Aimed to comprehensively investigate the presence of neural autoantibodies in the cerebrospinal fluid (CSF) and plasma of COVID-19 patients experiencing neurological complications during the Omicron wave in China. Forty consecutive COVID-19 patients with severe neurological complications and 15 disease controls (DC) were enrolled. Neural autoantibodies were detected using both the indirect immunofluorescence assay (IFA) on mouse brain tissue and the Brain-neuronal-antigen microarray. Our results indicated a significantly higher prevalence of neural autoantibodies in the CSF (62.16% vs. 0.0%) and plasma (38.71% vs. 13.33%) of COVID-19 patients compared to DC. Additionally, we identified 12 upregulated intrathecal IgG autoantibodies with differential levels between COVID-19 patients and DC, as well as 51 upregulated IgG autoantibodies in plasma. A high prevalence of anti-mGluR2 antibodies (13.33%) in COVID-19 patients was confirmed by cell-based assays. Western blot analysis showed these antibodies cross-react with both the nucleocapsid (N) and spike (S) proteins of SARS-CoV-2. Notably, strong binding to both the S protein's RBD-Fc and mGluR2 was observed, an association that was substantiated by bioinformatics analysis evaluating the similarity between SARS-CoV-2 proteins and the targeted antigens on the microarray. This finding hints at a potential cross-reactivity between anti-mGluR2 antibodies and the S protein in COVID-19 patients.

Inactivated coronavirus disease 2019 (COVID-19) vaccines showed impaired immunogenicity in some autoimmune diseases, but it remains unclear in primary biliary cholangitis (PBC). This study aimed to explore the antibody response to the inactivated COVID-19 vaccine in individuals with PBC, as well as to evaluate coverage, safety, and attitudes toward the COVID-19 vaccine among them. Two cohorts of patients with PBC were enrolled in this study. One cohort was arranged to evaluate the immunogenicity of the inactivated COVID-19 vaccine, another cohort participated in an online survey. The titers of the anti-receptor-binding domain (RBD)-specific immunoglobulin G (IgG), neutralizing antibody (NAb) toward severe acute respiratory syndrome coronavirus 2 wild-type, and NAb toward Omicron BA.4/5 subvariants were detected to assess antibody response from the vaccine. After booster vaccination for more than six months, patients with PBC had significantly lowered levels of anti-RBD-specific IgG compared to HCs, and the inhibition rates of NAb toward wild-type also declined in individuals with PBC. The detected levels of NAb toward Omicron BA.4/5 were below the positive threshold in patients with PBC and HCs. Laboratory parameters did not significantly correlate with any of the three antibodies. The online survey revealed that 24% of patients with PBC received three COVID-19 vaccines, while 63% were unimmunized. Adverse effect rates after the first, second, and third vaccine doses were 6.1%, 10.3%, and 9.5%, respectively. Unvaccinated patients with PBC were more worried about the safety of the vaccine than those who were vaccinated (P = 0.004). As a result, this study fills the immunological assessment gap in patients with PBC who received inactivated COVID-19 vaccines.

Objectives: Demyelination and immunocyte-infiltrated lesions have been found in neuro-Beh & ccedil;et 's disease (NBD) pathology. Lacking satisfying laboratory biomarkers in NBD impedes standard clinical diagnostics. We aim to explore the ancillary indicators for NBD diagnosis unveiling its potential etiology. Methods: 28 NBD with defined diagnosis, 29 patients with neuropsychiatric lupus erythematosus, 30 central nervous system idiopathic inflammatory demyelination diseases (CNS-IIDD), 30 CNS infections, 30 cerebrovascular diseases, and 30 noninflammatory neurological diseases (NIND) were retrospectively enrolled. Immunoglobulins (Ig) in serum and cerebral spinal fluid (CSF) were detected by immunonephelometry and myelin basic protein (MBP) by quantitative enzyme-linked immunosorbent assay. Results: IgA index is almost twice enhanced in NBD than NIND with an accuracy of 0.8488 in differential diagnosis, the sensitivity and specificity of which were 75.00 % and 90.00 % when the cutoff was > 0.6814. The accuracy of CSF Ig and quotient of Ig all exceed 0.90 in discerning NBD with damaged and intact blood -brain barrier (BBB). Clustering analyses divided NBD into two different phenotypes: one with BBB damage has lower Ig synthesis, the other with extra-synthesis in parenchymal sites but with intact BBB. MBP index is significantly correlated with kappa (KAP) index and lambda (LAM) index (r = 0.358, 0.575, P < 0.001), hinting the NBD pathogenesis of CNS demyelination in triggering excessive intrathecal Ig productions and humoral responses. Conclusions: IgA index acts as a potential diagnostic indicator in differentiating NBD from NIND and CNS-IIDD. Excessive immunoglobulin production induced by CNS inflammation and demyelination might be latent immunopathogenesis of NBD.

In women of childbearing age, extensive decidualization, shedding and remodeling of the endometrium during the menstrual cycle are fundamental for successful pregnancy. The role of prostaglandins (PGs) in menstruation has long been proposed in humans, and the rate-limiting enzyme cyclooxygenase was shown to play a key role in endometrial breakdown and shedding in a mouse menstrual-like model in our previous study. However, the specific types of PGs involved and their respective roles remain unclear. Therefore, our objective was to investigate the mechanism through which PGs regulate endometrial disintegration. In this study, the microscopy was observed by HE; the protein levels of prostaglandins E1 (PGE1), prostaglandins E2 (PGE2), prostaglandin F2α (PGF2α) and Prostaglandin I2 (PGI2) were detected by ELISA; the mRNA level of Pfgfr2, Vascular Endothelial Growth Factor(Vegf), Angiostatin and Hypoxia inducible factor-1α (Hif1α) were examined by real-time PCR; PTGFR Receptor (PTGFR), VEGF, Angiostatin and HIF-1α protein levels were investigated by western blotting; the locations of protein were observed by Immunohistochemistry; HIF-1α binding PTGFR promoter was detected by Chromatin Immunoprecipitation (ChIP) and real-time PCR. We found that the concentrations of PGE1, PGE2, and PGF2α all increased significantly during this process. Furthermore, Ptgfr mRNA increased soon after Progesterone (P4) withdrawal, and PTGFR protein levels increased significantly during abundant endometrial breakdown and shedding processes. PTGFR inhibitors AL8810 significantly suppressed endometrial breakdown and shedding, promoted Angiostatin expression, and reduced VEGF-A expressions and vascular permeability. And HIF-1α and PTGFR were mainly located in the luminal/gland epithelium, vascular endothelium, and pre-decidual zone. Interestingly, HIF-1α directly bound to Ptgfr promoter. Moreover, a HIF-1α inhibitor 2-methoxyestradiol (2ME) significantly reduced PTGFR expression and suppressed endometrial breakdown which was in accord with PTGFR inhibitor’s effect. Similar changes occurred in human stromal cells relevant to menstruation in vitro. Our study provides evidence that PGF2α/PTGFR plays a vital role in endometrial breakdown via vascular changes that are regulated by HIF-1α during menstruation.

BACKGROUND:Spermatogenesis is a highly regulated and complex process in which DNA methylation plays a crucial role. This study aimed to explore the differential methylation profiles in sperm DNA between patients with asthenospermia (AS) and healthy controls (HCs), those with oligoasthenospermia (OAS) and HCs, and patients with AS and those with OAS.RESULTS:Semen samples and clinical data were collected from five patients with AS, five patients with OAS, and six age-matched HCs. Reduced representation bisulfite sequencing (RRBS) was performed to identify differentially methylated regions (DMRs) in sperm cells among the different types of patients and HCs. A total of 6520, 28,019, and 16,432 DMRs were detected between AS and HC, OAS and HC, and AS and OAS groups, respectively. These DMRs were predominantly located within gene bodies and mapped to 2868, 9296, and 9090 genes in the respective groups. Of note, 12, 9, and 8 DMRs in each group were closely associated with spermatogenesis and male infertility. Furthermore, BDNF, SMARCB1, PIK3CA, and DDX27; RBMX and SPATA17; ASZ1, CDH1, and CHDH were identified as strong differentially methylated candidate genes in each group, respectively. Meanwhile, the GO analysis of DMR-associated genes in the AS vs. HC groups revealed that protein binding, cytoplasm, and transcription (DNA-templated) were the most enriched terms in the biological process (BP), cellular component (CC), and molecular function (MF), respectively. Likewise, in both the OAS vs. HC and AS vs. OAS groups, GO analysis revealed protein binding, nucleus, and transcription (DNA-templated) as the most enriched terms in BP, CC, and MF, respectively. Finally, the KEGG analysis of DMR-annotated genes and these genes at promoters suggested that metabolic pathways were the most significantly associated across all three groups.CONCLUSIONS:The current study results revealed distinctive sperm DNA methylation patterns in the AS vs. HC and OAS vs. HC groups, particularly between patients with AS and those with OAS. The identification of key genes associated with spermatogenesis and male infertility in addition to the differentially enriched metabolic pathways may contribute to uncovering the potential pathogenesis in different types of abnormal sperm parameters.

Endometrial damage repair disorder is the main reason of intrauterine adhesions (IUA) and thin endometrium (TA), which is caused by curettage or infection. Exosomal miRNAs derived from human umbilical cord mesenchymal stem cells (hucMSCs) were reported to play an important role in damage repair disorder, including endometrial fibrosis. In this study, we aimed to investigate the role of hucMSCs-derived exosomal microRNA-202-3p (miR-202-3p) in endometrial damage repair. We established rat endometrial injury model according to curettage to mimic women curettage abortion operation. The miRNA array analysis indicated that miR-202-3p was increased and matrix metallopeptidase 11 (MMP11) was decreased in the exosomes-treated rat uterine tissues. Bioinformatics analysis suggested that MMP11 is the target gene of miR-202-3p. We observed that the mRNA and protein of MMP11 were significantly decreased in exosome treatment group on day 3, and the components of extracellular matrix (ECM) COL1A1, COL3A1, COLVI and fibronectin (FN) protein were increased. And we found that when the injured human stromal cells were treated with miR-202-3p overexpression exosomes, the COLVI and FN were also upregulated in protein and mRNA expression level. For the first time MMP11 was proved to be the target gene of miR-202-3p by dual luciferase reporter system. At last, we found the state of stromal cells was better in miR-202-3p overexpression exosomes group compared to exosomes group, and miR-202-3p overexpression exosomes markedly upregulated the FN and collagen on day 3 after endometrial injury. We thought that miR-202-3p overexpression exosomes promoted endometrial repair by regulating ECM remodeling in early repair of damaged endometrium. Taken together, these experimental findings may provide a theoretical basis for understanding endometrial repair and an insight into the clinical treatment for IUA.

BackgroundThe immunogenicity of booster inactivated COVID-19 vaccines in patients with type 2 diabetes mellitus (T2DM) has remained unclear. Our study aims to investigate the antibody response to inactivated COVID-19 vaccine following booster vaccination in patients with T2DM. MethodsA total of 201 patients with T2DM and 102 healthy controls (HCs) were enrolled. The levels of anti-SARS-CoV-2 total antibodies, anti-receptor-binding domain (RBD)-specific IgG, neutralizing antibody (NAb) toward SARS-CoV-2 wild type (WT), and NAb toward SARS-CoV-2 Omicron BA.4/5 subvariant were measured to evaluate the vaccine-induced immunological responses. ResultsThe titers of anti-RBD-specific IgG (p = 0.018) and inhibition rates of NAb toward WT (p = 0.007) were significantly decreased in patients with T2DM compared to HCs after booster vaccination for more than 6 months. Both HCs and patients with T2DM showed poor resistance against BA.4/5 due to the detected inhibition rates being lower than the positive threshold. The levels of anti-RBD-specific IgG were positively associated with the proportions of CD3(+)CD4(-)CD8(-) T cells (p = 0.045), and patients with T2DM who had anti-RBD-specific IgG positivity showed higher proportions of CD3(+)CD4(-)CD8(-) T cells compared to those negative (p = 0.005). ConclusionsPatients with T2DM showed impaired antibody responses after booster vaccination for more than 6 months. Decreased anti-BA.4/5 responses give rise to the possibility of breakthrough infections for both patients with T2DM and HCs.

目的 利用小鼠人工诱导蜕膜化模型,探讨转录活化因子6(ATF6)在小鼠子宫内膜中的作用机制.方法 构建繁育野生型(Atf6+/+)和ATF6敲除型(Atf6-/-)小鼠,并构建小鼠人工诱导蜕膜化模型.雌鼠交配后见阴栓第1天记为Day 1.Day 4进行诱导蜕膜化,蜕膜化后48 h和72 h(即Day 6和Day 7)收集小鼠子宫进行相应指标检测,实时荧光定量PCR方法和蛋白质免疫印迹法检测Atf6 mRNA与蛋白质在蜕膜化子宫中的表达情况;免疫组织化学方法检测野生型(Atf6+/+)和ATF6敲除型(Atf6-/-)小鼠人工诱导蜕膜化模型中ATF6和催乳素(PRL)在子宫中的阳性信号表达情况.收集人子宫内膜生理周期中不同时相子宫内膜组织,进行石蜡包埋切片,免疫组织化学方法检测ATF6在子宫内膜中的阳性信号表达情况.结果 与小鼠未蜕膜子宫相比,Day 6和Day 7蜕膜化子宫中的Atf6 mRNA表达水平显著升高(P＜0.05),细胞核内的蛋白水平也显著增高(P＜0.05);Day 7时子宫大体形态学显示子宫较粗且均匀.野生型和敲除鼠诱导蜕膜化子宫HE染色显示,绝大部分子宫基质细胞变大变圆,表明诱导蜕膜化的子宫内膜蜕膜化充分;PRL免疫组化染色结果表明,在野生型(Atf6+/+)和敲除型(Atf6-/-)小鼠蜕膜化子宫内膜中,PRL染色阳性信号分布范围无显著性差异(P＞0.05).不同月经周期时相人子宫内膜组织的免疫组化结果提示,ATF6在月经期和增殖早期的表达水平较高,显著高于其余周期时相(P＜0.05).结论 ATF6能够参与子宫内膜蜕膜化,但不是蜕膜化发生的必要条件;ATF6可能通过参与子宫内膜的生理修复过程参与子宫内膜生理周期.

Objective:To investigate the effects of clock genes Cry1 and Bmal1 on the endometrial physiological repair process during menstruation-like period based on the mouse menses model.Methods:The menstruation-like physiological repair model in mice was established. The histopathology was observed by hematoxylin eosin staining（HE）. Immunohistochemical staining（IHC） was used to observe the expression mapping of CRY1 and BMAL1 proteins during menstruation-like period in mice. The expression levels of CRY1 and BMAL1 proteins were detected by Western blot.Results:After 24 to 48 hours of progesterone withdrawal, the general color of the uterus gradually faded, and the diameter of the uterus gradually decreased from thick to thin. HE results showed that most of the cells in the decidualized area underwent necrosis and collapse 24 hours after progesterone withdrawal. With the passage of time, the endometrial thickness increased, the surface of the uterine cavity appeared re-epithelialization, and the number of glands increased significantly. After 48 hours, the endometrium completely repaired and basically returned to normal uterine morphology. Immunohistochemical staining results showed that the positive staining of CRY1 and BMAL1 proteins was localized in the cytoplasm and nucleus, mainly concentrated in the nucleus, and the positive signals showed certain differences with the time of progesterone withdrawal. Western blot results showed that the expression of CRY1 protein in the endometrium significantly decreased 24 hours after progesterone withdrawal（P＜0.05）. At 40 hours after progesterone withdrawal, the expression level was relatively lower and the nadir point appeared. The expression of BMAL1 protein in the endometrium at 32 hours after progesterone withdrawal was significantly lower than that at 24 hours and 48 hours（P＜0.05）,and the nadir point appeared 8 hours earlier than that of CRY1 protein.Conclusions:Clock genes are regularly and cyclically expressed in the endometrium, which suggests that clock genes and their signaling pathways are involved in the physiological repair of endometrium.

AimsDiabetes mellitus (DM), one of the most common chronic diseases in China, is a risk factor for SARS-COV-2 infection and poor prognosis of COVID-19. The COVID-19 vaccine is one of the key measures to control the pandemic. However, the actual coverage of COVID-19 vaccination and associated factors remain unclear among DM patients in China. We conducted this study to investigate the COVID-19 vaccine coverage, safety, and perceptions among patients with DM in China. MethodsA cross-sectional study of a sample of 2200 DM patients from 180 tertiary hospitals in China was performed using a questionnaire developed through the Wen Juan Xing survey platform to collect information regarding their coverage, safety, and perceptions of COVID-19 vaccination. A multinomial logistic regression analysis model was performed to determine any independent relationships with COVID-19 vaccination behavior among DM patients. ResultsIn total, 1929 (87.7%) DM patients have received at least one dose COVID-19 vaccine, and 271 (12.3%) DM patients were unvaccinated. In addition, 65.2% (n = 1434) were booster vaccinated against COVID-19, while 16.2% (n = 357) were only fully vaccinated and 6.3% (n = 138) were only partially vaccinated. The prevalence of adverse effects after the first dose of vaccine, the second dose of vaccine, and the third dose of vaccine were 6.0%, 6.0%, and 4.3% respectively. Multinomial logistic regression analysis showed that DM patients complicated with immune and inflammatory diseases (partially vaccinated: OR = 0.12; fully vaccinated: OR = 0.11; booster vaccinated: OR = 0.28), diabetic nephropathy (partially vaccinated: OR = 0.23; fully vaccinated: OR = 0.50; booster vaccinated: OR = 0.30), and perceptions on the safety of COVID-19 vaccine (partially vaccinated: OR = 0.44; fully vaccinated: OR = 0.48; booster vaccinated: OR = 0.45) were all associated with the three of vaccination status. ConclusionThis study showed that higher proportion of COVID-19 vaccine coverage among patients with DM in China. The concern about the safety of the COVID-19 vaccine affected the vaccine behavior in patients with DM. The COVID-19 vaccine was relatively safe for DM patients due to all side effects were self-limiting.

Abstract Background The immunogenicity of booster inactivated COVID‐19 vaccines in patients with type 2 diabetes mellitus (T2DM) has remained unclear. Our study aims to investigate the antibody response to inactivated COVID‐19 vaccine following booster vaccination in patients with T2DM. Methods A total of 201 patients with T2DM and 102 healthy controls (HCs) were enrolled. The levels of anti‐SARS‐CoV‐2 total antibodies, anti‐receptor‐binding domain (RBD)‐specific IgG, neutralizing antibody (NAb) toward SARS‐CoV‐2 wild type (WT), and NAb toward SARS‐CoV‐2 Omicron BA.4/5 subvariant were measured to evaluate the vaccine‐induced immunological responses. Results The titers of anti‐RBD‐specific IgG (p = 0.018) and inhibition rates of NAb toward WT (p = 0.007) were significantly decreased in patients with T2DM compared to HCs after booster vaccination for more than 6 months. Both HCs and patients with T2DM showed poor resistance against BA.4/5 due to the detected inhibition rates being lower than the positive threshold. The levels of anti‐RBD‐specific IgG were positively associated with the proportions of CD3+CD4−CD8− T cells (p = 0.045), and patients with T2DM who had anti‐RBD‐specific IgG positivity showed higher proportions of CD3+CD4−CD8− T cells compared to those negative (p = 0.005). Conclusions Patients with T2DM showed impaired antibody responses after booster vaccination for more than 6 months. Decreased anti‐BA.4/5 responses give rise to the possibility of breakthrough infections for both patients with T2DM and HCs.