肾移植术后普乐可复应用的临床研究

目的研究肾移植术后应用普乐可复免疫抑制治疗的临床疗效和毒副作用。方法随机将62例肾移植受者分成两组：（1）FK506组（n=24）：应用FK506＋骁悉（MMF）＋强的松（Pred）治疗方案;（2）环孢素A（CsA）组（n=38）：应用CsA＋MMF＋Pred治疗方案。结果随访24个月,FK506组有2例（8.3%）、CsA组有9例（23.7%）发生急性排斥反应（P〈0.05）;两组肾功能恢复指标血清肌酐值无明显差异（P〉0.05）;FK506组和CsA组分别有3例（12.5%）和14例（36.8%）发生肝功能损害（P〈0.05）;分别有13例（54.2%）和22例（57.9%）血压升高（P〉0.05）;分别有4例（16.67%）和6例（15.79%）血糖升高（P〉0.05）。结论 FK506具有强效的免疫抑制作用,对于发生急性排斥反应、严重肝功能损害等肾移植术后患者由CsA切换成FK506治疗是安全有效的;应用时须根据血药浓度调整剂量,以减少不良反应的发生。     

免疫抑制剂FK506对大鼠移植性肝癌肿瘤生长和脾转移的影响

目的 观察在大鼠移植性肝癌模型中不同FK506剂量下肿瘤的大小和侵润以及脾脏转移情况,用以评价FK506对肝癌细胞增殖和转移的情况.方法在Wistar大鼠肝左叶种植walker-256细胞株成瘤的癌组织块以构建转移性肝癌模型,54只大鼠随机分为FK506低剂量组(0.5 mg/kg*d),正常剂量组(1.0 mg/kg*d)和空白对照组,测量肿瘤大小,通过病理组织学观察7、14、21 d脾脏转移发生率和中位数.结果与空白组相比,第14、21天两个剂量组肝癌体积明显增大(P<0.05);同时也出现了脾转移,第14天FK506各组转移中位数均不同.根据病理组织学,各组脾转移的发生率没有明显性差异.结论用药早期,脾脏还存在免疫功能,免疫抑制剂不能完全压制抗肿瘤效应.低剂量FK506也许有利于减少癌症脾转移,仍需进一步的探讨FK506低剂量长期使用在肝癌复发和转移的作用.     

Pharmacological preconditioning of primary rat cardiac myocytes by FK506

Pre-treatment with the immunosuppressant FK506 is shown to protect primary cardiocytes against a subsequent severe thermal or ischaemic stress. This effect is not observed with the related compounds cyclosporin A or rapamycin. It does not involve induction of the FK506 binding, heat inducible protein hsp56 or of the other heat shock proteins. In addition over-expression of hsp56 does not protect cardiac cells from severe stress in contrast to our previous results with hsp70 and hsp90. These results suggest the FK506 is acting via a novel mechanism to protect cardiac cells against cellular ischaemia which may not be related to its immunosuppressant action.     

Effect of a new cholecystokinin antagonist (FK 480) on gene expression of cholecystokinin and secretin in rat intestine

The effects of a new cholecystokinin (CCK) antagonist (FK 480; 0.1 mg/kg per day given by intragastric administration to rats for 3 days) on the expression of the CCK and secretin genes, plasma CCK immunoreactivity, and CCK content in the intestinal mucosa were examined. FK 480 increased the level of CCK mRNA in the intestine to 1.7 times the level in control rats, but did not affect the level of secretin mRNA. It did not increase plasma CCK immunoreactivity or CCK content in the intestinal mucosa. These results suggest that the ingested FK 480 directly increased CCK mRNA level in the intestine and produced a dissociation between the synthesis and release of CCK.     

Resistance to activated protein C (APCR) in children with venous or arterial thromboembolism

Resistance to activated protein C (APCR), in the majority of cases due to the point mutation Arg 506 Gln of the factor V gene, has emerged as the most important hereditary cause of venous thromboembolism. Using an activated thromboplastin time (aPTT) based method in the presence of APC together with a DNA technique based on the polymerase chain reaction, we investigated 37 children with venous (V: n = 19) or arterial (A: n = 18) thromboembolism and 196 age-matched healthy controls for the presence of this mutation. In the control group 10 children were detected to be heterozygous for the factor V Leiden mutation, indicating a prevalence of 5.1%. 10/19 children (52%) with venous thrombosis and 7/18 (38%) patients with arterial thromboembolism showed the common factor V gene mutation. Additional inherited coagulation disorders were found in 1/10 (V: 10%) and 2/7 (A: 28%) APC-resistant patients. Inherited coagulation disorders without APCR were diagnosed in 3/9 (V: 33%) and 2/11 (A: 18%) children. Furthermore, we diagnosed exogenous risk factors in 6/10 (V: 60%) and 2/7 (A: 28%) children with thrombosis and APCR. These data are evidence that APCR combined with exogenous reasons may play an important role in the early manifestation of thromboembolism during infancy and childhood.     

他克莫司对转化生长因子-β诱导的肾小管上皮细胞转分化的抑制作用及对核因子-κB活性的影响

目的:探讨他克莫司(FK506)对转化生长因子-β(TGF-β)诱导的肾小管上皮细胞转分化的作用,及其与核因子-κB(NF-κB)活性变化的关系.方法:以人类肾脏近曲小管上皮细胞株(Human kidney cell,HKC)为研究对象,分为以下4组:①阴性对照组;②TGF-β(8 ng/ml)组:③FK506(0.1、1、10、50 ng/ml)组:④TGF-β(8 ng/ml) FK506(0.1、1、10、50 ng/ml)组.应用形态学、间接免疫荧光法,免疫组化技术和酶联免疫吸附法观察FK506对TGF-β诱导的HKC细胞转分化的作用、细胞培养上清液纤连蛋白、Ⅰ型胶原的变化及FK506对HKC细胞NF-κB活性的影响.结果:FK506(1,10,50 ng/ml) TGF-β组比TGF-β组诱导的HKC细胞E-钙粘蛋白和细胞角蛋白表达有所增强,波形蛋白和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达比TGF-β组减弱(P＜0.05),同时HKC细胞NF-κB的活性比TGF-β组减弱(P＜0.05),培养上清液纤连蛋白及Ⅰ型胶原的浓度比TGF-β组降低(P＜0.05);FK506(1、10、50 ng/ml)使基础状态下HKC细胞NF-κB的活性明显下降(P＜0.05)和上清液中纤连蛋白及Ⅰ型胶原的水平明显降低(P＜0.05).结论:①FK506剂量依赖性地抑制TGF-β诱导的肾小管上皮细胞转分化和纤连蛋白及Ⅰ型胶原的形成及基础状态下和TGF-β诱导的HKC细胞NF-κB的表达.②FK506抑制TGF-β诱导的HKC细胞转分化很可能与NF-κB的表达下调有关,需要进一步研究.     

Prognosis of left anterior hemiblock in acute myocardial infarction

Four of 32 patients with left anterior hemiblock and an acute anterior wall myocardial infarction died. Left anterior hemiblock was present on admission in 24 patients, and subsequently appeared in 8. Of the 28 survivors, 21 are still alive an average of 2.8 years after the acute myocardial infarction.     

他克莫司和雷帕霉素对肝癌肝移植患者Foxp3+ Treg的影响及防治排斥反应的研究

目的 探讨免疫抑制剂他克莫司和雷帕霉素对肝癌肝移植受者Foxp3+ Treg产生的影响及其防治排斥反应的疗效.方法 自移植后第2个月到第12个月,每月采血,采用实时荧光定量PCR法检测他克莫司组和雷帕霉素组肝癌肝移植受者新鲜外周血单个核细胞中Foxp3 mRNA的表达水平,通过同期术后观察和实验室检查,比较两组受者间Foxp3 mRNA表达水平和急性排斥反应发生率的差异.结果 他克莫司组受者的外周血单个核细胞中Foxp3 mRNA表达水平(0.1032±0.0943)明显低于雷帕霉素组受者(1.2136±0.6738),差异有统计学意义(t=5.1610,P＜0.01);雷帕霉素组患者比同期他克莫司组受者术后急性排斥反应发生率明显减低,差异有统计学意义(x2=2.2222,P＜0.05).结论 他克莫司抑制了肝癌肝移植术后免疫耐受的诱导,而雷帕霉素可能参与了免疫耐受的诱导和维持;雷帕霉素对肝癌肝移植受者防治排斥反应的效果更好.     

EFFECTS OF NITRIC OXIDE SYNTHESIS INHIBITION ON FK506-INDUCED NEPHROTOXICITY IN RATS

This study was designed to evaluate the role of nitric oxide (NO) in FK506-induced nephrotoxicity by administering an inhibitor of NO synthesis, Nω-nitro-L-arginine methyl ester (L-NAME) to rats treated with FK506. After one week of treatment with FK506 (3.2 mg/kg/day, intramuscularly) and/or L-NAME (5 mg/100 mL of L-NAME in the drinking water), the arterial pressure, urinary NOx, and parameters for renal function were measured, and histological analysis of the kidney was made. In the L-NAME without FK506 group, L-NAME administration effectively inhibited urinary NOx excretion and increased mean arterial pressure (MAP) without any change in renal function. In the FK506 without L-NAME group, FK506 treatment showed increase in urinary NOx excretion and mild renal dysfunction. In the FK506 with L-NAME group, urinary NOx excretion was decreased by L-NAME administration and renal function was significantly worsened than FK506 without L-NAME group. The plasma creatinine, BUN and urinary N-acetyl-β-D-glucosaminidase increased 2-, 3-, and 3-fold, respectively and the creatinine clearance was reduced by 50% as compared with that in the FK506 without L-NAME group. Histological analysis revealed severe interstitial fibrosis and tubular atrophy in the FK506 + L-NAME treatment group. Thus, results suggest that NO synthesis is enhanced in the kidney during FK506-induced nephrotoxicity and that NO synthesis inhibition aggravates FK506-induced nephrotoxicity. NO may play a protective role attributable to the balance of vasoactive substances in FK506-induced nephrotoxicity.     

Proteomics reveals potential non-neuronal cholinergic receptor-effectors in endothelial cells

Aim:

The non-neuronal acetylcholine system (NNAS) in endothelial cells participates in modulating endothelial function, vascular tone, angiogenesis and inflammation, thus plays a critical role in cardiovascular diseases. In this study, we used a proteomic approach to study potential downstream receptor-effectors of NNAS that were involved in regulating cellular function in endothelial cells.

Methods:

Human umbilical vein endothelial cells were incubated in the presence of acetylcholine, oxotremorine, pilocarpine or nicotine at the concentration of 10 μmol/L for 12 h, and the expressed proteins in the cells were separated and identified with two-dimensional electrophoresis (2-DE) and LC-MS. The protein spots with the largest changes were identified by LC-MS. Biowork software was used for database search of the peptide mass fingerprints.

Results:

Over 1200 polypeptides were reproducibly detected in 2-DE with a pH range of 3–10. Acetylcholine, oxotremorine, pilocarpine and nicotine treatment caused 16, 9, 8 and 9 protein spots, respectively, expressed differentially. Four protein spots were identified as destrin, FK506 binding protein 1A (FKBP1A), macrophage migration inhibitory factor (MIF) and profilin-1. Western blotting analyses showed that treatment of the cells with cholinergic agonists significantly decreased the expression of destrin, FKBP1A and MIF, and increased the expression of profilin-1.

Conclusion:

A set of proteins differentially expressed in endothelial cells in response to cholinergic agonists may have important implications for the downstream biological effects of NNAS.