1.miR-218-5p regulates glycolysis in human non-small cell lung cancer A549 cells by targeting PDE7A
Haiying NIU ; Gang ZHAO ; Shanna SU ; Rongrong BAI ; Peijuan MU ; Dong ZHANG
Chinese Journal of Cancer Biotherapy 2024;31(6):592-597
Objective:To investigate the mechanism of miR-218-5p regulating the glycolytic process in human non-small cell lung cancer A549 cells by targeting phosphodiesterase 7A(PDE7A).Methods:A549 cells were routinely cultured,and miR-218-5p mimic,mimic-NC,PDE7A overexpression plasmid(PDE7A-oe)and PDE7A control plasmid(PDE7A-NC)were transfected into A549 cells using Lipo3000,and recorded as the miR-218-5p mimic group,the mimic-NC group,the PDE7A-oe group and the PDE7A-NC group.The transfection efficiency was verified by qPCR assay;the expressions of glycolysis key enzyme proteins were detected by WB assay;the 2-deoxyglucose and lactate contents in A549 cells of each transfection group were detected by glucose assay and lactate production assay;the target binding relationship between miR-218-5p and PDE7A was verified by dual-luciferase reporter gene assay,and the data from the TCGA database were used to analyze the expression level of PDE7A mRNA in lung cancer tissues.Results:miR-218-5p was successfully overexpressed in A549 cells(P<0.01).Overexpression of miR-218-5p significantly inhibited the expressions of PDE7A,HK2,PKM2 proteins(all P<0.01),glucose uptake and lactate production(both P<0.01)in A549 cells.Overexpression of PDE7A significantly promoted the expressions of PDE7A,HK2,and PKM2 proteins(all P<0.01),as well as glucose uptake and lactate production(both P<0.01)in A549 cells.miR-218-5p in A549 cells could directly bind to the 3′-UTR of PDE7A mRNA.Database data analysis showed that PDE7A mRNA was highly expressed in lung squamous cell carcinoma tissues(P<0.01).Conclusion:miR-218-5p targets PDE7A to regulate the expression levels of HK2 and PKM2 in A549 cells,which in turn inhibits the glycolytic process.miR-218-5p/PDE7A may be a potential target for clinical diagnosis and treatment of NSCLC.
2.Effects of HBV-induced exosomes on macrophage phenotype and function
Rongrong MU ; Shuang LI ; Haiyan YE ; Limin CHEN ; Yujia LI ; Shilin LI
Chinese Journal of Blood Transfusion 2022;35(5):488-493
【Objective】 To investigate the effect of exosomes produced by hepatitis B virus (HBV)-infected cells on the phenotype and function of macrophages. 【Methods】 The exosomes secreted by HepAD38 cells, which were capable of producing HBV and HepG2 cells, were collected by ultracentrifugation combined with immunosorbent method.The quality and purity of the extracted exosomes were verified by nanoparticle tracking analysis (NTA), scanning electron microscope and Western blot.The M0 THP-1 macrophages differentiated by PMA were stimulated by HepAD38 derived- or HepG2 derived exosomes.Total RNA and protein samples were collected at different time points after stimulation.Real-time fluorescence quantitative PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect cytokine mRNA and protein expressions, respectively.Meanwhile, neutral red assay was performed to analyze macrophage pinocytosis activity, and a commercial kit was used to measure reactive oxygen species (ROS) in THP-1 macrophages.Human reverse transcription chip detection was performed to obtain the microRNAs profile of the exosomes.And the effect of selected miRNA on macrophages was further confirmed by qRT-PCR. 【Results】 Compared with HepG2-derived exosomes, HepAD38-derived exosomes increased the mRNA and protein expressions of IL-1β, MCP-1 and TNFα significantly.However, no difference of pinocytosis capacity or ROS production was found between the HepAD38-derived exosomes group and HepG2-derived exosomes group.Human reverse transcription chip detection results were verified by KEGG analysis and qRT-PCR, and it was found that miR-6824-3p could also significantly increase the expression levels of IL-1β, MCP-1 and TNFα after high expression. 【Conclusion】 This study found that exosomes produced by HepAD38 cells may stimulate macrophages to produce inflammatory factors such as IL-1β, MCP-1 and TNFα through miR-6824-3p, thereby playing a role in HBV infection.
3.Zika virus infection and transfusion safety
Rongrong MU ; Haiyan YE ; Min XU ; Silin LI
Chinese Journal of Blood Transfusion 2021;34(4):424-427
Zika virus(ZIKV) is an arbovirus transmitted mainly by mosquitoes. ZIKV infection usually causes mild self-limiting disease, and most of the infected patients only developed relatively mild symptoms which were often easily overlooked. However, serious damage of the central nervous system is observed in a few of the infected patients, and microcephaly of embryos and fetuses in pregnant women infected by ZIKV has attracted wide attention in recent years. Although there is a high proportion of asymptomatic infection and a high incidence of viremia, ZIKV has not been included in the routine pathogen screening before blood donation in China yet, and there is a risk of transmission through blood transfusion. This article briefly introduces the biological characteristics, epidemic characteristics, and pathogenic mechanisms of ZIKV,, and discusses ZIKV infection and the safety of blood transfusion.
4.Continuing care of discharged patients with moderate or severe craniocerebral trauma
Dongmei MU ; Xiaoxia LIN ; Rongrong LU ; Bo YIN
Chinese Journal of Hospital Administration 2019;35(3):235-237
How to raise the quality of life of discharged patients with moderate or severe craniocerebral trauma, how to improve their social adaptive ability and reduce occurrence of complications constitute key researches in neurosurgery care. Using two-dimension code plus WeChat friends for continuing care of such patients, the hospital has significantly improved the functional state, activity of daily living, complications and family care ability, contributing to better outcomes of such patients care.
5.Influence of nursing intervention on cognition level for pressure sores and psychological state among accompany family members of patients with cerebral injury
Wei XIA ; Dongmei MU ; Rongrong YU
Chinese Journal of Practical Nursing 2010;26(27):55-56
Objective To survey the influence of nursing intervention on cognition level for pressure sores and psychological state among accompany family members of patients with cerebral injury. Methods 123 accompany family members of patients with cerebral injury were selected and took part in the investigation about cognition level for pressure sores, besides, SDS and SAS were also adopted to evaluate their psychological state. Later nursing intervention was given to increase their cognition level for pressure sores, then another evaluation was carried out. Results before and after the intervention were compared. Results The accompany family members lacked knowledge of pressure sore, showed serious anxiety and depression for nursing pressure sore before intervention. But their cognition level for pressure sore improved and anxiety and depression lightened after intervention. Conclusions Nursing intervention is effective to alleviate cognition level for pressure sore and psychological state among accompany family members of patients with cerebral injury, which is worthy of clinical application.
6.Efficacy and safety of pancreas freezing
Dawei QIU ; Lizhi NIU ; Feng MU ; Xiang PENG ; Liang ZHOU ; Haibo LI ; Rongrong LI ; Kecheng XU ; Jiazan NI ; Lingzhi JIANG ; Yize HU ; Zhuofang HAO
Chinese Journal of Pancreatology 2010;10(2):124-127
Objective To observe the blood biochemical and histological changes before and after pancreas freezing, to provide evidence for cryosurgery for pancreatic cancer. Methods Fifteen healthy pigs were divided into deep frozen group (n = 5), shallow frozen group (n = 5), non-frozen group (n = 3) and normal group (n = 2). After anesthesia and Iaparotomy, a probe of the Argon-Helium Surgical System was inserted into the pancreas, 100% and 10% argon output power were used in deep and shallow frozen group, respectively;and the temperature were - 130 ~ - 140℃ and - 110 ~ - 120℃, respectively;which results in an ice-ball with 15 ~ 20 mm in diameter. Then helium gas was inputted to increase the temperature to 10 ~ 20℃ for three minutes;then the whole process was repeated. A probe was inserted into the pancreas in the non-frozen group only and only laparotomy was performed in non-grozen group normal group and normal group. Serum amylase, IL-6, CRP levels before and after the experiment was determined;the pigs were sacrificed at day 7 and the pancreas was harvested for light microscope and electron microscope examination. Results The frozen pancreatic tissue became pitchy necrosis zone, and it could be distinguished from non-frozen tissue;there were obvious tissue necrosis in the center and para-center of frozen area, and the ultra-structure were destroyed and disappeared, mitochondria degranulation and rough endoplasmic reticulum degrannlation were observed. Serum amylase was elevated in 13 (86.7%) pigs and most returned to normal at 6th day. Serum IL-6 was slightly elevated in 5 (33.3%) pigs. There was no significant difference among all the groups in term of serum CRP. All the pigs were alive until the time of sacrifice. Conclusions Cryosurgery has affirmative fatal ablative effects on pancreatic tissue, and it is safe with no serious complications.

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