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Responsible Institution:National Health Commission of the People’s Republic of China
Sponsor:Chinese Medical Doctor Association
Honorary Editor-in-Chief:Wu Yiling
Editor-in-Chief:Jia Zhenhua
Managing Director:Ma Zhi
Editing:Editorial Board of Chinese Journal of Difficult and Complicated Cases
Publishing:Publishing House of Chinese Journal of Difficult and Complicated Cases
Address:238 Tianshan Street,Shijiazhuang,Hebei Province,China
Post Code:050035
Phone Number:0311-85901735
E-mail:ynbzz@163.com
Demestic Distributor: China Post Group Corporation Hebei Newspapers and Periodicals Distribution Bureau
 
    Current issue
Issue 8 in 2022    Publication date:2022-08-01
 
Wang Zhe ;Zhang Jing;Wu Rentong ;Zhang Chao;Chen Huaian;Miao Wenlong

Objective To investigate the effect of microRNA(miR)-489-3 p targeting chromosome 10 deleted phosphatase and tensin homolog gene(PTEN)/phosphatidylinositol 3 kinase(PI3 K)/protein kinase B(Akt) signaling pathway in the bladder Effects of cancer(BC) cell proliferation, apoptosis and invasion.Methods From June 2018 to July 2021, experiments were conducted at the First Affiliated Hospital of Hebei North University, using volcano plots to screen BC differentially expressed miRNAs from The Cancer Genome Atlas(TCGA-BLCA) database. The cancer tissues and adjacent tissues of 110 BC patients, purchased human bladder epithelial immortalized cells SV-HUC-1 and human BC cell lines 5637, J82, and T24 were used as the research objects, and real-time quantitative PCR was used to determine miR-489-3 p and PTEN mRNA expression. According to the relative expression of miR-489-3 p, BC patients were divided into 70 cases with low expression group(≤0.36) and 40 cases in high expression group(>0.36). changes in characteristics. The dual-luciferase experiment verified the targeting relationship between miR-489-3 p and PTEN. T24 cells with the lowest expression of miR-489-3 p were selected for transfection experiments. T24 cells were randomly divided into control group, miR-NC(negative control) group, miR-489-3 p mimics(mimetic) group, and miR-489-3 p mimics+pcDNA group and miR-489-3 p mimics+pc-PTEN group. MTT and colony formation assay, Transwell assay, and flow cytometry were used to detect cell proliferation, invasion and apoptosis, respectively; Western-blot was used to detect the protein expressions of PTEN, PI3 K and Akt.Results The significantly down-regulated miR-489-3 p in BC tissues was screened by volcano plot. The expression of miR-489-3 p in BC cancer tissues was lower than that in adjacent tissues, and the expression of PTEN mRNA was higher than that in adjacent tissues(t=186.168, 510.107,P<0.001); the expression of miR-489-3 p in BC cell lines was lower than that in SV-The expression of PTEN mRNA in HUC-1 cells was higher than that in SV-HUC-1 cells(F=883.826, 807.220, both P<0.001). The proportion of patients with low miR-489-3 p expression in TNM stage T3-4, lymph node metastasis, and poorly differentiated tumors was higher than that in the high expression group. Dual-luciferase experiments revealed that miR-489-3 p targets negatively regulate PTEN expression. The apoptosis rate and PI3 K and Akt protein expression levels in the miR-489-3 p mimics group were higher than those in the miR-NC group. The miR-489-3 p mimics+pc-PTEN group had lower apoptotic rate and PI3 K and Akt protein expression levels than the miR-489-3 p mimics group, and the OD values(24 h, 48 h, 72 h), the number of clones and the number of cell invasion were higher than those in the miR-489-3 p mimics group(P<0.01).Conclusion miR-489-3 p may target down-regulate PTEN expression and activate PI3 K/Akt signaling pathway, thereby inhibiting BC cell proliferation, invasion, and promoting apoptosis.

 
Yang Cheng;Zhang Yanfei;Guan Hongxin;Chen Yiming;Wei Liang;Zhong Chunlong;

Objective To observe the intervention effect of up-regulation of miR143 on intracranial aneurysm model rats through TLR4/NF κB signaling pathway.Methods From December 2020 to September 2021, the experiment was carried out in the Department of Neurosurgery, Dongfang Hospital Affiliated to Tongji University, and 60 SPF SD rats were randomly divided into blank group, model group, up-regulated group and down-regulated group, with 15 rats in each. Except for the blank group, the other rats established intracranial aneurysm models. After successful modeling, the model group was transfected with empty plasmid, the up-regulated group was transfected with miR143 overexpression plasmid, and the down-regulated group was transfected with miR143 silencing plasmid. The expression of miR143 gene was determined by miR143 lentivirus titration. The rate of intracranial aneurysm formation in rats was evaluated, and the serum transforming growth factor β1(TGF-β1), tumor necrosis factor-α(TNF-α), vascular endothelial growth factor(VEGF), matrix metalloproteinase-2(MMP-2), MMP-9 in each group were determined. levels and protein expression of TLR4/NF-κB signaling pathway in rat intracranial arteries.Results The expression of miR-143 in the model group was higher than that in the blank group(P<0.05), and the expression of miR-143 in the down-regulated group > the model group > up-regulated group(F/P=7.484/0.001). The serum levels of TGF-β1, TNF-α, VEGF, MMP-2, and MMP-9 in the model group were higher than those in the blank group(P<0.05). The levels of TGF-β1, TNF-α, VEGF, MMP-2, MMP-9 were down-regulated group > model group > up-regulated group(F/P=10.618/0.001, 8.087/0.001, 10.091/0.001, 13.827/0.001). The protein expressions of TLR4, NF-κB and Caspase-3 in the model group were higher than those in the blank group(P<0.05), and the protein expressions of TLR4, NF-κB and Caspase-3 were down-regulated group > model group > up-regulated group(F/P=16.171/0.001, 17.878/0.001, 12.078/0.001). Conclusion Up-regulation of miR-143 gene may act on the TLR4/NF-κB signaling pathway, regulate the expression of TLR4, NF-κB, and Caspase-3 proteins, which can effectively protect rat brain tissue and effectively inhibit the occurrence and development of rat cerebral aneurysms.

 
Gan Kunning;Zhang Bo;He Hui;Cui Xiangrong;Xiong Kai;

Objective To analyze the serum levels of bone alkaline phosphatase(BALP), procollagen type I(PCⅠ) and bone morphogenetic protein 2(BMP-2) in postmenopausal fracture patients and their correlation with the degree of osteoporosis.Methods A total of 171 postmenopausal fracture patients admitted to Nanning Second People's Hospital from February 2019 to February 2020 were selected as the research group. According to bone mineral density(BMD), they were divided into normal subgroup of 55 cases, decreased subgroup of 67 cases, and loose subgroup 49 cases. In addition, 177 postmenopausal women without fractures who underwent physical examination in the hospital during the same period were selected as the control group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the serum BALP, PCⅠ and BMP-2 levels of subjects and compared between groups; Spearman method was used to analyze the correlation between serum BALP, PCⅠ and BMP-2 levels and bone metabolism-related indexes; The predictive value of serum BALP, PCI and BMP-2 in postmenopausal fractures was analyzed by ROC curve; the influencing factors of postmenopausal fractures were analyzed by multivariate Logistic regression analysis.Results Compared with the control group, the serum levels of BALP and PCI in the study group were increased, and the level of BMP-2 was decreased(t/P=4.173/<0.001, 11.693/<0.001, 10.038/<0.001); normal subgroup, decreased subgroup and The serum levels of BALP and PCI in the loose subgroup increased in turn, while the level of BMP-2 decreased in turn(F/P=9.943/<0.001, 83.323/<0.001, 63.602/<0.001). Analysis by Spearman method showed that serum BALP and PCI in postmenopausal fracture patients were positively correlated with osteocalcin N-terminal middle molecular fragment(N-MID), β-linked degradation product(β-CTX), and osteocalcin(OC)(BALP:r/P=0.424/0.001, 0.536/<0.001, 0.622/<0.001; PCI: r/P=0.503/<0.001, 0.510/<0.001, 0.497/0.006), serum BMP-2 and N-MID, β-CTX and OC were negatively correlated(r/P=-0.597/<0.001,-0.614/<0.001,-0.544/<0.001); ROC curve analysis showed that serum BALP, PCI, BMP-2 and their combination predicted menopause The AUC of posterior fractures was 0.734, 0.901, 0.845, and 0.963, and the combined predictive value of the three was higher than that of the individual indicators(Z=8.046, 3.377, 5.165, all P<0.001). High β-CTX, high OC, high BALP, high PCI and low levels of BMP-2 were risk factors for postmenopausal fractures [OR(95%CI)=2.223(1.230-3.987),1.498(1.114-2.014),3.401(1.713-6.754),3.560(1.616-7.843),5.222(1.956-13.941),3.168(1.986-5.294)].Conclusion The serum levels of BALP and PCⅠin postmenopausal fracture patients increased, while the level of BMP-2 decreased, and the three were related to the degree of osteoporosis.

 
Zhang Li;Wang Liang;Wang Jun;Cheng Yanrong;Wang Lin;

Objective To investigate the gene expression and clinical significance of microprocessor complex subunit 8(DGCR8) and ribonuclease Ⅲ(DICER1) in thyroid cancer tissue.Methods A total of 90 patients with thyroid cancer who underwent surgery in the Department of Endocrinology, Xi'an High-tech Hospital Affiliated to Northwestern University from January 2015 to January 2017 were selected as the research objects. The gene expression levels of DGCR8 and DICER1 in thyroid cancer tissues and adjacent tissues were detected by real-time fluorescence quantitative PCR. Pearson correlation analysis was performed to analyze the correlation between the gene expressions of DGCR8 and DICER1, and to analyze the relationship between the gene expressions of DGCR8 and DICER1 with clinicopathological characteristics and prognosis. Multivariate COX regression analysis was used to analyze the risk factors of recurrence in patients with thyroid cancer.Results Compared with adjacent tissues, the gene expression of DGCR8 in thyroid cancer tissues was increased, while the gene expression of DICER1 was decreased(t/P=26.458/<0.001,15.019/<0.001). Compared with AJCC stage I-II thyroid cancer, the gene expression of DGCR8 in lesions of patients with AJCC stage III-IV was increased, and the gene expression of DICER1 was decreased(t/P=7.160/<0.001, 17.441/<0.001). Pearson correlation analysis showed that the gene expressions of DGCR8 and DICER1 in thyroid cancer tissues were significantly negatively correlated(r=-0.662,P<0.001). The 3-year recurrence rate of thyroid cancer patients with high gene expression of DGCR8 and low gene expression of DICER1 was higher than that of patients with low gene expression of DGCR8 and high gene expression of DICER1(χ2/P=13.580/<0.001, 27.889/<0.001). The median tumor-free survival time of patients with high gene expression of DGCR8 and low gene expression of DICER1 was shorter than that of patients with low gene expression of DGCR8 and high gene expression of DICER1(Z/P=3.425/<0.001, 3.589/<0.001). High gene expression of DGCR8, low gene expression of DICER1 and AJCC stage III-IV were independent risk factors for postoperative recurrence of thyroid cancer [OR(95%CI)=2.537(1.411-4.463),1.659(1.119-2.576),1.628(1.032-2.497)].Conclusion The increased gene expression of DGCR8 and the decreased gene expression of DICER1 in thyroid cancer tissue are related to the AJCC staging of thyroid cancer patients, which can assist in the clinical evaluation of the prognosis of thyroid cancer patients.