血浆miR-106b、miR-146a表达水平与癫痫患儿脑电图参数、Th17细胞和凋亡分子的相关性分析

摘要 目的：分析血浆miR-106b、miR-146a表达特点及其与脑电图参数、辅助性T细胞17（Th17）和凋亡分子的相关性以及诊断癫痫的价值。方法：选择2018年1月至2020年10月我院收治的癫痫患儿75例作为癫痫组,检测受试者血浆miR-106b、miR-146a表达,外周血Th17细胞占比、血清B细胞淋巴瘤/白血病-1（Bcl-1）、BCL2-Associated X蛋白（Bax）、Survivin、半胱氨酸天冬酰胺酶（Caspase-3）水平和脑电图参数α、β、 δ、θ波功率。分析miR-106b、miR-146a与Th17细胞占比、Bcl-1、Bax、Survivin、Caspase-3以及α、β、 δ、θ波功率的相关性,受试者工作特征（ROC）曲线分析miR-106b、miR-146a诊断癫痫的价值。结果：癫痫组血浆miR-106b、miR-146a表达、Th17细胞占比、Bax、Caspase-3水平高于对照组（P＜0.05）,α波功率、θ波功率、Bcl-1、Survivin水平低于对照组（P＜0.05）。miR-106b、miR-146a表达与Th17细胞占比、Bax、Caspase-3呈正相关（P＜0.05）,与α波功率、θ波功率、Bcl-1、Survivin呈负相关（P＜0.05）。联合miR-106b和miR-146a诊断癫痫的曲线下面积（AUC）为0.975,高于单独miR-106b和miR-146a诊断的0.884、0.835。结论：癫痫患儿血浆miR-146a、miR-106b表达增高,miR-146a、miR-106b高表达与脑电图异常、Th17细胞功能障碍以及神经细胞凋亡有关,miR-146a、miR-106b有望成为癫痫诊断的新生物学标志物。     

miR-125b在急性加重期慢性阻塞性肺疾病患者血浆中的表达及与肺功能和炎症细胞因子的关系

目的:探讨微小RNA-125b(miR-125b)在急性加重期慢性阻塞性肺疾病(COPD)患者血浆中的表达及其与肺功能、炎症细胞因子的关系。方法:选择2016年11月至2019年1月应急总医院收治的69例急性加重期COPD患者作为急性加重组,并于同期随机选取58例稳定期COPD患者作为稳定组和50例健康体检者作为对照组。采用实时荧光定量PCR法检测各组血浆miR-125b表达水平;采用肺功能检测仪测定肺功能,包括用力肺活量(FVC)、第1秒用力呼气量(FEV_1)、FEV_1/FVC;采用酶联免疫吸附法测定血清炎症细胞因子,包括白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、高敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)。结果:急性加重组血浆miR-125b表达水平高于稳定组和对照组,差异有统计学意义(P0.05)。急性加重组、稳定组肺功能指标FVC、FEV_1、FEV_1/FVC低于对照组,且急性加重组低于稳定组,差异有统计学意义(P0.05)。急性加重组、稳定组血清炎症细胞因子IL-6、IL-8、hs-CRP、TNF-α水平高于对照组,且急性加重组高于稳定组,差异有统计学意义(P0.05)。Pearson相关分析结果显示,急性加重期COPD患者血浆miR-125表达水平与FVC、FEV_1、FEV_1/FVC呈负相关(P0.05),与IL-6、IL-8、hs-CRP、TNF-α呈正相关(P0.05)。结论:miR-125b在急性加重期COPD患者血浆中异常表达,并与肺功能及炎症细胞因子密切相关,可作为临床辅助诊断及评估患者病情严重程度的参考指标。     

miR-126 在膀胱癌患者尿液中的表达及临床意义

目的:探讨miR-126在膀胱癌患者尿液中的表达与临床病理特征的关系,评估miR-126的肿瘤标志物诊断价值。方法:收集48例初发膀胱尿路上皮癌患者与32例健康对照者晨尿,提取尿液总RNA,通过实时荧光定量PCR技术检测各样本中的miR-126的表达水平,并经受试者工作曲线(ROC)分析其诊断价值。结果:膀胱癌患者尿液中的miR-126表达水平相对健康对照组明显上调(P0.01),其表达水平在不同病理级别之间存在显著差异(P均0.05),且低级别组表达水平略高于高级别组,与肿瘤大小、数目以及淋巴转移也有一定的相关性(P0.05),而与患者的年龄、性别、TNM分期等均无相关性(P0.05)。通过ROC曲线分析尿液中miR-126诊断膀胱肿瘤的曲线下面积(AUC)为0.861,当最佳切点定在7.475时,miR-126诊断膀胱肿瘤的敏感性和特异性分别为75.0%、81.2%。结论:膀胱癌患者尿液中miR-126的表达差异能够反映病情进展程度,其表达水平对膀胱肿瘤的早期诊断及病情评估具有一定的价值。     

儿童白血病患者血浆循环DNA定量分析

目的:通过对儿童白血病患者血浆循环DNA定量检测,分析血浆循环DNA含量用于儿童白血病诊断的价值.方法:用血液微量DNA抽提试剂盒提取血浆循环DNA,以SYBR GreenI荧光染色法分别检测46例儿童急性白血病患者、20例健康对照标本血浆循环DNA含量;利用受试者工作特征(ROC)曲线评价血浆循环DNA对儿童白血痛的诊断价值.结果:儿童白血病患者血浆循环DNA含量为(30.04±15.13)ng/mL,高于对照组(12.22±7.10)ng/mL,二者差异有统计学意义(P＜0.05);循环DNA含量16.31 ng/mL为诊断儿童白血病最佳临界值,敏感性和特异度分别为86.96%和75.00%,ROC曲线下面积(AUC)为0.879.结论:血浆循环DNA定量检测有可能成为一种新的用于儿童白血病诊断方法.     

NGF蛋白及前列腺F2α受体与子宫腺肌病痛经的相关性研究

目的:研究NGF蛋白及前列腺F2α受体(PTGFR)与子宫腺肌病痛经的相关性。方法:选择我院2016年7月～2017年7月收治的36例子宫腺肌病痛经患者,按视觉模拟评分法(VAS)将痛经程度分为11例轻度组、14例中度组、11例重度组。同期选择36例月经正常者作为对照组。比较各组血清NGF蛋白和血浆PTGFR水平,分析二者和痛经评分之间的相关性及其单独和联合检测时诊断子宫腺肌病痛经的敏感性、特异性及受试者工作特征(ROC)曲线下面积。结果:子宫腺肌病痛经组血清NGF蛋白和血浆PTGFR水平均显著高于对照组(P0.05)。轻度痛经组血清NGF蛋白及血浆PTGFR水平均显著低于中度组和重度组(P0.05)。血清NGF蛋白和血浆PTGFR水平和子宫腺肌病痛经评分均呈显著正相关,r分别为0.812,0.884(P0.05)。ROC曲线分析结果显示血清NGF蛋白联合血浆PTGFR蛋白检测诊断子宫腺肌病痛经的ROC曲线下面积明显大于血清NFG蛋白及血浆PTGFR水平单独检测(P0.05)。结论:NGF蛋白和PTGFR可能参与了子宫腺肌病痛经的发生和发展,二者联合检测有助于诊断子宫腺肌病痛经。血清NGF蛋白水平及血浆PTGFR蛋白水平和子宫腺肌病痛经程度呈正相关。     

血浆microRNA与HIV感染及治疗关系的研究

本次研究分析了Ⅰ型艾滋病病毒(HIV-1)感染者血浆中miRNA表达特点,探讨了血浆miRNA作为HIV-1感染及治疗监测生物学标志物的潜在可行性。选取2017年7月-2018年7月期间在昆明市第三人民医院就诊的HIV-1感染未经抗病毒治疗患者50例作为未治疗组,HIV-1感染经抗病毒治疗患者50例作为治疗组,选取同期本院健康体检者50例作为对照组,应用qPCR技术检测三组对象血浆miR-29a、miR-122,miR-155的相对表达量。结果显示,与健康对照组相比HIV-1感染未经抗病毒治疗组血浆miR-29a、miR-155、miR-122表达均升高,差异有统计学意义(P<0.05);未治疗组与治疗组相比较血浆miR-29a、miR-155和miR-122表达均升高,差异有统计学意义(P<0.05)。对未治疗组的研究显示,血浆miR-29a、miR-155表达水平与HIV-1RNA病毒载量呈正相关(P<0.05),miR-122与HIV-1RNA病毒载量无显著相关(P>0.05),miR-29a、miR-155表达水平与CD4;T细胞计数呈负相关(P<0.05)。本研究提示HIV-1感染后血浆miRNA有变化,血浆miR-29a、miR-122,miR-155与HIV-1感染相关,可能是HIV-1感染的潜在生物学标志物。而抗病毒治疗可以诱导miR-29a、miR-155和miR-122表达降低,这些miRNA可能具有成为监测HIV-1感染抗病毒治疗效果观察指标的潜在价值。未治疗组血浆miR-29a、miR-155与HIV-1RNA呈正相关,表明其与HIV-1病毒复制相关。     

肾病综合征患者血浆leptin、IL-4、IL-18及TNF-α水平变化及其临床意义

摘要 目的：探究肾病综合征患者血浆瘦素（leptin）、白介素-4（IL-4）、白介素-18（IL-18）及肿瘤坏死因子-α（TNF-α）水平变化及其临床意义。方法：选择2018年2月至2019年10月我院诊治的110例肾病综合征患者作为观察组,选择同期在我院体检的110名健康者作为对照组。检测并比较两组的血浆leptin、IL-4、IL-18、TNF-α水平及24 h蛋白尿、血浆白蛋白水平,采用Pearson法分析血浆leptin、IL-4、IL-18和TNF-α与24 h蛋白尿、血浆白蛋白的相关性,采用受试者工作特征曲线（ROC）分析各指标对肾病综合征的诊断价值。结果：与对照组相比,观察组的血浆leptin、IL-4、IL-18及TNF-α水平明显升高（P<0.05）。与对照组相比,观察组的24 h蛋白尿明显升高,血浆白蛋白水平明显下降（P<0.05）。血浆leptin、IL-4、IL-18和TNF-α水平与24 h蛋白尿均呈正相关性,而与血浆白蛋白均呈负相关性（P<0.05）。血浆leptin、IL-4、IL-18和TNF-α联合检测对肾病综合征的诊断价值明显高于各项指标单独检测。结论：肾病综合征患者血浆leptin及炎症因子水平明显升高,并且与患者24h蛋白尿以及血浆白蛋白具有相关性,血浆leptin和IL-4、IL-18、TNF-α联合检测对肾病综合征具有较高的诊断价值。     

血浆外泌体miR-1290、miR-1307表达水平与卵巢癌患者临床病理特征的关系及其诊断价值分析

摘要 目的：探讨卵巢癌患者血浆外泌体中微小RNA（miR）-1290和 miR-1307的表达,分析两者与临床病理参数的关系及诊断价值。方法：选取2019年1月至2021年1月于靖江市人民医院诊治的60例卵巢癌患者为卵巢癌组,另分别选取同期52例良性卵巢肿瘤患者和50例女性健康体检者为良性肿瘤组和健康对照组。采用实时荧光定量PCR（qRT-PCR）法检测miR-1290、miR-1307在各组血浆外泌体中的表达,分析miR-1290、miR-1307与卵巢癌患者临床病理特征的关系,采用受试者工作特征（ROC）曲线分析血浆外泌体miR-1290、miR-1307对卵巢癌的诊断效能。结果：与良性肿瘤组和健康对照组相比,卵巢癌组的血浆外泌体miR-1290、miR-1307表达水平较高（P＜0.05）。卵巢癌组的miR-1290表达水平与FIGO分期、分化程度及淋巴结转移有关,miR-1307表达水平与FIGO分期、淋巴结转移有关（P＜0.05）;卵巢癌患者血浆外泌体miR-1307表达与miR-1290表达呈显著正相关（r＝0.478,P＜0.05）。血浆外泌体miR-1290、miR-1307联合应用诊断卵巢癌的曲线下面积（AUC）（0.95CI）为0.726（0.526～0.926）,高于两指标单独应用的0.724（0.466～0.987）、0.845（0.750～0.933）。结论：卵巢癌患者血浆外泌体miR-1290、miR-1307表达水平升高,两者联合检测对卵巢癌的早期诊断具有较高的价值,有望成为新的卵巢癌的分子标志物。     

miR-29b对人子宫内膜癌RL95-2细胞凋亡的影响

为了探讨miR-29b对人子宫内膜癌(endometrial cancer, EC) RL95-2细胞凋亡的影响及机制,本研究将RL95-2细胞转染miR-29b mimics和miR-NC后,采用MTT法检测细胞活力;运用Annexin V-PI法检测细胞凋亡;采用Western blotting检测Caspase-3蛋白表达;使用流式细胞术检测ROS水平。研究结果显示:与miR-NC组比较,miR-29b mimics过表达miR-29b组在培养24 h和48 h后的RL95-2细胞活力显著降低(p0.05)。与miR-NC组比较,miR-29b mimics组的RL95-2细胞凋亡率显著增加(p0.05)。与miR-NC组比较,miR-29b mimics组的RL95-2细胞Cleaved caspase-3蛋白表达水平显著增加(p0.05)。此外,miR-29b mimics组的RL95-2细胞ROS水平明显低于miR-NC组(p0.05)。本研究的初步结论表明:miR-29b可能通过调控ROS水平促进人RL95-2细胞凋亡。     

活动期肺结核患者外周血miR-29a的表达及其临床意义

目的分析活动期肺结核患者外周血miR-29a的表达情况及其临床意义。方法收集2015年1月至2016年12月绍兴市立医院诊断的45例未经抗结核治疗的活动性肺结核患者(活动组),45例其他肺部疾患者(其他肺疾病组)和45例健康体检者(健康对照组)的外周血,通过实时定量PCR技术检测各组研究对象血清中miR-29a的表达水平并进行ROC曲线分析。采用ELISA法检测各组研究对象血清IFN-γ表达水平;采用流式细胞术测定各组研究对象外周血T细胞亚群水平。分析活动性结核患者血清miR-29a表达水平与IFN-γ以及T细胞亚群水平之间的相关性。结果活动组患者血清miR-29a表达水平均显著高于其他肺疾病组以及健康对照组(t=7.005、4.307,P0.01)。ROC曲线分析表明miR-29a的曲线下面积为0.874,诊断活动性肺结核的敏感度为66.7%,特异度为93.3%。活动组患者血清中IFN-γ的表达水平显著高于其他肺疾病组和健康对照组(t=16.001、14.771,P0.01)。活动组患者外周血CD_3~+、CD_4~+细胞水平以及CD_4~+/CD_8~+比值显著低于其他肺疾病组和健康对照组(t=7.109、5.601,P0.01;t=3.133、4.455,P0.01;t=2.949、2.500,P0.01),而CD+8细胞水平显著高于其他肺疾病组和健康对照组(t=2.375、2.908,P0.01)。活动组患者血浆miR-29a水平与IFN-γ水平呈负相关(r=-0.8542,P0.01)。活动组患者血浆miR-29a水平与外周血CD_3~+、CD_4~+细胞水平以及CD_4~+/CD_8~+比值呈负相关(r=-0.8412、-0.8856、-0.7746,P0.01),同时与CD_8~+细胞水平呈正相关(r=0.8336,P0.01)。结论活动期肺结核患者外周血miR-29a表达水平显著升高,影响患者免疫应答功能,可作为诊断活动期肺结核的临床标志物。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Chemokines and cytokines: axis and allies in asthma and allergy

Allergic asthma can be precipitated by many factors. For the atopic person, fungus, pollen, dust mites, cockroach antigens, and diesel exhaust are all agents that may trigger an allergic attack. Cytokines and chemokines are integral mediators of fungal asthma. From the earliest time points, they recruit and activate the cells required for the clearance of fungus as well as being critical factors involved in the immunopathology of this disease. In the final analysis, it is clear that these mediators can act to the benefit or the detriment of the host.     

Oxidation and nitration of ribonuclease and lysozyme by peroxynitrite and myeloperoxidase

In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier.