IntroductionAmyotrophic lateral sclerosis (ALS) is a rare, devastating neurodegenerative disease that affects upper and lower motor neurons. To date, no effective treatment or reliable biomarker for ALS has been developed. In recent years, many factors have been proposed as possible biomarkers of ALS; however, no consensus has been reached. Therefore, a reliable biomarker is urgently needed. Eosinophils may play a crucial role in healthy humans and diseases, and serve as a biomarker for many chronic diseases.MethodsRoutine blood test results were collected from 66 healthy controls and 59 patients with ALS. The percentages and total numbers of each cell population were analyzed, and the correlation between these indicators and patient ALS functional rating scale–revised (ALSFRS-R) score or disease progression rate (ΔFS score) was analyzed.ResultsCompared to healthy controls, the number of blood leukocytes, neutrophils, monocytes, and basophils was significantly decreased in patients with ALS (p = 0.002, p = 0.001, p = 0.049, and p < 0.0001, respectively). There was an increase in the number of eosinophils (p < 0.0001), but no difference in the number of lymphocytes between patients with ALS and healthy controls was found (p = 0.563). Compared to healthy controls, the percentage of neutrophils was decreased and the percentage of lymphocytes and eosinophils was increased in patients with ALS (p = 0.01, p = 0.012, and p = 0.001, respectively). There was no difference between patients with ALS and healthy controls in the percentage of monocytes and basophils (p = 0.622 and p = 0.09, respectively). However, only the percentage and number of eosinophils had a correlation with the ΔFS score. Further multivariate analysis revealed a significant correlation between the disease duration, eosinophil count and percentage, and the disease progression rate (p < 0.0001, p = 0.048, and p = 0.023, respectively). The neutrophil-to-eosinophil ratio (NER), lymphocyte-to-eosinophil ratio (LER), and monocyte-to-eosinophil ratio (MER) were significantly lower in patients with ALS than in healthy controls. However, only the LER was significantly correlated with the ΔFS score.ConclusionThese observations implicate neutrophils, lymphocytes, and eosinophils as important factors, and increasing eosinophil counts were negatively correlated with the ΔFS score in patients with ALS.

BACKGROUND:Metastasis is the leading cause of mortality in patients with breast cancer (BC). Studies demonstrate that circular RNAs (circRNAs) were involved in BC progression, while the molecular mechanisms remain largely unclear.METHODS:The microArray circRNA profiles were used to explore the differential expression circRNAs in BC and paracancerous normal tissues, and the quantitative reverse transcription-polymerase chain reaction was used to validate their expression level in clinical samples and cell lines. Nuclear/cytosolic fractionation and fluorescence in situ hybridization (FISH) assays were performed to examine circRRM2 (hsa_circ_0052582) subcellular location. The scratch wound healing and transwell assays were conducted to evaluate the impact of circRRM2 on BC cell migration and invasion. We predicted miRNAs that might bind with cricRRM2 and the downstream target genes using bioinformatics analysis and explored their expression levels and prognostic value in BC. FISH, RNA immunoprecipitation, Co-immunoprecipitation, Western blot, and rescue experiments were implemented to figure out circRRM2 function and underlying mechanisms in BC.RESULTS:The present study revealed several aberrant circRNAs in BC tissues and observed that circRRM2 was upregulated in tumor tissues of 40 patients with BC. High circRRM2 was significantly associated with advanced N stage in patients with BC. Gain- and loss- of function experiments revealed that circRRM2 promoted the migration and invasion of cells and functioned as an oncogene in BC. Mechanism studies showed that circRRM2 competed with miR-31-5p/miR-27b-3p to upregulate the IGF2BP1 expression. Furthermore, IGF2BP1 upregulated the circRRM2 level via interacting with MYC, which functioned as the transcriptional factor of circRRM2. Thus, the positive feedback loop that was composed of circRRM2/IGF2BP1/MYC was identified.CONCLUSION:This study confirms that upregulated circRRM2 functions an oncogenic role in BC metastasis. The positive feedback loop of circRRM2/IGF2BP1/MYC enforces the circRRM2 expression, which might offer a potential target for BC treatment.

Targeted therapy with tumour-associated macrophages (TAMs) has emerged as a new paradigm for immunotherapy of cervical cancer. Nocardia rubra cell wall skeleton (Nr-CWS) for external use is an immunotherapeutic agent. In this study, we aimed to explore the effects of Nr-CWS on TAMs and the potential mechanisms. Cervical tissue samples were collected before and after Nr-CWS treatment from patients with high-risk HPV infection and cervical intraepithelial neoplasia (CIN). The effect of Nr-CWS on macrophages in vivo was examined by immunohistochemistry and double-labeling immunofluorescence histochemistry. In vitro experiments were performed using a TAM model established by THP-1 cells under Nr-CWS treatment. We found that Nr-CWS treatment significantly reduced the numbers of total macrophages and M2 macrophages, increased the proportion of M1 macrophages and decreased the proportion of M2 macrophages in cervical tissues. After Nr-CWS treatment in vitro, the expression levels of the M1 macrophage markers were increased, while the expression levels of the M2 macrophage markers were decreased. Nr-CWS treatment also activated STAT1 pathways but inhibited STAT6 pathways. These results indicated that Nr-CWS may improve local immune response and reverse immunosuppression by regulating the M2 to M1 polarization of TAMs via STAT1/STAT6 pathways.

Background: Chronic allograft dysfunction (CAD) is the leading cause of graft loss among kidney transplant recipients (KTRs). Bile acids (BAs) play an important role in regulating inflammatory process, which is the major contributor to the development of CAD. The aim of this study was to evaluate the association between BAs metabolic dys-regulation and CAD in KTRs.Material/Methods: Fifteen serum BA species were determined in 43 healthy controls (HCs) and 131 KTRs by UPLC-MS/MS. KTRs were grouped into stable renal function (STA) and CAD1 and CAD2 groups based on eGFR levels. Circulating CYP7A1, CYP7B1, CYP27A1, and SLCO2B1 mRNA levels were determined by RT-PCR.Results: Total BA concentrations were comparable among the 4 groups. However, KTRs showed significantly different BAs profiling compared to HCs. KTRs with severe CAD (CAD2) had significantly lower unconjugated BAs and secondary BAs (SBAs) compared to the other 3 groups. KTRs had significantly lower SBAs/primary BAs (PBAs) ratios than HCs, which were comparable among the 3 KTR groups. Conjugated/unconjugated BAs ratios in-creased significantly with the deterioration of allograft function, which was further confirmed by correlation analysis. Differential correlation network analysis revealed that perturbations in intraclass and interclass BA co -regulation existed during CAD progression. Moreover, relative gene expressions of CYP7B1 and CYP27A1 were positively correlated with eGFR.Conclusions: BA species profiling, but not total BA concentrations, was significantly altered in KTRs with CAD. The shifts from unconjugated BAs toward conjugated BAs, SBAs toward PBAs, and distinct pairwise BAs coregulation patterns were the main characteristics of KTRs with CAD.

列举了医疗设备入网的分类,从医疗机构管理分工、医疗设备入网技术、网络安全和数据安全、诊疗信息共享和资源利用等方面分析了医疗设备入网不规范的原因以及产生的影响,并详细阐述了医疗设备入网各阶段的规范化措施,旨在有效预防医疗设备入网的不安全行为的发生,提升医护人员的人机交互体验,提升为患者提供的诊断与治疗服务效率,为医疗资源合理化利用提供依据,为临床和科研提供大量有价值的临床数据.

介绍了5G、VR技术的特点以及二者结合的优势;以河北医科大学第一医院为例,介绍了其5G网络建设,详细阐述了基于5G+VR技术的探视系统的架构及应用情况.

The Nocardia rubra cell wall skeleton (Nr-CWS) for external use is an immune enhancer, which has been widely used in human cervix diseases such as cervical erosion, but the mechanism of Nr-CWS enhancing immunity is still unclear. The purpose of this study was to explore the effect and mechanism of Nr-CWS on the local immune status of cervical tissue in patients with high-risk human papillomavirus (HR-HPV) infection and cervical precancerous lesion, cervical intraepithelial neoplasia (CIN). The recruited patients with HR-HPV infection and CIN were treated with Nr-CWS. The specimens were taken from these patients before and after local application of Nr-CWS respectively. The normal control specimens were tested simultaneously. Serial section analysis of immunohistochemistry and co-expression analysis were performed to characterize populations of T cells and the expressions of programmed cell death-1 (PD-1) and programmed cell death-ligand 1 (PD-L1). The levels of cytokines in local cervical tissue were also detected. Nr-CWS significantly increased T cells including CD4 + , CD8 + T cells, and reduced the expression of PD-L1 in the patients’ local cervical tissues. Co-expression analyses showed that the proportions of PD-1 + CD4 + cells in CD4 + T cells and PD-1 + CD8 + cells in CD8 + T cells decreased after Nr-CWS application. Furthermore, the increase in the number of immune cells was accompanied by increased pro-inflammatory cytokines interleukin-12 (IL-12), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and decreased suppressive cytokine IL-10. The results indicate that Nr-CWS, as an immunotherapeutic agent for HR-HPV infection and CIN, plays an immune promoting role related to the upregulation of T cell subsets and the inhibition of PD-1/PD-L1 pathway.

目的 探讨IL-17A对人子宫内膜异位症在位内膜细胞增殖的影响.方法 免疫细胞化学方法检测IL-17A受体在正常和子宫内膜异位症在位内膜上皮细胞及基质细胞中的表达;MTT法观察不同浓度IL-17A对正常和子宫内膜异位症在位上皮和基质细胞增殖的影响.结果 正常和子宫内膜异位症在位上皮细胞和基质细胞均表达IL-17A;但IL-17A对正常子宫内膜细胞增殖无影响;250 μg/ml IL-17A对子宫内膜异位症在位内膜细胞增殖影响显著(P＜0.05).结论 250 μg/ml IL-17A能显著促进子宫内膜异位症在位内膜细胞的增殖,为进一步证实IL-17A可能在子宫内膜异位症的发生作用提供实验基础.

Oocytes reconstructed by spindle transfer (ST) are prone to chromosome abnormality, which is speculated to be caused by mechanical interference or premature activation, the mechanism is controversial. In this study, C57BL/6N oocytes were used as the model, and electrofusion ST was performed under normal conditions, Ca2+ free, and at room temperature, respectively. The effect of enucleation and electrofusion stimulation on MPF activity, spindle morphology, γ-tubulin localization and chromosome arrangement was compared. We found that electrofusion stimulation could induce premature chromosome separation and abnormal spindle morphology and assembly by decreasing the MPF activity, leading to premature activation, and thus resulting in chromosome abnormality in oocytes reconstructed via ST. Electrofusion stimulation was an independent factor of chromosome abnormality in oocytes reconstructed via ST, and was not related to enucleation, fusion status, temperature, or Ca2+. The electrofusion stimulation number should be minimized, with no more than 2 times being appropriate. As the electrofusion stimulation number increased, several typical abnormalities in chromosome arrangement and spindle assembly occurred. Although blastocyst culture could eliminate embryos with chromosomal abnormalities, it would significantly decrease the number of normal embryos and reduce the availability of embryos. The optimum operating condition for electrofusion ST was the 37°C group without Ca2+.

MicroRNAs (miRNAs) have been reported as key gene regulators, and they control many fundamental biological processes. Previously, we demonstrated that miR-214 had a protective effect against myocardial apoptosis and myocardial fibrosis. In this study, we sought to investigate the expression of miR-214 in L6 skeletal myoblast (SKM), the regulatory effect of miR-214 on hydrogen peroxide (H2O2) induced cell apoptosis and the underlying mechanisms of the antiapoptotic effect. MiR-214 expression was up-regulated by H2O2 in a dose and time-dependent manner in L6 SKMs. To investigate the regulatory effects of miR-214 on L6 SKM, both gain-of-function and loss-of-function approaches were applied. The results showed that miR-214 improved cell survival and inhibited cell apoptosis, and blockage of miR-214 abrogated the protective effect on cell survival and resistance to apoptosis. Phosphatase and tensin homolog (PTEN) was negatively regulated by miR-214, and PTEN inhibitor obviously reversed the effect of miR-214 blockage on enhancing cell apoptosis. In addition, miR-214 up-regulated antiapoptotic protein Bcl-2, down-regulated proapoptotic protein Bax, prevented release of cytochrome c and inhibited caspase-3 activation. In summary, H2O2-induced injury increases miR-214 expression in L6 SKM, and miR-214 contributes to the protection of L6 SKM against apoptosis via lowering PTEN and subsequently inhibiting the mitochondrial-mediated caspase-dependent apoptotic signaling pathway.

目的 研究分析PD-1抑制剂免疫治疗肺癌的有效护理措施.方法 选取2019年4月～2020年4月我院收治的采用PD-1抑制剂免疫治疗的非小细胞肺癌患者200例,将所选患者分为两组,对照组应用常规护理方案,观察组应用针对性护理,对比分析两组患者护理效果.结果 观察组患者MLHFQ评分显著低于对照组,观察组患者护理满意度显著高于对照组,差异有统计学意义(P<0.05).结论 对应用PD-1抑制剂免疫治疗的非小细胞肺癌患者采用相应针对性护理能帮助提高患者生活质量水平和护理满意度,值得推广.

目的 使用秀丽隐杆线虫作为模式生物,观察 rsa 基因对 mbk-2突变型线虫胚胎发育和虫卵孵化的影响,初步探讨 mbk-2的在胚胎发育中的作用及机制.方法 以 RNAi 技术定向干扰 mbk-2突变型线虫 rsa 基因的表达,并对 rsa沉默后线虫的虫卵孵化水平及胚胎发育情况进行分析,检测其在线虫生长发育、生殖能力等方面的变化.结果 与 L4440喂养的 mbk-2基因突变组相比,rsa-1和 rsa-2RNAi细菌处理 mbk-2基因突变组虫卵未孵化率显著升高,达到41.8%和49.2%(P<0.05),且 rsa-2处理组显著高于 rsa-1组(P<0.05);rsa-1和 rsa-2RNAi细菌处理的N2野生型线虫虫卵未孵化率分别为3.9%和5.7%,显著低于rsa-1和rsa-2RNAi细菌处理的mbk-2突变型线虫(P<0.05).rsa-2RNAi处理 mbk-2突变型线虫组,线虫胚胎发育异常显著增加.结论 mbk-2是线虫发育的一个重要因子,rsa是 mbk-2的重要修饰基因,两者相互作用调节线虫的胚胎发育和虫卵孵化.

Objective(s): To investigate and test the hypotheses that TGF-β1 enhanced myocardial differentiation through Wnt/β-catenin pathway with rat bone marrow mesenchymal stem cells (BMSCs).Materials and Methods: Lentiviral vectors carrying the TGF-β1 gene were transduced into rat BMSCs firstly. Then several kinds of experimental methods were used to elucidate the related mechanisms by which TGF-β1 adjusts myocardial differentiation in rat BMSCs. Results: Immunocytochemistry revealed that cTnI and Cx43 expressed positively in the cells that were transduced with TGF-β1. The results of Western blot (WB) test showed that the levels of intranuclear β-catenin and total β-catenin were all significantly decreased. However, the cytoplasmic β-catenin level was largely unchanged. Moreover, the levels of GSK-3β were largely unchanged in BMSCs, whereas phosphorylated GSK-3β was significantly decreased in BMSCs. When given the activator of Wnt/β-catenin pathway (lithium chloride, LiCl) to BMSCs transducted with TGF-β1, β-catenin was increased, while phosphorylated β-catenin was decreased. In addition, cyclinD1, MMP-7, and c-Myc protein in BMSCs transducted with Lenti-TGF-β1-GFP were significantly lower. Conclusion: These results indicate that TGF-β1 promotes BMSCs cardiomyogenic differentiation by promoting the phosphorylation of β-catenin and inhibiting cyclinD1, MMP-7, and c-Myc expression in Wnt/β-catenin signaling pathway.

目的 探讨过表达GATA-4大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向心肌细胞分化的作用.方法 全骨髓法分离大鼠BMSCs,经多次传代纯化后,借助细胞穿透肽VP22的载体作用,采用脂质体转染法将GATA-4(pVP22-GATA-4/myc-His)基因瞬时转染SD大鼠BMSCs,于转染后第3天加入适宜浓度的5-氮杂胞苷(5-azacytidine,5-aza)诱导1d,同时观察细胞形态变化.转染2d后,采用免疫印迹法检测GATA-4、myc的表达.于诱导后21d采用免疫细胞化学及免疫印迹法检测GATA-4、心肌肌钙蛋白T(cardiac troponin T,cTnT)的表达,以鉴定细胞分化.结果 转染2d后,免疫印迹法检测结果显示实验组有GATA-4、myc表达.诱导后21d,免疫印迹法、免疫细胞化学法检测结果显示实验组GATA-4、cTnT表达水平显著高于空质粒转染组和空白对照组(P＜0.05).结论 过表达GATA-4基因能够促进经5-aza诱导的BMSCs向心肌细胞分化.

目的 观察姜黄素在治疗糖尿病及其并发症过程中对整合素β3、白细胞抑制因子(leukemia inhibitory factor,LIF)表达水平的影响.方法 将雌性SD大鼠30只随机分为3组进行灌胃治疗,分别为:正常组10只,选用食用玉米油灌胃10周2.5mL/kg;糖尿病模型组10只,选用食用玉米油灌胃10周2.5 mL/kg;姜黄素治疗组10只,选用姜黄素化合物混悬液灌胃10周每日250 mg/kg.10周后,按雌雄比例3∶2合笼,次日晨查雌鼠阴道,发现阴栓者计为妊娠第1天,雌性大鼠均于妊娠第4天取材.分别检测血糖、胰岛素,采用HE染色观察子宫内膜上皮的形态结构,运用免疫组织化学和免疫印迹实验法分析各组大鼠子宫内膜整合素β3、LIF表达水平.结果 糖尿病组与正常组比较,其血糖和胰岛素明显升高,子宫内膜整合素β3和LIF表达量明显减少,姜黄素治疗组与糖尿病组比较,其血糖和胰岛素明显降低,子宫内膜整合素β3和LIF表达量明显升高.结论 姜黄素可降低血糖和胰岛素,增加子宫内膜整合素β3和LIF蛋白的表达,最终改善子宫内膜容受性.

目的 探讨"雨课堂"教学平台在组织学与胚胎学教学中的应用效果.方法 选取2018级护理学专业两个大班为研究对象,研究组在教学过程中应用"雨课堂"教学平台,对照组采取传统教学方法 .课程结束后对两组学生的期末理论成绩进行统计学分析,并通过问卷调查进行"雨课堂"使用效果评价.结果 研究组学生期末理论考试成绩显著高于对照组;问卷调查显示学生对雨课堂的使用效果评价较高.结论 将"雨课堂"教学平台应用于实际教学中,可显著提高教学效果,推动组织学与胚胎学教学的信息化改革.

目的 通过用四氧化锇、磷钨酸、钼酸铵和醋酸双氧铀4种染液分别对外泌体负染色,并对染色效果进行观察和对比分析,以期选出对外泌体染色效果最好的负染液.方法 用差速离心法提取外泌体后,采用漂浮法分别用四氧化锇、磷钨酸、钼酸铵、醋酸双氧铀4种不同染液对外泌体进行负染色,透射电镜观察,从外泌体的杯托状或圆盘状结构清晰度、膜边缘锐利与否、染色背景干净程度、反差效果等方面对比4种染液的效果.结果 四氧化锇的染色背景出现大量泡状结构,外泌体结构不明显.磷钨酸对外泌体的染色效果不如醋酸双氧铀和钼酸铵,外泌体的胞膜边缘不清晰,染色背景有杂质.钼酸铵对外泌体的染色效果也较好,其囊泡的膜结构较清楚,外泌体的杯托状结构可较清楚见到,但是反差较弱.醋酸双氧铀染色下,外泌体典型的杯托状或圆盘状结构清晰可见,膜边缘锐利,染色背景干净,反差效果佳.结论 醋酸双氧铀对外泌体的染色效果最好,是最合适的负染染液.

高校实验室作为人才培养、科学研究、生产试验、技术开发的重要基地[1],其重要性是不言而喻的.同时实验室的建设和管理状况更是一所院校综合实力的重要体现[2].由此可见,实验室的规范管理与科学建设对于医学院校发展有着举足轻重的作用.医学实验室是因其独有的专业性而存在的实验场所,其实验内容及实验方法复杂多样,涉及安全隐患较多,安全管理难度大.因此,规范化的管理对医学[3]实验室尤为重要.随着社会的现代化发展及新时期下对医学生全面素质教育的要求,医学院校实验室的管理工作更应该与时俱进、动态化、现代化.

组织芯片是将数十个至上千个小块组织整齐地排列在同一张载玻片上而制成的微缩组织切片.根据组织芯片放置组织的多少,分为多组织片、组织阵列片(最多可含有60个直径2 mm左右的组织)和组织微阵列片(最多含1 000个直径0.6 mm左右的组织);按组织来源不同,又可分为人类组织芯片、动物组织芯片;人类组织芯片又可分为人类疾病组织芯片、正常组织芯片和胚胎组织芯片.组织芯片具有节省材料、信息量大、对比性强,可按不同的需要进行排列组合设计等特点[1].

Radiation therapy is important for the comprehensive treatment of intracranial tumors. However, the molecular mechanisms underlying the pathogenesis of delayed cognitive dysfunction are not well-defined and effective treatments or prevention measures remain insufficient. In the present study, 60 adult male Wistar rats were randomly divided into three groups, which included a control, whole brain radiotherapy (WBRT) (single dose of 30 Gy of WBRT) and nimodipine (single dose of 30 Gy of WBRT followed by nimodipine injection intraperitoneally) groups. The rats were sacrificed 7 days or 3 months following irradiation. At 3 months, the Morris water maze test was used to assess spatial learning and memory function in rats. The results demonstrated that the WBRT group demonstrated a significantly impaired cognitive performance, decreased numbers of hippocampal Cornu Ammonis (CA)1 neurons and upregulated expression of caspase-3 in the dentate gyrus compared with those in the control and nimodipine groups. Reverse transcription-quantitative polymerase chain reaction analysis demonstrated that the WBRT group exhibited increased ratio of B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax)/Bcl-2 compared with that in control and nimodipine groups on day 7 following irradiation. However, the WBRT group exhibited decreased levels of brain-derived neurotrophic factor (BDNF) compared with that in control and nimodipine groups at 3 months following brain irradiation. The levels of growth-associated protein 43 and amyloid precursor protein between the nimodipine group and WBRT group were not statistically significant. The present study demonstrated that neuron apoptosis may lead to delayed cognitive deficits in the hippocampus, in response to radiotherapy. The cognitive impairment may be alleviated in response to a calcium antagonist nimodipine. The molecular mechanisms involved in nimodipine-mediated protection against cognitive decline may involve the regulation of Bax/Bcl-2 and BDNF in the hippocampus.