N⁶-methyladenosine (m⁶A) modification plays a critical role in cell cycle regulation, while the mechanism of m⁶A in regulating mitosis remains underexplored. Here, we found that the total m⁶A modification level in cells increased during mitosis by the liquid chromatography-mass spectrometry/mass spectrometry and m⁶A dot blot assays. Silencing methyltransferase-like 3 (METTL3) or METTL14 results in delayed mitosis, abnormal spindle assembly, and chromosome segregation defects by the immunofluorescence. By analyzing transcriptome-wide m⁶A targets in HeLa cells, we identified polo-like kinase 1 (PLK1) as a key gene modified by m⁶A in regulating mitosis. Specifically, through immunoblotting and RNA pulldown, m⁶A modification inhibits PLK1 translation via YTH N⁶-methyladenosine RNA binding protein 1, thus mediating cell cycle homeostasis. Demethylation of PLK1 mRNA leads to significant mitotic abnormalities. These findings highlight the critical role of m⁶A in regulating mitosis and the potential of m⁶A as a therapeutic target in proliferative diseases such as cancer.
Humans ; Polo-Like Kinase 1 ; Cell Cycle Proteins/metabolism* ; Proto-Oncogene Proteins/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; Mitosis/physiology* ; HeLa Cells ; Adenosine/genetics* ; Methyltransferases/metabolism* ; RNA, Messenger/metabolism* ; RNA-Binding Proteins/metabolism*

Objective Preliminarily discussing the effects of VEGF and SOX2 on metastasis and progno-sis of squamous cell lung carcinoma(SCLC).Methods Using immunohistochemical method to detect 47 cases of pulmonary squamous cell carcinoma and 30 cases of para-carcinoma tissue in the expression of VEGF and SOX2, using statistical methods to analyze the expression of VEGF and SOX2 and the relationship between clinical parame-ters and its relationship with prognosis. Results The positive expression rates of VEGF,SOX2 in SCLC tissues were higher than adjacent normal tissues.There was a significant correlation between VEGF and SOX2 expression. VEGF expression was associated with lymph node metastases and TNM stage.SOX2 expression was associated with tumor differentiation and lymph node status.Survival analysis indicated that VEGF(+)/SOX2(+)patients had the worst prognosis.Furthermore,multivariate analysis suggested that tumor diameter and lymph node metastases were independent prognostic factors for SCLC. Conclusion The expression of VEGF was positively correlated with lymph node metastasis and clinical stage,the expression of SOX2 was positively correlated with tumor differentia-tion degree and lymph node metastasis,and high expression of VEGF and SOX2 may indicate lung squamous can-cer patients with poor prognosis.

Objective To summarize the assessment standards of organ donation after cardiac death (DCD) donor lungs application and donor lung function maintenance experience.Method From Jan.2013 to June 2015,139 cases of DCD donors were subjected to rigorous assessment and effective donor lung function maintenance,and 11 donor lungs for lung transplantation were obtained.The donor lung cold ischemia time was (526.8-± 12.6) min (312 to 675 min).Double lung transplantation was performed on 9 cases,and 2 cases received single lung transplantation.Result Perioperatively,1 lung transplant recipient died of pulmonary infection.The survived 10 recipients had no rejection after operation,and obtained good quality of life during discharge to the final follow-up.Condusion The effect of donor lung transplantation using organ donation is satisfactory.The assessment standards and functional maintenance of donor lung are important factors to guarantee the success of lung transplantation.

Objective To investigate the effect of hydrochloric acid (HCl) stimulation and mechanical stretch on epithelial-mesenchymal transition (EMT) and hyaluronan (HA) production in human lung epithelial cells. Methods Human lung epithelial cell line BEAS-2B was cultured in vitro, which was divided into phosphate-buffer saline (PBS) + static group, HCl + static group, PBS + stretch group, and HCl + stretch group respectively in the logarithmic phase. The BEAS-2B cells in two stretching groups were challenged by cyclic stretch with 20% amplitude, frequency of 0.33 Hz, sine wave of the FX-5000T system for 48 hours. The morphology changes in cells before and after stretch were observed with inverted microscope. The protein expressions of epithelial markers E-cadherin and cytokeratin-8 (CK-8) as well as mesenchymal markers vimentin and α-smooth muscle actin (α-SMA) were determined by Western Blot. The secretion of HA was determined by enzyme linked immunosorbent assay (ELISA). Results ① It was shown by microscopic observation that BEAS-2B cells displayed cobblestone morphology, linked closely and cell polarity in PBS + static group, which did not change obviously after HCl stimulation alone. Given purely mechanical stretch after 48 hours, the cells morphology changed from cobblestone shape into long spindle, and increased intercellular space obviously. Double hit of HCl and stretch changed the cells morphology more significantly. ② It was shown by Western Blot that compared with the PBS + static group, HCl alone or combined with purely mechanical stretch after 48 hours, the expressions of E-cadherin and CK-8 were decreased, while those of vimentin and α-SMA were increased, and it was more pronounced in HCl + stretch group [the expression quantity (gray value) as base 1 in PBS + static group, E-cadherin: 0.16±0.08 vs. 1, CK-8: 0.10±0.03 vs. 1, vimentin: 3.35±0.38 vs. 1, α-SMA: 3.10±0.45 vs. 1, all P < 0.01]. ③ It was shown by ELISA that both HCl stimulation and stretch could induce BEAS-2B cells secreting HA as compared with PBS + static group (μg/L: 55.763±0.687, 63.005±0.493 vs. 49.876±1.867), and the production of HA increased more remarkably after double hit (μg/L: 78.220±1.085 vs. 49.876±1.867, P < 0.01). Conclusions Both HCl and mechanical stretch could induce EMT and increase HA secretion in human lung epithelial cells in vitro. Double hit of HCl stimulation and mechanical stretch induced EMT apparently, and further increased the production of HA.

Objective To summarize the evaluation standard of cardiac death organ donation from Chinese first category donor heart application and experience of donor heart function maintenance.Method From Jan.2013 to Jan.2015,donor hearts for heart transplantation were obtained in 18 cases of Chinese first category cardiac death organ donors from 109 cases of organ donation donors through rigorous assessment and effective donor heart function maintenance.The diagnosis of brain death was based on the diagnosis of brain death criteria (adult) by the neurological department of internal medicine,department of neurosurgery,and intensive care unit with brain death qualification.Organ donation work followed the basic principles of voluntary,free,fair,equitable and technical access.Result Eighteen case of heart transplantations were all operated with double lumen venous anastornosis.The cold ischemia time was (125.5+ 18.7) min (61-60 min),and cardiopulmonary bypass time was (130.4+ 12.5) min (99-193 min).In 18 heart transplantations,16 cases survived,and 2 eases died of acute right heart failure.During the follow-u period,the quality of life in the survival recipients was satisfactory.Conclusion Using cardiac death organ donation from Chinese first category donor heart transplantation has achieved good results.The accurate assessment of donor hearts and effective donor heart function maintenance are the key factors to guarantee the success of heart transplantation.