颅内细菌性和病毒性感染与外周血CD14关系的研究

为了解CD14在颅内感染患者与正常人外周血中表达水平的临床意义,用流式细胞仪检测20例正常人、30例病毒性脑炎病人、10例化脓性脑炎病人的外周血(PB)中CD14的表达水平,并进行统计学处理及比较.结果,正常人PB 中CD14为3.825±0.760,病脑患者PB中CD14为6.025±2.183,化脑患者PB中CD14为6.227±2.580,正常人与两组患者CD14比较均有显著性差异.CD14在颅内感染患者外周血中表达水平的升高,提示其在颅内感染的发病过程中起作用.     

Role of β2 integrins in glomerular leucocytes in Henoch-Schoenlein purpura nephritis: an age-matched comparative study with immunoglobulin A nephropathy

SUMMARY: A comparative immunohistological study was performed for the glomerular deposition of complements (C1q and C3c), fibrin/fibrinogen‐related antigen (FRA), the expression of intercellular adhesion molecule‐1 (ICAM‐1), and the infiltration of leucocytes bearing β₂ integrins (leucocyte function associated antigen‐1 (LFA‐1), complement receptor 3 (CR3) and complement receptor 4 (CR4)) on renal biopsy specimens from 49 cases with Henoch‐Schoenlein purpura nephritis (HSPN), and 49 age‐matched cases with immunoglobulin A nephropathy (IgAN). the glomerular expression of ICAM‐1 was signifcantly correlated with the glomerular infiltration of leucocyte function associated antigen (LFA)‐1⁺ leucocytes in both diseases, and with that of CR3⁺ leucocytes in HSPN. the expression of ICAM‐1 was closely localized with the infiltration of LFA‐1⁺ leucocytes in the study with double immunostaining. the incidence and intensity of glomerular deposition of FRA were significantly higher in HSPN than in IgAN (P< 0.001), and those of C3c were significantly lower in HSPN than in IgAN (P< 0.001). the glomerular deposition of FRA was significantly correlated with the glomerular infiltration of CR4⁺ leucocytes in HSPN (P<0.05) but not in IgAN. In contrast, the glomerular deposition of C3c was significantly correlated with the glomerular infiltration of CR4⁺ leucocytes in IgAN (P<0.05), but not in HSPN. Studies with double immunostaining revealed a close association of CR4⁺ leucocytes with FRA deposition in HSPN and with C3c deposition in IgAN, respectively. the number of glomerular leucocytes bearing β₂ integrins was significantly correlated with urinary protein at the time of renal biopsy in both diseases. These results suggested the differential roles of β₂ integrins in the induction of glomerular injury in HSPN and IgAN. the ICAM‐1/LFA‐1 interaction may commonly be involved in the glomerular infiltration of leucocytes in both diseases. the ICAM‐1/CR3 interaction may be involved only in HSPN. Complement receptor 4 may function as a fibrin/fibrinogen receptor in HSPN, while CR4 may function as a complement receptor in IgAN.     

sCD14、Ang2、CRP与急诊创伤骨折伴多发伤患者病情转归关系探究

目的 探讨可溶性白细胞分化抗原14（soluble cluster of differentiation antigen 14,sCD14）、血管生成素2（angiopoietin 2,Ang2）、C反应蛋白（C-reactive protein,CRP）与急诊创伤骨折伴多发伤患者病情转归的关系及意义。 方法 选取创伤骨折伴多发伤患者324例,根据患者出院时病情转归情况分为良好组（275例）、不良组（49例）,比较2组一般资料、sCD14、Ang2、CRP水平,应用Pearson分析sCD14、Ang2、CRP与损伤严重程度评分（injury severity score,ISS）关系,采用Cox回归分析急诊创伤骨折伴多发伤患者病情转归的相关影响因素,采用受试者工作特征曲线（receiver operating characteristic,ROC）分析sCD14、Ang2、CRP对病情转归预测价值。 结果 不良组ISS评分高于良好组（P＜0.05）;不良组sCD14、Ang2、CRP高于良好组（P＜0.05）;sCD14（r=0.785）、Ang2（r=0.778）、CRP（r=0.842）与ISS评分呈正相关（P＜0.05）;sCD14、Ang2、CRP均是预后相关独立危险因素（P＜0.05）;sCD14、Ang2、CRP预测病情转归的ROC下面积（area under the curve,AUC）依次为0.813、0.757、0.749;挑选出预测敏感度最高（sCD14）、特异度最高（Ang2）的两个指标进行sCD14+Ang2的联合ROC分析显示,两者联合预测病情转归的AUC为0.935,大于任一单一指标（P＜0.05）。 结论 sCD14、Ang2、CRP与急诊创伤骨折伴多发伤患者病情严重程度及病情转归有关,均可作为预测病情转归的标志物,但联合检测sCD14、Ang2能提高预测可靠性,为临床诊疗及护理提供更准确的参考信息。     

活动性肺结核患者血清Fad D9重组蛋白、sCD14-ST及单核细胞P糖蛋白水平及其临床意义

目的探讨活动性肺结核(APTB)患者血清Fad D9重组蛋白、可溶性白细胞分化抗原14亚型(sCD14-ST)、单核细胞P糖蛋白(Pgp)表达及其临床意义。 方法选取96例APTB患者(APTB组),按照肺部病灶有无空洞和病灶肺叶数分亚组。同期选择体检健康且经γ干扰素释放试验检测阴性者(健康对照组,HC组)及阳性者(结核潜伏感染组,LTBI组)各48例。比较各组血清Fad D9重组蛋白、sCD14-ST、Pgp水平。评估Fad D9重组蛋白、sCD14-ST、Pgp诊断APTB和鉴别APTB、LTBI的价值。 结果Fad D9重组蛋白、sCD14-ST、Pgp水平APTB组＞LTBI组＞HC组,且有空洞者高于无空洞者,病灶肺叶≥3个者高于＜3个者(P＜0.05)。Fad D9重组蛋白、sCD14-ST、Pgp对APTB诊断和APTB、LTBI鉴别有良好效能(P＜0.05)。 结论APTB患者血清Fad D9重组蛋白、sCD14-ST及Pgp明显升高,且与肺部病灶的严重程度有关。     

The sodium channels of the neuroblastoma x glioma 108 CC 15 hybrid cell change their sensitivity for volatile and local anesthetics upon continuous passage

Summary We have studied the ion flux through the sodium channels of low passage number (<50p.) and high passage number (>150p.) neuroblastoma x glioma hybrid cells using [¹⁴C] guanidinium and specific neurotoxins to induce channel opening and closing. The sodium channels of low passage number hybrid cells could be opened by veratridine alone, suggesting the presence of voltage dependent channels in agreement with electrophysiological studies reported in the literature. The sodium channels of the high passage number hybrid cells, however, needed the synergistic action of veratridine and scorpion toxin for activation suggesting that these channels are silent. The [¹⁴C] guanidinium ion flux through the sodium channels of the high passage number hybrid cells was inhibited by significantly lower concentrations of the volatile anesthetics (halothane, isoflurane and enflurane) and the lcoal anesthetics (tetracaine and bupivacaine) than the comparable flux through the sodium channels of the low passage number hybrid cells.Dedicated to Prof. Dr. E. Wecker on the occasion of his 65th birthday.     

Compartmentation of cardiac adenine nucleotides and formation of adenosine

Summary After prelabeling the adenine nucleotides (ATP, ADP, AMP) of isolated perfused guinea pig hearts with either¹⁴C-adenine or¹⁴C-adenosine for 35 min, labeled adenosine, inosine, hypoxanthine and cyclic 35-AMP (cAMP) were continuously released into the cardiac perfusate. Determination of the specific activities (SA) of the adenine nucleotides, cAMP, and their breakdown products (adenosine, inosine, hypoxanthine) in tissue and perfusate revealed: Under steady state conditions the SA of adenosine and cAMP in the perfusate were of the same order of magnitude and proved to be many times higher than the SA of the respective precursor adenine nucleotides. This difference was observed regardless whether adenine or adenosine was used as prelabeling substance. The SA of inosine and hypoxanthine in the perfusate were constantly lower than the SA of adenosine. Cardiac ischemia of 6 min, which resulted in a markedly increased formation of adenosine, led to a pronounced decrease in the SA of adenosine released from the heart.Our findings provide evidence that at least two different adenine nucleotide compartments of the heart serve as precursors for the formation of adenosine and cAMP, one characterized by a high, the other by a lower SA. Under normoxic conditions adenosine and cAMP released into the cardiac perfusate are derived mainly from a nucleotide fraction of high SA, which appears to be rather small. During ischemia a second compartment of much lower SA in addition contributes to the formation of adenosine.A preliminary report of part of this work appeared in Biochemistry and Pharmacology of Myocardial Hypertrophy, Hypoxia and Infarction Vol. 7 of Recent advances in studies on cardiac structure and metabolism. (P. Harris, R. J. Bing, A. Fleckenstein, eds.), pp. 171–175. München: Urban & Schwarzenberg 1976A preliminary report of part of this work appeared in Biochemistry and Pharmacology of Myocardial Hypertrophy, Hypoxia and Infarction Vol. 7 of Recent advances in studies on cardiac structure and metabolism. (P. Harris, R. J. Bing, A. Fleckenstein, eds.), pp. 171–175. München: Urban & Schwarzenberg 1976     

Serum 27E10 antigen: a new potential marker for staging HIV disease.

MRP8 and MRP14 are myeloic related proteins expressed by most circulating and emigrated neutrophils and monocytes. Their composite molecule MRP8/14 (27E10 antigen) was shown to exhibit striking antimicrobial properties. The aim of the present study was to assess the value of MRPs as markers for detection of the different stages of HIV infection (Centres for Disease Control and Prevention, 1993). By employing the ELISA technique we measured serum concentrations of these proteins in samples from 122 HIV patients at the various stages of disease, and the results were compared with those for healthy controls. Serum levels of the heterodimeric molecule 27E10 were significantly increased (P < 0.001) in patients with CDC stages II and III, with the highest levels being in patients with stage III and acute ongoing opportunistic infections. For the single component MRP14, significantly raised levels (P < 0.05) were only found in HIV stage III individuals with acute clinical events. Similar associations were not found for MRP8 alone. Increase was not related to CD4+ cell count. There was a significant correlation between 27E10 antigen serum concentrations and levels of neopterin in patients with HIV stages II and III without acute concurrent illness. Patients being treated with Zidovudine showed no statistically significant variation in levels of 27E10 and its single components MRP8 and MRP14 compared with untreated patients. These findings suggest that elevation of MRP14 levels occurs in HIV+ individuals at later stages post-HIV infection, after the onset of opportunistic infections. 27E10 antigen is concluded to be a potential marker for the different stages of HIV disease.     

Age-dependent cerebral metabolic effects of unilateral nucleus basalis magnocellularis ablation in rats

To investigate the age-dependent functional importance of cholinergic neocortical inputs, and to explore whether cortical cholinergic denervation in aged animals might better model the cerebral metabolic changes of Alzheimer's disease, the effects of unilateral ablation of the nucleus basalis magnocellularis (NBM) on cerebral glucose metabolism were studied in young and aged rats. Regional cerebral metabolic rates for glucose (rCMRglc) were determined, using the [14C]deoxyglucose method, in 48 brain regions of 3- and 24-month old Fischer-344 rats at 3, 7, 14 and 28 days after stereotaxic injection of ibotenate into the right NBM, and in sham-operated animals at 3 and 14 days later. For both ages the peak effect of unilateral NBM ablation occurred 3 days later: in young rats, rCMRglc was significantly reduced (compared to the contralateral side) in all 24 anterior cortical areas examined (mean decline 20%), whereas in aged animals, only 9 of 24 areas showed a significant decline in glucose utilization, and the magnitude of rCMRglc reduction (9%) was smaller. Near complete recovery of rCMRglc occurred by 7 days in young and old rats. We conclude that the basalocortical cholinergic projection plays a smaller role in neocortical function of aged rats, possibly because its tonic activity is reduced. Both young and aged rats undergo cortical metabolic normalization after unilateral NBM ablation; hence the NBM-lesioned aged rat is not a better model of the progressive decline in rCMRglc that occurs in Alzheimer's disease.     

14-3-3σ调控p27抑制Rat1-Akt细胞增殖

目的： 研究腺病毒介导14-3-3·σ（Ad-14-3-3·σ）对Akt过表达Rat1-Akt细胞增殖的影响，并探讨其作用是否通过调控p27而实现。 方法： 通过5-溴-2′脱氧尿嘧啶（BrdU）实验检测Ad-14-3-3·σ对Rat1-Akt细胞增殖的影响，并通过激酶分析法和免疫荧光实验探讨Ad-14-3-3·σ对p27磷酸化水平及其在细胞内定位的影响。 结果： Ad-14-3-3σ转染的细胞BrdU阳性率（45%）低于PBS处理组（100%）或Ad-β-gal转染的对照组细胞（98%）。14-3-3σ可降低磷酸化p27的水平和减少Akt介导的p27在胞浆中的定位。 结论： 转染14-3-3σ基因能抑制Akt过表达细胞株Rat1-Akt增殖，14-3-3σ通过降低Akt激酶磷酸化p27的活性，阻断Akt介导的p27胞浆错位，从而发挥其抑制Rat1-Akt细胞增殖。     

S100A8, S100A9 and the S100A8/A9 heterodimer complex specifically bind to human endothelial cells: identification and characterization of ligands for the myeloid-related proteins S100A9 and S100A8/A9 on human dermal microvascular endothelial cell line-1 cells

The natural ligands of the S100 EF hand proteins S100A8 and A9 [myeloid-related proteins 8 and 14] have long been searched for in order to further the understanding of the role of the S100A8/A9-expressing monocyte subpopulation in progressing inflammatory processes. We demonstrate that S100A8, S100A9 and the S100A8/A9 heterodimeric complex bind to human dermal microvascular endothelial cell line (HMEC)-1 with an increasing binding capacity progressing from S100A8 < or = S100A9 < or = S100A8/A9. Similar results were obtained in the apolipoprotein E knockout mouse model, where preferably recombinant S100A9 but no S100A8 bound to the endothelium of the aorta ascendens. The binding of the S100A8/A9 heterodimer complex to activated HMEC-1 is specific as demonstrated by a dose-responding and satiable binding curve and the competition of FITC-labeled versus unlabeled protein. The protein character of the binding site was proven by treatment with trypsin. S100A8/A9 binding to HMEC-1 is inducible by lipopolysaccharide and tumor necrosis factor-alpha, and in the presence of calcium. A 163-kDa protein was isolated from a cell lysate of activated HMEC-1 cells using an affinity-chromatography protocol. The endothelial cell-associated ligand proteins isolated by the use of the S100A9 monomer and the S100A8/A9 dimer were subjected to mass spectrometry for protein identification. Clearly, alpha(2)-macroglobulin was identified as a binding partner for the S100A9 monomer, whereas no protein could be identified from the database for the ligand of the S100A8/A9 dimer.