China ; Inventions

Glycine/analogs & derivatives* ; Humans ; Vasculitis, Leukocytoclastic, Cutaneous

[Abstract] Objective: To detect the expression of miR-126 in oral squamous cell carcinoma (OSCC) and to analyze its correlation with clinicopathological features and prognosis of patients, as well as to explore the effect of miR-126 over-expression on the malignant biological behaviors of Tca8113 cells. Methods: A total of 62 pairs of cancer and para-cancerous tissue specimens from OSCC patients who were surgically treated in the First Affiliated Hospital of Zhengzhou University from June 2016 to June 2018 were collected for this study; in addition, human tongue squamous carcinoma Tca8113 cell line and human mouth keratinocyte HOK cell line were also selected for this study. The expression of miR-126 in cancer tissues and cells was detected by qPCR, and the relationship between miR-126 expression and clinicopathological features and prognosis of the patients was analyzed. miR-126 mimics and miR-NC plasmids were respectively transfected into Tca8113 cells by liposome transfection technology. Cell proliferation, apoptosis, migration and invasion were detected by MTT method, Flow cytometry and Transwell chamber method, respectively; and the expressions of apoptosis, migration and invasion related proteins were detected by Western blotting. Results: The expression level of miR-126 in OSCC tissues and Tca8113 cells was significantly lower than that in para-cancerous tissues and HOK cells (all P<0.01). The expression of miR-126 was associated with TNM stage and lymph node metastasis (all P<0.05), and patients with high miR-126 expression had significantly better overall survival rate than patients with low expression (P<0.05). After transfection with miR-126 mimics, the cell proliferation, migration and invasion ability significantly decreased (P<0.05 or P<0.01) while the apoptosis rate significantly increased in Tca8113 cells (P<0.01), the expression levels of Bcl-2, N-cadherin and vimentin in Tca8113 cells significantly decreased (all P<0.01), and expression levels of Bax and E-cadherin significantly increased (all P<0.01). Conclusion: miR-126 is low expressed in OSCC tissues and Tca8113 cells. Up-regulation of miR-126 inhibits cell proliferation, migration and invasion and promotes apoptosis of Tca8113 cells.

[摘 要] 目的：探究蛋白酪氨酸磷酸酶D（PTPRD）去甲基化通过PI3K/Akt/mTOR通路对胃癌细胞增殖、迁移及化疗耐药性的影响。方法：体外培养胃癌细胞MKN-74、MKN-45和人胃黏膜上皮细胞GES-1并检测PTPRD表达。常规培养MKN-45细胞及耐药MKN-45/5-FU细胞，分别转染PTPRD空载体（NC组、NC/5-FU组）、PTPRD过表达腺病毒（PTPRD组、PTPRD/5-FU组）、shRNA空载体（sh-NC组、sh-NC/5-FU组）、shRNA-PTPRD慢病毒（sh-PTPRD组、sh-PTPRD/5-FU组）和PTPRD过表达腺病毒 + 10 μmol/L 740Y-P处理（PTPRD + 740Y-P组、PTPRD + 740Y-P/5-FU组）。MTT法、划痕愈合实验检测各组细胞的增殖活力和迁移能力，细胞自噬实验检测细胞的自噬水平，WB法检测细胞中EMT和PI3K/Akt/mTOR通路相关蛋白的表达。采用0、2.5、5、10、20、40 μmol/L的5-aza处理MKN-45细胞，qPCR法、MTT法检测细胞中PTPRD mRNA表达和细胞增殖活力。结果：PTPRD mRNA和蛋白在胃癌细胞中均呈低表达（P < 0.05）。与MKN-45组相比，PTPRD组自噬体与自噬溶酶体数量、PTPRD、上皮钙黏素（E-cadherin）、BAX蛋白表达均增加（均P < 0.05），细胞增殖活力、细胞迁移率、p-PI3K、波形蛋白（vimentin）、p-Akt、p-mTOR蛋白表达均降低（均P < 0.05），sh-PTPRD组细胞增殖活力、细胞迁移率、p-PI3K、vimentin、p-Akt、p-mTOR蛋白表达均增加（均P < 0.05），自噬体与自噬溶酶体数量、PTPRD、E-cadherin、BAX蛋白表达均减少（均P < 0.05）；与PTPRD组相比，PTPRD + 740Y-P组细胞增殖活力、细胞迁移率、p-PI3K、vimentin、p-Akt、p-mTOR蛋白表达均增加（均P < 0.05），自噬体与自噬溶酶体数量、PTPRD、E-cadherin、BAX蛋白表达减少（均P < 0.05）。随着5-aza浓度的增加，MKN-45细胞中PTPRD mRNA表达增加、细胞增殖活力均降低（均P < 0.05）。与MKN-45/5-FU组相比，PTPRD/5-FU组细胞迁移率、细胞增殖活力均降低（均P < 0.05），sh-PTPRD/5-FU组细胞迁移率、细胞增殖活力均增加（均P < 0.05）；与PTPRD/5-FU组相比，PTPRD + 740Y-P/5-FU组细胞迁移率、细胞增殖活力均增加（均P < 0.05）。结论：PTPRD在胃癌细胞中呈低表达状态，PTPRD去甲基化可能通过抑制PI3K/Akt/mTOR信号通路抑制胃癌细胞增殖、迁移并增强其对化疗的敏感性。

[摘 要] 目的：探讨甲基转移酶样因子3（METTL3）在食管鳞状细胞癌（ESCC）组织和细胞中的表达水平及其对ESCC细胞糖酵解和增殖能力的影响和潜在的分子机制。方法：基于TCGA数据库分析METTL3在ESCC细胞中的表达及可能的富集通路。收集2021年1月至2021年6月间在北川医学院附属医院外科手术切除的34例ESCC组织及相应癌旁组织，采用免疫组化法验证ESCC组织中METTL3的表达。采用CCK-8法和平板克隆形成实验检测干扰METTL3后ESCC细胞增殖能力的变化，利用比色法检测干扰METTL3后ESCC细胞总RNA中m6A的表达水平，采用甲基化RNA免疫沉淀定量PCR（MeRIP-qPCR）检测METTL3对葡萄糖转运蛋白4（GLUT4）基因mRNA的m6A修饰水平的影响，采用WB和qPCR等技术探索METTL3参与ESCC细胞糖酵解的生物学机制。结果：METTL3在ESCC组织以及细胞中均呈高表达（均P<0.001）。干扰METTL3表达后，ESCC细胞的增殖能力明显减弱、细胞内总RNA的m6A修饰水平显著降低（均P<0.001）。此外，干扰METTL3可显著抑制KYSE150和TE-1细胞中GLUT4基因mRNA的m6A修饰水平（均P<0.01），并通过下调GLUT4的表达抑制葡萄糖的摄取以及乳酸的释放（均P<0.01），最终下调mTORC1通路活性并抑制ESCC细胞的增殖；在干扰METTL3的ESCC细胞同时联合运用mTORC1通路抑制剂显示有协同的抗癌作用。结论：METTL3介导的m6A修饰通过调控GLUT4-mTORC1信号轴影响ESCC细胞的糖酵解及增殖。

Objective:To investigate evaluation methods to predict the outcome of nervous system development in high-risk infants. Methods:From March, 2015 to March, 2016, 336 high-risk infants were enrolled. They were assessed by General Movements (GMs) Quality Assessment, 0~1 Years Old 20 Items Neuromotor Assessment and Gesell Developmental Schedules. Results:A total of 236 infants finishied the study. GMs Quality Assessment showed that 203 cases were normal and 33 cases were abnormal in the writhing movements stage; 218 cases were normal and 18 cases were abnormal in the fidgety movemonts stage. 0~1 Years Old 20 Items Neuromotor Assessment showed that 202 cases were normal and 34 cases were abnormal. Gesell Developmental Schedules showed that 12 cases were abnormal. Conclusion:The combination of GMs Quality Assessment, 0~1 Years Old 20 Items Neuromotor Assessment and Gesell Developmental Schedules could better predict the nervous system development of high-risk infants.

Diabetic nephropathy (DN) is a common microvascular complication of type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM)，which is also the main cause of chronic kidney disease (CKD) and end-stage renal disease (ESRD). However, the treatment methods are limited at present. More and more evidences have indicated that inflammatory response is involved in the pathogenesis and progression of DN. Several anti-inflammatory strategies that target specific inflammatory mediators (transcription factors, pro-inflammatory cytokines, chemokines, adhesion molecules) and intracellular signaling pathways have shown benefits in the DN rodent model. The mechanisms related to inflammation in the development and progression of DN were summarized and new strategies to prevent or treat DN based on inflammation were briefly discussed in this review.

Cervical spondylotic myelopathy is the most serious subtype of cervical spondylosis and the most common cause of spinal cord injury. At present, it is considered that mechanical compression and ischemic changes caused by spinal stenosis constitute the pathophysiological basis of spinal stenosis, and the dynamic instability of cervical spine is also an important factor causing spinal cord injury. Its clinical manifestations and physical examinations are complex and varied, and often need to be differentiated from some geriatric diseases. In addition to the abnormal long bundle sign caused by upper motor neuron injury, about 51.9% of cervical spondylotic myelopathy patients also have root lesions. Magnetic resonance imaging is the preferred method of examination in patients with suspected cervical spondylotic myelopathy. Compared with magnetic resonance imaging, diffusion tensor imaging is much more sensitive in detecting early patients. It is suggested that, patients with moderate to severe cervical spondylotic myelopathy should receive surgery, and patients with mild cervical spondylotic myelopathy should consider conservative treatment within three years from the beginning of diagnosis.

[摘 要] 目的：探讨circ_0000615在胆管癌细胞中的表达及其对胆管癌细胞增殖、迁移和侵袭能力的影响和可能的调控机制。方法：通过qPCR检测circ_0000615在正常胆管上皮HIBEC细胞和胆管癌CCLP-1、QBC939、TFK-1和RBE细胞中的表达水平。双荧光素酶报告基因实验验证circ_0000615与miR-432-5p的靶向关系。将si-NC、si-circ_0000615（si-circ-1、si-circ-2）、inhibitor-NC和inhibitor-miR-432-5p转染至CCLP-1和QBC939细胞，转染细胞分为si-NC组、si-circ-1组、si-circ-2组、si-NC+inh-NC组、si-NC+inh-miR-432-5p组和si-circ-1+inh-miR-432-5p组，通过CCK-8、EdU和Transwell实验检测各转染组胆管癌细胞的增殖、迁移和侵袭能力。结果：在胆管癌细胞中circ_0000615呈高表达而miR-432-5p呈低表达（P<0.05或P<0.01）；双荧光素酶报告基因实验证实circ_0000615与miR-432-5p之间存在靶向关系（P<0.01）；敲减circ_0000615可明显抑制CCLP-1、QBC939细胞的增殖、侵袭和迁移能力（P<0.05或P<0.01）；下调miR-432-5p可部分逆转敲减circ_0000615对胆管癌CCLP-1和QBC939细胞增殖、迁移和侵袭的抑制作用（P<0.05或P<0.01）。结论：circ_0000615通过靶向miR-432-5p调控胆管癌细胞的增殖、侵袭和迁移。

[摘 要] 目的：明确康莱特注射液（KLTi）通过调控胆固醇代谢对肺腺癌A549细胞恶性生物学行为的抑制作用。方法：构建A549细胞体外培养模型，设置空白对照组（CON组）、KLTi组、顺铂（DDP）组及KLTi+DDP组，分别给予对应药物干预，采用CCK-8法检测不同分组的药物干预对A549细胞增殖的影响，并确定IC50值用于后续实验；使用细胞划痕实验、平板克隆形成实验、Transwell侵袭实验观察不同分组药物对A549细胞恶性生物学行为的影响；流式细胞术检测不同分组药物对A549细胞晚期凋亡水平的影响；WB法检测各组细胞上皮间质转化（EMT）相关蛋白表达，ELISA法检测促炎因子释放水平。采用比色法检测细胞胆固醇含量水平的组间差异，借助WB法检测胆固醇代谢相关膜通道蛋白ATP结合盒转运蛋白A1（ABCA1）及功能蛋白ATP柠檬酸裂解酶（ACLY）、肽基脯氨酰异构酶B（PPIB）表达水平差异。结果：KLTi及DDP对A549细胞抑制作用具有时间及剂量依赖性（均P<0.05），最终选择2 mg/mL KLTi、3 μg/mL DDP作为干预剂量，按分组加药干预48 h后显示，KLTi单用或联合DDP均可抑制A549细胞克隆形成、迁移、侵袭能力且促进其晚期凋亡，KLTi+DDP组的效果更加明显（P<0.05或P<0.01）； KLTi单用或联合DDP可通过调控E-cadherin、vimentin、snail蛋白表达从而影响A549细胞EMT进程（P<0.05或P<0.01），同时下调IL-6及IL-8的释放水平（P<0.05或P<0.01）。KLTi单用及联合DDP均可以明显降低A549细胞胆固醇含量（P<0.05或P<0.01），并且对ABCA1、ACLY、PPIB表达具有调控作用，联合组的效果更加明显（P<0.05或P<0.01）。结论：KLTi可能通过调控胆固醇代谢水平及相关通道蛋白抑制肺腺癌A549细胞增殖、迁移、侵袭等恶性生物学行为及EMT进程。