新钙灵对兔血清中若干骨形成因素相关性的分析

用于儿童的钙剂直接移用于年老者补钙效果并不佳。试图探讨血清中Ca、BGP、CT、AKPP、BUN六个有关骨形成因素的相关性。本文结果如下：（1）报告兔血清中以上六个因素的正常平均值；（2）BGP与Ca、CT、AKP均有程度不等的相关性；BGP与兔体重有密切相关，体重大于2.5kg，多数BGP形成减少；根据BGP形成多少，血清中GBP：Ca、BGP：P等比值也有差异。认为BGP是血清中骨形成因素的主要因素：（3）给予家兔小剂量新钙灵后，血清BGP与有关因子的比值，和BGP形成低下的兔血清中BGP的相关性相符。活性钙能使BGP：CT王相关，此因素分别又与其他因素呈现负相关，不利BGP、CT存留，达不到理想效果。     

慢性肾衰竭兔血清对主动脉平滑肌细胞增殖及核因子κB活化的作用

目的 研究慢性肾衰竭兔血清对其主动脉平滑肌细胞增殖和核因子κB （NF-κB）活化的影响,并探讨其作用的可能机制。 方法 建立慢性肾衰竭兔模型,采集血清。采用原代培养的兔主动脉平滑肌细胞,用不同浓度的慢性肾衰竭兔血清刺激不同时间,四甲基偶氮噻唑盐（MTT）法检测细胞增殖;Hoechst33342染色观察细胞凋亡;Western印迹检测慢性肾衰竭血清对主动脉平滑肌细胞增殖细胞核抗原（PCNA）、NF-κB p65蛋白表达的影响;免疫荧光观察NF-κB p65核转位。 结果 （1）在较低浓度（≤10%）时,慢性肾衰竭兔血清对平滑肌细胞的增殖有明显的促进作用,且呈浓度、时间依赖性;但血清浓度继续增加后,对平滑肌细胞的促增殖作用却明显减弱,与正常血清组差异有统计学意义（P < 0.05）。（2）Hoechst33342染色表明,慢性肾衰竭血清在低浓度时（≤10%）细胞凋亡率和正常血清组差异无统计学意义（P > 0.05）,但高浓度（>10%）时,平滑肌细胞凋亡率增加,与正常血清组差异有统计学意义（P < 0.01）。（3）慢性肾衰竭血清刺激24 h后,低浓度促进PCNA、NF-κB p65蛋白表达,高浓度抑制PCNA、NF-κB p65表达,与正常血清组差异有统计学意义（P < 0.01）。（4）10%慢性肾衰竭血清与平滑肌细胞孵育24 h后免疫荧光显示,NF-κB p65发生了核转位。 结论 不同浓度的慢性肾衰竭兔血清可导致平滑肌细胞增殖或凋亡,其促增殖作用可能与其活化了NF-κB有关。本研究为防治慢性肾衰竭加速性动脉粥样硬化提供理论依据。     

Direct identification of the murine pluripotential stem cell using rabbit anti-mouse brain serum.

Previous studies have shown that antisera prepared in rabbits against mouse brain tissue (RAMBS) contain activity against the murine bone marrow colony-forming unit (CFU-s) or pluripotential hemotopoietic stem cell. In the present study, the F(ab')2 portion of RAMBS was examined for its potential efficacy in the identification of the mouse CFU-s when used in an indirect immunofluorescence-labeling technique. After separation of mouse bone marrow cells by a discontinuous bovine serum albumin density gradient, fluorescent cells were observed only in those bands which, by the splenic colony-forming assay, demonstrated CFU-s. Furthermore, the quantity of CFU-s demonstrated by the spleen colony-forming assay approximated the number of fluorescent cells observed in the corresponding band. It appears that the F(ab')2 portion of RAMBS used in an immunofluorescent assay may provide a method for the direct quantitation and identification of the CFU-s content of murine bone marrow.     

Radioimmune assay of heterologous serum gamma-globulin in patients receiving rabbit antihuman thymocyte globulin.

A radioimmune assay (RIA) method for detecting heterologous serum rabbit gamma-globulin (RG) and antibody to this protein is described. The methodology is used for monitoring serum levels of rabbit globulin in patients receiving rabbit ATG (RATG). In 7 cardiac recipients receiving RATG, maximum serum levels of RG were achieved 1-3 days after administration of final dose. RG half-life subsequent to peak serum levels was rapid (X = 36 hr) in 4 patients and prolonged (X = 18 days) in 3 patients. Patient antibody to rabbit gamma-globulin was detectable only in those patients with short RG half-life. Antirabbit antibody titers in these patients were extremely low and barely detectable by RIA.     

Doxorubicin treatment of rabbit renal VX-2 carcinoma: nephrotoxicity,serum parameters and weight

Serum electrolytes, creatinine, urea, protein, albumin, bilirubin and glucose were examined every 4 days until time of death in rabbits with VX-2 carcinoma implanted in one kidney. The rabbits were treated with doxorubicin, nephrectomy or combinations thereof and observed for up to 1 year. Rabbits treated with doxorubicin only showed a slight creatinine rise initially, but over time creatinine reached almost the same concentration as that in nephrectomized rabbits receiving equivalent doses of doxorubicin. Creatinine concentrations increased significantly above the normal range following nephrectomy combined with doxorubicin. Doxorubicin nephrotoxicity in rabbits occurs at lower doses than previously reported. In all rabbits the parameters except creatinine remained stable within the established normal ranges, except for the last 4 days before time of death in the animals with metastatic disease. Weight loss was the best parameter for making a prognosis for an individual rabbit, since peak weight was noted 16–20 days before death. In experimental work with VX-2 carcinoma, weight is thus the most important indicator of the time at which rabbits not responding to treatment can be put to death to avoid unnecessary suffering before the end of the experiment.     

Growth of rabbit aortic smooth-muscle cells cultured in media containing diabetic and hyperlipemic serum.

Smooth-muscle cell cultures were grown from thoracic aortas of normal and diabetic rabbits. The effect of diabetic rabbit serum on the growth of these cultures was studied both in the first, rapid-growth phase and the following, more "stationary" phase of growth. Control experiments were carried out on normal sera to which glucose had been added. The concentrations of cholesterol, phospholipid, and triglyceride were same in both normal and diabetic sera. Media containing diabetic serum stimulated the growth of cultures significantly in both phases (2p less than 0.01). This occurred in experiments utilizing cells from normal as well as from diabetic rabbits. Control media containing normal serum with added glucose had no such effect. The growth-promoting effects of diabetic serum and of hyperlipemic serum from nondiabetic rabbits were of the same order of magnitude. Autoradiographic studies showed that the number of 3H-thymidine-labeled cells increased significantly after culture in diabetic serum (2p less than 0.005). Cells cultured from the very beginning in diabetic serum or normal serum with added glucose were significantly larger than cells grown in control serum (2p less than 0.05 and 2p less than 0.01, respectively). Cells grown in hyperlipemic serum were significantly smaller than those grown in normal serum (2p less than 0.01). These results indicate that diabetic serum contains a factor or factors that stimulate the arterial medial cell to excessive growth. This factor is not glucose, insulin, or lipid. The results may be of relevance for the understanding of human diabetic macroangiopathy.     

血清酶谱检查在家兔急性肠系膜缺血性疾病诊断中的作用

目的研究急性肠系膜缺血后各时段酶谱变化规律,为临床早期诊治该病提供帮助。方法将36只家兔分为动脉阻断、静脉阻断、动静脉阻断3组,每组12只。每组动物分别于阻断前、阻断后1、3、6、9h抽静脉血,检验血清内丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、谷酰转酞酶(γ-GT)、肌酸激酶(CK)、肌酸激酶同工酶(CK—MB)、乳酸脱氢酶(LDH)水平。同时观察肠管的颜色变化。结果各组家兔肠管在血管被阻断3h后开始发生坏死,同时ALT、AST、ALP、γ-GT、LDH均有明显升高。结论血清酶谱检查有助急性肠系膜缺血性疾病的诊断。     

Inducible osteonecrosis in a rabbit serum sickness model: Deposition of immune complexes in bone marrow

We established inducible osteonecrosis in a rabbit serum sickness model. Osteonecrosis with marrow necrosis could be induced by the intravenous injection of horse serum in two doses separated in time by a period of three weeks. In this model, osteonecrosis could be successfully produced in rabbit femoral metaphysis. The incidence of marrow necrosis was 45% (9 of 20 rabbits) and trabecular necrosis occurred in 6 of 20 rabbits (30%) at 7 days after the second injection of the horse serum. In bone marrow of the femoral metaphysis, extravasation of erythrocytes and the formation of microthrombi in arterioles were often observed in an early stage of the present model and both findings correlate well each other (p = 0.0001). Immune complexes could be demonstrated using immunohistochemistry in bone marrow of the femoral metaphysis as well as in glomeruli of the kidney. Extravasation of erythrocytes in bone marrow of the femoral metaphysis was observed in 8 of 12 (67%) cases with immune complex deposition in the sinusoidal space of the femoral metaphysis and in 12 of 21(57%) cases with immune complex deposition in glomeruli of the kidney. Immune complex deposition both in the sinusoidal space of femoral bone marrow (p = 0.0385) and in glomeruli of the kidney (p = 0.0209) closely related to extravasation of erythrocytes and microthrombi in arterioles in the early stage of this model. Early microcirculatory injury (extravasation of erythrocytes and microthrombi in arterioles) adjacent to osteonecrosis could be induced by immune complex deposition in femoral bone marrow and might be predictable characteristics for the inducible osteonecrosis in the present serum sickness model. The important findings in this study were that early microcirculatory injury was closely related to the deposition of immune complexes in femoral bone marrow, and that early microcirculatory injury associated with immune complex deposition was located close to osteonecrotic regions.     

不同浓度骨痹舒片含药血清对体外培养软骨细胞增殖的影响

目的：观察不同浓度骨痹舒片含药血清对体外培养的兔关节软骨细胞增殖的影响。方法：获取1月龄兔关节软骨细胞,随机分为空白组（正常兔血清）及骨痹舒片含药血清组,每组再分5%、10%、15%、20%4个浓度,共8组。分别于培养第1、3、5、7、9天用MTT法检测软骨细胞的增殖情况。结果：骨痹舒片含药血清组细胞呈血清浓度依赖型增殖。同一时间点各组间比较差异均有统计学意义（P〈0.05）,以20%组最佳;空白血清组中5%、10%组与20%组相比在各时间点差异均有统计学意义（P〈0.05）,15%组与20%组相比在第1、3、5、7天各时间点相比差异无统计学意义（P〉0.05）,在第9天时差异有统计学意义（P〈0.05）,以20%组最佳。20%骨痹舒片含药血清组与20%空白血清组相比,在第1、3、5、7天时差异有统计学意义（P〈0.05）,第9天差异无统计学意义（P〉0.05）。结论：20%的骨痹舒片含药血清能显著促进软骨细胞的增殖,并将细胞的指数增长期提前至第3天。     

Improvement in rejection of human decay accelerating factor transgenic pig-to-primate renal xenografts with administration of rabbit antithymocyte serum

BACKGROUND: Survival in pig-to-baboon kidney xenotransplantation is currently limited by acute humoral xenograft rejection (AHXR). We hypothesized that the administration of rabbit antithymocyte serum (RATS) would delay or prevent AHXR as compared with a cyclophosphamide (CyP)-based immunosuppressive regimen. METHODS: Nine baboons received life-supporting heterotopic single-kidney transplants from human decay accelerating factor transgenic pigs. Immunosuppression consisted of GAS (a galactosyl alpha-1,3-galactose analog), cyclosporine, and steroids. Group 1 (n=2) was also treated with CyP and a rapamycin derivative (RAD), group 2 (n=4) received RATS and RAD, and group 3 (n=3) received only RATS. Animals were maintained until death or sacrifice because of uncontrollable rejection or other complications. Graft histopathology was assessed at the study endpoint. RESULTS: Mean survival was 28+/-11.3 days, 23+/-2.5 days, and 20+/-2.5 days for groups 1, 2, and 3 (not significant). Graft rejection was the cause of death in both CyP-treated animals. One RATS-treated animal died of rejection; the others died of infections or bleeding. Two RATS-treated animals developed posttransplant lymphoproliferative disorder, and one died of cytomegalovirus pneumonitis. Histopathology revealed severe AHXR in group 1 kidneys, involving 100+/-0% of the tissue examined. In contrast, AHXR was reduced in groups 2 and 3, involving 21+/-14% and 18+/-28%, respectively, of the tissue examined (P<0.01). CONCLUSIONS: Substitution of RATS for CyP was well tolerated and resulted in reduced severity of AHXR in this model. Complications seen in RATS-treated animals may be preventable through the use of standard prophylaxis for infections. Our data suggest that further studies are warranted to explore the use of antilymphocyte agents in xenotransplantation.