Morphologic characterization of osteoblast-like cell cultures isolated from newborn rat calvaria

Summary Two methods for harvesting osteoblast-like cell populations from newborn (10 days) rat calvaria were compared. The first one consisted in culturing the periosteum-free bones and then trypsinizing the cells on the bone surface. The second one involved the migration of the osteoblasts on glass fragments before trypsinization. Since the plating efficiency, the proportion of alkaline phosphatase-positive cells, the population doubling time, and the calcium deposition were more adequate, the second method was used to further characterize the behavior of the cultures. During the first week of culture, the cells featured shapes similar to those observedin vivo on the surface of periosteum-free calvaria. They formed multilayers and, in the presence of ascorbic acid, synthetized an organic matrix containing exclusively type I collagen. Later, small amounts of type III collagen appeared. The cells were embedded in the matrix and progressively acquired the morphologic phenotype of osteocyte-like cells. The matrix mineralized in the presence of β-glycerophosphate. The technique of dropinoculation (high concentration of cells in a small volume of medium) promoted the multilayer formation and the achievement of large mineralized plates (about 1 cm²) in 3 weeks of culture.     

细胞内钙信号的变化调节血管平滑肌细胞增殖

目的探讨细胞内钙信号的变化对大鼠血管平滑肌细胞(VSMC)增殖作用的影响及其对细胞内信号转导机制的变化。方法以培养的大鼠VSMC为模型,用雷尼丁(RY)剌激VSMC内贮Ca2 释放入胞浆,用3H亮氨酸及3H胸腺嘧啶掺入量作为反应VSMC增殖的指标,加入不同的细胞内信号转导阻断剂,观察对RY效应的影响。结果与对照组相比,RY浓度依赖性地促进细胞内游离钙浓度的增高,差异显著(P<0.05或0.01)。RY剌激组蛋白核酸合成速率明显增高,与对照组相比差异显著(P<0.01);尼卡地平(Nicardipine),蛋白激酶C抑制剂(H7),钙调素激酶(CaMPK)抑制剂(W7)和丝裂素活化蛋白激酶(MAPK)抑制剂(PD98059)能明显抑制RY介导的VSMC蛋白核酸合成速率增高,与RY剌激组相比差异显著(P<0.01)。结论细胞内钙信号的变化明显促进VSMC增殖,但其效应可能通过Ca2 、PKC、MAPK来介导。钙离子拮抗剂可抑制血管平滑肌细胞增殖。     

动态旋转系统构建组织工程化血管模型中血管内皮细胞分泌功能监测

目的 比较静态和动态旋转系统中构建组织工程化血管模型中血管内皮细胞分泌功能。方法新型可降解材料聚羟基丁酸酯(PHB),用胶原包埋,形成多孔状PHB 胶原管形支架。分离传代、分化人脐静脉内皮细胞,接种于PHB管型支架内腔面,分别在静态、动态旋转系统中培养14 d后,测定血管内皮细胞分泌一氧化氮(NO)、前列环素(PGI2)水平。结果 在动态旋转系统构建组织工程化血管模型中血管内皮细胞分泌NO、PGI2水平显著高于静态系统,在11 d NO分别为(120.52±3.83)μmmol／L、(80.98±5.98)μmmol／L,PGI2分另9为(20.48±1.52)μg／L,(16.59±1.29)μg／L,与静态系统相比,差异有显著性(P<0.05)。结论胶原包埋PHB支架有利于细胞的黏附和生长,可作为组织工程化血管的支架材料。在动态旋转系统构建组织工程化血管模型中血管内皮细胞,具有与正常血管类似的"生理功能"。     

ABO血型不合的异基因造血干细胞移植--2例报告附文献复习

探讨ABO血型不合的异基因造血干细胞移植 (Allo -HSCT)的造血问题。方法 :对 2例患者分别进行异基因外周血造血干细胞移植 (Allo -PBSCT)及脐血造血干细胞移植 (CBSCT) ,用血细胞分离机或 6 %羟乙基淀粉去除移植物中的红细胞 ,定期检测受者血型 ,根据情况输注血细胞。结果 :2例患者全部获得造血重建 ,未出现溶血反应。结论 :ABO血型不合可以进行Allo -HSCT ,去除供者移植物中的红细胞能有效地防止急性溶血反应的发生 ,定期检测受者血型 ,及时调整输血方案有重要的临床意义。     

An alternative mechanism of actions for neuroleptic and antidepressant drugs

It has earlier been proposed by the author that the aetiology of schizophrenic symptomatology may be due to the presence of abnormally connected interhemispheric fibres which link specialised functions in the brains of schizophrenics that are not connected in normal subjects, and that the neuroleptic drugs may produce their action through a local anaesthetic-like effect in suppression of conduction in these fibres. This line of thought has been extended here to consider the possible mechanism of action of the neuroleptic drugs in more detail, as well as that of the tricyclic antidepressant drugs which are derivatives of the phenothiazine group. Pharmacological similarities with the local anaesthetics both structurally and functionally have been considered, as well as the effects that these drug groups may have in common with the lithium salts. It has been suggested that these drugs all produce their primary effect on cell membranes, though not necessarily at the synapse, that the time course of their clinical effect may correlate with their incorporation into various cell membranes within the CNS, and that they may thus bring about a fundamental alteration in cell membrane microstructure. The possible role of electroconvulsive therapy has also been considered. The corollary of this argument is that the affective disorders may be genetically determined diseases of cell membrane microstructure.     

Siebenj?hrige erfahrungen mit der ?dintraoperativen maschinellen autotransfusion? und ?pr?operativen plasmapherese? bei chirurgischorthop?dischen eingriffen

Zusammenfassung Bericht über ?dintraoperative maschinelle autotransfusion?, kombiniert mit ?pr?operativer plasmapherese?. Mit maschineller autotransfusion kann intraoperativer blutverlust durch rückführung eigener erythrozyten halbiert werden. In kombination mit pr?operativer plasmapherese werden weitere fremdblutkonserven eingespart. übertragung von infektionskrankheiten und erzeugung irregul?rer antikōrper kōnnen verhindert werden. Bei der derzeitigen AIDS-situation wird das infektionsrisiko auf null reduziert.      

Successful Liver and Kidney Transplantation From Cadaveric Donors With Left-Sided Bacterial Endocarditis

Bacterial infections are frequent in cadaveric organ donors and can be transmitted to the transplantation recipient, which could have devastating consequences for the recipients if adequate preventive measures are not adopted.
From the 355 consecutive brain dead cadaveric organ donors procured at our center in the last four years, 2000–2003, four of them (1.1%) had bacterial endocarditis as cause of death. The bacteria responsible for the endocarditis were Staphylococcus epidermidis, coagulase-negative Staphylococcus , Staphylococcus hominis and Streptococcus viridans , respectively. We performed five kidney and two liver transplantations on seven recipients. All donors and recipients received antibiotic treatment against the germ causing the respective endocarditis.
Infection by the bacteria responsible for the endocarditis in the respective donors was not transmitted to any of the recipients. Six of the seven recipients were alive with normal-functioning grafts after between 13 and 24 months' follow-up. Transplantectomy was performed on one kidney recipient due to thrombosis of the renal vein of the graft not related to the endocarditis.
Liver and kidney transplantation from donors dying from bacterial endocarditis can be performed without causing the transmission of infection to the recipient or the dysfunction of the graft.     

LRIG1下调原代星形细胞瘤细胞增殖的机制

目的观察LRIGl蛋白表达对表皮生长因子（EGF）促肿瘤细胞增殖作用的影响，探讨LRIGl抑制肿瘤细胞增殖的机制。方法原代培养19例星形细胞瘤细胞，用原位杂交检测LRIGl的表达，用噻唑蓝（MTT）法观察EGF对培养细胞的促增殖作用，并分析培养细胞LRIGl表达和EGF干预后的细胞生长率的关系。结果（75．3±11．6）％的培养细胞表达LRIGl蛋白；EGF促进培养细胞增殖（38．0±14．8）％，EGF促细胞增殖的细胞生长率与LRIGl表达程度呈负相关。结论LRIGl蛋白可能通过抑制EGF—EGFR信号抑制肿瘤细胞的增殖。     

明胶降解物对大鼠真皮成纤维细胞增殖及Ⅰ型胶原蛋白分泌的影响

目的 探讨组织工程心瓣膜常用人工基质材料--明胶发生降解后的产物对大鼠真皮成纤维细胞增殖及Ⅰ型胶原蛋白分泌的影响. 方法 利用酶组合技术制备明胶降解物（gelatin hydrolysate, GH）,采用超滤的方法分离出低分子量明胶降解物组分（gelatin hydrolysate fraction, GHF）,用于5只SD大鼠真皮成纤维细胞的体外培养.采用CCK-8活细胞计数试剂盒（Cell Counting Kit-8）检测细胞的增殖情况,ELISA试剂盒检测细胞I型胶原蛋白的分泌情况. 结果 ①GHF对细胞增殖作用的浓度效应：在0.125～1.0 mg/ml浓度范围内,GHF对细胞的促增殖作用呈现先增强后减弱的趋势,在0.25 mg/ml时,细胞增殖率最大,达到（47.54±16.35）%;②GH与GHF对细胞增殖影响比较：GH与GHF均具有促进细胞增殖的作用,且在浓度为0.25 mg/ml时,促细胞增殖作用无显著差异[（27.04±15.21）% vs （39.22±15.55）%, P=0.177];③GH与GHF对细胞分泌Ⅰ型胶原蛋白影响比较：GHF组第3天时Ⅰ型胶原分泌量为（28.06±5.60）pg/105,与空白对照的（18.24±4.52）pg/105相比,有显著差异（P=0.006）,而GH组直至第6天时Ⅰ型胶原蛋白分泌量,与空白对照相比,无统计学意义（P=0.103）. 结论 分子量大小不同的明胶降解物GH与GHF均具有促进大鼠真皮成纤维细胞增殖的作用,且分子量较小的GHF还具有促进细胞I型胶原分泌的功能.     

The Hemodynamic Mechanisms of Lung Injury and Systemic Inflammatory Response Following Brain Death in the Transplant Donor

Brain-dead donors are the major source of lungs for transplantation. Brain death is characterized by two hemodynamic phases. Initially, massive sympathetic discharge results in a hypertensive crisis. This is followed by neurogenic hypotension. Up-regulation of pro-inflammatory mediators occurs in all organs and lung injury develops; this can adversely affect graft function post-transplantation. The mechanisms of the systemic and lung inflammation are unknown. We hypothesized that the hemodynamic changes are responsible for these inflammatory phenomena. Brain death was induced by intra-cranial balloon inflation in rats. This resulted in hypertensive crisis, followed by hypotension. There was a significant increase in blood neutrophil CD11b/CD18 expression and pro-inflammatory cytokine levels in serum and bronchoalveolar lavage, compared with control animals. Rupture of the capillary-alveolar membrane was demonstrated by electron microscopy. Elimination of the hypertensive response by α-adrenergic antagonist pre-treatment prevented inflammatory lung injury, reduced the systemic inflammatory markers and preserved capillary-alveolar membrane integrity. Correction of the neurogenic hypotension with noradrenaline ameliorated the systemic inflammatory response and improved oxygenation. We conclude that the sympathetic discharge triggers systemic and lung inflammation, which can be further enhanced by neurogenic hypotension. Management of the brain-dead donor with early anti-inflammatory treatment and vasoconstrictors is warranted.