Additional file 1: Figure S1. of Quantitation of next generation sequencing library preparation protocol efficiencies using droplet digital PCR assays - a systematic comparison of DNA library preparation kits for Illumina sequencing

Comparison of the overall yields for libraries prepared with the Truseq Nano kit with either the Sanger adaptors (original Illumina adaptors, in pink) and the modern Illumina adaptors (in blue). Figure S2 Bar charts showing the stepwise DNA library preparation yields of the different kits tested. Initial DNA input: 500 ng. Except where mentioned otherwise all libraries were prepared using the original Illumina Paired end adaptor (Sanger adaptors) [6, 22]. The most critical steps correspond to the adaptor ligation for which the yield varies from 3.50 to 100 % depending on the kit tested. Figure S3 Bar charts showing the comparison of the overall DNA library preparation yields of the different kits tested depending on the initial DNA input. Although higher DNA inputs lead to slightly higher adaptor ligation yields, the final PCR yield appears much greater when the initial DNA input is low. Figure S4 Bioanalyzer traces of 3 libraries prepared with the PhiX amplicons of 3 different sizes. The initial input sample contained a equimolar ratio of the 3 amplicons whereas this ratio varies in the final libraries presented here depending on the kit used (Truseq Nano in red, SureSelect in blue and KAPA hyper in green). Figure S5 Enzymatic shearing using the fragmentase provided with the KAPA HyperPlus kit. A) Tunability and robustness of the fragmentase treatments depending on the GC content of the DNA sample, DNA input and the incubation time. B) KAPA HyperPlus libraries GC contents and their correlation with the theoretical values. (PPTX 367 kb

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

Additional file 1 of The effect of metformin on senescence of T lymphocytes

Additional file 1: Supplementary Figure 1. The composition of T cell subsets at different age groups. (a)The proportion of lung cancer patients at different ages from Sun Yat-sen University First Affiliated Hospital from 2017-2023 (n=4498). The frequency of senescent T cells (b), Teff cells (CD3+CD45RA+CCR7-) (d) and Tem cells (CD3+CD45RO+CCR7-) (e) in CD8+T cells and CD4+T cells at different age groups. (c) Pie charts depicting the events of Tn (CD3+CD45RA+CCR7+), Tcm (CD3+CD45RO+CCR7+), Tem and Teff cell subsets of CD8+ and CD4+T cells from different age groups. Expressed as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001; Mann– Whitney test (two-tailed) and nonpaired Student’s t-test. Tn, naïve T cell; Teff, effector T cell; Tem, effector memory T cell; Tcm, central memory T cell. Young, young-age group; Middle, middle-age group; old, elderly group

Effect of resveratrol on albuminuria and renal hyperfiltration in diabetic rats.

<p>The levels of blood glucose (A), body weight (B), albuminuria (C), kidney weight (D), and CCr (E) in each group of rats. *<i>P</i><0.05 vs CON; #P<0.05 vs DN. CON, control; DN, diabetic nephropathy; RSV, resveratrol. UAER, urine albumin excretion rate; CCr, creatinine clearance rate. </p

Effect of resveratrol on VEGF-induced cellular junction disruption and hyperpermeability in cultured mouse glomerular endothelial cells.

<p>Cultured mouse glomerular endothelial cells were pre-incubated with or without resveratrol (25μM) for 24 hours, and then treated with or without VEGF (50ng/mL) for 3 hours. (A) Immunofluorescence study of Claudin-5 and ZO-1 in each group of cells. (B) In vitro vascular permeability assay in each group of cells. *<i>P</i><0.05 vs CON; #P<0.05 vs VEGF. CON. control; RSV, resveratrol. </p

Effect of resveratrol on Flk-1 expression in cultured mouse glomerular endothelial cells.

<p>(A) Western blot analysis of Flk-1 expression in cultured mouse glomerular endothelial cells treated with resveratrol in various concentrations for 24 hours. *<i>P</i><0.05 vs NG+M; #P<0.05 vs HG. &<i>P</i><0.05 vs HG+R5; P<0.05 vs HG+R10. (B) Western blot analysis of Flk-1 expression in cultured mouse glomerular endothelial cells treated with resveratrol (25μM) for various time periods. *P<0.05 vs NG+M; #P<0.05 vs HG. &P<0.05 vs HG+R12; <i>P</i><0.05 vs HG+R24. (C) Western blot analysis of Sirt1 and Flk-1 expression in the lentivirus-infected cultured mouse glomerular endothelial cells treated with or without resveratrol (25μM) for 24 hours. *<i>P</i><0.05 vs Scramble shRNA. (D) Western blot analysis of Sirt1 and Flk-1 expression in the plasmids-transfected cultured glomerular endothelial cells. *<i>P</i><0.05 vs pCruzHA. NG+M, normal glucose plus mannitol; HG, high glucose; R, RSV, resveratrol. </p

Effect of resveratrol on VEGF expression in cultured mouse podocytes.

<p>(A) Western blot analysis of VEGF expression in cultured mouse podocytes treated with resveratrol in various concentrations for 24 hours. *<i>P</i><0.05 vs NG+M; #P<0.05 vs HG. &<i>P</i><0.05 vs HG+R5; P<0.05 vs HG+R10. (B) Western blot analysis of VEGF expression in cultured mouse podocytes treated with resveratrol (25μM) for various time periods. *P<0.05 vs NG+M; #P<0.05 vs HG. &P<0.05 vs HG+R12; <i>P</i><0.05 vs HG+R24. (C) ELISA analysis of VEGF concentrations in the cultured podocytes media treated with or without resveratrol (25μM) for 24 hours. *<i>P</i><0.05 vs NG+M; #P<0.05 vs HG. (D) Western blot analysis of Sirt1 and VEGF expression in the lentivirus-infected cultured podocytes treated with or without resveratrol (25μM) for 24 hours. *<i>P</i><0.05 vs Scramble shRNA. NG+M, normal glucose plus mannitol; HG, high glucose; R, RSV, resveratrol. </p

Effect of resveratrol on renal angiogenesis in diabetic rats.

<p>(A) Immunohistochemistry of Sirt1 expression in normal glomeruli of human, rat and mouse. (B) Western blot analysis of VEGF, Flk-1, Ang-1, and Tie-2 expression in the kidneys from each group of rats. (C) Realtime PCR analysis of renal Ang-2 mRNA transcription in each group of rats. *<i>P</i><0.05 vs CON; #P<0.05 vs DN. P, podocyte; M, mesangial cell; E, endothelial cells; CON, control; DN, diabetic nephropathy; RSV, resveratrol. </p

Effects and Safety of Calcimimetics in End Stage Renal Disease Patients with Secondary Hyperparathyroidism: A Meta-Analysis

<div><h3>Purpose</h3><p>Secondary hyperparathyroidism (SHPT) is one of the most common abnormalities of mineral metabolism in patients with chronic kidney disease. We performed a meta-analysis to determine the effect and safety of cinacalcet in SHPT patients receiving dialysis.</p> <h3>Methods</h3><p>The meta-analysis was performed to determine the effect and safety of cinacalcet in SHPT patients receiving dialysis by using the search terms ‘cinacalcet’ or ‘mimpara’ or ‘sensipar’ or ‘calcimimetic’ or ‘R586’ on MEDLINE and EMBASE (January 1990 to February 2012).</p> <h3>Results</h3><p>Fifteen trials were included, all of which were performed between 2000 and 2011 enrolling a total of 3387 dialysis patients. Our study showed that calcimimetic agents effectively ameliorated iPTH levels(WMD, −294.36 pg/mL; 95% CI, −322.76 to −265.95, <em>P</em><0.001) in SHPT patients and reduced serum calcium (WMD, −0.81 mg/dL; 95% CI, −0.89 to −0.72, <em>P</em><0.001) and phosphorus disturbances(WMD, −0.29 mg/dL; 95% CI, −0.41 to −0.17, <em>P</em><0.001). The percentage of patients in whom there was a 30% decrease in serum iPTH levels by the end of the dosing was higher in cinacalcet group than that in control group(OR = 10.75, 95% CI: 6.65–17.37, <em>P</em><0.001). However, no significant difference was found in all-cause mortality and all adverse events between calcimimetics and control groups(OR = 0.86, 95% CI: 0.46–1.60, <em>P</em> = 0.630; OR = 1.30, 95% CI: 0.78–2.18, <em>P</em> = 0.320, respectively). Compared with the control therapy, there was a significant increase in the episodes of hypocalcemia (OR = 2.46, 95% CI: 1.58–3.82, <em>P</em><0.001), nausea (OR = 2.45, 95% CI: 1.29–4.66, <em>P</em> = 0.006), vomiting(OR = 2.78, 95% CI: 2.14–3.62, <em>P</em><0.001), diarrhea(OR = 1.51, 95% CI: 1.04–2.20, <em>P</em> = 0.030) and upper respiratory tract infection (OR = 1.79, 95% CI: 1.20–2.66, <em>P</em> = 0.004)in calcimimetics group.</p> <h3>Conclusions</h3><p>Calcimimetic treatment effectively improved biochemical parameters of SHPT patients receiving dialysis without increasing all-cause mortality and all adverse events.</p> </div

Forest plot of iPTH of patients treated with calcimimetics and control therapy.

<p>Studies are identified by name of the first author and year of publication. Mean differences (MDs) are pooled using the fixed-effect model and shown on a scale of −500 to 500.</p