宫内注射脂多糖对围产期大鼠肺Toll样受体4信号转导通路的影响

目的 观察宫内注射脂多糖(LPS)对围产期大鼠肺内天然免疫相关的Toll样受体4(TLR4)信号转导通路的影响,探讨天然免疫在宫内感染中的免疫调节能力及对肺发育的影响.方法 将30只孕17d的SD大鼠随机分为LPS组和生理盐水对照组,LPS组宫内注射LPS 10μl(40μg/ml),对照组宫内注射等体积的灭菌生理盐水.分别留取胎龄18、20、22 d(E18、E20、E22)的胎鼠肺组织、胎盘组织标本以及生后1、3、7d(P1、P3、P7)新生鼠肺组织标本,HE染色观察病理改变,RT-PCR技术检测TLR4、髓样分化因子88(MyD88)和白介素1 β(IL-1β)mRNA表达,免疫组织化学技术检测肺组织TLR4、MyD88的表达分布情况.实验数据采用单因素方差分析和q检验进行统计学分析.结果 (1)LPS组孕鼠胎盘组织有大量中性粒细胞浸润,宫内感染模型建立成功;(2)在E18、E20和E22时,LPS组胎鼠肺组织无明显病理学改变,以后逐渐出现改变,于P7时可见肺泡数量减少,肺泡腔变大,间隔变薄,但未见明显结构紊乱;(3)LPS组TLR4、MyD88和IL-1β mRNA水平于E20和E22均高于对照组,差异有统计学意义(P<0.05),且均于E22表达达高峰,后缓慢下降;(4)免疫组织化学结果显示E18时两组肺组织内均未见明显TLR4和MyD88阳性染色,后均逐渐表达增加,且主要在细支气管和肺泡上皮细胞表达.结论 (1)宫内注射LPS可导致胎鼠和早产鼠肺组织TLR4、MyD88表达在一定范围内增加,后逐渐回复正常水平,同时肺组织的病理改变和炎症反应较为温和,推测在围产期胎肺天然免疫系统可以调节LPS诱导的炎症反应强度;(2)该实验在一定程度上证实宫内感染激活的信号转导通路是MyD88依赖性途径.     

脓毒症患儿前炎症因子、Toll样受体信号途径因子和负性调节因子变化初探

目的 探讨Toll样受体(TLRs)信号途径负性调节因子在小儿脓毒症异常炎症反应发病机制中的可能作用.方法 以脓毒症患儿10例、严重全身性感染患儿13例为研究对象,采用实时荧光定量PCR检测TLRs途径传导分子、负性调节因子及前炎症细胞因子mRNA表达;ELISA法检测前炎症细胞因子水平.结果 (1)脓毒症患儿前炎症因子IL-1β、IL-6、TNF-a mRNA表达及蛋白水平明显高于对照组(P<0.01);(2)脓毒症患儿TLRs信号传导途径分子TLR2、TLR4、MyD88、TRAF6、IRAK4、TAB2及TAK1mRNA表达明显增高(P<0.01);(3)脓毒症患儿TLRs负性调节因子SIGIRR、DAP12和FLN29 mRNA表达增高,严重脓毒症组表达低于脓毒症组(P<0.01).结论 TLRs信号传导途径传导分子及负性调节因子异常表达可能是脓毒症全身性炎症反应综合征的原因之一.     

系膜细胞来源的IL-13对系膜细胞促炎性细胞因子基因表达的调控作用

目的：探讨脂多糖(LPS)对体外培养的人肾小球系膜细胞(HMC)表达IL-13的调节作用及IL-13对HMC促炎性细胞因子、趋化因子和促纤维化因子基因表达的影响,以探讨IL-13对肾小球疾病状态下系膜细胞炎症反应的抑制作用。方法：HMC分为实验组和对照组,实验组予以不同浓度的LPS(1 μg/ml,10 μg/ml,100 μg/ml)刺激或用不同浓度的IL-13(1 ng/ml, 10 ng/ml, 100 ng/ml)预处理。应用RT-PCR和ELISA检测系膜细胞IL-13 mRNA 和蛋白表达;应用核酸酶保护法检测系膜细胞TNF-α,IL-1α,IL-1β,MCP-1,IL-8和TGF-β1 mRNA表达。结果：①未予任何刺激的对照组,系膜细胞不表达IL-13 mRNA和蛋白;LPS可呈剂量依赖性的方式促进系膜细胞IL-13 mRNA的表达和蛋白分泌。②正常培养状态下,系膜细胞可组成型表达TNFα,IL-1β,IL8和TGF-β1,而不表达IL-1α和MCP-1 mRNA;LPS刺激后上述炎症因子表达显著上调;IL-13可呈剂量依赖性地抑制LPS诱导的系膜细胞炎症性细胞因子的基因表达。结论：IL-13是系膜细胞的自分泌因子;IL-13可抑制LPS诱导的系膜细胞促炎性细胞因子、趋化因子和促纤维化因子的表达,提示自分泌和旁分泌的IL-13对于肾小球疾病状态下肾脏系膜细胞的炎症反应具有抑制作用。     

Peroxisome proliferator‐activated receptor γ 2 mutation may cause a subset of ulcerative colitis

Aim: Previous studies suggest the homeostasis between acquisition of tolerance to the indigenous microflora and protective immune responses appears to be disrupted in inflammatory bowel disease (IBD). Some experimental studies indicate peroxisome proliferator‐activated receptor γ (PPARγ) has been implicated as a regulator of intestinal inflammatory responses. In addition, the toll‐like receptor (TLR)‐4 can regulate expression of PPARγ in colonic epithelial cells. We attempted to demonstrate whether the functional imbalance between TLRs and PPARγ could lead to the onset and some polymorphisms of those genes could contribute to susceptibility to IBD. Methods: RT‐PCR analysis were performed to detect TLR4 and PPARγ mRNA associated with those of P65 of NFκB, TNFα, MyD88, NOD2/CARD15, TLR‐2,5,9, in the diseased colonic mucosa in ulcerative colitis (UC; n = 13) and Crohn's disease (CD; n = 7) compared with normal controls (n = 18). Consequently, we genotyped UC (n = 29) and CD (n = 10) compared with normal controls (n = 134) for the prevalence of suspicious mutations. Results: In a subset of UC patients who were revealed to carry PPARγ Pro12Ala mutation later, impaired expression of normal PPARγ mRNA was noted in the diseased mucosa accompanied with upregulations of MyD88 TLR‐4, 5, 9, P65 and TNFα in mRNA levels. The prevalence of PPARγ Pro12Ala mutation was more frequently found in UC patients compared with CD patients and normal controls (P < 0.05). Conclusions: These findings suggested that imbalances between TLRs and PPARγ in response to luminal bacteria could lead to colonic inflammation in some UC patients. Alternative explanations will be needed for the onset of the rest of UC and CD.     

Young patients with both type 1 diabetes mellitus and asthma have a unique IL-12 and IL-18 secretory pattern

Rachmiel M, Bloch O, Shaul AA, Ben‐Yehudah G, Bistritzer Z, Weintrob N, Ofan R, Rapoport MJ. Young patients with both type 1 diabetes mellitus and asthma have a unique IL‐12 and IL‐18 secretory pattern. Background: The expression of the regulatory cytokines interleukin (IL)‐12 and IL‐18 in patients with both Th1‐ and Th2‐mediated diseases, type 1 diabetes mellitus (T1DM) and asthma, is unknown. Objective: To investigate the in vivo and in vitro IL‐12 and IL‐18 secretion patterns in patients with both T1DM and asthma. Methods: Peripheral blood mononuclear cells (PBMC) were collected from 44 patients. Mean age 19.4 ± 4.7 yr (10.5–28 yr), divided into four paired groups: T1DM and asthma, asthma only, T1DM only, and healthy controls. T‐cell proliferative response was assessed. IL‐12 and IL‐18 serum levels and expression by PBMC following in vitro stimulation by lipopolysaccharide (LPS) were determined by enzyme‐linked immunosorbent assay (ELISA). Results: Patients with T1DM and asthma had higher serum levels of both IL‐12 and IL‐18 compared to controls: 146.2 ± 69.2 and 109.7 ± 34.6 pg/mL, p = 0.038 and 436.1 ± 117.9, 320.2 ± 99.1 pg/mL, p = 0.028, respectively. Stimulated IL‐12 secretion was significantly lower in these patients compared to those with one disease only: 809 ± 426.4, 2111.6 ± 2214.3, 3188.1 ± 2692.9 pg/mL and after 48 h: 956.3 ± 489.3, 2429.8 ± 2394.6, 3874.5 ± 2820.3 pg/mL, respectively, p < 0.03 for all. The IL‐18/IL‐12 serum ratio was also significantly higher in patients with both diseases compared to those with asthma only, p = 0.017. Conclusion: Patients with both T1DM and asthma display a different pattern of IL‐12 and IL‐18 expression compared to patients with one disease only and controls.     

Prevalence of microalbuminuria in young patients with type 1 and type 2 diabetes mellitus

This study was designed to determine the prevalence of microalbuminuria and the associated risk factors in patients with childhood-onset diabetes mellitus (DM). One hundred and sixty-three patients (141 with type 1 DM [DM1] and 22 with type 2 DM [DM21), aged 8 to 28 years, were evaluated for albumin excretion rate and HbA(1c). The mean duration of DM was 8.1 +/- 3.4 and 5.5 +/- 3.9 years in DM1 and DM2, respectively. Persistent microalbuminuria and macroalbuminuria were observed in 11.3% and 2.8% of patients with DM1, and 18.2% and 4.5% in patients with DM2, respectively. In DM1, the duration of DM, age of onset, and HbA(1c) levels were significant predictors of microalbuminuria. Our observations suggest that screening for microalbuminuria should be started from early adolescence in patients with DM1 and DM2.     

High tissue factor in lungs and plasma associates with respiratory morbidity in preterm infants

Aim: In preterm infants, inflammation and intra‐alveolar fibrin formation characterize respiratory distress syndrome (RDS). Tissue factor (TF) is a link between inflammation and coagulation pathways. We investigated the relationship between TF and cytokines in preterm infants to gain information of the role of TF in the inflammatory response. Methods: We measured TF in plasma and in tracheal aspirates and analysed TF on monocytes by flow cytometry and 13 cytokines from plasma, in 56 preterm infants (birthweight 600–1500 g) during their first week. Results: Plasma TF increased and peaked on day 3 and correlated with both RDS and inversely with paO2/FIO2. On day 1, TF in tracheal aspirates was 10‐fold higher than in plasma and correlated with plasma TF (4888 vs. 506 pg/mL, R = 0.692, p = 0.013, n = 12). Of main pro‐inflammatory cytokines, plasma TF correlated post‐natally with IL‐8 and IL‐6 but not with IL‐1 or TNF‐α. Conclusions: Respiratory morbidity associates with high TF in lungs and plasma. In sick newborn infants, upregulation of TF may be mediated by IL‐6 and IL‐8. High TF and pro‐inflammatory cytokines may together participate in the pathogenesis of pulmonary and extrapulmonary injury in preterm infants through pro‐inflammatory mechanisms.     

Surfactant lavage decreases systemic interleukin‐1β production in meconium aspiration syndrome

Background: Surfactant lavage has been used to remove meconium debris in meconium aspiration syndrome (MAS), but the influence of surfactant lavage on pro‐inflammatory cytokines and cellular apoptosis is unclear. The aim of this study was to investigate the response of pro‐inflammatory cytokine and the influence on alveolar cellular apoptosis using therapeutic bronchoalveolar lavage with diluted surfactant to treat MAS. Methods: Twelve newborn piglets were anesthetized, intubated via tracheostomy, and artificially ventilated. MAS was induced by intratracheal instillation of 3–5 mL/kg of 20% human meconium. The piglets were then randomly assigned to a surfactant lavage group (n= 6) or a control group (n= 6). Piglets in the lavage group received bronchoalveolar lavage with 30 mL/kg diluted surfactant (5 mg/mL) in two aliquots. Cardiopulmonary parameters were monitored continuously. Serum was obtained hourly to measure concentrations of pro‐inflammatory cytokines, including interleukin (IL)‐Iβ, IL‐6, and tumor necrosis factor α. Lung tissue was histologically examined after experiments, and terminal deoxynucleotidyl transferase‐mediated nick‐end labeling assay for apoptotic cell death was also performed. Results: The animals in the lavage group displayed significantly better gas exchange and lower serum concentrations of IL‐1β than the animals in the control group (P < 0.05). The number of apoptotic cells in lung tissues was significantly lower in the lavage group than the control group, and also in the nondependent than the dependent site. Conclusion: Therapeutic surfactant lavage improves oxygenation, decreases production of systemic pro‐inflammatory cytokine IL‐1β, and alleviates the severity of lung cell apoptosis in newborn piglets with experimentally‐induced MAS.     

Toll样受体信号途径活化在川崎病免疫发病机制中的作用

目的探讨Toll样受体(TLR s)信号途径在川崎病(KD)免疫发病机制中的作用。方法急性期KD患儿16例,正常同年龄对照组16例。KD患儿分别于静脉丙种球蛋白(IVIG)治疗前后直接取血备检,未加任何体外丝裂原刺激培养。采用逆转录-聚合酶链反应(RT-PCR)及荧光定量PCR检测外周血单个核细胞TLR s 1~10,MD-2,MyD88,IL-1,βIL-6及IL-8 mRNA的表达;流式细胞术分别检测单核/巨噬细胞表面TLR s 2、4及共刺激分子CD80、CD86的表达。结果(1)急性期KD患儿TLR4 mRNA及蛋白表达均显著高于正常同年龄对照组[Real-tim e PCR(325.22±50.34)vs.(2.20±0.23),P<0.01);流式细胞术检测(15.96±5.94)%vs.(3.21±0.62)%,P<0.01],其他TLR表达无明显改变;(2)TLR4传导途径相关因子MD-2及MyD88亦明显增高(P<0.01),IVIG治疗后有不同程度下降;(3)急性期KD患儿组单核/巨噬细胞表面共刺激分子及前炎症细胞因子表达亦明显增高(P<0.01)。结论急性期KD患儿TLR4及其相关分子MD-2、MyD88异常增高,提示TLR4异常活化可能是KD免疫功能紊乱的始动因素之一。     

Microalbuminuria in adolescents with insulin-dependent diabetes mellitus

Two hundred ten adolescents aged 12 to 18 years with insulin-dependent diabetes mellitus were screened for microalbuminuria (albumin excretion rate of 15 to 300 micrograms/min). Sixteen (7.6%) showed persistent microalbuminuria (mean albumin excretion rate of 70.9 +/- 56.2 micrograms/min). There were no significant differences between those with and without microalbuminuria with respect to age, sex, disease duration, and blood pressure over the previous 9 months and hemoglobin A1c level measured over the preceding 3 years. Within the group with microalbuminuria, there was no correlation between albumin excretion rate and blood pressure. However, there was a significant positive correlation between log albumin excretion rate and mean hemoglobin A1c values measured over the preceding 3 years. Our findings suggest that when microalbuminuria has developed, poorer metabolic control is associated with a higher albumin excretion rate. An actual rise in systemic blood pressure may not always precede the development of microalbuminuria.     

Microalbuminuria and abnormal ambulatory blood pressure in adolescents with type 2 diabetes mellitus

OBJECTIVE: To determine whether risk factors for cardiovascular disease and diabetic nephropathy, as evidenced by abnormalities of ambulatory blood pressure (ABP), dyslipidemia, and microalbuminuria (MA), are present in adolescents with type 2 diabetes mellitus (T2DM). STUDY DESIGN: We enrolled 26 minority adolescents recently diagnosed with T2DM and 13 obese control subjects without diabetes mellitus. ABP monitoring was performed, and a 24-hour urine, a fasting lipid profile, blood urea nitrogen, creatinine, homocysteine, and hemoglobin A 1 c levels were obtained. The patients with T2DM underwent echocardiograms. RESULTS: Forty percent of the patients with T2DM had MA (> or = 30 mg of microalbumin/day), compared with none of the control subjects ( P < .05). There were no significant differences between patients with T2DM who had MA and patients with T2DM who didn't have MA in demographics, characteristics, casual BP, echocardiographic findings, and hemoglobin A 1 c levels. Average daytime systolic BP was greater in patients with T2DM with MA than patients without MA (129 versus 121 mm Hg, P = .03) and compared with the control subjects (113 mm Hg, P = .01). Patients with MA had an average daytime systolic BP load that was higher than patients without MA (37.1 versus 5.1%, P = .008) and compared with the control subjects (2.6%, P < .001). CONCLUSION: As in adults, adolescents with T2DM exhibit abnormalities of ABP, dyslipidemia, and microalbuminuria.     

Enhanced expression of cytokines/chemokines in cerebrospinal fluids in mumps meningitis in children

Background: The mumps virus is frequently the causative agent in aseptic meningitis and mumps has still prevailed in Japan. We compared data obtained from patients with mumps meningitis and patients with aseptic meningitis caused by other viruses in order to identify mumps meningitis‐specific cytokine/chemokine alterations in cerebrospinal fluide (CSF). Methods: We elucidated the cytokine/chemokine network based on the cytokine/chemokine profiles in CSF from children with mumps meningitis and meningitis due to other viral infections using multiplex cytokine measurement. Seventeen cytokines/chemokines, namely interleukin (IL)‐1β, IL‐2, IL‐4, IL‐5, IL‐6, IL‐7, IL‐8, IL‐10, IL‐12 (p70), IL‐13, IL‐17, interferon (IFN)‐γ, tumor necrosis factor (TNF)‐α, granulocyte colony‐stimulating factor (G‐CSF), granulocyte monocyte colony‐stimulating factor (GM‐CSF), monocyte chemoattractant protein‐1 (MCP‐1) and macrophage inflammatory protein‐1β (MIP‐1β), were measured simultaneously in CSF supernatants from eight children with mumps meningitis, 11 children with other types of viral meningitis and eight children with fever without neurological complications such as convulsion. Results: We found that IL‐8, IL‐10, IL‐12, IL‐13 and IFN‐γ showed a statistically significant increase in CSF from mumps meningitis when compared to other types of viral meningitis and fever without neurological complications. Conclusion: Mumps meningitis may induce a distinct immunological response when compared with other types of viral meningitis.     

Different cytokine signatures in children with localized and invasive adenovirus infection after stem cell transplantation

Haveman LM, de Jager W, van Loon AM, Claas ECJ, Prakken BJ, Bierings M. Different cytokine signatures in children with localized and invasive adenovirus infection after stem cell transplantation. Pediatr Transplantation 2010: 14:520–528. © 2010 John Wiley & Sons A/S. Abstract HAdV infection is a dangerous complication after pediatric SCT. In this study, we aimed at determining the cytokine profile in plasma samples in case of HAdV infection after SCT to gain more knowledge about the HAdV‐specific immune response. In this prospective study, 47 pediatric SCT recipients were included in three yr. By using particle‐based MIA, 17 different cytokines were analyzed in 41 plasma samples of patients with a localized HAdV infection (presence of HAdV in feces, urine or throat detected by culture) and patients with invasive HAdV infection (HAdV viremia in blood, detected by PCR). In patients with invasive HAdV infection, but not in patients with localized HAdV infection, the pro‐inflammatory cytokines IL1β, IL6, IL8, IL12, IFNγ, TNFα, and also IL17, MIP1α, OSM, and IP10 were produced. The simultaneous release of the cytokines IL1β, IL17, IL18, OSM, MIP1α, and IP10 was related to invasive HAdV infections. We also show that cytokine signatures can be helpful to differentiate invasive HAdV infection from GvHD and EBV infections. In conclusion, after SCT, children with invasive HAdV infection have a different cytokine profile compared with patients with a localized HAdV infection.     

Elevated tricuspid regurgitation velocity in congenital hemolytic anemias: Prevalence and laboratory correlates

Elevated tricuspid valve regurgitation jet velocity (TRV ≥ 2.5 m/s) is associated with mortality among adults with sickle cell disease (SCD), but correlative biomarkers are not studied according to treatment exposure or genotypes. To investigate the associations between biomarkers and TRV elevation, we examined the relationship between TRV and hemolytic, inflammatory, and cardiac biomarkers, stratified by disease‐modifying treatments and SCD genotype. In total, 294 participants with SCD (mean age, 11.0 ± 3.7 years) and 49 hereditary spherocytosis (HS; mean age, 22.9 ± 19.75 years) were included for comparison and enrolled. TRV was elevated in 30.7% of children with SCD overall: 18.8% in HbSC/HbSβ⁺‐thalassemia, 28.9% in untreated HbSS/HbSβ⁰‐thalassemia, 34.2% in HbSS/HbSβ⁰‐thalassemia hydroxyurea‐treated, and 57% in HbSS/HbSβ⁰‐thalassemia chronic transfusion treated. TRV was elevated in 10.7% and 27.8% in HS children and adults, respectively. In children with SCD, elevated TRV was correlated with hemoglobin (odds ratio [OR] = 0.78, P = 0.004), lactate dehydrogenase (LDH; OR = 2.52, P = 0.005), and N‐terminal pro‐brain natriuretic peptide (NT‐pro BNP; OR = 1.003, P = 0.004). In multivariable logistic regression, adjusting for genotype, sex, hemolytic index, and treatment, hemoglobin concentration remained the only significant variable associated with elevated TRV in untreated HbSS/HbSβ⁰‐thalassemia participants. TRV was not associated with inflammatory markers, other markers of hemolysis, or NT‐pro BNP in untreated HbSS/HbSβ⁰‐thalassemia. Neither hemoglobin nor LDH was associated with TRV in HbSC/HbSβ⁺‐thalassemia. These results suggest that elevated TRV is influenced by the degree of anemia, possibly reflecting sickling as part of the disease pathophysiology. Prospective studies should monitor hemoglobin concentration as children with SCD age into adulthood, prompting initiation of TRV screening and monitoring.     

Prevalence of retinopathy and microalbuminuria in pediatric type 2 diabetes mellitus

BACKGROUND: There has been a dramatic rise of type 2 diabetes mellitus (DM2) in the US pediatric population, yet the presence of retinopathy and microalbuminuria in this cohort has not yet been described. AIM: To assess the presence of retinopathy and microalbuminuria in a cohort of predominantly minority adolescents (African American and Caribbean Hispanic) with DM2. Patients and Methods: Forty pediatric patients with DM2 were examined between July, 2001 and June, 2003 for the presence of retinopathy. Data were also collected regarding microalbuminuria, body mass index, HbA1c, and family history of DM. RESULTS: Of the 40 patients examined, one patient (2.5%) had retinopathy, whereas nine (27.3%) had microalbuminuria. CONCLUSIONS: In young predominantly minority adolescents with DM2, retinopathy is rare, while microalbuminuria is common.     

Toll-like receptors and agonist responses in the developing fetal sheep lung

Toll-like receptors (TLRs) are pattern recognition molecules that initiate innate immune responses. Intra-amniotic exposure of fetal sheep to pro-inflammatory stimuli causes pulmonary inflammation and induced lung maturation. We examined TLR ontogeny and fetal lung responsiveness to three different TLR agonists. We cloned ovine TLRs 2, 3, and 4 and found 83-88% homology between these ovine and human TLRs. Lung TLR2 and 4 mRNAs increased throughout late gestation to 50% of adult level in the term newborn lamb. Doses of 10 mg of PAMCysK4 (TLR2 agonist), poly I:C dsRNA (TLR3 agonist), or E. coli O55:B5 lipopoysaccharide (LPS) (TLR4 agonist) were given by intra-amniotic injection 2 d or 7 d before operative delivery of preterm lambs at 123 d (n = 4-7/group). The TLR4 agonist induced lung inflammation and maturation, whereas the TLR2 agonist gave less consistent responses. Intra-amniotic LPS increased TLR2 mRNA expression primarily in the inflammatory cells and TLR4 mRNA diffusely in multiple cell types. The TLR3 agonist had no effects, and TLR3 mRNA in the fetal lung did not change after LPS exposure. We conclude that TLR2 and TLR4 mRNAs increase through gestation and expression of TLR2 and TLR4 are induced by LPS in the fetal sheep lung.     

Serum and urinary cytokine homeostasis and renal tubular function in children with type 1 diabetes mellitus

AIM: To explore the relationships between tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL-6) and urinary N-acetyl-beta-D-glucosaminidase (NAG) and the function of renal proximal tubules in children with type 1 diabetes mellitus (DM1). METHODS: Fifty-six children with DM1 and 35 healthy controls were analyzed. We measured NAG (A and B isoforms) in urine as well as serum TNFalpha and urinary IL-6. RESULTS: The children with DM1 with microalbuminuria (group A) had significantly higher urinary IL-6 and serum TNFa than the children without microalbuminuria (group B). The diabetic patients with no sign of nephropathy showed significantly higher TNFalpha and NAG and its A and B isoforms in urine compared to the healthy group. Additionally, groups A and B both showed a positive significant correlation between serum TNFalpha and urinary isoform B. CONCLUSIONS: From our pilot results it appears that TNFalpha might be a sensitive marker of damage to the renal proximal tubules occurring prior to microalbuminuria. Conversely, the increase in NAG and its isoform B activity in patients with no clinical sign of diabetic nephropathy may indicate the onset of microalbuminuria.     

B7同源体3经Toll样受体2依赖性机制增强肺炎链球菌脑膜炎小鼠的炎性反应和病理损伤

目的 探讨B7同源体3(B7 - H3)对肺炎链球菌(SP)脑膜炎大鼠的炎性反应和血脑屏障完整性的影响及其机制.方法 经小鼠侧脑室穿刺注入SP悬液,制备脑膜炎模型.野生型鼠分5组,分别为PBS组、SP组、SP+B7 -H3组、SP+同型单抗组、SP+B7 - H3阻断性单抗组.Toll样受体2基因剔除(TLR2-KO)鼠分3组:PBS组、SP组、SP+ B7 - H3蛋白组.术后6h、18h、30 h,麻醉其下眼眶采血法收集血清,取脑组织,制备脑组织匀浆.ELISA检测其血清TNF-α、IL-6、IL-1β和单核细胞趋化因子蛋白-1(MCP-1)水平以及脑组织匀浆中血清蛋白( Albumin)和IgG水平.结果 1.ELISA检测野生型鼠血清SP组、SP+ B7 - H3组注射18 h后TNF-α、IL-6和MCP-1的表达及30 h后TNF-α、IL-6、IL-1β和MCP-1的表达均较PBS组显著升高(Pa＜0.05);SP+同型单抗组与SP组比较,各时间点各炎性因子的表达差异均无统计学意义(Pa＞0.05);SP +B7- H3组注射18h后MCP-1的表达及30 h后TNF-α和IL-6表达均显著高于SP组[(42.010±3.883) ng·L-1vs(29.620±3.830) ng· L-1;(37.550±3.232)ng· L-1vs (24.570±2.377) ng·L-1;(66.160±5.766) ng·L-1vs (48.630±4.418) ng·L-1,Pa＜0.05];SP+ B7 - H3阻断性单抗组注射30h后,TNF-α和IL-6的表达均显著低于SP组[(18.680±1.798) ng·L-1vs(24.570±2.377) ng·L-1;( 37.180±3.150) ng·L- 1vs (48.630±4.418) ng· L-1,Pa＜0.05].2.ELISA检测脑组织匀浆:SP组、SP+ B7 - H3组注射18h、30h后Albumin、IgG的表达较PBS组均显著升高(Pa＜0.05);SP+同型单抗组与SP组比较,各时间点Albumin、IgG的表达差异均无统计学意义(Pa＞0.05);SP+ B7 - H3组注射18 h、30 h后脑组织匀浆Albumin的表达及30 h后脑组织匀浆IgG的表达均显著高于SP组[(59.090±4.184) μg· g-1vs ( 35.450±4.256) μg·g-1;( 59.890±4.701)μg·g-1 vs (43.790±3.508) μg·g-1;(36.220±2.775)μg·g-1 vs(25.440±2.620)μg·g-1,Pa＜0.05].3.SP+ B7 - H3阻断性单抗组注射后18 h、30 h Albumin的表达及30 h IgG的表达显著低于SP组[(20.590±1.720) μg·g-1vs(35.450±4.256) μg·g-1;(28.650±3.063) μg· g-1vs(43.790±3.508)μg·g-1;(17.380±1.595) μg·g-1vs(25.440±2.620) μg·g-1,Pa＜0.05].3.TLR2-KO鼠血清和脑组织匀浆的ELISA检测显示:SP +B7-H3组较SP组各监测指标的表达在任何时间点的差异均无统计学意义(Pa＞0.05).结论 B7 - H3通过TLR2依赖性机制促进SP诱导的脑膜炎小鼠的炎性反应和血脑屏障的破坏.     

Influenza A H1N1 pdm09‐associated myocarditis during zanamivir therapy

A 9‐year‐old girl developed influenza A H1N1 pdm09‐associated myocarditis and pericarditis 2 days after starting zanamivir therapy. The virus was detected in the respiratory tract but not in the serum or pericardial effusion. The virus sampled from the respiratory tract had normal susceptibility to neuraminidase inhibitors. Although no differences in interferon‐γ, interleukin (IL)‐1β, and tumor necrosis factor‐α were observed between the plasma and pericardial effusion, some inflammatory cytokines or chemokines (IL‐6 and IL‐8) and vascular endothelial growth factor were remarkably elevated in the pericardial effusion compared with the plasma. This suggested that the influenza virus, after infecting the respiratory tract, affected the myocardium, causing myocarditis to gradually develop, which might have been followed by an autoreactive pericarditis causing increased pericardial effusion. Therefore, influenza‐associated myocarditis should be considered when influenza patients have respiratory and cardiac involvement, even during treatment with a neuraminidase inhibitor.