviates from the feeding dose. NIH/3T3 mouse fibroblast cells were cultured on the electrospun films. The morphology and proliferation of the cells were studied. The results implied that the cell compatibility of poly(l-lactide-co-trimethylene carbonate) copolymer was much better than that of the PLLA homopolymer.

Inflammatory myofibroblastic tumor (IMT) is a very rare benign tumor composed of myofibroblastic spindle cells of uncertain etiology, which can occur at any age and affect any organ system. More and more cases of IMT in children have been described in pediatric literature in recent years. However, this tumor occurring intraabdominally in children has rarely been reported in Taiwan. Here we present a 1-year-9-month-old boy who had fever and abdominal pain only for 2 days, symptoms mimicking acute abdomen. After imaging study, a huge tumor nearly 10 cm in diameter was incidentally found over the right abdomen with unknown origin and nature. After surgical removal of the tumor, IMT was confirmed by the pathological findings. It is very difficult to make an accurate preoperative diagnosis on this tumor according to past experience, so the role of pathological diagnosis with immunohistochemical study becomes important. This case illustrates that IMT should be considered as a possible cause of intra-abdominal mass in children who have fever of unknown origin.

Objective: To study the clinical features of juvenile primary fibromyalgia syndrome (FMS) and to evaluate outcome after treatment., Methods: Six patients with juvenile primary FMS were registered in department of rheumatology and their clinical data were assessed, including degree of pain (visual analog scale, VAS), fatigue, depression, anxiety, sleep disturbances, arthrodynia, subjective joint swelling, abdominal pain, irritable bowel symptoms, urinary urgency, dysmenorrhea, morning stiffness, paresthesias, illness changes with weather, feeling worse with exercise, laboratory examination and outcome of treatment., Results: Abdominal pain was the first symptom in 5 of the cases with juvenile primary FMS, diffuse aching and left knee pain were the first symptoms in one patient. All the 6 patients were misdiagnosed prior to their rheumatological evaluation. Diffuse aching, fatigue, sleep disturbances, illness changes with weather and feeling worse with exercise existed in all the 6 patients (100%), the mean pain score was 8.8 and the mean initial tender points (TP) count was 13.7. Arthrodynia, subjective joint swelling, abdominal pain, irritable bowel symptoms and urinary urgency existed in 5 of the 6 patients (83%). Dysmenorrhea existed in 4 (67%), depression in 3 (50%), morning stiffness in 2 (33%), paresthesias in 2 (33%) and anxiety in 2 (33%), respectively. The results of laboratory examination were normal and the outcomes of treatment were good., Conclusion: Juvenile primary FMS may not be a rare disease and the clinicians should pay more attention to it for avoiding misdiagnosis.