Is polyethylene glycol loxenatide 100 µg the preferred glucagon-like peptide-1 receptor agonist for type 2 diabetes mellitus? A meta-analysis and trial sequential analysis.

OBJECTIVE: This study aimed to systematically evaluate the efficacy, safety and optimal dose of polyethylene glycol loxenatide (PEX168) for treating type 2 diabetes mellitus (T2DM). MATERIALS AND METHODS: Clinical trials of PEX168 for T2DM were identified in 8 databases, with a build time limit of January 2023. Included studies were subjected to meta-analysis and trial sequential analysis (TSA). RESULTS: On the efficacy endpoint, the meta-analysis showed that PEX168 100 µg significantly reduced 0.86% glycated hemoglobin type A1c (HbA1c) (MD -0.86, 95% CI -1.02 - -0.70,  p<0.00001), 1.11 mmol/L fasting plasma glucose (FPG) (MD -1.11, 95% CI -1.49 - -0.74, p<0.00001) and 1.91 mmol/L 2h postprandial glucose (PPG) (MD -1.91, 95% CI -3.35 - -0.46, p=0.01) compared with placebo. The TSA showed that all these benefits were conclusive. On safety endpoints, total adverse events (AEs), gastrointestinal (GI) AEs, serious AEs, and hypoglycemia were comparable to placebo for PEX168 100 µg (p>0.05). In the dose comparison, the HbA1c, FPG, and 2h PPG of PEX168 200 µg were comparable to 100 µg (p>0.05), while GI AEs were significantly higher than 100 µg (RR=2.84, 95% CI 1.64-4.93,  p=0.0002). CONCLUSIONS: PEX168 100 µg can significantly lower blood glucose and does not increase the risk of total AEs, GI AEs, and hypoglycemia, which may be a preferred glucagon-like peptide-1 receptor agonist for type 2 diabetes mellitus.

[CRISPR/Cas9-mediated microRNA-21 knockout increased imatinib sensitivity in chronic myeloid leukemia cells].

Objective: To observe the effects of miR-21 knockout on proliferation and drug resistance in K562/G01 cells, and to preliminarily explore the mechanism of imatinib sensitivity by knocking out miR-21 in K562/G01 cells. Methods: Using CRISPR/Cas9 to knock out the miR-21 gene in K562/G01 cells, and single-cell-derived clones of miR-21 knockout were obtained by genomic DNA PCR screening, Sanger sequencing, and real-time PCR. We used MTT and cell colony formation assays to assess the cell proliferation, and determined imatinib sensitivity by MTT assay and Annexin-Ⅴ-APC/7-AAD double staining flow cytometry. Using western blot, we examined the potential mechanisms affecting imatinib sensitivity by knocking out miR-21 in K562/G01 cells. Results: Three miR-21 knockout K562/G01 single-cell-derived clones were successfully constructed. The mutation efficiency mediated by CRISPR/Cas9 was 7.12%-8.11%. MiR-21 knockout inhibited the proliferation of K562/G01 cells; the clone formation rates of WT and 1#, 2#, 6# K562/G01 single-cell clones were (57.67±8.25) %, (26.94± 5.36) %, (7.17±2.11) %, (31.50±3.65) %, respectively. MiR-21 knockout increased the sensitivity of K562/G01 cells to imatinib, IC(50) of imatinib in WT, and 1#, 2#, 6# K562/G01 single-cell clones were (21.92±1.36) µmol/ml, (3.98±0.39) µmol/ml, (5.38±1.01) µmol/ml, (9.24±1.36) µmol/ml. After the knockout of miR-21, the activation of PI3K/Akt signaling molecules was inhibited, while the expression of P210(B)CR-ABL and p-P210(BCR-ABL) was downregulated; however, the expression of PTEN was not affected. Conclusion: The knockout of miR-21 can suppress cell proliferation and improve sensitivity to imatinib in K562/G01 cells, which may be achieved by inhibiting the PI3K/AKT signaling pathway and BCR-ABL expression.

The effect of maternal infection on cognitive development and hippocampus neuronal apoptosis, proliferation and differentiation in the neonatal rats.

Many epidemiological reports stated a strong association between maternal infection and development of cerebral palsy, which is a major cause of cognitive impairment. The pathophysiological mechanism of intrauterine inflammation is complex. Recently, it was demonstrated that inflammation has a modulating effect on adult neurogenesis. In this study, we discovered the effect of maternal infection to hippocampal neuronal apoptosis, proliferation and differentiation, and cognitive development in the developing brains of neonatal rats. Morris water maze test was used to assess learning and memory. Terminal deoxynucleotidyl transferase dUTP nick end labeling assay was used to determine neuronal apoptosis, immunostaining was conducted to assess neurogenesis, and Western blot for extracellular signal-regulated kinase (ERK), cyclic AMP responsive element-binding protein (CREB) and brain-derived neurotrophic factor (BDNF) expression in the hippocampus. Results demonstrated that maternal infection increased neuronal apoptosis and significantly impaired spatial learning and memory ability. Maternal infection significantly increased cell proliferation, accompanied by an increased expression of ERK (P3-P7), CREB (P3-P7) and BDNF (P3). On P28, there was no significant difference of cell survival and differentiation in two groups. These results suggest that variation in ERK activity and subsequent expression of its downstream targets, including CREB and BDNF might contribute, at least partially, to modulation of inflammation related cell proliferation, survival and differentiation. Maternal infection increased hippocampal neuronal apoptosis and affected cell proliferation and differentiation in neonatal rats, which may be regarded as an etiological factor in cognitive development impairment.

Effects of calcitonin gene-related peptide on apoptosis of peripheral blood mononuclear cells from patients with systemic lupus erythematosus.

BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune disease with abnormal apoptosis and autoantibody production. Calcitonin gene-related peptide (CGRP) is a neuropeptide produced by the central and peripheral nervous systems and by endocrine cells. It can influence cell death in thymocytes and cultured smooth muscle cells, and inhibits the production of interleukin (IL)-2, which inhibits apoptosis. AIMS: To investigate the effects of calcitonin gene-related peptide (CGRP) on apoptosis of peripheral blood mononuclear cells (PBMCs) from patients with SLE. METHODS: The percentage of apoptosis of PBMCs from patients with SLE and healthy blood donors were examined using annexin-V/propidine iodide staining 48 h after culturing with CGRP and/or its receptor antagonist CGRP(8-37) at various concentrations. IL-2 activity in culture supernatants was determined using the incorporation of 3H-TdR by the IL-2-dependent cell line CTLL and evaluated by ELISA. RESULTS: The percentage of spontaneous apoptosis of PBMCs from patients with SLE was higher than that of healthy blood donors (34.18 +/- 2.86 vs. 18.23 +/- 0.81, P < 0.001). CGRP, at all tested concentrations, had no effect on apoptosis of PBMCs from healthy blood donors, but significantly inhibited apoptosis of PBMCs from patients with SLE at concentrations of >or= 0.5 x 10(-8) mol/L (0.5 x 10(-8) mol/L: 32.01 +/- 3.98 vs. 34.18 +/- 2.86, P < 0.05; 1.0 x 0(-8) mol/L: 26.76 +/- 2.69 vs. 34.18 +/- 2.86, P < 0.001; 1.5 x 10(-8) mol/L: 25.97 +/- 2.65 vs. 34.18 +/- 2.86, P < 0.001), and the effect plateaued at 1.0 x 10(-8) mol/L, at which level inhibition was not significantly increased with increasing concentration. However, CGRP, at any concentration tested, could not reduce the rate of apoptosis of PBMCs from patients with SLE to the normal range. There were significant positive relationships between the effect of CGRP, on apoptosis of PBMCs and SLE Disease Activity Index (0.5 x 10(-8) mol/L: r(s) = 0.328, P < 0.05; 1.0 x 10(-8) mol/L: r(s) = 0.431, P < 0.01; 1.5 x 10(-8) mol/L: r(s) = 0.419, P < 0.01). CGRP(8-37) itself had no effect on apoptosis, but was able to block the effects of CGRP on PBMCs (0.5 x 10(-8) mol/L: 32.01 +/- 3.98 vs. 33.12 +/- 2.37, P < 0.05; 1.0 x 10(-8) mol/L: 26.76 +/- 2.69 vs. 34.73 +/- 2.32, P < 0.001; 1.5 x 10(-8) mol/L: 25.97 +/- 2.65 vs. 35.25 +/- 3.37, P < 0.001). There was no effect of CGRP on IL-2 production. CONCLUSIONS: These results indicate that CGRP plays an important role in the apoptosis of PBMCs from patients with SLE via its receptor; low plasma levels of CGRP may cause accelerated apoptosis. This regulation does not seem to be related to IL-2.