LPS及IL-1ra对系膜细胞产生一氧化氮及增殖的影响

目的:观察脂多糖(LPS)对体外培养的系膜细胞(GMC)产生NO的影响及这一过程对GMC增殖的影响。方法:用培养的第一、二代SD大鼠肾小球系膜细胞进行实验,分别加入LPS或LPS加白细胞介素-1受体拮抗剂(IL-1ra),24h后用经典的Griess方法测定培养上清中亚硝酸盐含量,用[³H]-TdR掺入率测定GMC的增殖,狭缝和Northern杂交测定iNOSmRNA表达。结果:LPS刺激后,GMC出现iNOSmRNA的表达及亚硝酸盐增多[(0.64±0.25)nmmol/10⁴细胞vs(0.12±0.06)nmmol/10⁴细胞],GMC增殖(3735±1177.9vs1785±280.6);LPS加IL-1ra组,iNOSmRNA表达比LPS组减少40%,上清亚硝酸盐含量更高[(3.28±0.33)nmmol/10⁴细胞],GMC增殖被抑制(818±77.27)。结论:LPS诱导GMCiNOSmRNA表达及亚硝酸盐增多,并有GMC增殖。IL-1ra部分拮抗iNOSmRNA的表达,但亚硝酸盐产量更高,GMC增殖被抑制。     

糖尿病患者外周血白细胞介素18分泌及基因表达

目的　探讨糖尿病 (DM)患者血浆白细胞介素 18(IL 18)水平及外周血单个核细胞 (PBMCs)IL 18mRNA表达量变化。方法　选取 10名健康志愿者及 30例DM患者 (Ⅱ型糖尿病 15例 ,Ⅰ型糖尿病 15例 )。应用IL 18特异的酶联免疫吸附法(ELISA)测定IL 18血浆水平 ,同时采用半定量逆转录多聚酶链反应 (RT PCR)技术 ,分析PBMCsIL 18mRNA表达量。结果　Ⅱ型糖尿病患者血浆IL 18水平及PBMCsIL 18mRNA表达量与正常对照组差异无统计学意义 [(2 73.1± 99.4 6 )vs (2 4 8.86± 82 .6 8)and(0 .731± 0 .12 9)vs (0 .715± 0 .116 ) ,P >0 .0 5 ];Ⅰ型糖尿病患者血浆IL 18水平及PBMCsIL 18mRNA表达量较正常对照组增高 ,差异有显著统计学意义 [(36 0 .17± 12 5 .4 8)vs (2 4 8.86± 82 .6 8)and (0 .935± 0 .132 )vs (0 .715± 0 .116 ) ,P <0 .0 5 ]。结论　Ⅰ型糖尿病患者外周血IL 18的高表达 ,提示其可能介导本病发病过程 ;IL 18可能不参与Ⅱ型糖尿病的发病过程     

CD40L单克隆抗体对HSP患儿PBMC及单核细胞株THP-1产生炎症因子的影响

目的　观察CD4 0配体单克隆抗体 (CD4 0LMcAb)对HSP(亨诺 许兰紫癜 )患儿PBMC及单核细胞株THP 1产生炎症因子的影响。方法　采用细胞培养、流式细胞技术及ELISA法分别检测正常对照、过敏性紫癜患儿的PBMC及单核细胞株THP 1表达膜蛋白分子CD4 0L、CD4 0的阳性率、产生炎症因子IL 1、TNF α、IL 6水平以及CD4 0LMcAb加入细胞培养体系后上述指标的变化。结果　与正常比较 ,HSP患儿的PBMC表达CD4 0L[(8.2 6± 4 .15 ) % ,对照 (0 .5 4± 0 .5 8) % ]显著增高、CD4 0表达无差异 ;PBMC培养上清中炎症因子IL 1[(10 0 0 .4± 5 74 .5 )pg ml,对照 (2 4 6 .8± 2 0 7.1)pg ml]、TNF α[(978.7± 2 0 5 .8)pg ml,对照 (45 2 .4± 2 4 8.5 )pg ml]的水平也显著高于对照 ,CD4 0LMcAb可使增高的IL 1[(16 4 .7± 12 7.9)pg ml]、TNF α[(6 74 .7± 2 6 9.2 )pg ml]降至正常水平。THP 1自发表达CD4 0 ,低浓度产生IL 1、TNF α。与正常比较 ,HSP患儿的PBMC培养上清诱导其表达CD4 0 ,产生IL 1,TNF α增高 ,CD4 0LMcAb同样具有抑制THP 1表达CD4 0、分泌IL 1、TNF α的作用。结论　CD4 0LMcAb能抑制炎症因子的产生。     

新生儿脐血单核细胞在脂多糖刺激下分泌IL-6及出现IL-6mRNA表达的实验研究

目的 :通过观察LPS对新生儿脐血单个核细胞 (MC)分泌IL 6及表达IL 6mRNA基因的影响 ,探讨严重细菌感染时新生儿机体防御反应机制。方法 :取肝素抗凝剂脐血 ,用密度离心分离法分离MNC ,以RPMI16 40培养液调整细胞浓度为1× 10 6 ml- 1 ,将细胞悬液铺于 2 4孔培养板上 ,依次加入不同浓度脂多糖 (LPS)培养 36h或同一浓度LPS(1μg ml)培养不同时间 ,收集培养上清液及细胞 ,分别用ELISA和RT PCR方法测定IL 6及IL 6mRNA表达情况。结果 :①脐血MNC在LPS刺激 3、6、12、18、2 4、36h后IL 6分泌水平逐步增高 ,6h以后增加尤为明显 ,与其他各时间点比较有非常显著的差异 (P <0 0 0 1)。LPS刺激组与无LPS对照组相同时间点比较 ,6h内IL 6变化水平无差异 ,6h以上各点有显著差异 (P <0 0 1)。RT PCR方法检测显示LPS刺激后 3h即可见IL 6mRNA基因表达。②脐血MNC受不同浓度LPS刺激时 ,IL 6分泌水平随LPS浓度递增。③全部脐血MNC均检测到IL 6mRNA基因表达。结论 :LPS能诱导新生儿脐血MNCIL 6mRNA基因转录 ,从而促使IL 6合成、分泌 ,该作用呈时间、剂量依赖性变化。     

核因子κB顺式诱饵元件对哮喘大鼠模型治疗作用的观察

目的　探讨核因子κB(NF κB)顺式诱饵元件治疗支气管哮喘的可行性。方法　通过从气道局部给药的方式 ,将NF κB顺式诱饵寡核苷酸 (N ODN)或无序诱饵ODN(S ODN)转染至哮喘大鼠肺部。运用原位组织杂交、反转录PCR、免疫组织化学、酶联免疫吸附试验及肺部病理学检测和气道反应性测定等方法检测NF κB顺式诱饵ODN对哮喘大鼠肺组织白细胞介素 5 (IL 5 )、诱导型一氧化氮合酶 (iNOS)mRNA和蛋白质表达及肺部炎症反应、气道反应性的影响。结果　ODN转染组大鼠肺部均有明显的FITC着色 ,说明此种局部滴入ODN的方法是可行的。经NF κB顺式诱饵ODN转染的哮喘大鼠 (N ODN组 )IL 5、iNOSmRNA表达均较未治疗的哮喘大鼠 (A组 )明显降低 (N ODNvsA ,0 .17±0 .0 3vs 0 .2 4± 0 .0 5 ,0 .2 4± 0 .0 5vs 0 .77± 0 .10 ,P <0 .0 5 ) ;蛋白质表达亦较A组减少 (40 .0± 10 .8vs80 .0± 2 5 .8,0 .19± 0 .0 4vs 0 .37± 0 .0 6 ,P <0 .0 5 ) ;且其肺部嗜酸粒细胞数量显著减少 (10 .0 0± 2 .94vs 16 .2 5± 4 .6 5 ,P <0 .0 5 ) ,气道反应性降低 (PC50 1.4± 0 .4vs0 .4± 0 .3,P <0 .0 5 )。而无序诱饵ODN转染的哮喘大鼠与未治疗的哮喘大鼠相比 ,上述指标差异无显著性 (P值均 >0 .0 5 )。结论　NF κB顺式诱饵元件整体给     

白细胞介素-4对大鼠气道平滑肌细胞的促增生作用

气道平滑肌增殖是支气管哮喘的特征性病理改变。本研究用体外培养的大鼠气道平滑肌细胞 (ASMC) ,观察了白细胞介素 4(IL 4 )对其增殖的影响。实验用四唑盐比色法 (MTT法 )和3 H TdR掺入法。MTT检测结果以OD值表示 ,加入IL 4的 2 4h组和 48h组分别为 0 32 3± 0 0 2 6 (x—±s,下同 )和 0 4 5 3± 0 0 48,比相应时间对照组的 0 191±0 0 18和 0 335± 0 0 6 3明显增加 ,(P均 <0 0 0 1) ;3 H TdR掺入也得到类似结果 ,IL 4 2 4h组 3 H掺入量为 76 5 8± 6 34 ,高于对照组的 6 0 6 0± 6 71counts min(P <0 0 1) ;用MTT检测法还观察到作用 2 4h ,IL 4 +胸腺肽组为 0 30 8± 0 0 0 7、IL 4 +地塞米松组为 0 2 45± 0 0 0 8比IL 4组 0 32 4± 0 0 14降低 (分别P <0 0 5及P <0 0 0 1) ,说明两种药物均可抑制IL 4的促增殖作用。实验表明IL 4可能还通过促进ASMC的增殖参与哮喘发病 ;而抑制IL 4对ASMC的促增殖作用可能是胸腺肽和地塞米松治疗哮喘的机制之一。     

乙型肝炎患者外周血单个核细胞凋亡的检测及意义

目的研究激活诱导细胞死亡 (AICD)现象在乙型肝炎慢性化和重型化机制中的意义。方法分离 2 0例慢性 /慢性重型乙型肝炎病人与 10例健康献血员外周血单个核细胞 (PBMC) ,在 PHA的刺激下培养 72 h后收集细胞 ,经流式细胞仪检测凋亡。结果 PBMC凋亡率乙型肝炎组明显高于正常对照组 [(2 5 .48± 14.0 7) % vs(11.45± 5 .2 7) % ,P<0 .0 1];慢性乙型肝炎组 [(30 .5 7± 13.43) % ]明显高于正常对照组 [(11.45± 5 .2 7) % ,P<0 .0 1]和慢性重型乙型肝炎组 [(13.5 9± 6 .44 ) % ,P<0 .0 1];PBMC凋亡率乙型肝炎 HBe Ag(+ )组明显高于正常对照组 [(2 9.5 0± 12 .5 4) % vs(11.45± 5 .2 7) % ,P<0 .0 1]。结论 AICD可能是形成 HBV慢性感染免疫耐受的一个重要机制。     

低氧性肺动脉高压大鼠肺组织硫化氢的变化

本文旨在探讨低氧对大鼠内源性硫化氢体系的变化。采用生化反应方法测定血浆中硫化氢的含量和肺组织中硫化氢合酶的活性 ,采用定量竞争性RT PCR的方法检测肺组织中胱硫醚 γ 裂解酶 (CSE)mRNA的含量。结果显示 ,与对照组相比 ,低氧组大鼠的血浆硫化氢含量明显减少 [(1 92 2± 2 2 1 ) μmol Lvs (30 1 6± 32 4 ) μmol L ,P <0 0 1 ) ],肺组织中硫化氢合酶的活性明显下降 [(0 1 2 7± 0 0 2 3)vs (0 2 78± 0 0 99)nmol mgwettissue·min ,P <0 0 1 ],CSEmR NA的含量明显减少 [(1 0 2± 0 1 5 )× 1 0 - 6 fmolvs (2 1 7± 0 2 2 )× 1 0 - 6 fmol,P <0 0 1 ]。以上研究表明 ,低氧对大鼠的内源性硫化氢体系有抑制作用     

肠血管活性多肽调节多器官功能衰竭时肠黏膜肥大细胞的活性

目的　探讨肠血管活性多肽 (vasoactiveintestinalpeptide ,VIP)对多器官功能衰竭 (multi pleorganfailure ,MOF)时肠黏膜肥大细胞 (intestinalmucosalmastcells,IMMC)活性的调节及其病理生理意义。方法　酵母多糖腹腔注射法制作大鼠MOF模型 ,注射酵母多糖后 ,经尾静脉输入VIP(剂量为2 0pmol g体重或 0 .2pmol g体重 ) ,观察动物肠、肝、肾、肺等重要器官的组织病理改变及谷丙转氨酶ALT、肌酐Cr、血氧分压PO2 等功能指标变化 ,测定动物外周血和小肠组织组胺水平 ,透射电镜观察IMMC超微结构变化。结果　大剂量VIP输注后 ,与MOF对照组相比 ,大鼠各重要器官病变明显加重 ,ALT升高 [(180 .6 0± 2 3.4 0 )U Lvs (331.34± 35 .0 0 )U L],Cr升高 [(10 2 .35± 17.3)U Lvs 2 0 .5 0U L],PO2 降低 [(12 .5 4± 2 .6 0 )kPavs (7.4 4± 2 .17)kPa],P <0 .0 1,小肠组胺水平明显升高 [(8.4 0± 1.79)ng g体重vs (14 .30± 2 .70 )ng g体重 ,P <0 .0 1],IMMC脱颗粒现象明显改善。而小剂量VIP输注后 ,大鼠各重要器官病变明显减轻 ,ALT下降 6 2 .35 % ,Cr下降 6 3.2 0 % ,PO2 升高 38.30 % ,P <0 .0 1,小肠组胺水平明显降低 [(8.4 0± 1.79)ng g体重vs (4.70± 0 .4 5 )ng g体重 ,P <0 .0 1],IMMC脱颗粒现象更明显。     

狼疮性肾炎患者外周血IL-18水平及其基因表达

为了探讨白细胞介素 18(IL 18)在狼疮性肾炎 (LN )发生、发展中的作用。我们采用逆转录多聚酶链反应 (RT PCR )及酶联免疫吸附 (ELISA )法测定 16例正常人及 18例LN患者外周血单个核细胞 (PBMC )IL 18mRNA表达量及其血浆水平。结果提示LN患者PBMCIL 18mRNA表达量及血浆IL 18水平均较正常对照组显著增高 [IL 18mRNA表达量为 :1 2 6 2± 0 189vs0 84 4± 0 15 5 ,P <0 0 0 1;IL 18血浆水平为 :(82 2 0 9± 5 32 77)pg/mlvs (2 39 5 7± 75 0 6 )pg/ml,P <0 0 0 1]。且WHOIV型LN增高较非IV型LN更为显著 [IL 18mRNA表达量为 :1 32 9± 0 2 1vs 1 138± 0 15 2 3,P <0 0 5 ;IL 18血浆水平为 :(1135 5 4± 5 15 34)pg/mlvs (5 0 8 6 5± 341 36 )pg/ml,P <0 0 1]。另外 ,血浆IL 18水平与肾组织活动指数 ,肾小管间质损害程度呈等级相关 (r分别为 :0 6 10和 0 4 99,P均 <0 0 5 ) ,也与血清肌酐 (Scr) ,血清内生肌酐清除率 (Ccr)及 2 4h尿蛋白排泄量 (2 4hUPQ )呈直线相关 (r分别为 :0 898、 0 6 2 8和 0 5 37,P均 <0 0 5 )。本研究认为循环IL 18表达和分泌增高可能参与LN的免疫发病过程 ,并与狼疮活动有一定的关系     

纳米水平分子成像与诊疗一体化研究进展与挑战

分子成像能以非侵入性的方式重现活体细胞的生理功能和生物学过程,提高疾病的早期和特异性诊断水平。纳米颗粒/材料具有物理性质可控性高、易于表面修饰、血液循环时间长和可功能化等优点,在疾病诊断与治疗中显示出巨大潜力。但如何阐明纳米材料多功能间的内在联系、解决其代谢及安全性等关键机制难题、实现纳米颗粒/材料多功能性到临床多功能...     

Association between functional alterations of senescence and senility and disorders of gait and balance

OBJECTIVES:

Declines in cognition and mobility are frequently observed in the elderly, and it has been suggested that the appearance of gait disorders in older individuals may constitute a marker of cognitive decline that precedes significant findings in functional performance screening tests. This study sought to evaluate the relationship between functional capacities and gait and balance in an elderly community monitored by the Preventive and Integrated Care Unit of the Hospital Adventista Silvestre in Rio de Janeiro, RJ, Brazil.

METHODS:

Elderly individuals (193 females and 90 males) were submitted to a broad geriatric evaluation, which included the following tests: 1) a performance-oriented mobility assessment (POMA) to evaluate gait; 2) a mini-mental state examination (MMSE); 3) the use of Katz and Lawton scales to assess functional capacity; 4) the application of the geriatric depression scale (GDS); and 5) a mini-nutritional assessment (MNA) scale.

RESULTS:

Reductions in MMSE, Katz and Lawton scores were associated with reductions in POMA scores, and we also observed that significant reductions in POMA scores were present in persons for whom the MMSE and Katz scores did not clearly indicate cognitive dysfunction. We also demonstrated that a decline in the scores obtained with the GDS and MNA scales was associated with a decline in the POMA scores.

CONCLUSIONS:

Considering that significant alterations in the POMA scores were observed prior to the identification of significant alterations in cognitive capacity using either the MMSE or the Katz systems, a prospective study seems warranted to assess the predictive capacity of POMA scores regarding the associated decline in functional capacity.     

男性乳腺发育症临床病理分析及雌,孕激素受体检测

对１１３例男性乳腺发育症进行临床病理分析。同时检测其中３０例乳腺组织中雌激素受体和孕激素受体分布情况，结果发现两者阳性率分别为８０．０％和８３．３３％。结合文献讨论了男性乳腺发育症的发生与高血清激素浓度及乳腺组织高受体水平的关系。     

Alterations of regular and mature monocytes are distinct,and dependent of intensity and duration of exercise

Circulating monocytes comprise functionally distinct regular (CD14^bright+) and mature (CD141^low+) cells. Cell surface receptors were determined by three colour flow cytometry in 8 healthy control subjects. Compared to regular monocytes, mature monocytes had lower levels of the high affinity Fcy receptor 1 (CD64), complement receptor 3 (CDllb), CD45RO and higher levels for HLA-DR, LFA-1 (CD11a/CD18), interleukin-2 receptor (CD25), CD45RA and the Fc receptor 3 (CD16). Both regular and mature monocytes were measured before and up to three hours after four different types of exercise (Ex) in endurance trained athletes (n=9-16). Immediately after anaerobic exercise of I min with a maximal lactate concentration (la_max) of I2.3 (SD I.4) mmol · l^–1 and exhaustive exercise of 24 (SD 8) min with a maximal lactate concentration (la_max) of 7.4 (SD 2.6) mmol· l^–1 mature monocytes increased more than regular monocytes. Exhaustive endurance exercise of 87 (SD 21) min [la_max 3.7 (SD I.0)] led to a similar increase of regular and mature monocytes. 15–33 min after a 100km run regular monocytes increased significantly, whereas mature monocytes decreased. Up to three hours after the end of all exercises mature monocytes fell below pre-exercise values. In conclusion, duration and intensity of exercise alter distinct maturation stages of monocytes differently. It is probable that the avidity of adhesion molecules like LFA-1 to their endothelial ligands is increased to enable the firm attachment to the endothelium.     

Triptorelin and cetrorelix induce immune responses and affect uterine development and expressions of genes and proteins of ESR1, LHR,and FSHR of mice

Context: GnRH immunity can reduce the expression of pituitary GnRH levels, and cause the changes in reproductive behaviors. It is unclear whether triptorelin (TRI) and cetrorelix (CET) immunity influences uterine development and expression of follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), and estradiol receptor 1 (ERS1) in the uterus.

Objective: The study investigated the effects of active immunity of GnRH agonist and antagonist on uterine development, microstructures, expression of hormone receptors mRNAs, and proteins in uteri.

Materials and methods: One hundred and five mice were assigned into CET, TRI, and control groups (CG). Mice in CET-1, CET-2, and CET-3 (n?=?15) were subcutaneously injected with 10, 20, and 40?μg CET antigens for seven days, respectively. Mice in TRI-1, TRI-2, and TRI-3 were injected with 10, 20, and 40?μg TRI antigens for seven days, respectively. The qPCR and Western blot were implemented to determine expressions of ESR1, LHR and FSHR mRNAs, and proteins.

Results: Compared with CG, the uterine weights of CET-1, CET-2, and CET-3 increased by 42.86, 62.86, and 10.00% on day 35 (p?p?p?p?p?Conclusions: CET immunity promoted the uterine development, improved EET and UWT, and also promoted the expressions of ESR1 and FSHR protein levels. It lessened the LHR protein levels. TRI immunity blocked EET and UWT, inhibited uterine growth and development. The efficacy of CET immunity was more obvious than TRI.     

韧带和肌腱的生物力学和力学生物学研究

国际韧带和肌腱研讨会(The International Symposium on Ligaments and Tendons,ISI&T)于2000年在美国佛罗里达州奥兰多市首次召开。研讨会的宗旨是引起对韧带和肌腱研究的重视,并为生物工程师、生物学家、临床医师提供一个可以分享、评论、讨论韧带和肌腱最新研究成果的论坛。从2000年起,国际韧带和肌腱研讨会已经开展了15届;每届研讨会上涌现了大量令人振奋的关于当前韧带和肌腱研究热点和未来挑战的讨论。多年来,韧带和肌腱领域内的研究数量大幅增加,研究质量不断提升。为纪念《医用生物力学》杂志创刊30周年,本文总结过去30年里韧带和肌腱研究的主要进展,包括组织力学、力学生物学、损伤与治愈机制、组织修复和再生。     

Morphology and cytochemistry of leucocytes and thrombocytes of six species of fish

Cytochemical reactions of blood leucocytes and thrombocytes from six species of fish, rainbow trout (Onchorhynchus mykiss), coho salmon (Onchorhynchus kisutch), white sturgeon (Acipenser transmontanus), goldfish (Carassius auratus), striped bass (Morone saxatulis), and channel catfish (Ictalurus punctatus) were determined. Because the staining reactions were generally similar to the reactions found in mammalian leucocytes with similar morphological features, it is reasonable to classify fish leucocytes using the same terminology as is used for mammalian leucocytes. However, in some species leucocytes with features similar to mammalian eosinophils or basophils were not found. In goldfish leucocytes were found that had segmented nuclei and unstained, moderately refractile cytoplasmic granules. These cells were classified as segmented, granular leucocytes. Although these cells do not appear similar to any mammalian or avian leucocyte, the pattern of positive cytoplasmic alkaline phosphatase staining and negative granular staining is similar to that of equine eosinophils.     

Pathology and genetics of phaeochromocytoma and paraganglioma

Phaeochromocytoma and paraganglioma (PHEO/PGL) are rare tumours with an estimated annual incidence of 3 per million. Advances in molecular understanding have led to the recognition that at least 30–40% arise in the setting of hereditary disease. Germline mutations in the succinate dehydrogenase genes SDHA, SDHB, SDHC, SDHD and SDHAF2 are the most prevalent of the more than 19 hereditary genetic abnormalities which have been reported. It is therefore recommended that, depending on local resources and availability, at least some degree of genetic testing should be offered to all PHEO/PGL patients, including those with clinically sporadic disease. It is now accepted that that all PHEO/PGL have some metastatic potential; therefore, concepts of benign and malignant PHEO/PGL have no meaning and have been replaced by a risk stratification approach. Although there is broad acceptance that certain features, including high proliferative activity, invasive growth, increased cellularity, large tumour nests and comedonecrosis, are associated with an increased risk of metastasis, it remains difficult to predict the clinical behaviour of individual tumours and no single risk stratification scheme is endorsed or in widespread use. In this review, we provide an update on advances in the pathology and genetics of PHEO/PGL with an emphasis on the changes introduced in the WHO 2017 classification of endocrine neoplasia relevant to practising surgical pathologists.     

壬基酚与卵清蛋白的结合与鉴定

目的：合成壬基酚与卵清蛋白的偶联物。方法：在磷酸盐（PBS，pH=8．0）缓冲液中利用甲醛通过曼尼希反应连接壬基酚与卵清蛋白（OVA）；通过抗体芯片技术与紫外扫描鉴定偶联物。结果：壬基酚与OVA偶联成功，壬基酚单克隆抗体对偶联物的识别浓度小于2．68μg／ml。结论：本方法可用于壬基酚与卵清蛋白的偶联，且方法简单易实现。