支架及无支架条件下构建组织工程软骨的研究进展

支架材料的研究是组织工程的研究热点之一,软骨细胞复合培养给组织工程软骨的构建带来希望,单一的材料诸如胶原、透明质酸、壳聚糖、纤维蛋白凝胶等已经证明可以与软骨细胞复合培养,两种或者多种材料复合可以提高材料的性能,更好地用于组织工程软骨的构建,并满足软骨缺损修复的需要;同样,在无支架情况下应用软骨细胞聚集培养、沉淀培养的方法,可以构建组织工程软骨,并给软骨缺损的修复带来新希望,但目前的研究较少。两者是组织工程软骨构建的两个主要方向。     

17 β雌二醇通过丝裂原活化蛋白激酶抑制前列腺癌细胞系PC3增殖

目的探讨丝裂原活化蛋白激酶在17β雌二醇(E2)抑制前列腺癌PC3细胞生长中的作用。方法检测不同浓度E2作用不同时间后PC3细胞生长抑制率。流式细胞仪分析细胞周期分布,TUNEL染色检测凋亡。Western blot检测ERK1/2,JNK和p38活性。RT-PCR法检测雌激素受体(ER)α、ERβ、P21WAF1和cyclinD1的表达。结果E2抑制PC3细胞增殖,并且可以激活ERK1/2、JNK和p38。处理因素作用48h后,对照组、E2、E2 PD98059、E2 SP600125、E2 SB203580组细胞的凋亡率分别为(0.9±0.1)%;(23.0±1.4)%;(30.0±1.2)%;(10.6±0.8)%和(14.6±0.7)%,(P<0.05)。E2使细胞阻滞在G1期,PD98059、SP600125、SB203580分别预处理1h后细胞分别进一步阻滞在G1期;轻度阻滞在G1期和进入S期。RT-PCR发现PC3细胞表达ERα和ERβ,E2、E2 PD98059、E2 SP600125、E2 SB203580组中cyclinD1、P21WAF1基因表达分别为对照组的(0.42±0.03)、(3.13±0.02)倍;(0.21±0.03)、(3.08±0.05)倍;(0.43±0.01)、(1.31±0.04)倍;(2.81±0.02)、(3.14±0.02)倍(P<0.05)。结论E2激活JNK增加P21WAF1基因表达,并激活p38抑制cyclinD1表达,使细胞阻滞在G1期,JNK和p38通路还介导E2引起PC3细胞凋亡。同时E2又可激活ERK1/2,轻度拮抗上述作用。     

非功能性胰岛细胞瘤16例临床分析

目的探讨非功能性胰岛细胞瘤的诊断和治疗方法。方法对近16年间收治的非功能性胰岛细胞瘤16例临床资料回顾性分析。其中位于胰头部4例,胰体尾部12例,瘤体长径平均12cm。结果行肿瘤摘除术6例,胰十二指肠切除术3例,行包括肿瘤在内的远端胰腺及脾脏切除3例,保留脾脏的胰体尾切除术3例,因肿瘤广泛侵浸未能切除1例。治愈性切除率为93.75%(15/16),全组无手术死亡。随访资料显示:11例良性肿瘤患者全部存活,已2～16年。5例恶性肿瘤患者中,1例因肿瘤广泛侵犯未能切除者,术后5个月死于多器官衰竭;1例胰体尾切除并左肝切除者存活2年;胰十二肠切除者2例,分别存活1年和6年,胰体尾切除加脾切除1例已存活12年8个月。结论钡餐,B超,CT,MRI,以及ERCP对该病有诊断价值,手术切除是治疗非功能性胰岛细胞瘤的主要方法,效果良好。     

当归对衰老小鼠脑组织细胞凋亡及胸腺指数的影响

目的：研究当归的抗衰老作用及作用机制。方法：用D—半乳糖诱导小鼠制成衰老模型，同时给予不同剂量的当归水煎剂，观察当归对脑组织细胞凋亡及胸腺指数的影响。结果：当归能明显抑制小鼠脑组织细胞凋亡，提高胸腺指数。结论：当归具有抗衰老作用。     

冬凌草甲素对Raji细胞的增殖抑制作用及其机制

目的探讨冬凌草甲素对B淋巴细胞白血病细胞株Raji细胞的增殖抑制作用及其机制。方法以不同浓度的冬凌草甲素作用于体外培养的Raji细胞，MTT法检测细胞生长抑制率，应用流式细胞仪、Hoechst 33258荧光染色法观察细胞凋亡，TRAP-PCR-ELISA法检测细胞凋亡前后的端粒酶活性。结果冬凌草甲素可显著降低Raji细胞的端粒酶活性，抑制细胞的生长，诱导细胞发生凋亡，并呈现出明显的剂量-效应与时间-效应关系。结论冬凌草甲素对B淋巴细胞白血病具有明显的体外抗白血病作用，降低细胞端粒酶活性，诱导细胞发生凋亡，可能是其重要作用机制。     

益气复智颗粒对多发脑梗死性痴呆模型大鼠脑组织细胞凋亡的影响

目的 观察益气复智颗粒对多发脑梗死性痴呆模型大鼠脑皮质形态学、细胞凋亡的影响。方法 采用颈内动脉注射血栓的方法，复制多发梗死性痴呆大鼠模型，观察益气复智颗粒12．42g/kg分别于手术前、手术前后、手术后灌胃对实验动物脑皮质形态学、细胞凋亡的影响。结果 益气复智颗粒能使脑缺血后脑内神经细胞凋亡数目下降。结论 益气复智颗粒具有较好的保护脑神经元，阻断脑缺血致神经细胞死亡病理过程的作用。     

大鼠移植胰腺冷缺血再灌注后细胞凋亡的变化

目的　探讨移植胰腺冷缺血再灌注后胰腺细胞凋亡的变化过程。方法　 6 5只SD大鼠随机分成 7组 :假手术组 ,冷缺血 2h组 ,冷缺血 2h再灌注 1、3、6、9、12h组。通过HE染色后光镜及电镜观察各组的胰腺组织的病理变化 ;采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测凋亡细胞的分布及计数。结果　胰腺移植后早期即可观察到细胞凋亡的典型改变 ,胰腺冷缺血再灌注后发生凋亡的高峰期为再灌注后 3h[AI为 (9.4 6± 2 .91) % ,P <0 .0 1) ,再灌注 6h较 3小时细胞凋亡有所减少 [AI为 (5 .74± 1.6 6 ) % ,P <0 .0 1],再灌注 9h及 12h细胞凋亡进一步减少 [AI分别为 (3.6 0± 1.6 4 ) %及 (3.2 6± 1.35 ) % ,P <0 .0 5 ]。结论　细胞凋亡是胰腺移植后的早期事件 ,移植胰腺冷缺血再灌注后早期主要的死亡方式是凋亡 ;移植胰腺冷缺血再灌注后的凋亡高峰发生在再灌注后 3h。     

Growth fraction in human brain tumors defined by the monoclonal antibody Ki-67

Summary The monoclonal antibody Ki-67, which reacts with cells in the active part of the cell cycle, was used to evaluate immunocytochemically the growth fraction in 22 primary brain neoplasms. The percentage of labelled cells reflected the histological grade of malignancy of each neoplasms. High percentage of Ki-67-positive cells were observed in one choroid plexus carcinoma (60%), one primary melanoma of meninges (40%), three medulloblastomas (40%–50%), one anaplastic astrocytoma and six glioblastomas (10%–40%). One ependymoma had 7% positive cells. Rare positive cells (1%) were present in one pilocytic astrocytoma and one ganglioglioma. Except one negative case, the meningiomas (five cases) had values of positivity ranging from 1% to 6%. Two acoustic schwannomas were negative. These results suggest that immunocytochemical staining with the Ki-67 may be a useful method for measuring the growth fraction in brain neoplasms.Supported in part by Associazione Italiana Ricerca sul Cancro and Ministero Italiano della Pubblica Istruzione     

Monoclonal antibody production to human and bovine 2′:3′-cyclic nucleotide 3′-phosphodiesterase (CNPase): high-specificity recognition in whole brain acetone powders and conservation of sequence between CNP1 and CNP2

Monoclonal antibodies against human and bovine 2′:3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) were generated by fusing FOX-NY myeloma cells with spleen cells from RBF/Dn mice previously immunized with the purified brain antigens. The enzyme isolated from bovine brain was quite basic, with an isoelectric point of 9.71 and both the bovine and human enzymes consisted of a closely spaced doublet at approximately 44 and 46 kDa on SDS-PAGE. Six monoclonals were identified as strongly recognizing the enzyme on both ELISA plates and on immunoblots of whole brain protein. Four monoclonals very weakly cross-reacted with guinea pig myelin basic protein. In contrast with two previous reports, some of our monoclonal antibodies did immunostain 2 or 3 protein bands in peripheral nerve, two bands closely corresponding to those immunostained in central nervous system (CNS) myelin, the Wolfgram protein fraction and in acetone powders of whole brain. Each of the 6 monoclonals reacting strongly on immunoblots recognized the enzyme in from 2 to 5 of the species examined (human, bovine, rat, mouse and rabbit). In addition, all 6 monoclonals that immunostained the enzyme in whole brain, myelin and Wolfgram protein immunoblots recognized both CNP1 (44 kDa) and CNP2 (46 kDa). The two closely spaced protein bands observed on SDS-PAGE and previously stained on immunoblots of CNS CNPase using polyvalent rabbit anti-bovine CNPase antisera, and now different monoclonal antibodies, appear to be immunologically related and to contain highly conserved sequences.