Homocysteine level and gestational diabetes mellitus: a systematic review and meta-analysis.

AIMS: It is reported that elevated homocysteine (Hcy) level represents an independent risk factor for gestational diabetes mellitus (GDM). However, the relationship between Hcy level and GDM remains controversial. Our study aimed to systematically review available literature linking Hcy to GDM for a comprehensive understanding of the relationship between circulating Hcy level and GDM in humans. METHODS: PubMed, The Cochrane Library, and Web of Science were searched for studies published up to January 2021. Manual searches of references of the relevant studies were also conducted. Standard mean difference (SMD) with 95% confidence interval (95%CI) were calculated to evaluate the relationship between Hcy level and GDM using the Review Manager 5.3 and Stata 12.0. RESULTS: Of 106 references reviewed, 12 studies with a total of 712 GDM patients contributed to the present meta-analysis. Hcy level was significantly elevated in women with GDM compared with those without GDM (SMD = 0.55; 95% CI: 0.25-0.85, p = .0003). In the subgroup meta-analyses, this evidence was more consistent among women with Hcy sampling during the second trimester (SMD = 0.76, 95% CI: 0.34-1.18, p = .0004) and with average age ≥30 years (SMD = 0.69, 95% CI: 0.25-1.12, p = .002). CONCLUSION: The evidence indicated that the level of circulating Hcy was significantly elevated among women with GDM compared with those with normal glucose tolerance, especially with mean age ≥30 years and in the second trimester.

NF-κB-dependent transcriptional upregulation of cyclin D1 exerts cytoprotection against hypoxic injury upon EGFR activation.

Apoptosis of neural cells is one of the main pathological features in hypoxic/ischemic brain injury. Nuclear factor-κB (NF-κB) might be a potential therapeutic target for hypoxic/ischemic brain injury since NF-κB has been found to be inactivated after hypoxia exposure, yet the underlying molecular mechanisms of NF-κB inactivation are largely unknown. Here we report that epidermal growth factor receptor (EGFR) activation prevents neuron-like PC12 cells apoptosis in response to hypoxia via restoring NF-κB-dependent transcriptional upregulation of cyclin D1. Functionally, EGFR activation by EGF stimulation mitigates hypoxia-induced PC12 cells apoptosis in both dose- and time-dependent manner. Of note, EGFR activation elevates IKKß phosphorylation, increases IκBα ubiquitination, promotes P65 nuclear translocation and recruitment at cyclin D1 gene promoter as well as upregulates cyclin D1 expression. EGFR activation also abrogates the decrease of IKKß phosphorylation, reduction of IκBα ubiquitination, blockade of P65 nuclear translocation and recruitment at cyclin D1 gene promoter as well as downregulation of cyclin D1 expression induced by hypoxia. Furthermore, NF-κB-dependent upregulation of cyclin D1 is instrumental for the EGFR-mediated cytoprotection against hypoxic apoptosis. In addition, the dephosphorylation of EGFR induced by either EGF siRNA transfection or anti-HB-EGF neutralization antibody treatment enhances hypoxic cytotoxicity, which are attenuated by EGF administration. Our results highlight the essential role of NF-κB-dependent transcriptional upregulation of cyclin D1 in EGFR-mediated cytoprotective effects under hypoxic preconditioning and support further investigation of EGF in clinical trials of patients with hypoxic/ischemic brain injury.

[Expression and role of CXC chemokine 5 in liver cancer cells].

OBJECTIVE: To explore the expression of CXC chemokine 5 (CXCL5) in liver cancer cells and its effect on cell proliferation, migration and invasion. METHODS: Real-time (RT)-PCR and enzyme-linked immunosorbent assay (ELISA) were used to detect the mRNA and protein levels of CXCL5 in 4 liver cancer cell lines with different metastatic potentials (in ascending order: HepG2, SMMC7721, MHCC97L and MHCC97H). HepG2 with a low expression of CXCL5 was treated with CXCL5. There were four groups: 0 nmol/L CXCL5, 0.1 nmol/L CXCL5, 1.0 nmol/L CXCL5 and 10.0 nmol/L CXCL5. Cell proliferation was evaluated by cell counting kit-8 (CCK-8) assay. Transwell chambers and basement membrane matrix (Matrigel) were used to observe the cellular migration and invasion. Statistical analysis was performed with SPSS 16.0. Statistical comparison of the results was made by analysis of variance (ANOVA). RESULTS: The relative mRNA expression levels of CXCL5 in HepG2, SMMC7721, MHCC97L and MHCC97H were 0.002% ± 0.000%, 0.005% ± 0.000%, 1.030% ± 0.070% and 0.980% ± 0.190% (F = 33.88, P < 0.01) while their protein levels 14.3 ± 0.4, 25.7 ± 1.4, 82.8 ± 3.2 and 98.9 ± 1.7 respectively (F = 447.08, P < 0.01). The CCK-8 results showed that cell proliferation increased with the treatment of CXCL5, but no significant difference existed (F < 1.00, P > 0.05), cell numbers of migration of 0, 0.1, 1.0, 10.0 nmol/L CXCL5 groups were 29 ± 3, 56 ± 16, 113 ± 7 and 130 ± 15 (F = 51.94, P < 0.01), while cell numbers of invasion 17.3 ± 1.8, 33.0 ± 3.2, 65.7 ± 4.4 and 94.3 ± 3.5 respectively (F = 104.13, P < 0.01). CONCLUSIONS: Liver cancer cells with high metastatic potential have a higher expression of CXCL5. And exogenous CXCL5 can increase the proliferation, migration and invasion of liver cancer cells with low metastatic potential. Thus CXCL5 may be associated with the metastasis of liver cancer.

[Effect of let-7c on the proliferation of human hepatocellular carcinoma cell HCCLM3].

OBJECTIVE: To investigate let-7c's effect on the proliferation of human hepatocellular carcinoma cell HCCLM3 by transient transfection and the mechanism inside. METHODS: Lipofectamine 2000 was used to transfect miRNAs into HCCLM3 cells. The cells were divided into three groups, let-7c group: let-7c was transfected, negative control group: negative control miRNA was transfected, blank control group: nothing was transfected. The proliferation of HCCLM3 cells was evaluated using Cell Counting Kit-8 (CCK-8). The cell cycles of each group were assayed by flow cytometry. Western blot and Real time PCR were used to analyze the protein and mRNA expressions of cyclin D1. Statistical analysis was performed with SPSS 17.0. RESULTS: The absorbances of let-7c group were 0.70 ± 0.05, 0.77 ± 0.09 at 48 h and 72 h after transfection, lower than that of blank control group (0.97 ± 0.10, 1.21 ± 0.12) and negative control group (0.91 ± 0.07, 1.12 ± 0.09), 48 h: F = 14.431, P < 0.05, 72 h: F = 21.146, P < 0.05. The flow cytometry at 72 h after transfection revealed that let-7c increased the percentage of cells in G1 phase. The percentage of blank control group was 43.53% ± 0.86%, the negative control group was 44.82% ± 0.77%, and the let-7c group was 54.52% ± 0.13%, F = 240.739, P < 0.05. let-7c suppressed expressions of cyclin D1 at both protein and mRNA levels. The protein levels of cyclin D1 were 0.48 ± 0.09, 0.47 ± 0.06 and 0.23 ± 0.06 (F = 11.316, P < 0.05) in blank control group, negative control group and let-7c group, respectively. The mRNA levels were 1.03% ± 0.29%, 1.01% ± 0.11% and 0.63% ± 0.14% (F=6.315, P < 0.05) in the above three groups, respectively. CONCLUSION: Let-7c can inhibit proliferation of HCCLM3 cells and increase the proportion of cells in G1 phase. The mechanism may be that let-7c represses the expressions of cyclin D1 at both protein and mRNA levels.

Visfatin level and gestational diabetes mellitus: a systematic review and meta-analysis.

It is reported that elevated visfatin level is associated with gestational diabetes mellitus (GDM). However, the relationship between visfatin level and GDM remains controversial. The aim of our study was to systematically review available literature linking visfatin to GDM for a comprehensive understanding of the relationship between circulating visfatin level and GDM in human. PubMed, The Cochrane Library and Web of Science were searched for studies published up to July 2020. Standard mean difference with 95% confidence interval was calculated to evaluate the relationship between visfatin level and GDM using the Review Manager 5.3 and Stata 12.0. The evidence indicated that no significant difference was observed in the level of circulating visfatin between the women with GDM and normal glucose tolerance, suggesting circulating visfatin level is not independently related to GDM. Nevertheless, visfatin is involved in the development of GDM in obese women.