Growth differentiation factor-15/adiponectin ratio as a potential biomarker for metabolic syndrome in Han Chinese

AimsGrowth differentiation factor-15 (GDF-15) and adiponectin are adipokines that regulate metabolism. This study aimed to evaluate the roles of GDF-15, adiponectin, and GDF-15/adiponectin ratio (G/A ratio) as biomarkers for detecting metabolic syndrome (MS).Materials and methodsThis cross-sectional study included 676 participants aged 20–70 years in Jurong, China. The participants were divided into four groups based on sex and age (<40 and ≥40 years). MS was defined according to the modified National Cholesterol Education Program Adult Treatment Panel III criteria. Receiver operating characteristic curves were used to evaluate the performance of GDF-15, adiponectin, and the G/A ratio in predicting MS.ResultsThe prevalence of MS was 22.0% (149/676). Logistic regression analysis indicated that the G/A ratio and adiponectin levels, but not GDF-15 levels, were correlated with MS [odds ratio; 95% CI 1.010 (1.006–1.013) and 0.798 (0.735–0.865), respectively] after adjusting for confounding factors. The G/A ratio displayed a significant relationship with MS in each subgroup and with each MS component in both men and women; however, adiponectin concentrations were significantly associated with MS and all its components only in men (all P <0.05). The area under the curve (AUC) of the G/A ratio and the adiponectin level for MS was 0.758 and 0.748, respectively. The highest AUC was 0.757 for the adiponectin level in men and 0.724 for the G/A ratio in women.ConclusionsThis study suggests that the G/A ratio and adiponectin are potential biomarkers for detecting MS in women and men, respectively

Charakterisierung von resistenten Zellen

Current chemotherapy with doxorubicin fails to eradicate anaplastic thyroid cancer or even to stop tumour progress. It is hypothesized that cancer initiation, progression and metastasis are driven by a small population of cancer stem cells (CSCs), which are also responsible for drug resistance. The aim of the present work was (1) to identify whether the putative cancer stem cells exist in anaplastic thyroid carcinoma cells, (2) to generate stable doxorubicin resistant anaplastic thyroid carcinoma cell line, (3) to prove whether or not drug resistance is partly due to cancer stem cells which could expel chemotherapeutic drugs and (4) to detect whether the ABC transporter inhibitors can reverse drug resistance. To test these hypotheses, anaplastic thyroid cell lines were characterized by FACS for their content of cancer stem cells, their in vitro sphere-forming capacity and their expression of multidrug resistance transporters of the ABC gene family which may confer drug resistance to the cells. Cells were treated with doxorubicin in short-term and long-term culture up to 6 months to establish a resistant cell line. The survival of cancer and cancer stem cells and the differential expression of transporters were analyzed. This work demonstrated that anaplastic thyroid cancer cell lines (C643/ HTh74/ SW1736) consisted of 0.2 – 0.6 % side population (SP) cells, which enabled the exclusion of the Hoechst dye that otherwise binds to the DNA in the major fraction of non-SP cells. SP cells highly expressed stem cell marker Oct4 and ABCG2 and multi-drug-resistant 1 (MDR1) transporters of the ABC gene family which were characterized as cancer stem cells. HTh74 anaplastic thyroid cancer cells were treated with doxorubicin in short-term and long-term culture up to 6 months to establish a resistant cell line, designated HTh74R. The survival of cancer and cancer stem cells and the differential expression of transporters were analyzed. Treatment with doxorubicin killed the large majority of cancer cells derived from anaplastic thyroid carcinoma cell lines. This conferred a growth advantage to cancer stem cells which in turn overgrew the culture. HTh74R cells consisted of about 70% SP cells, expressed high levels of ABCG2, MDR1 and Oct4 and were more clonogenic than HTh74 wild type cells. Inhibitors of ABCG2 and/or MDR1 (verapamil and fumitremorgin C) sensitized HTh74R to doxorubicin. They potentiated the doxorubicin-induced apoptosis by activating the caspase- dependent pathway in resistant anaplastic thyroid carcinoma cells and thus partly resolved drug resistance. In conclusion, the present work suggests that the failure of doxorubicin to eradicate all anaplastic thyroid carcinoma cells is mainly due to resistance of CSCs to the chemotherapeutic drug although there is a smaller fraction of resistant cells that do not express drug- exporting ABC transporters. ABC transporter inhibitors can sensitize the effect of doxorubicin to resistant cells, which is mediated by a caspase- dependent pathway. Further therapeutic strategies have to be developed that target not only the main population of cancer cells but also to eradicate CSCs that are responsible for tumour progression and recurrence.Die gegenwärtig in der Behandlung des anaplastischen Schilddrüsenkarzinoms empfohlene Chemotherapie mit Doxorubicin führt weder zu einer ausreichenden Zerstörung der Tumormasse noch kann sie die Tumorprogression aufhalten. Nach einer mittlerweile weitverbreiteten Hypothese ist die Tumorentstehung, die Wachstumsprogression und die Metastasierung auf eine kleine Population von sogenannten Karzinomstammzellen (CSCs, cancer stem cells) zurückzuführen, die auch für die Chemotherapeutika – Resistenz verantwortlich sind. Das Ziel der gegenwärtigen Arbeit war (1) mögliche Karzinomstammzellen in anaplastischen Schilddrüsenkarzinomen zu identifizieren und zu charakterisieren, (2) eine stabile Doxorubicin-resistente anaplastische Schilddrüsencarcinom-Zelllinie herzustellen, (3) zu analysieren, ob Karzinomstammzellen Chemotherapeutika aus der Zelle ausschleusen können und daher für Chemotherapieresistenz in diesen Karzinomzellen verantwortlich sind und (4) zu untersuchen, ob Hemmer der ABC- Transporter die Chemotherapieresistenz rückgängig machen können. Diese Arbeit wies nach, dass anaplastische Schilddrüsenkarzinomzelllinien (C643 / HTh74/ SW1736) 0,2 – 0,6 % sogenannter side population - Zellen aufweisen, die die Ausschleusung des Hoechst Farbstoffes aus CSC ermöglichen, während bei nicht- CSC- Zellen der Farbstoff an die DNA bindet. HTh74 side population-Zellen exprimierten den Stammzellmarker Oct4 und ABCG2 und multiple drug resistance 1 (MDR1)-Transporter der ABC Gene-Familie. HTh74 - Zellen wurden mit Doxorubicin in einer Kurzzeit- und einer Langzeitkultur bis zu 6 Monaten behandelt mit dem Ziel einer Etablierung einer resistenten Zelllinie HTh74R. Das Überleben der Karzinom – und Karzinomstammzellen und die differentielle Expression der Transporter wurden analysiert. Die Behandlung mit Doxorubicin zerstörte die große Mehrheit von Karzinomzellen der anaplastischen Schilddrüsenkarzinomzelllinien. In der Folge kam es zu einem Wachstumsvorteil der Karzinomstammzellen, die die Kultur überwuchsen. HTh74R – Zellen bestanden aus etwa 70% side population - Zellen, die eine höhere Expression von ABCG2, MDR1 und Oct4 und ein ausgeprägteres klonales Wachstum im Vergleich zu HTh74 – Zellen aufwiesen. Hemmstoffe der ABCG2- und/oder MDR1 – Transporter (Verapamil und Fumitremorgin C) machten die resistenten HTh74R – Zellen wieder empfindlich gegenüber Doxorubicin und verminderten so deren Resistenz. Verapamil potenzierte die durch Doxorubicin induzierte Apoptose über eine Aktivierung caspase - abhängiger Stoffwechselwege in den resistenten anaplastischen Schilddrüsenkarzinomzellen. Zusammenfassend lässt sich sagen, dass die unzureichende chemotherapeutische Wirkung von Doxorubicin bei anaplastischen Schilddrüsenkarzinomzellen im Wesentlichen auf eine Resistenz von Karzinomstammzellen zurückzuführen ist, wobei eingeschränkt gesagt werden muss, dass eine kleinere Anzahl resistenter Zellen die Chemotherapeutika – exportierenden ABC - Transporter nicht exprimierten. Hemmstoffe der ABC - Transporter führen folglich dazu, dass Doxorubicin in den resistenten Zellen wieder wirksam wurde. Dieser Effekt wird im Wesentlichen über den caspase - abhängigen Stoffwechselweg vermittelt. Künftige therapeutische Strategien müssen darauf zielen, nicht nur die Hauptpopulation der Karzinomzelllinien sondern auch die Karzinomstammzellen zu vernichten, die für die Tumorprogression und für die Rezidive verantwortlich sind

Analysis of Regulatory T Cell Subsets and Their Expression of Helios and PD-1 in Patients with Hashimoto Thyroiditis

Hashimoto thyroiditis (HT) is an autoimmune disease that presumably arises consequent to loss of immune tolerance to autoantigen in thyroid. Regulatory T cells (Tregs) are considered to play a vital role in maintaining the immune balance, as they own intensive suppressive function. This study was undertaken to analyze numbers of Tregs and their expressions of Helios and PD-1 in HT patients. It also aimed to explore the relationship of these with thyroid function and specific autoantibodies. Peripheral blood mononuclear cells (PBMCs) were extracted from blood of 20 healthy controls (HC) and 42 HT patients with varying thyroid functions (10 overt hypothyroidism, 12 subclinical hypothyroidism, and 20 euthyroidism). We performed flow cytometry analysis in PBMCs to detect CD4+CD25+Foxp3+Tregs and their subsets, including CD45RO+Foxp3high activated Treg cells (aTregs), CD45RO-Foxp3low resting Tregs cells (rTregs), and CD45RO+Foxp3low secreting Treg cells (sTregs), as well as the expression of Helios and PD-1 on these cells. The results showed that the percentage of Tregs, aTregs was significantly lower in HT patients and it showed inverse correlation to thyroid function states, in comparison with these in healthy controls. In addition, patients with HT showed decreased expression of Helios in aTregs, while having increased expression of PD-1 in Tregs and sTregs. The levels of Tregs, aTregs, and Helios expressing aTregs were all negatively correlated with antithyroid antibodies. In conclusion, the deficiency of Tregs frequency and aberrant expressions of Helios and PD-1 may possibly contribute to thyroid immune damage in HT

Image_1_Association of subclass distribution of insulin antibody with glucose control in insulin-treated type 2 diabetes mellitus: a retrospective observational study.tif

ObjectiveTo examine the distribution and effects of the subclass of insulin antibodies on glucose control and side events in patients with type 2 diabetes treated with premixed insulin analog.MethodsA total of 516 patients treated with premixed insulin analog were sequentially enrolled from the First Affiliated Hospital of Nanjing Medical University from June 2016 to August 2020. Subclass-specific insulin antibodies (IAs) (IgG1-4, IgA, IgD, IgE, and IgM) were detected in IA-positive patients by electrochemiluminescence. We analyzed glucose control, serum insulin, and insulin-related events between IA-positive and IA-negative groups, as well as among patients with different IA subclasses.ResultsOverall, 98 of 516 subjects (19.0%) were positive for total IAs after premixed insulin analog therapy; of these participants, 92 had subclass IAs, and IgG-IA was the predominant subclass, followed by IgE-IA. IAs were associated with serum total insulin increase and local injection-site reactions but not glycemic control and hypoglycemia. In the subgroup analysis in patients with IA-positive, the IgE-IA and IA subclass numbers were more associated with increased serum total insulin levels. Additionally, IgE-IA might be correlated more strongly with local responses and weakly with hypoglycemia, while IgM-IA might be correlated more strongly with hypoglycemia.ConclusionWe concluded that IAs or IA subclasses might be associated with unfavorable events in patients receiving premixed insulin analog therapy, which can be used as an adjunctive monitoring indicator in clinical insulin trials.</p

Data from: Molecular mechanisms of 2, 3′, 4, 4′, 5-pentachlorobiphenyl-induced thyroid dysfunction in FRTL-5 cells

Polychlorinated biphenyls (PCBs) can severely interfere with multiple animals and human systems. To explore the molecular mechanisms underlying 2, 3′, 4, 4′, 5- pentachlorobiphenyl (PCB118)-induced thyroid dysfunction, Fischer rat thyroid cell line-5(FRTL-5) cells were treated with either different concentrations of PCB118 or dimethyl sulfoxide (DMSO). The effects of PCB118 on FRTL-5 cells viability and apoptosis were assessed by using a Cell Counting Kit-8 assay and apoptosis assays, respectively. Quantitative real-time polymerase chain reaction was used to quantify protein kinase B (Akt), Forkhead box protein O3a (FoxO3a), and sodium/iodide symporter (NIS) mRNA expression levels. Western blotting was used to detect Akt, phospho-Akt (p-Akt), FoxO3a, phospho-FoxO3a (p-FoxO3a), and NIS protein levels. Luciferase reporter gene technology was used to detect the transcriptional activities of FoxO3a and NIS promoters. The effects of the constitutively active Akt (CA-Akt) and dominant-negative Akt (DN-Akt) plasmids on p-Akt, p-FoxO3a, and NIS levels were examined in PCB118-treated FRTL-5 cells. The effects of FoxO3a siRNA on FoxO3a, p-FoxO3a, and NIS protein levels were examined in the PCB118-treated FRTL-5 cells. The effects of pcDNA3 (plsmid vectors designed for high-level stable and transient expression in mammalian host)-FoxO3a on NIS promoter activity were examined in the PCB118-treated FRTL-5 cells. Our results indicated that relatively higher PCB118 concentrations can inhibit cell viability in a concentration- and time-dependent manner. Akt, p-Akt, and p-FoxO3a protein or mRNA levels increased significantly in PCB118-treated groups and NIS protein and mRNA levels decreased considerably compared with the control groups. FoxO3a promoter activity increased significantly, whereas NIS promoter activity decreased. These effects on p-FoxO3a and NIS could be decreased by the DN-Akt plasmid, enhanced by the CA-Akt plasmid, and blocked by FoxO3a siRNA. The overexpressed FoxO3a could reduce NIS promoter activity. Our results suggested that PCB118 induces thyroid cell dysfunction through the Akt/FoxO3a/NIS signaling pathway