Toll-like receptor 9 signaling mediates the anti-inflammatory effects of probiotics in murine experimental colitis

BACKGROUND & AIMS: We tested whether the attenuation of experimental colitis by live probiotic bacteria is due to their immunostimulatory DNA, whether toll-like receptor (TLR) signaling is required, and whether nonviable probiotics are effective. METHODS: Methylated and unmethylated genomic DNA isolated from probiotics (VSL-3), DNAse-treated probiotics and Escherichia coli (DH5 alpha) genomic DNA were administered intragastrically (i.g.) or subcutaneously (s.c.) to mice prior to the induction of colitis. Viable or gamma-irradiated probiotics were administered i.g. to wild-type mice and mice deficient in different TLR or in the adaptor protein MyD88, 10 days prior to administration of dextran sodium sulfate (DSS) to their drinking water and for 7 days thereafter. RESULTS: Intragastric and s.c. administration of probiotic and E. coli DNA ameliorated the severity of DSS-induced colitis, whereas methylated probiotic DNA, calf thymus DNA, and DNase-treated probiotics had no effect. The colitis severity was attenuated to the same extent by i.g. delivery of nonviable gamma-irradiated or viable probiotics. Mice deficient in MyD88 did not respond to gamma-irradiated probiotics. The severity of DSS-induced colitis in TLR2 and TLR4 deficient mice was significantly decreased by i.g. administration of gamma-irradiated probiotics, whereas, in TLR9-deficient mice, gamma-irradiated probiotics had no effect. CONCLUSIONS: The protective effects of probiotics are mediated by their own DNA rather than by their metabolites or ability to colonize the colon. TLR9 signaling is essential in mediating the anti-inflammatory effect of probiotics, and live microorganisms are not required to attenuate experimental colitis because nonviable probiotics are equally effective.     

Furlow术式及改良兰式修复浅Ⅱ度腭裂的临床研究

目的对比Furlow术式及改良兰式对浅Ⅱ度腭裂的修复效果,评价这两种手术方式对于修复此类腭裂的优劣。方法 50例浅Ⅱ度腭裂患者随机分成两组,A组使用Furlow术式,B组使用改良兰式,对比两组患者的术后软腭延长长度、手术时间、术中出血量、术后3 d平均体温及术后语音改善的程度。结果两种术式在术后软腭延长长度方面,Furlow术式优于改良兰式,前者为(0.794±0.198)cm,后者为(0.118±0.076)cm,差异有统计学意义(t=15.928,P<0.001)。手术时间、术中出血量、术后3 d平均体温及术后语音改善程度,2种术式差异无统计学意义(P>0.05)。结论两种术式均可用于浅Ⅱ度腭裂的修复,但Furlow术式在延长软腭方面优势明显,对于不同类型腭裂可选用个体化修复方案。     

吗啡成瘾性大鼠脑内核团神经细胞[Ca 2+]i变化的研究

目的 研究吗啡成瘾对大鼠脑内与成瘾相关核团神经细胞[Ca 2 ]i的影响.方法 将50只SD大鼠随机分成吗啡成瘾组和对照组,观察成瘾后的戒断症状;应用对Ca2 敏感的探针Fluo-3与Ca2 络合后被激光激发发出荧光的特性,利用激光共聚焦显微镜观察伏核、海马、内侧额前皮质神经细胞[Ca 2 ]i变化,并用图像分析软件进行处理.结果 吗啡成瘾大鼠脑内伏核、海马及内侧额前皮质神经细胞[Ca 2 ]i和对照组比较明显增高(P<0.01).结论 长期使用吗啡可明显增高伏核、海马和内侧额前皮质神经细胞[Ca 2 ]i.     

XT-4000i血液分析仪检测异型淋巴细胞的实验研究

目的通过XT-4000i血液分析仪白细胞分类检测通道（DIFF通道）检测高荧光淋巴细胞（highfluorescence lymphocytes,HFL）区域即HFL计数与人工镜检、EB病毒壳抗原（EBV-VCA）检测结果比较,探讨XT-4000i血液分析仪对异型淋巴细胞检出意义。方法 XT-4000i血液分析仪对230份标本采用DIFF通道检测HFL,与显微镜下计数异型淋巴细胞比较,并用血清作免疫酶标法检测EBV-VCA。结果 XT-4000i血液分析仪对异型淋巴细胞检出的假阳性率为13.6%,假阴性率为2%。结论 XT-4000i血液分析仪对异型淋巴细胞的检出有一定的有效性,对临床诊断效率的提高有一定帮助。     

冠心病介入术后氧自由基的生成及其对脂质过氧化和纤溶的影响

目的:探讨冠心病介入治疗术后氧自由基的生成及其对患者体内脂质过氧化和纤溶状况的影响。方法:48例接受经皮冠脉介入治疗(PCI)术的冠心病患者分别于术前,术后即刻、30 min、1 h、24 h和一周采用ELISA法测定血浆中丙二醛、D-二聚体、血管假性血友病因子相关抗原(vWF Ag)浓度。结果:PCI术后短时间内(0～24 h)血浆中丙二醛、D-二聚体,vWFAg水平均明显升高(P<0．05)。结论:PCI术后短时间内即有大量的氧自由基生成,凝血系统的激活和继发性纤溶功能亢进。     

Impact of genetic variations in the WRN gene on age related pathologies and mortality

Mutations in the WRN gene lead to the Werner syndrome (WS), which resembles premature aging. Here, we hypothesize that genetic variations in the WRN gene may also influence aging-trajectories in the population at large. To test this hypothesis, we assessed the impact of the i1-C/T, L1074F and C1367R polymorphisms in the WRN gene on the occurrence of cardiovascular pathologies, on cognitive performance and on the risks of all-cause, cardiovascular and cancer mortalities in the population-based Leiden 85-plus Study. This prospective follow-up study includes 1,245 participants aged 85 years and older, with a total follow-up of 5,164 person-years. At baseline the risks of myocardial infarction, myocardial ischemia, intermittent claudication, arterial surgery and stroke dependent on the i1-C/T, L1074F and C1367R polymorphisms, did not vary between the different genotypes. Also no differences in cognitive functioning were observed, except for attention, where carriers of the 1367R allele performed worse compared to the 1367C homozygotes (94.2 (4.35) versus 84.8 (1.84), p=0.04). Mortality risks, calculated separately for all SNPs, were similar between the different genotype carriers of the i1-C/T, L1074F and C1367R polymorphisms, showing no evidence of altered survival. In conclusion, the i1-C/T, L1074F and C1367R polymorphisms in the WRN gene do not influence the aging-trajectories and survival in the population at large.     

Complement receptors type 1 (CR1, CD35) and 2 (CR2, CD21) cooperate in the binding of hydrolyzed complement factor 3 (C3i) to human B lymphocytes

The C3b-binding receptor, CR1/CD35, supports CR2/CD21-mediated activation of complement by human B lymphocytes, possibly by associating with CR2 to promote or stabilize the binding of hydrolyzed C3 (C3i), the primary component of the AP convertase, C3i-Bb. To evaluate this hypothesis, we examined the uptake kinetics and binding equilibria for C3i dimer interaction with human blood cells in the absence and presence of CR1- and CR2-blocking mAb. C3i displayed dual uptake kinetics to B lymphocytes, comprising of rapid binding to CR1 and slower binding to CR2. The forward rate constants (k(1)) for CR1 and CR2, operating independently, differed ca. 9-fold (k(1)=193+/-9.4 and 22.2+/-6.0 x 10(3) M(-1)s(-1), respectively). Equilibrium binding of C3i to B lymphocytes was also complex, varying in strength by ca. 13-fold over the C3i concentration range examined. The maximum association constant (K(a, max)=109+/-27.2 x 10(7) l/mole) was ca. 9- and 6-fold greater, respectively, than those for CR1 or CR2 acting alone (K(a)=13.2+/-5.3 and 18.5+/-3.5 x 10(7) l/mole). The high avidity of the CR1-CR2 complex for C3i is consistent with its rates of C3i uptake and release being determined by CR1 and CR2, respectively.     

The antiplatelet activity of Danggijakyaksan by inhibition of phospholipase C

We investigated the effects of the traditional Korean prescription, Danggijakyaksan (DJS) on antiplatelet activity in human platelet suspensions. The effect of oriental medicinal prescriptions, Danggijakyaksan consisting of 6 herbes of <i>Paeoniae Radixi> (2 g), <i>Poria Cocosi> (1.33 g), <i>Angelicae Sinensis Radixi> (1 g), <i>Cnidii Rhizomai> (1 g), <i>Atractylodis Macrocephalae Rhizomai> (1.33 g) and <i>Alismatis Rhizomai> (1.66 g), was studied. In this study, the mechanism involved in the antiplatelet activity of DJS in human platelet suspensions was investigated. Danggijakyaksan did not significantly affect the thromboxane synthetase activity of aspirin-treated platelet microsomes and DJS (20 and 40 µg/mL) significantly inhibited [³H]arachidonic acid (AA) released in collagen-activated platelets but not in unactivated-platelets. Nitric oxide (NO) production in human platelets was measured by a chemiluminesence detection method in this study. Danggijakyaksan did not significantly affect nitrate production in collagen (10 µg/mL)-induced human platelet aggregation. On the other hand, various concentrations of DJS (10, 20, and 40 µg/mL) dose-dependently inhibited [³H]inositol monophosphate (IP) formation stimulated by collagen (10 µg/mL) in [³H]myoinositol-loaded platelets at different incubation times (1, 2, 3, and 5 min). It is concluded that the antiplatelet activity of DJS may possibly be due to the inhibition of phospholipase C activity, leading to reduced phosphoinositide breakdown, followed by the inhibition of thromboxane A2 formation, and then inhibition of [Ca²⁺]i mobilization of platelet aggregation stimulated by agonists.     

Epitope mapping and characterization of a novel CD4-induced human monoclonal antibody capable of neutralizing primary HIV-1 strains

Human immunodeficiency virus (HIV-1) enters target cells by binding its gp120 exterior envelope glycoprotein to CD4 and one of the chemokine receptors, CCR5 or CXCR4. CD4-induced (CD4i) antibodies bind gp120 more efficiently after CD4 binding and block the interaction with the chemokine receptor. Examples of CD4i antibodies are limited, and the prototypes of the CD4i antibodies exhibit only weak neutralizing activity against primary, clinical HIV-1 isolates. Here we report the identification of a novel antibody, E51, that exhibits CD4-induced binding to gp120 and neutralizes primary HIV-1 more efficiently than the prototypic CD4i antibodies. The E51 antibody blocks the interaction of gp120-CD4 complexes with CCR5 and binds to a highly conserved, basic gp120 element composed of the beta 19-strand and surrounding structures. Thus, on primary HIV-1 isolates, this gp120 region, which has been previously implicated in chemokine receptor binding, is accessible to a subset of CD4i antibodies.