Inhibitory effect of Cyrtomium falcatum on melanogenesis in α-MSH-stimulated B16F10 murine melanoma cells

Objective:To explore the anti-melanogenic potential of Cyrtomium falcatum.Methods:The effects of Cyrtomium falcatum crude extract and its solvent fractions on tyrosinase activity,melanin content,and the expressions of melanogenesis-related genes and proteins were analyzed inα-melanocyte-stimulating hormone(α-MSH)-stimulated B16F10 cells.Results:α-MSH treatment significantly increased tyrosinase activity,and extracellular and intracellular melanin content,as well as the expression levels of tyrosinase,microphthalmia-associated transcription factor(MITF),tyrosinase-related protein(TRP)-1,and TRP-2 in B16F10 cells.Treatment with Cyrtomium falcatum crude extract and its solvent fractions reduced tyrosinase activity and extracellular and intracellular melanin content and downregulated the expression levels of tyrosinase,MITF,TRP-1,and TRP-2 in a dose-dependent manner.Conclusions:Cyrtomium falcatum has potential anti-melanogenesis effects and can be used as a potential source material in cosmeceutical industry for the research and development of novel lead molecules with whitening properties.

安罗替尼通过调控miR-16-5p/PD-1轴抑制人非小细胞肺癌细胞系增殖并促进凋亡

目的 探讨安罗替尼对非小细胞肺癌细胞增殖和凋亡的影响及其分子机制。方法 将非小细胞肺癌细胞系A549和H1299分别采用安罗替尼、miR-16-5p激动剂和/或PD-1过表达载体进行处理。CCK-8实验和EDU实验检测细胞增殖;流式细胞术检测细胞凋亡;RT-qPCR检测miR-16-5p相对表达量;Western blot检测程序性细胞死亡-1蛋白(PD-1)的表达。双荧光素酶报告实验确定miR-16-5p和PD-1的靶向关系。用A549细胞构建裸鼠成瘤模型,检测安罗替尼对体内肿瘤生长的影响。结果 安罗替尼在A549和H1299细胞中以剂量依赖的方式显著增加miR-16-5p表达,同时降低PD-1表达,并且抑制细胞增殖,促进细胞凋亡(P<0.05)。miR-16-5p过表达可抑制细胞增殖,促进细胞凋亡(P<0.05)。miR-16-5p能靶向PD-1,且负调控PD-1表达。siRNA下调PD-1表达后明显抑制细胞增殖,并促进细胞凋亡(P<0.05)。过表达PD-1则可逆转安罗替尼介导的miR-16-5p对A549和H1299细胞的促增殖和抗凋亡作用(P<0.05)。体内实验证实,安罗替尼能够明显抑制肿瘤生长(P<0.05)。结论 安罗替尼可有效抑制非小细胞肺癌细胞增殖,促进细胞凋亡,减少体内肿瘤生长,其作用机制与miR-16-5p/PD-1信号轴相关。

苍附导痰汤对肥胖型PCOS大鼠内分泌激素及miRNA-16和PDCD-4表达的影响

目的:探讨苍附导痰汤对肥胖型多囊卵巢综合征(PCOS)大鼠内分泌激素及微小RNA(miRNA)-16和程序性细胞死亡因子4(PDCD-4)表达的影响。方法:将50只SPF级雌性SD大鼠随机分为5组,每组10只。阳性对照组大鼠灌胃格华止(0.43 g/kg),低剂量组大鼠灌胃低剂量苍附导痰汤(1.42 g/kg),高剂量组大鼠灌胃高剂量苍附导痰汤(5.68 g/kg),模型组和正常组大鼠灌胃等量生理盐水,各组均持续给药14 d。比较各组大鼠体质量,内分泌激素水平,miRNA-16和PDCD-4 mRNA表达,以及PDCD-4蛋白表达。结果:模型组、阳性对照组、低剂量组和高剂量组大鼠体质量均高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠体质量均低于模型组(P<0.05),且呈剂量依赖性。模型组、阳性对照组、低剂量组和高剂量组大鼠血清E_(2)均水平低于正常组,而血清T、FSH和LH水平均高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠血清E_(2)水平均高于模型组,而血清T、FSH和LH水平均低于模型组(P<0.05),且呈剂量依赖性。模型组、阳性对照组、低剂量组和高剂量组大鼠卵巢miRNA-16表达均低于正常组,而PDCD-4 mRNA表达高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠卵巢miRNA-16表达均高于模型组,而PDCD-4 mRNA表达低于模型组(P<0.05),且呈剂量依赖性。模型组、阳性对照组、低剂量组和高剂量组大鼠卵巢PDCD-4蛋白相对表达量均高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠卵巢PDCD-4蛋白相对表达量均低于模型组(P<0.05),且呈剂量依赖性。结论:苍附导痰汤可调节肥胖型PCOS大鼠内分泌激素水平,其机制可能与上调miRNA-16表达及下调PDCD-4表达有关。

血清SDC-1、HDAC6、CC16水平联合检测对慢性阻塞性肺疾病患者预后不良的预测价值

目的:探讨血清多配体蛋白聚糖1(SDC-1)、组蛋白去乙酰化酶6(HDAC6)、克拉拉细胞分泌蛋白16(CC16)水平联合检测对慢性阻塞性肺疾病(COPD)患者预后不良的预测价值。方法:选取2020年11月至2023年11月该院收治的147例COPD患者进行横断面研究,设为研究组;选取同期于该院体检的147名健康者设为对照组。比较两组、不同COPD病情程度患者、不同预后COPD患者血清SDC-1、HDAC6、CC16水平;采用受试者工作特征(ROC)曲线分析入院时血清SDC-1、HDAC6、CC16水平单项及联合检测预测COPD患者预后不良的价值。结果:研究组血清SDC-1、HDAC6水平均高于对照组,血清CC16水平低于对照组,差异有统计学意义(P<0.05)。重度COPD患者血清SDC-1、HDAC6水平高于中重度、中度、轻度患者,且中重度高于中度、轻度患者,中度高于轻度患者;重度COPD患者血清CC16水平低于中重度、中度、轻度患者,且中重度低于中度、轻度患者,中度低于轻度患者,差异均有统计学意义(P<0.05)。预后不良COPD患者入院时血清SDC-1、HDAC6水平高于预后良好患者,血清CC16水平低于预后良好患者,差异均有统计学意义(P<0.05)。ROC曲线分析结果显示,入院时血清SDC-1、HDAC6、CC16水平联合检测预测COPD患者预后不良的曲线下面积(AUC)为0.938,高于三者单项检测诊断(AUC=0.774、0.771、0.716)。结论:入院时血清SDC-1、HDAC6、CC16水平联合检测预测COPD患者预后不良的价值高于三者单项检测。

Human Papillomavirus 16 E6,E7 siRNAs Inhibit Proliferation and Induce Apoptosis of SiHa Cervical Cancer Cells

Objective： To evaluate the effects of HPV16 E6/E7 siRNAs on cervical cancer SiHa cells. Methods： The expressions of the E6, E7, p53 and Rb genes were assayed by RT-PCR and Western-bloting respectively. The proliferation and apoptosis of the cells were evaluated by MTT and flow cytometry. Results： HPV 16 E6 and E7 oncogenes were selectivly downregulated by HPV 16 E6 and E7 siRNAs, which sustained at least 96 h by single dose siRNA. Furthermore, reduction of E6 and E7 oncogenes expression upregulated the expressions of P53 and RB protein and induced apoptosis in SiHa cells. Conclusion： Introduction of HPV16 E6/E7 siRNA might be a potentially potent and specific approach to inhibit proliferation and induce apoptosis of SiHa cervical cancer cells.

干扰素诱导蛋白16的生物学机制研究进展

干扰素诱导蛋白16(interferonγ-inducible protein 16,IFI16)是人类PYHIN(pyrin and HIN domain-containing protein)家族(也称干扰素诱导蛋白P200家族)成员之一,在人体器官组织中广泛存在,参与细胞周期调控、细胞衰老、细胞凋亡及免疫反应等多种生物过程。不同生理及病理状态下,IFI16的含量及定位均会发生改变,近年来研究发现,其在抗病毒、肿瘤、炎性疾病及其他多种疾病发生发展过程中可能发挥重要作用。该文对其机制及其在疾病中的研究现状进行综述,以期为深入研究IFI16提供参考。

Suberoylanilide hydroxamic acid upregulates reticulophagy receptor expression and promotes cell death in hepatocellular carcinoma cells

BACKGROUND Hepatocellular carcinoma(HCC)is a common clinical condition with a poor prognosis and few effective treatment options.Potent anticancer agents for treating HCC must be identified.Epigenetics plays an essential role in HCC tumorigenesis.Suberoylanilide hydroxamic acid(SAHA),the most common histone deacetylase inhibitor agent,triggers many forms of cell death in HCC.However,the underlying mechanism of action remains unclear.Family with sequence similarity 134 member B(FAM134B)-induced reticulophagy,a selective autophagic pathway,participates in the decision of cell fate and exhibits anticancer activity.This study focused on the relationship between FAM134B-induced reticulophagy and SAHA-mediated cell death.AIM To elucidate potential roles and underlying molecular mechanisms of reticulophagy in SAHA-induced HCC cell death.METHODS The viability,apoptosis,cell cycle,migration,and invasion of SAHA-treated Huh7 and MHCC97L cells were measured.Proteins related to the reticulophagy pathway,mitochondria-endoplasmic reticulum(ER)contact sites,intrinsic mitochondrial apoptosis,and histone acetylation were quantified using western blotting.ER and lysosome colocalization,and mitochondrial Ca^(2+)levels were characterized via confocal microscopy.The level of cell death was evaluated through Hoechst 33342 staining and propidium iodide colocalization.Chromatin immunoprecipitation was used to verify histone H4 lysine-16 acetylation in the FAM134B promoter region.RESULTS After SAHA treatment,the proliferation of Huh7 and MHCC97L cells was significantly inhibited,and the migration and invasion abilities were greatly blocked in vitro.This promoted apoptosis and caused G1 phase cells to increase in a concentration-dependent manner.Following treatment with SAHA,ER-phagy was activated,thereby triggering autophagy-mediated cell death of HCC cells in vitro.Western blotting and chromatin immunoprecipitation assays confirmed that SAHA regulated FAM134B expression by enhancing the histone H4 lysine-16 acetylation in the FAM134B promoter region.Further,SAHA disturbed the Ca^(2+)homeostasis and upregulated the level of autocrine motility factor receptor and proteins related to mitochondria-endoplasmic reticulum contact sites in HCC cells.Additionally,SAHA decreased the mitochondrial membrane potential levels,thereby accelerating the activation of the reticulophagy-mediated mitochondrial apoptosis pathway and promoting HCC cell death in vitro.CONCLUSION SAHA stimulates FAM134B-mediated ER-phagy to synergistically enhance the mitochondrial apoptotic pathway,thereby enhancing HCC cell death.

Mushroom Tyrosinase Inhibition and Antimelanogenesis Activities of <i>Bacopa monnieri</i>(L.) Methanol Extract in B16F10 Melanoma Cells

Melanocytes that form stratum basale of skin epidermis express tyrosinase enzyme, which catalyzes initial two rate-limiting steps in the biotransformation of tyrosine into dark pigment called melanin. Even today, Tyrosinase inhibitors are among the promising candidates in cosmetic industry for skin-lightening formulations. Overexpression of tyrosinase causes excess melanin biosynthesis and deposition resulting in dark skin color. Moreover, localized overexpression of tyrosinase cause variety of hyperpigmentation disorders like melanoma, melasma, chloasma, dark patches, liver patches, etc. There has been a renewed interest in the natural products as main ingredients in the formulation of safe products for skin-whitening and treatment options for hyperpigmentation disorders. In the present communication, the results of our investigations on tyrosinase inhibition, modulation of intracellular tyrosinase and melanin levels in cultured B16F10 melanoma cells by Bacopa monnieri (L.) methanol extract (BME) are presented and discussed as safe option for skin lightening and to treat hyperpigmentation disorders. BME showed 11%, 29%, 54% and 80% inhibition of mushroom tyrosinase activity at an initial 100, 200, 400 and 600 μg of extract. Treatment of α-melanocyte stimulating hormone (α-MSH) stimulated cultured murine melanoma B16F10 cells with 100 μg/ml of the extract showed a decrease in the levels of cellular melanin and cellular tyrosinase content by 22% and 46% respectively. The cytotoxicity studies by MTT assay revealed that the LC50 of the BME is ≥1000 μg/ml in cultured mouse melanoma B16F10 and HEK293 cells.

Multinucleated giant cells of bladder mucosa are modified telocytes:Diagnostic and immunohistochemistry algorithm and relation to PDL1 expression score

BACKGROUND Multinucleated giant cells(MGCs)in bladder carcinomas are poorly studied.AIM To describe the function,morphogenesis,and origin of mononuclear and MGCs in urothelial carcinoma(UC)of the bladder in Bulgarian and French patients.METHODS Urothelial bladder carcinomas(n=104)from 2016-2020 were analyzed retrospectively using immunohistochemical(IHC)and histochemical stain examination.Giant cells in the bladder stroma were found in 35.6%of cases,more often in highgrades.RESULTS We confirm that MGCs in the mucosa in UC of the bladder were positive for both mesenchymal and myofibroblast markers(vimentin,smooth muscle actin,Desmin,and CD34)and the macrophage marker CD68.Furthermore,IHC studies revealed the following profile of these cells:Positive for p16;negative for epithelial(CK AE1/AE3 and GATA-3),vascular(CD31),neural(PS100 and CKIT),cambial,blastic(CD34-blasts and C-KIT),and immune markers(IG G,immunoglobulin G4,and PD-L1);no proliferative activity,possess no specific immune function,and cannot be used to calculate the Combined Positive Score scale.CONCLUSION In conclusion,the giant stromal cells in non-tumor and tumor bladder can be used as a characteristic and relatively constant,although nonspecific,histological marker for chronic bladder damage,reflecting the chronic irritation or inflammation.Likewise,according to the morphological and IHC of the mono-and multinucleated giant cells in the bladder,they are most likely represent telocytes capable of adapting their morphology to the pathology of the organ.

The associated regulators and signal pathway in rIL-16/CD4 mediated growth regulation in Jurkat cells

IL-16 is a ligand and chemotactic factor for CD4+ T cells. IL-16 inhibits the CD3 mediated lymphocyte activation and proliferation. The effects of IL-16 on the target cells are dependent on the cell type, the presence of co-activators etc. To understand the regulation function and mechanism of IL-16 on target cells, we used a 130 a.a. recombinant IL-16 to study its effects on the growth of Jurkat T leukemia cells in vitro. We found that the rIL-16 stimulated the proliferation of Jurkat cells at low dose (10^-9M), but inhibited the growth of the cells at higher concentration (10^-5M). Results showed that 10^-5 M of rIL-16 treatment induced an enhanced apoptosis in Jurkat cells. The treatment blocked the expression of FasL, but up-regulated the c-myc and Bid expression in the cells. Pre-treatment of PKC inhibitor or MEK1 inhibitor markedly increased or decreased the rIL-16 induced growth-inhibiting effects on Jurkat cells, respectively.The results suggested that the rIL-16 might be a regulator for the growth or apoptosis of Jurkat cells at a dose-dependent manner. The growth-inhibiting effects of rIL-16 might be Fas/FasL independent, but,associated with the activation of PKC, up-regulated expression of c-Myc and Bid, and the participation of the ERK signal pathway in Jurkat cells.

CC16在肺部疾病中的研究进展

Clara细胞分泌蛋白16(CC16)作为主要分布于呼吸道Clara细胞分泌的主要物质,有着多种生物学功能。在呼吸道保护作用中起着重要作用,作为生物学标志物,能预测多种肺部疾病发展,作为治疗药物,能延缓多种肺部疾病加重。目前针对其在肺部疾病的研究仍是热点。本文从CC16来源及结构、功能等方面进行总结,就其在肺部疾病中研究进展做一综述。

血清CXCL16、ESM-1水平对脑小血管病患者认知障碍的诊断价值

目的探讨血清CXC趋化因子配体16(CXCL16)、内皮细胞特异性分子1(ESM-1)水平对脑小血管病(CSVD)患者认知障碍的诊断价值。方法选取2019年10月至2022年10月期间我院收治的131例CSVD患者,根据患者入院时的蒙特利尔认知评估量表(MoCA)评分结果,将CSVD患者分为认知正常组(60例)和认知障碍组(71例)。Spearman法分析血清CXCL16、ESM-1表达水平与MoCA评分的相关性,Logistic回归分析影响CSVD患者发生认知障碍的相关因素,受试者工作特征(ROC)曲线分析血清CXCL16和ESM-1对CSVD患者认知障碍的诊断价值。结果认知障碍组患者血清CXCL16表达水平高于认知正常组,血清ESM-1表达水平低于认知正常组(P<0.05);血清CXCL16表达水平与MoCA评分呈负相关(r=-0.767,P<0.05),血清ESM-1表达水平与MoCA评分呈正相关(r=0.723,P<0.05);认知障碍组年龄≥60岁、有高血压史、脑梗死的患者所占比例高于认知正常组(P<0.05);年龄、高血压史、脑梗死及CXCL16为CSVD患者发生认知障碍的危险因素,ESM-1为保护因素(P<0.05);血清CXCL16、ESM-1以及二者联合诊断CSVD患者发生认知障碍的AUC分别为0.748、0.671、0.851,二者联合检测优于血清CXCL16、ESM-1各自单独检测(Z二者联合-CXCL16=3.284、Z二者联合-ESM-1=4.111,P=0.001、<0.001),对CSVD并发认知障碍具有较高的诊断价值。结论血清CXCL16、ESM-1的表达水平与CSVD并发认知障碍的发生密切相关,二者联合对CSVD并发认知障碍具有较高的诊断价值。

Exogenous p16 gene therapy combined with X-ray irradiation suppresses the growth of human glioma cells

In this study, we infected human glioma U251 cells with a replication-defective recombinant adenovirus carrying the p16 gene. This adenovirus constructed was able to transfect exogenous p16 into the human glioma cells efficiently, and direct a high level of p16 protein expression. Tumor-inhibition experiments demonstrated that treatment with the adenovirus-p16 significantly inhibited the growth of glioma cells in vitro as well as the in vivo development of tumors in nude mice bearing a brain glioma. The combination of adenovirus-p16 gene treatment and X-ray irradiation resulted in a greater inhibition of tumor growth. Adenovirus-mediated p16 gene therapy conferred a significant antitumor effect against human glioma cells both in vitro and in vivo, and that there was a synergistic effect when X-ray irradiation was also used.

血清CC16、sRAGE水平与ARDS患者疾病转归的相关性及预测价值研究

目的 探讨血清Clara细胞分泌蛋白16(CC16)、可溶性晚期糖基化终产物受体(sRAGE)水平变化与急性呼吸窘迫综合征(ARDS)患者疾病转归的相关性及预测价值。方法 选取2021年1月至2023年1月灵宝市第二人民医院收治的127例ARDS患者为研究对象,根据ARDS严重程度将患者分为轻度组(39例)、中度组(52例)和重度组(36例)。比较不同严重程度患者及不同疾病转归患者血清CC16、sRAGE水平、氧合指数(PaO_(2)/FiO_(2))、急性生理学及慢性健康状况Ⅱ(APACHEⅡ)评分、序贯器官衰竭评估(SOFA)评分,分析血清CC16、sRAGE水平与PaO_(2)/FiO_(2)、APACHEⅡ评分、SOFA评分及疾病转归的相关性。采用logistic回归分析ARDS患者病死的危险因素,受试者工作特征(ROC)曲线分析血清CC16、sRAGE水平及二者联合检测对ARDS患者病死的预测价值。结果 ARDS不同程度患者入院时血清CC16、sRAGE水平及APACHEⅡ、SOFA评分比较:轻度组<中度组<重度组,PaO_(2)/FiO_(2)比较:轻度组>中度组>重度组,差异均有统计学意义(P<0.05);病死患者入院时血清CC16、sRAGE水平及APACHEⅡ、SOFA评分均高于生存患者,而PaO_(2)/FiO_(2)低于生存患者,差异均有统计学意义(P<0.05);患者入院时血清CC16、sRAGE水平与PaO_(2)/FiO_(2)均呈负相关(P<0.05),与APACHEⅡ、SOFA评分及疾病转归均呈正相关(P<0.05);入院时血清CC16、sRAGE水平为ARDS患者病死的危险因素,且两指标联合预测ARDS患者病死的ROC曲线下面积大于各单项指标(P<0.05)。结论 ARDS患者血清CC16、sRAGE表达上调,其水平与疾病转归密切相关,二者联合检测可作为临床早期预测转归情况的有效指标。

HPV16 E6 gene mutation promotes the proliferation of cervical cancer cells by regulating the expression of BDNF/TrKB

Objective:Study on the mechanism of HPV16 E6 gene mutation promoting the proliferation of cervical cancer cells by influencing the expression of BDNF/TrkB.Methods:The expression levels of HPV16 E6 T350G,BDNF,TrkB and p53 mRNA in cervical cancer tissue samples and CINII cervical tissues were detected by Real-time PCR.HPV16 E6 T350G lentivirus(pLV5-HPV16 E6 T350G)and empty vector(pLV5-vector)were designed and constructed,and transfected with HCerEpiC cells,the expression levels of HPV16 E6 T350G,BDNF,TrKB and p53 mRNA were detected by Real-time PCR,and the expression levels of BDNF,TrKB,PI3K,pPI3K,AKT and pAKT protein were detected by western blot;cell proliferation was detected by MTT experiments.Results:Compared with cinii cervical tissue,HPV16 E6 T350G,BDNF and TrkB mRNA expression levels were all positive,while p53 mRNA expression was negative.After overexpression of HPV16 E6 T350G in HCerEpiC cells,it can up-regulate the expression levels of BDNF and TrKB protein and mRNA,and activate the PI3K/AKT signaling pathway which is the downstream of BDNF/TrKB,and reduce p53 protein expression levels;HPV16 E6 T350G overexpression can enhance the proliferation capacity of HCerEpiC cells.Conclusion:Overexpression of HPV16 E6 T350G can promote the proliferation of cervical cancer cells,which may be related to the upregulation of BDNF/TrKB expression,the activation of PI3K/AKT signaling pathway,and the decrease of p53 expression.

CD16+ Cells and Costimulatory Molecules of Lymphocyte Activation Present inside Human Kidney Grafts and in Blood Circulation

Background: We studied the expression of important costimulatory molecules of lymphocyte activation and the presence of CD16+ cells on aspiration biopsies of kidney transplants, measured three soluble factors and when indicated tested their robustness in diagnosing acute rejection. Methods: Fine-needle aspiration biopsies were performed either on days seven or 14 - 30 post-transplantation among stable kidney transplants and on the day of acute rejection diagnosis, while a sample of peripheral blood was collected simultaneously. The cyto preparations were studied by the enzymatic avidin biotin complex staining. The immunocytochemistry was directed to CD16, CD28, CD152, ICOS, CD40, CD154, CD26 and CD27. We performed the analysis in the peripheral blood by ELISA for soluble(s) CD16, CD26, and CD154. Results: The group of acute rejection cases showed a significant up-regulated expression of CD16, CD26, ICOS and CD40 as compared to the group of stable cases. Both sCD16 and sCD154 were significantly higher in the blood samples of the group with acute rejection. Thymoglobulin down-regulated CD154 and sCD16. CD16, CD26 and ICOS exhibited very high sensitivity and specificity for acute rejection diagnosis. Conclusions: The presence of CD16+ cells inside the graft emerged as a distinct player in acute rejection, confirming other previous reports whereas we first document that in human kidney transplants, ICOS and CD26 are significantly up-regulated and both reached positive predictive values for acute rejection ≥ 80%. The other costimulatory molecules, with the exception of CD40, though widely known, did not show robust association with immune events.

血清CCL5及CC16水平联合检测对支气管哮喘患儿沙丁胺醇雾化吸入治疗疗效的预测价值

目的探讨血清趋化因子配体5(CCL5)、Clara细胞分泌蛋白16(CC16)水平检测对支气管哮喘(BA)患儿沙丁胺醇雾化吸入治疗疗效的预测价值。方法选择2019年10月至2022年10月该院收治的急性发作期BA患儿207例为研究对象,所有BA患儿均给予常规治疗联合吸入用硫酸沙丁胺醇溶液雾化治疗4周,根据治疗效果分为有效组和无效组。采用酶联免疫吸附试验检测两组治疗前血清CCL5和CC16水平,比较两组临床资料,采用多因素Logistic回归分析沙丁胺醇雾化吸入治疗BA临床疗效的影响因素,采用受试者工作特征(ROC)曲线分析血清CCL5、CC16水平对沙丁胺醇雾化吸入治疗BA临床疗效的预测价值。结果治疗4周后,有效161例(有效组),无效46例(无效组)。有效组治疗前血清CCL5水平低于无效组,血清CC16水平高于无效组,差异均有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,血清CCL5水平升高是沙丁胺醇雾化吸入治疗BA患儿临床有效的独立危险因素(P<0.05),而哮喘控制测试评分、呼气峰值流速、血清CC16水平升高是其保护因素(P<0.05)。血清CCL5、CC16单独及联合检测预测沙丁胺醇雾化吸入治疗BA临床疗效的曲线下面积(AUC)分别为0.798、0.868和0.930,联合检测的AUC大于单独检测(P<0.05)。结论血清CCL5水平升高是沙丁胺醇雾化吸入治疗BA患儿临床疗效的独立危险因素,血清CC16水平升高是其保护因素,二者联合检测能够较好地预测其临床疗效。

Comparison of DNA double-strand breaks induced by ^(16)O^(8+) in deproteinized DNA and intact cells

The yield of DNA double-strand breaks (DSBs) is sure to be influenced by theenvironment around DNA molecule. Inverse pulsed-field gel electrophoresis (PIGE) has beenapplied to compare the sensitivity of B16 cells and their DNA in DSBs induced by 75 MeV/u16O8+ beam. Results show that the percentages of DNA released from the plug(PR) in bothkinds of tile samples increase with the dose and approach a similar quasi-threshold of about81%. A simple new equation was presented to calculate the break level of DNA molecules.Within a certain dose, the relationship between the break level and the dose is linear. Theyield of DSBs in deproteinized DNA was 1.11 DSBs/100 Mbp/Gy, while that in intact cells was0.60DSBs/100Mbp/Gy. It is testified that deproteinized DNA is more sensitive to oxygen ionsirradiation than intact cells.

LncRNA FGD5-AS1靶向miR-16-5p/CREB1轴减轻缺氧/复氧诱导大鼠H9c2心肌细胞凋亡的机制研究

目的:探究长链非编码RNA(LncRNA)FGD5反义RNA1(FGD5-AS1)对缺氧/复氧(H/R)诱导的大鼠H9c2心肌细胞凋亡的影响以及对微小RNA-16-5p/cAMP响应元件结合蛋白1(miR-16-5p/CREB1)轴的调节机制。方法:将H9c2细胞分为对照组和H/R组(缺氧6 h,复氧6 h),然后将H/R组细胞分别进行转染,分为oe-NC组、oe-FGD5-AS1组、oe-FGD5-AS1+miR-16-5p mimic-NC组、oe-FGD5-AS1+miR-16-5p mimic组。实时荧光定量逆转录聚合酶链式反应法(RT-qPCR)检测细胞中FGD5-AS1、miR-16-5p、CREB1的mRNA水平;蛋白免疫印迹(Western Blot)法测定裂解凋亡蛋白酶-3(cleaved Caspase-3)、B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白表达;CCK-8法测定细胞存活率;流式细胞术检测细胞凋亡率;双荧光素酶活性实验分别验证miR-16-5p和FGD5-AS1、CREB1的靶向关系。结果:与对照组比较,H/R组细胞中FGD5-AS1和CREB1的mRNA水平、细胞存活率、Bcl-2蛋白表达降低(P<0.05),miR-16-5p mRNA水平、细胞凋亡率及cleaved Caspase-3、Bax蛋白表达升高(P<0.05);与H/R组和oe-NC组比较,转染过表达FGD5-AS1基因的H9c2细胞中FGD5-AS1和CREB1的mRNA水平、细胞存活率、Bcl-2蛋白表达增高,凋亡率及miR-16-5p、cleaved Caspase-3、Bax水平下降(P<0.05)。双荧光素酶活性实验验证了FGD5-AS1、CREB1均与miR-16-5p有结合位点。结论:过表达FGD5-AS1可能通过靶向下调miR-16-5p表达,上调CREB1表达,抑制H/R诱导的H9c2心肌细胞凋亡。

环氧合酶-2反义寡核苷酸对鳞状细胞癌Colo-16细胞基质金属蛋白酶-2表达的影响

目的:研究环氧合酶(COX)-2反义寡核苷酸(antisense oligodeoxynucleotide,AsODN)对人皮肤鳞状细胞癌细胞株Colo-16细胞基质金属蛋白酶-2(metalloproteinase-2,MMP-2)表达的影响。方法:免疫细胞化学法检测Colo-16细胞中COX-2蛋白表达;免疫荧光显微镜观察转染情况;蛋白免疫印迹(Western blot)及半定量反转录聚合酶链反应法(RT-PCR)检测Colo-16细胞MMP-2蛋白和MMP-2mRNA表达水平。结果:Colo-16细胞中COX-2蛋白阳性表达,并分布在胞质、胞核;Colo-16细胞胞质和胞核可见荧光标记的COX-2无义寡核苷酸、反义寡核苷酸;Western blot及RT-PCR检测结果显不COX-2反义寡核苷酸作用Colo-16细胞48 h后,MMP-2表达显著下调(P<0.05)。结论:COX-2反义寡核苷酸能有效下调Colo-16细胞MMP-2表达,提示COX-2反义寡核苷酸可能通过抑制MMP-2表达来阻断皮肤鳞状细胞癌的浸润、转移。