Asprosin is a white adipose tissue-derived hormone that increases abnormally in mammals with insulin resistance. However, the role of asprosin in polycystic ovary syndrome (PCOS), a disease partly characterized by insulin resistance, and its potential connection with type 2 diabetes mellitus (T2DM) and PCOS has not been thoroughly elucidated to date. To investigate the association of asprosin with metabolic profiles, sex-related hormones, or inflammation in females with T2DM or PCOS, plasma asprosin and metabolic indicators were measured in 66 healthy females, 53 female patients with T2DM, and 41 patients with PCOS. Spearman’s correlation analysis and binary logistic regression analysis models were used. Plasma asprosin was significantly higher in T2DM females than in healthy subjects (P<0.001) and was positively correlated with fasting blood glucose (FBG), hemoglobin A1c (HbA1c), and HOMA-IR (P<0.05). Asprosin in PCOS subjects was also higher than in healthy subjects (P<0.001) but lower than in T2DM subjects (P<0.05), and it was positively correlated with FBG, HbA1c, HOMA-IR, LDL-c, APOB, APOE, and testosterone (P<0.05). The BMI-categorized subgroups of PCOS subjects also showed correlations of asprosin with metabolic profiles and sex-related hormones. Binary logistic regression analysis revealed that plasma asprosin level acted as an independent risk factor for T2DM or PCOS. These findings suggest the correlation of plasma asprosin level with glucose metabolism, lipid metabolism, sex-related hormones, and inflammation in females, supporting asprosin as a potential predictive factor for females with metabolic-related diseases. This trial is registered with ChiCTR-ROC-17010719 .
To study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia.coli.In cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice.The COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62∓0.39)% vs (8.81∓1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95∓0.59)% vs (8.85∓0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36∓0.06, significantly greater than that of COTD (6.01∓0.07) and revertant (6.29∓0.06) strains (P<0.05); the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25∓0.05 vs 5.87∓0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively.OmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.
Targeted therapies have been proven as promising in the treatment of breast cancer and have improved survival and quality of life in advanced breast cancer. We previously identified a novel peptide SA12 which showed significant activity in the inhibition of proliferation and induction of apoptosis in SKBr-3 cells.The present study investigated the potential antitumor role of SA12 in breast cancer cell lines MDA-MB-231 and MCF-7 through Cell Counting Kit-8 assay and colony formation assay, and examined the cell cycle distribution using flow cytometry analysis. Furthermore, the expression of cell cycle-related genes cyclin D1, CDK4, and tumor suppressor gene p16 were examined by real-time polymerase chain reaction and Western blot to explore the molecular mechanism.We determined that peptide SA12 suppressed the proliferation of MDA-MB-231 and MCF-7 cell lines through the G0/G1 phase cell cycle arrest. Moreover, the expressions of cell cycle-associated genes cyclin D1 and CDK4 were downregulated and the expression of tumor suppressor gene p16 was upregulated after treatment with SA12. MECP2 was required for the enhanced expression of p16 gene induced by SA12, which further inhibits CDK4/CDK6 activation and arrests the cell cycle progression from G0/G1 to S phase.We concluded that SA-12 inhibits the proliferation of MCF-7 and MDA-MB-231 cells through G0/G1 cell cycle arrest. Cell cycle related genes cyclin D1, CDK4, and p16 participate in the process, and MECP2 is essential for the enhanced expression of p16 gene induced by SA-12.
Urease B is an important virulence factor that is required for Helicobacter pylori to colonise the gastric mucosa. Mouse monoclonal antibodies (mAbs) that inhibit urease B enzymatic activity will be useful as vaccines for the prevention and treatment of H. pylori infection. Here, we produced murine mAbs against urease B that neutralize the enzyme's activity. We mapped their epitopes by phage display libraries and investigated the immunogenicity of the selected mimotopes in vivo. The urease B gene was obtained (GenBank accession No. DQ141576) and the recombinant pGEX-4T-1/UreaseB protein was expressed in Escherichia coli as a 92-kDa recombinant fusion protein with glutathione-S-transferase (GST). Five mAbs U001-U005 were produced by a hybridoma-based technique with urease B-GST as an immunogen. Only U001 could inhibit urease B enzymatic activity. Immunoscreening via phage display libraries revealed two different mimotopes of urease B protein; EXXXHDM from ph.D.12-library and EXXXHSM from ph.D.C7C that matched the urease B proteins at 347-353 aa. The antiserum induced by selected phage clones clearly recognised the urease B protein and inhibited its enzymatic activity, which indicated that the phagotope-induced immune responses were antigen specific. The present work demonstrated that phage-displayed mimotopes were accessible to the mouse immune system and triggered a humoral response. The urease B mimotope could provide a novel and promising approach for the development of a vaccine for the diagnosis and treatment of H. pylori infection.
Collective cell behavior in response to mechanical injury is central to various regenerative and pathological processes. Using a double-stranded locked nucleic acid probe for monitoring real-time intracellular gene expression, we examined the spatiotemporal response of epithelial cells during injury-induced collective migration and compared to the blocker assay with minimal injury as control. We showed that cells ∼150 μm from the wound edge exhibit a gradient in response to mechanical injury, expressing different genes depending on the wounding process. While release of contact inhibition is sufficient to trigger the migratory behavior, cell injury additionally induces reactive oxygen species, Nrf2 protein, and stress response genes, including heat shock protein 70 and heme oxygenase-1, in a spatiotemporal manner. Furthermore, we show that Nrf2 has an inhibitory role in injury-induced epithelial-mesenchymal transition, suggesting a potential autoregulatory mechanism in injury-induced response. Taken together, our single-cell gene expression analyses reveal modular cell responses to mechanical injury, manipulation of which may afford novel strategies for tissue repair and prevention of tumor invasion in the future.
Background: Increasing evidence indicates that the gut microbiota contributes to the occurrence and development of metabolic diseases. However, little is known about the effects of commonly used antidiabetic agents on the gut microbiota. In this study, we investigated the roles of dipeptidyl peptidase-4 inhibitors (DPP-4i) and α-glucosidase inhibitor in modulating the gut microbiota.
Abstract Background Thyrotropin-secreting pituitary neuroendocrine tumors (PitNETs) are rare pituitary adenomas that are occasionally accompanied by hypersecretion of other anterior pituitary hormones, such as growth hormone (GH) and prolactin (PRL). The clinical, biochemical, and pathological characteristics may represent diverse circumstances. Case presentation In this report, a 33-year-old female diagnosed with a TSH PitNET co-secreting GH presented no obvious clinical symptoms. The main characteristics were elevated thyroid-stimulating hormone (TSH), free tri-iodothyronine (FT3), and free thyroxine (FT4) levels accompanied by slightly elevated GH and insulin-like growth factor-1 (IGF-1) levels. Magnetic resonance imaging (MRI) detected a pituitary macroadenoma (18 × 16 × 16 mm) with cavernous sinus and suprasellar invasion. Immunohistochemistry revealed diffuse positivity for TSH, strong immunoreactivity for GH, and sporadic positivity for PRL. The electron microscope and double immunofluorescence staining confirmed a plurimorphous plurihormonal adenoma producing TSH, GH, and PRL. After preoperative somatostatin receptor ligand (SRL) treatment and transsphenoidal surgery, the patient achieved temporary clinical and biochemical remission. However, 3 months after surgery, the patient was suspected of having Hashimoto’s thyroiditis due to higher thyroglobulin antibody (TGAb), thyroid peroxidase antibody (TPOAb), and thyroid receptor antibody (TRAb) and an enlarged thyroid nodule. During follow-up, thyroid function and TSH slowly transformed from transient hyperthyroidism to hypothyroidism. They were maintained in the normal range by L-T4. Conclusion In the TSH PitNET, the positive immunohistochemistry for TSH, GH, and PRL translated into hormonal overproduction with TSH and GH.
In this study, healthy young males were randomized into groups with moderate intensity training (n = 24), high intensity training (n = 24) and utmost intensity training (n = 24). At the end of 8-week training period, HRV measurements demonstrated a marked increase of RMSSD (P = 0.003), PNN50 (P = 0.002), HF (P = 0.002), SDNN (P = 0.002) and LF (P = 0.003) in the moderate intensity group and a decreasing tendency in LFn and LF/HF; however, in the utmost intensity group HFn (P = 0.012) decreased prominently while its LF (P = 0.032), LFn (P = 0.039) and LF/HF (P = 0.015) increased significantly. Nevertheless marked changes were not found in the above indexes of the high intensity group. While resting HR of the three groups declined significantly at the end of 8 weeks (P was 0.001, 0.0001 and 0.001 respectively); RMSSD, PNN50, HF, LF and SDNN were significantly higher in the moderate intensity group than in the other two groups (P P = 0.012) was significantly lower but its LFn and LF/HF were markedly higher (P was 0.025 and 0.015 respectively); LF/HF of both the high and utmost intensity group was significantly higher (P was 0.033 and 0.037 respectively). Despite a significant reduction of plasma NE in all the three groups at the end of 8-week training period (P was 0.016, 0, 0.031 respectively), plasma NE level of moderate and high intensity group was much lower than that of the utmost intensity group (P was 0.001, 0 respectively). Utmost and moderate endurance training results in altered sympathetic and parasympathetic balance towards sympathetic dominance and parasympathetic dominance respectively; whereas high intensity endurance training almost has no effect on ANS function. CPT and HUTT reveal the potential danger posed by utmost intensity endurance training.
Post-COVID syndrome is a multiorgan disease characterised by persistent symptoms 12 weeks or more following SARS-CoV2 infection. However, the pathophysiology remains unknown and is likely multifactorial due to the heterogeneity of clinical manifestations. The present study aimed to understand the changes and contribution of the systemic immune response in patients with post-COVID syndrome.
Methods
Observational study of hospitalised and non-hospitalised participants 3–16 months post-COVID at a single centre (Dundee, UK). Stabilised peripheral blood was processed for mRNAseq. In a participant subset, immunophenotyping of peripheral blood immune cells was carried out using mass cytometry and Maxpar Direct Immune Profiling kit comprising 35 cell-surface markers. Cytobank platform was used for manual gating for mass cytometry data and dimensionality reduction was carried out using the tSNE-CUDA algorithm.
Results
92 post-COVID participants were included (age 56±12.5 years (mean±SD), 46.7% male). Differences in immunophenotype were identified between those initially hospitalised (n=11) or not (n=7); significantly higher proportions of transitional monocytes (Mann-Whitney, p=0.0441), total CD8 αβ cells (p=0.0114) including effector memory (p=0.0185) and terminal effector (p=0.0083) subpopulations, as well as CD4 terminal effector cells (Unpaired T-test, p=0.0330) were found in those hospitalised. Notably, the hospitalised group had significantly more males (Fishers exact test, p=0.0294) and were older (Unpaired T-test, p=0.0005). 28 significantly differentially expressed genes (adjusted pvalue<0.05; Wald test with Benjamini-Hochberg correction) were identified between those initially hospitalised (n=49) or not (n=30). Genes relating to neutrophil activity including neutrophil elastase, MPO, azurocidin-1, defensin alpha 3 (DEFA3) and DEFA4 were upregulated in those who were hospitalised, in addition to lactotransferrin, BPI and CEACAM8, expressed in both neutrophils and monocytes. Those who were hospitalised were more likely to experience dyspnoea/fatigue beyond 12 weeks following acute infection (Fishers exact test, p=0.0021). Further, those with ongoing dyspnoea/fatigue (n=13) had increased neutrophils and a significant increase in T-regulatory cells (Unpaired T-test, p=0.0059), compared to those without (n=7).
Conclusion
Post-COVID, an increase in T-cell subsets and neutrophil-associated genes is associated with a more severe initial infection leading to hospitalisation. Neutrophil and T-regulatory cells are further associated with ongoing symptoms post-COVID, suggesting a role for these cell types in post-COVID syndrome.
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