Neuroinflammation is a critical mechanism underlying various forms of sensorineural hearing loss (SNHL). Contrary to the conventional view that cochlea is an immune-privileged organ shielded by the blood-labyrinth barrier, it is now evident that it hosts a diverse immune landscape dominated by cochlear-resident macrophages (CoRMs). These CoRMs share a common developmental origin with central nervous system microglia. Despite this phylogenetic homology, CoRMs have evolved distinct phenotypic and functional profiles to adapt to the unique peripheral sensory environment of cochlea. Recent single-cell RNA sequencing studies have further shown a remarkable spatiotemporal heterogeneity among CoRMs subsets, which are strategically distributed across specific anatomical niches, such as the basilar membrane, stria vascularis, spiral ligament, osseous spiral lamina, spiral ganglions, and spiral limbus.​ This spatial and functional diversification may underlie their dual capacity to act as sentinels that maintain cochlear homeostasis and promote repair or as saboteurs that propagate neuroinflammatory injury. However, the functional diversity of CoRMs across regional compartments and the impact of dynamic subpopulation shifts on hearing loss progression remain poorly defined, representing a significant knowledge gap. This review systematically addresses this gap by synthesizing the developmental origins, spatial distribution, and functional heterogeneity of CoRMs. By integrating cutting-edge single-cell transcriptomics and fate-mapping studies, we elucidated the subset-specific roles within cochlear niches, proposing a dual guardian-saboteur paradigm. Furthermore, we explore novel macrophage-targeted therapeutic strategies—including pharmacological modulators, genetic interventions, and biomaterial engineering, designed to precisely regulate CoRMs function for hearing preservation. Our study provides a mechanism-oriented framework that bridges fundamental neuroimmunology with translational applications, offering insights into macrophage subpopulation functions and guiding future research toward targeted therapies for hearing disorders. Cochlear resident macrophages (CoRMs) are found in distinct anatomical locations, showing high spatiotemporal heterogeneity and performing divergent roles in auditory health and disorders. This graphical abstract illustrates the ontogenic origins and injury-response timelines of CoRMs (temporal heterogeneity), maps the niche-specific distribution of CoRM subsets across cochlear compartments (spatial heterogeneity), and highlights precision therapeutic strategies. By enhancing the protective functions of “Sentinel” subsets or inhibiting the inflammatory damage of “Saboteur” subsets, these approaches aim to prevent or halt irreversible hearing loss.

Background: Type 1 diabetes (T1D) is an autoimmune disorder characterized by destruction of insulin-producing β-cells. While conventional insulin therapy manages hyperglycemia, it fails to halt autoimmunity. Oral immunotherapy targeting autoantigens like GAD65 offers potential for antigen-specific tolerance; however, its efficacy is limited by gastrointestinal degradation and poor mucosal uptake. Lactococcus lactis (L. lactis), a food-grade delivery vector, enables sustained antigen release and intestinal tract immune modulation, yet the differential transcriptomic mechanisms underlying mucosal versus systemic immune responses remain uncharacterized. Methods: Non-obese diabetic (NOD) mice were randomized into control and GAD65 groups, receiving oral PBS or the GAD65 recombinant L. lactis vaccine, respectively. Fasting blood glucose was monitored weekly. GAD65-specific IgA and IgG, along with immune tolerance-related factors, were quantified using ELISA. Lymphocyte subsets were analyzed by flow cytometry, alongside RNA sequencing and transcriptional profiling. Results: The study demonstrated that the orally administered GAD65-L. lactis vaccine could significantly induce GAD65-specific IgA antibody and TGF-β cytokine and alleviate hyperglycemia and diabetes symptoms in NOD mice. Our study facilitated the induction of GAD65-specific regulatory T cells within both intestinal lamina propria lymphocytes (LPLs) and splenic lymphocytes. Notably, antigen-specific tolerance was mainly observed in intestinal LPLs. Crucially, the immune responses elicited by the vaccine demonstrated significant disparities between intestinal LPLs and splenic lymphocytes, with intestinal LPLs exhibiting unique local immune tolerance transcriptomic profiles. Conclusions: Our findings have enhanced the comprehension of the mechanisms by which oral vaccines influence the interplay between mucosal and systemic immune responses, thereby establishing a foundational framework for the design of oral vaccines. This understanding is instrumental in advancing antigen-specific immune tolerance strategies for autoimmune diseases such as Type 1 Diabetes (T1D).

Background: Our previous study found dementia as a significant risk factor for delirium development in elderly patients with hip fracture. However, the causal relationship between dementia and delirium remains unclear. Methods: To assess the causal relationship between delirium and dementia, we conducted a bidirectional Mendelian randomization (MR) analysis. Inversevariance weighted (IVW), weighted median, MR Egger, weighted mode, and simple mode were employed to conduct the MR analysis. Heterogeneity was assessed using the Cochran Q statistic in MR-Egger and IVW methods. Horizontal pleiotropy was examined via the MR pleiotropy residual sum and outliers (MR-PRESSO) and MR-Egger intercept tests. Results: The forward MR analysis revealed a significant association between unclassified dementia (1.604 (1.326-1.941), p = 1.12 x 10(-6)), Alzheimer's disease (1.259 (1.128-1.405), p = 4.10 x 10(-5)), and dementia with Lewy bodies (1.121 (1.026-1.225), p = 0.011) with an increased risk of delirium. In the reverse MR analysis, delirium was also suggested to increase the risk of unclassified dementia (1.133 (1.066-1.204), p = 6.31 x 10(-5)) and vascular dementia (1.246 (1.075-1.444), p = 0.003). These significant results were further validated in the multivariable MR analysis. No evidence of heterogeneity or horizontal pleiotropy was observed (p > 0.05). Limitations: (1) Limited to European populations. (2) Sample population overlap between delirium and dementia. (3) Not all dementia subtypes were causally associated with delirium. Conclusions: This study provides genetic evidence supporting a causal relationship between dementia and delirium, indicating that dementia may influence the risk of delirium while delirium may also increase the risk of dementia.

An imbalance between M1 and M2 macrophage polarization is critical in osteoarthritis (OA) development. We investigated the effect of M2 macrophage-derived extracellular vesicles (M2-EVs) to reprogramme macrophages from the M1 to M2 phenotype for OA treatment. M1 macrophages and mouse OA models were treated with M2-EVs. Proteomic analysis was performed to evaluate macrophage polarization in vitro. The OA models were as follows: destabilization of the medial meniscus (DMM) surgery-induced OA and collagenase-induced OA (CIOA). Hyaluronic acid (HA) was used to deliver M2-EVs. M2-EVs decreased macrophage accumulation, repolarized macrophages from the M1 to M2 phenotype, mitigated synovitis, reduced cartilage degradation, alleviated subchondral bone damage, and improved gait abnormalities in the CIOA and DMM models. Moreover, HA increased the retention time of M2-EVs and enhanced the efficiency of M2-EVs in OA treatment. Furthermore, proteomic analysis demonstrated that M2-EVs exhibited a macrophage reprogramming ability similar to IL-4, and the pathways might be the NOD-like receptor (NLR), TNF, NF-κB, and Toll-like receptor (TLR) signaling pathways. M2-EVs reprogrammed macrophages from the M1 to M2 phenotype, which resulted in beneficial effects on cartilage and attenuation of OA severity. In summary, our study indicated that M2-EV-guided reprogramming of macrophages is a promising treatment strategy for OA.

Background Rheumatoid arthritis (RA) is a common autoimmune inflammatory disease. Currently, a complete cure for RA is still unavailable. Mendelian randomization (MR) has emerged as a valuable tool for identifying potential therapeutic targets or drug repurposing opportunities for certain diseases. Therefore, our aim was to identify novel effective targets or drug repurposing opportunities for RA and analyze their mechanisms and potential side effects. Methods A MR integrating the identified druggable genes was used to evaluate the causal effects of druggable gene cis-expression quantitative trait loci (cis-eQTLs) on RA, while additional RA cohort was employed for validation. Colocalization analysis was performed to determine the probability of shared causal variants between the identified targets and RA. The protein-protein interaction network analysis was conducted to explore associations between the identified druggable genes and current RA drug targets. The MR and colocalization analyses were used to assess the potential side effects of the identified targets in RA treatment. Results Nine druggable genes ( TYK2 , PTPN22 , ATP2A1 , APOM , RXRB , NOTCH4 , HLA-DRA , CCR6 , and CTLA4 ) showed significant MR results in both the training cohort ( p <1.99E-05) and validation cohort ( p <0.0025). Colocalization analysis indicated that cytotoxic T-lymphocyte-associated protein 4 (CTLA4) and RA (PP.H4.abf=0.98), as well as C-C motif chemokine receptor 6 (CCR6) and RA (PP.H4.abf=0.99), shared the same causal variant. Hence, these two genes were identified as the final therapeutic targets. Furthermore, CTLA4 and CCR6 interacted with the current RA drug targets. Subsequent MR analysis revealed that genetically proxied activation of CTLA4 and inhibition of CCR6 might decrease the risk of hypothyroidism but increase the risk of malignant skin neoplasm. Additionally, genetically proxied activation of CTLA4 may also reduce the risk of type 1 diabetes. Conclusions This study supports the idea that targeting the activation of CTLA4 and the inhibition of CCR6 may reduce the risk of RA with fewer side effects, and highlights the potential of CTLA4 and CCR6 as promising druggable targets for RA treatment.

Study ObjectiveTo explore how to improve the treatment, and prognosis of ovarian pregnancy (OP).PatientsA total of 111 OP patients, one of the patients suffered from OP twice.ResultsIn this study, 112 OP cases confirmed by postoperative pathology were retrospectively analyzed. Common risk factors for OP were previous abdominal surgery (39.29%) and intrauterine device use (18.75%). We modified the ultrasonic classification into four types: gestational sac type, hematoma type I, hematoma type II, and intraperitoneal hemorrhage type. Among these four types, the proportion of patients who underwent emergency surgery as initial treatment after admission was 68.75%, 10.00%, 92.00%, and 81.36%, respectively. The treatment for hematoma type I patients was often delayed. The rate of OP rupture was 86.61%. All methotrexate therapy for OP patients failed. All these 112 cases underwent surgery treatment finally. The surgical procedures were pregnancy ectomy and ovarian reconstruction by laparoscopy or laparotomy. No significant differences were observed in the operation time or intraoperative blood loss between laparoscopy and laparotomy. Laparoscopy showed less influence on patients regarding length of hospital stay and postoperative fever than laparotomy. Further, 49 patients who desired fertility were followed up over 3 years. Among them, 24 (48.98%) experienced spontaneous intrauterine pregnancy.ConclusionAmong the four modified ultrasonic classifications, hematoma type I was associated with more delays in surgical time. Laparoscopic surgery was a better choice for OP treatment. The reproductive prognosis of OP patients was promising.

Methods for the detection of adulterated milk are essential for assessing the quality of goat milk products. We hypothesized that goat milk oligosaccharides could provide a basis for this purpose and compared the levels of α3'-galactosyllactose (α3'-GL) and N-acetylhexaminyllactose (NHL) between goat milk and bovine milk oligosaccharides using reverse-phase high-performance liquid chromatography. The α3'-GL was detected to be three times more abundant in goat milk than in bovine milk, whereas NHL showed the opposite trend. Linear relationships were established between the relative proportions of α3'-GL and NHL levels for different ratios of bovine and goat milk, with a minimum detection limit of 2% bovine milk. The new method was validated by analyses of adulterants in eight commercially available goat dairy products. Overall, the degree of adulteration in goat milk products can be determined based on the relative proportions of α3'-GL and NHL.

In this study, we obtained all the homogeneous LBGP70 (arabinogalactan-protein complex), LBGP70-OL (sugar chain of LBGP70), and LBGP70-OL-I (backbone part of LBGP70-OL) fraction from crude Lycium barbarum polysaccharides (LBP) by stepwise precipitation. The structural features of LBGP70-OL were investigated by a series of analytical techniques and chemical methods, which was identified as a highly branched polysaccharide with an average of 9 branches per 10 sugar backbone units. The backbone was only →6)-β-Galp-(1→ residues, substituted at the C3 position. The side chains contained β-Galp-(1→, α-Araf-(1→, α-Araf-(1 → 3)-β-Araf-(1 → 3)-β-Araf-(1→, α-Araf-(1 → 3)-β-Gap-(1 → 3)-β-Galp-(1→, α-Araf-(1 → [5)-β-Araf-(1]4 → 3)-β-Galp-(1→. LBGP70-OL had the highest anti-ageing activity, with its side chain and backbone exhibiting a synergistic effect. LBGP70-OL exerted the anti-ageing activity by attenuating SA-β-Gal activity, preventing cell cycle arrest, increasing antioxidant enzyme activity, protecting the cell membranes from oxidative damage, and regulating the senescence-related genes Sirt1, NAMPT, and Prx1. These results lay the fundamental for further studies on the structure-function relationships of LBP.

Human milk oligosaccharides (HMOs) are the second most abundant carbohydrates in colostrum. In this study, we performed a quantitative analysis of 13 oligosaccharides in 99 colostrum samples obtained from mothers living in Northwest China. The analysis combined liquid chromatography-mass spectrometry (LC-MS) with 2-amino-N-(2-aminoethyl)benzamide (AEAB) labeling and nonsecretors accounted for 17%. Compared with healthy secretor mothers, those with gestational diabetes mellitus presented lower levels of sialylated oligosaccharides, especially 3'-sialyllactose. Colostrum from mothers with pregnancy-induced hypertension had higher levels of fucosylated oligosaccharides, but the difference was not significant, and hypothyroidism appeared to have no effect on HMOs. Most HMOs (especially 6'-sialyllactose) were more abundant in colostrum from mothers who underwent vaginal delivery than a C-section. These findings show that the concentration of total or individual HMOs is affected by multiple factors. These findings provide a reference for evaluating variations in HMO expression among different populations and potential guidance for providing personalized clinical nutrition.

Galactooligosaccharides are composed mainly of galactosyl lactose, which is important for infant growth and as a functional food additive. Although galactosyl lactose is abundant in goat milk, its complex structure has hindered the separation and analysis of its isomers. In this study, 5 isomers of goat milk galactosyl lactose were separated by HPLC: β6′-galactosyl lactose (β6′-GL), α6′-galactosyl lactose (α6′-GL), β4′-galactosyl lactose (β4′-GL), α3′-galactosyl lactose (α3′-GL), and β3′-galactosyl lactose (β3′-GL). This composition differs from that of commercial galactooligosaccharide products, which comprise mainly β-configuration oligosaccharides. The isomers were then qualitatively and quantitatively compared at different lactation stages using online HPLC-mass spectrometry. Relative quantitative analysis showed that the total content of the 5 galactosyl lactose isomers was highest in transitional goat milk. Specifically, β3′-GL was the main isomer in colostrum and α3′-GL was the main isomer in transitional and mature milk. β6′-Galactosyl lactose and β4′-GL tended to increase and then decrease during lactation. Moreover, α3′-GL content was 2 times higher than in colostrum and 10 times higher in transitional milk than in mature milk; in contrast, for β3′-GL, the values were 5 and 2 times higher, respectively. Absolute quantitative analysis revealed that β3′-GL was the most abundant isomers in colostrum (32.3 mg/L), and α3′-GL was the most abundant in transitional milk (88.1 mg/L) and mature milk (36.3 mg/L). These findings provide an important quantitative basis for understanding the relationship between structure and function of galactosyl lactose in goat milk, as well as its exploitation as a functional food.

Exosomes derived from mesenchymal stromal cells (MSCs) have emerged as novel drug and gene delivery tools. Current study aimed to elucidate the potential therapeutic role of human placental MSC (hPLMSC)-derived exosomes carrying AntagomiR-4450 (EXO-AntagomiR-4450) in intervertebral disc degeneration (IDD) progression. Initially, the differentially expressed miRNAs related to IDD were identified by microarray analysis, which provided data predicting the interaction between microRNA-4450 (miR-4450) and zinc finger protein-121 (ZNF121) in IDD. Next, miR-4450 and ZNF121 were elevated or silenced to determine their effects on the damage of nucleus pulposus cells (NPCs) treated with tumor necrosis factor alpha (TNF-alpha). The therapeutic effects of EXO-AntagomiR-4450 on NPCs were verified both in vitro and in vivo (15-week-old C57BL/6 male mice); especially gait analysis and fluorescent molecular tomography were used in live mice with IDD. Our results revealed that miR-4450 was highly expressed, while ZNF121 was poorly expressed in IDD patients and NPCs treated with TNF-alpha. Furthermore, miR-4450 was identified to specifically target ZNF121. In addition, the inhibition of miR-4450 exerted an alleviatory effect on the inflammation, apoptosis, and damage of the NPCs by upregulating ZNF121 (allP < 0.05). Moreover, EXO-AntagomiR-4450 retarded damage of NPCs in vitro, alleviated IDD damage, and ameliorated gait abnormality in vivo (allP < 0.05). hPLMSC-derived exosomes could be a feasible nanovehicle to deliver inhibitory oligonucleotides like AntagomiR-4450 in IDD.

Background Exosomes derived from mesenchymal stem cells (MSCs) have emerged as novel drug and gene delivery tools. Current study aimed to elucidate the potential therapeutic role of human placental MSC (hPLMSC)-derived exosomes carrying antagomiR-4450 (EXO-antagomiR-4450) in intervertebral disc degeneration (IDD) progression. Methods Initially, the differentially expressed miRNAs related to IDD were identified by microarray analysis which provided data predicting the interaction between miR-4450 and ZNF121 in IDD. Next, miR-4450 and ZNF121 were elevated or silenced to determine their effects on the damage of NPCs treated with TNF-α. The therapeutic effects of EXO-antagomiR-4450 on nucleus pulposus cells (NPCs) were verified both in vitro and in vivo, especially gait analysis and fluorescent molecular tomopraphy were used in live IDD mice. Results Our results revealed that miR-4450 was highly expressed, while ZNF121 was poorly expressed in IDD patients and NPCs treated with TNF-α. Furthermore, miR-4450 was identified to specifically target ZNF121. Additionally, the inhibition of miR-4450 exerted an alleviatory effect on the inflammation, apoptosis and damage of the NPCs by up-regulating ZNF121. Moreover, EXO-antagomiR-4450 retarded damage of NPCs in vitro, alleviated IDD damage and ameliorated gait abnormality in vivo. Conclusion hPLMSC-derived exosomes could be a feasible nanovehicle to deliver inhibitory oligonucleotides like antagomiR-4450 in IDD.

Thyroid dysfunction is a frequently found endocrine disorder among reproductively aged women. Subclinical hypothyroidism is the most common condition of thyroid disorders during pregnancy and is defined as manifesting a thyro id-stimulating hormone concentration exceeding the trimester-specific reference value, with a normal free thyroxine concentration. Here, we evaluated the prospective association between spontaneous miscarriage and first-trimester thyroid function. We conducted a case–control study (421 cases and 1684 controls) that was nested. Thyroid-st imulating hormone (TSH), free thyroxine (FT4), thyroid-peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb) status were measured. We found that higher TSH was relat ed to spontaneous miscarriage (OR 1.21; 95% CI, 1.13–1.30, P < 0.001). Compared with women with TSH levels of 0.4–<2.5 mIU/L, the risk of miscarriage was increased in wom en with TSH levels of 2.5–<4.87 mIU/L (OR 1.47; 95% CI, 1.16–1.87) and TSH greater th an 4.87 mIU/L (OR 1.97; 95% CI, 1.22–3.18). After controlling for the confounding facto r, TPOAb positivity status and FT4, the results were similar. The present study showed that higher TSH was associated with miscarriage in early pregnancy. In fact, TSH levels betwee n 2.5 and 4.87 mIU/L increased the risk for miscarriage, with TSH greater than 4.87 mIU/L increasing the risk even further.

Background Graves' disease (GD) is a common autoimmune disease characterized by genetic and environmental factors. Fc gamma receptors (Fc gamma Rs) are involved in several autoimmune disorders through recognizing immunoglobulin (Ig) G antibodies and mediating immune response. The study on the expression of Fc gamma Rs in GD patients is scarce. The purpose of this study was to evaluate the expression of three different types of Fc gamma Rs in patients with active and remissive GD. Methods Blood samples of patients and healthy subjects were collected to analyze the percentage of Fc gamma RI (CD64), Fc gamma RII (CD32), and Fc gamma RIII (CD16) on peripheral blood mononuclear cells (PBMCs) and monocytes by flow cytometry and Western blotting. CD32 isotypes were also examined in cases and controls by real-time PCR. Results The cell percentages expressed CD32 and protein expressions of CD32 on PBMCs, and monocytes from patients with active GD were significantly reduced compared to controls and patients with remissive GD. In particular, the expression of CD32B on PBMC was also decreased in active GD patients. However, the cell percentages expressed CD16 and CD64 from PBMCs and monocytes were comparable between three groups. Besides, the percentages of CD14(+)CD32(+) cells were negatively correlated with TRAb titers in active GD patients (r = -0.5825, P 0.001). Conclusion These results suggested that CD32 may act as a novel marker for active GDs. The expression of monocytic CD32, in particular CD32B, in GD patients might play a crucial role in maintaining Fc gamma Rs function and be a therapeutic target in GD patients.

Introduction and aim: This study aimed to confirm the association of the transmembrane 6 superfamily member 2 (TM6SF2) E167K variant with non-alcoholic fatty liver disease (NAFLD) and the degree of steatosis, as well as the additive effect of body mass index (BMI) or the patatin-like phospholipase domain-containing protein 3 (PNPLA3) I148M and TM6SF2 E167K variants in NAFLD. Materials and methods: A total of 158 NAFLD patients and 158 matched controls were recruited. Steatosis was classified as mild, moderate and severe by FibroScan. Associations between the TM6SF2 E167K variant and NAFLD as well as clinical parameters were evaluated. Results: Although the frequency of the T allele was low in the Chinese population (MAF = 7.4%), there was still a significant association between the E167K variant and NAFLD (odds ratio = 3.379, 95% confidence interval: 1.500-7.612, P = 0.003). In particular, the TM6SF2 genotype was also associated with the degree of steatosis (P= 0.023). The TM6SF2 variant was associated with increased alanine aminotransferase (ALT) but no other clinical parameters, such as aspartate aminotransferase (AST), alkaline phosphatase (ALP) and lipids. Notably, we also found that an additive effect of the TM6SF2 E167K and PNPLA3 I148M variants in NAFLD. Furthermore, we did not identify an association between the TM6SF2 E167K variant and NAFLD in the non-obese population. Conclusion: The TM6SF2 E167K variant was associated with NAFLD in northeast China, and there was an interaction between the PNPLA3 I148M and TMS6F2 E167K variants in NAFLD. (C) 2019 Fundacion Clinica Medica Sur, A.C. Published by Elsevier Espana, S.L.U.

Background Pregnant women with subclinical hypothyroidism are associated with an increased risk of spontaneous abortion. This study aims to investigate the mechanisms underlying the effects of maternal subclinical hypothyroidism during early pregnancy on abortion in the uterus, focusing upon the LIF/STAT3 signaling pathway. Methods One hundred five Wistar rats were randomly divided into three groups (35 rats in each group): control (CON) group, subclinical hypothyroidism (SCH) group and overt hypothyroidism (OH) group. We examined the weight of rat uteri, rat placenta and embryos. We also determined the number of implantation sites and the embryo absorption rates. The protein and mRNA expressions of TSHR, TR-α, TR-β, LIFR, gp130, JAK1, p-STAT3 and STAT3 were measured by immunohistochemical staining, real-time PCR and Western blotting. Results The weights of rat uteri, rat placenta and embryos were significantly reduced in the SCH and OH groups. The number of implantation sites was significantly decreased in the SCH and OH groups, while embryo absorption rates were significantly increased. The mRNA and protein expressions of TSHR were upregulated in the SCH and OH groups, while TR-α and TR-β showed no difference when compared between the three groups. The expression levels of LIFR, gp130, JAK1 and p-STAT3 were significantly higher in the SCH and OH groups. Conclusions Clinical and subclinical hypothyroidism during early pregnancy might cause adverse pregnancy outcomes. Implantation failure in rats with subclinical hypothyroidism was associated with abnormal LIF/STAT3 signaling.

PURPOSE:Growth arrest-specific protein 6 (Gas6) is a vitamin K-dependent protein that plays an important role in the pathogenesis of autoimmune diseases. The purpose of this study was to explore the expression of Gas6 and its effects on autoimmune thyroiditis (AIT).METHOD:A total of 24 male NOD.H-2h4 mice were randomly assigned to three groups: (1) a control group supplied with regular water; (2) a sodium iodide (NaI) group supplied with 0.005% sodium iodide water; and (3) a group treated with recombinant mouse Gas6 (rmGas6) after iodine supplementation (NaI + Gas6 group). The severity of lymphocytic infiltration in the thyroid was measured through histopathology. Serum levels of tumor necrosis factor α (TNF-α), interleukin (IL) 6 and IL-1β, as well as anti-thyroglobulin antibody (TgAb) titers were measured using an enzyme-linked immunosorbent assay. In addition, the expression of Gas6, Caspase 3, TAM receptors (Axl and MerTK), nuclear factor κB (NF-κB) and I-kappa-B α (IκB-α) were measured by Western blotting. Finally, the proportions of T cells were determined in the splenocytes of NOD.H-2h4 mice by flow cytometry.RESULTS:The mRNA and protein expression of Gas6 was significantly lower in the NaI group compared to the control group. Serum levels of TgAb, TNF-α, IL-6 and IL-1β were also significantly higher in the NaI group but were dramatically reduced after rmGas6 injection. The prevalence of thyroiditis and the infiltration of lymphocytes were significantly lower in the NaI + Gas6 group compared to the NaI group. The protein expression of cleaved-Caspase 3, phosphorylation of MerTK, and NF-κB and IκB-α in the thyroid gland were significantly reduced after rmGas6 administration. The proportion of Th1, Th2 and Th17 cells in splenocytes were also significantly reduced after rmGas6 treatment, whereas there was a dramatic increase in the proportion of Treg cells.CONCLUSION:Gas6 exerts an anti-inflammatory effect in a mouse model of AIT and may therefore be a potential therapeutic target.

An increasing number of studies suggest that subclinical hypothyroidism (SCH) is associated with complications of gestation, including spontaneous abortion (SA). However, the underlying mechanism is not clear. MicroRNA (miRNA) has been demonstrated to be closely related to gynecological reproductive diseases. We determined miRNA expression in patients with SCH, SCH with SA (SCH + SA), and in those with SA as well as healthy controls (HCs), and analyzed whether dysregulation in several miRNAs was specific to these cohorts. An Agilent Human miRNA array was used to explore miRNA levels in pooled serum samples as a pilot study, followed by a validation of selected miRNAs by real-time polymerase chain reaction in SCH (N = 24), SA (N = 19), SCH + SA (N = 21), and HC cohorts (N = 18). The relative expression of miR-940 was elevated in the SCH + SA group compared with SCH, SA, and HC groups. In addition, miR-486-5p was upregulated in the SCH + SA group compared with SA and HC groups, without a difference noted between SCH + SA and SCH groups. Further analysis suggested that miR-940 or miR-486-5p may be potential predictive biomarkers for the early diagnosis of SA in patients with SCH.

PURPOSE:Myeloid-related protein (MRP) family plays an important role in the promotion of cell proliferation and the production of inflammatory cytokines. We investigated the expression of MRP6, MRP8 and MRP14 in thyroid tissues, serum, and peripheral blood monocular cells (PBMCs) in patients with autoimmune thyroid diseases (AITD).METHOD:The expression of MRP6, MRP8, and MRP14 was investigated using immunohistochemical staining and quantitative real-time polymerase chain reaction in the thyroid glands of 7 patients with Graves' disease (GD), 8 with Hashimoto's thyroiditis (HT), and 7 healthy controls (HC). The serum levels of MRP8/MRP14 complex and MRP6 were investigated in 30 patients with GD, 36 with HT, and 30 with HC. The mRNA expression of MRP proteins in PBMCs was also explored. PBMCs from each group were incubated with MPRs and their effect on Toll-like receptor 4(TLR4) expression and their effect on the levels of the pro-inflammatory cytokines in supernatant were analyzed upon incubating with TLR4 and signaling pathways inhibitors.RESULTS:Serum levels of MRP8/MRP14 and MRP6 were up-regulated in patients with AITD. In addition, mRNA expression of MRP proteins in PBMCs and the thyroid gland was markedly elevated in AITD patients. MRP6 and MPR8 promoted the secretion of TNF-α and IL-6 in cultured PBMCs, and this elevation was more pronounced in AITD patients; we also found that this up-regulation was regulated by TLR4/phosphoinositide 3-kinase/nuclear factor-κB signaling pathway.CONCLUSION:The expression of MRP proteins was elevated in AITD patients. Therefore, an MRP-TLR4 dependent signaling may play an important role in the pathogenesis of AITD.