骨骼肌卫星细胞梗死心肌移植的细胞因子分泌及血管再生作用

目的　研究骨骼肌卫星细胞梗死心肌移植的胰岛素样生长因子(IGF)-1、碱性纤维母细胞生长因子(bFGF)分泌及血管再生的作用。方法　骨骼肌卫星细胞经结扎的冠状动脉左前降支远端灌注移植入梗死区,2、4、8周后取标本,应用免疫组化学方法检测细胞因子表达及梗死区血管密度。结果　骨骼肌卫星细胞移植2、4、8周后梗死区IGF-1和bFGF表达分别为81.68±3.34、96.87±7.78、90.43±7.36及81.87±3.58、65.66±4.57、74.20±6.41,明显高于对照组(P＜0.01);同时,移植2、4、8周后梗死区血管密度高于对照组(P＜0.05)。结论　骨骼肌卫星细胞梗死心肌移植除有心肌再生外,尚可通过细胞因子分泌对梗死心肌起到积极作用。     

骨髓单个核细胞自体移植促进心脏血管再生

目的探讨骨髓单个核细胞自体移植于梗死心肌后实现血管再生的能力。方法结扎45只5月龄日本大耳白兔的冠状动脉左前降支建立心肌梗死模型，随机分为骨髓单个核细胞自体移植组(5-溴-2’-脱氧尿嘧啶核苷标记的自体骨髓单个核细胞直接心肌注射至梗死周边区域)，心肌梗死未治疗组及假手术组，分别在细胞移植后2、4、8周时处死，应用免疫组织化学方法检测骨髓单个核细胞在梗死心肌组织中的分化、毛细血管密度以及细胞因子的表达。结果骨髓单个核细胞在梗死心肌组织中分化为血管内皮细胞和血管平滑肌细胞。在骨髓单个核细胞移植早期，心肌白细胞介素-1β(IL-1β)、碱性纤维母细胞生长因子(bFGF)、血管内皮生长因子(VEGF)表达水平显著增高。细胞移植后4、8周骨髓单个核细胞移植组毛细血管密度明显高于心肌梗死未治疗组及假手术组。结论骨髓单个核细胞自体移植于梗死心肌区能促进血管再生，改善侧支循环。     

急性心梗后骨髓细胞心肌内移植促进心肌细胞再生的实验研究

目的 研究急性梗死心肌内移植骨髓细胞的促进心肌再生作用。方法 局部注射将骨髓细胞移植入大鼠急性心肌梗死模型的急性心肌梗死区域，1、2、4、8周后处死动物，取心肌标本行组织形态学检查和梗死面积测量。结果 梗死区心肌标本观察到带荧光的骨髓细胞，部分已分化成肌源性细胞，电镜观察到不同分化阶段的新生心肌细胞。与成熟心肌细胞以闰盘相联。骨髓细胞移植组梗死心肌面积明显小于对照组。结论 在梗死心肌内移植的骨髓细胞具有心肌再生能力，并能缩小梗死心肌面积。     

骨髓细胞梗死心肌移植对心肌细胞结蛋白的影响

目的　探讨骨髓细胞梗死心肌移植对心肌细胞结蛋白表达的影响。方法　体外提取、纯化骨髓细胞、羟酸乙酸盐琥珀酸脂 (CFSE)荧光标记。骨髓细胞经局部注射移植入心肌梗死区域 ,1、2、4、8周后取心肌标本 ,行组织形态学检查 ,免疫组织化学检测心肌细胞结蛋白表达。结果　移植后 4周和 8周 ,电镜观察到不同分化阶段的幼稚心肌细胞 ,并与成熟心肌细胞以闰盘相连接。移植组和对照组梗死区心肌细胞结蛋白表达都明显高于正常区域心肌细胞 (P <0 .0 1) ,移植组术后 1、2、4、8周 ,梗死区心肌细胞结蛋白表达 (灰度值 )分别为 114 .0 6± 4.80、12 0 .3 1± 5 .10、118.3 7± 4.5 1、12 1.3 5± 3 .86。对照组梗死区心肌细胞结蛋白排列紊乱 ,未见肌小节显示。 1周、2周时移植组梗死区所见与对照组基本相同 ,但 4周和 8周时部分心肌细胞结蛋白已恢复排列并重见肌小节。结论　骨髓细胞梗死心肌移植对梗死区心肌组织结构起到保护作用 ,使部分心肌细胞结蛋白的分布恢复正常 ,但对梗死区心肌细胞结蛋白的表达量无影响。     

细胞因子对关节软骨细胞增殖的实验研究

目的　探讨细胞因子对关节软骨细胞增殖的影响。方法　应用关节软骨细胞体外培养 ,分别加入IL - 1β,TNF -α,TGF - β1,IL - 1β和TGF - β1,TNF - β和TGF - β1,采用MTT法观察软骨细胞增殖情况 ,瑞氏染色观察细胞形态及分裂指数测定。结果　细胞因子作用软骨细胞 2 4h ,与对照组比较 ,1ng/ml、 10ng/mlTGF - β1组 ,10ng/mlIL - 1β和 1ng/mlTGF - β1组 ,高于对照组。细胞因子作用软骨细胞 4 8h ,与对照组比较 ,2 0ng/mlIL - 1β组 ,1ng/ml、 10ng/mlTNF -α组 ,0 1、 1、 10ng/mlTGF - β1,10ng/mlIL - 1β和 1ng/mlTGF - β1组 ,1ng/mlTNF -α和 1ng/mlTGF - β1组 ,高于对照组。IL - 1β,TNF -α作用软骨细胞 2 4h ,形态发生改变。培养 4 8h ,分裂指数测定 10ng/mlTGF - β1组显著高于对照组。结论　TGF - β1对体外培养关节软骨细胞有明显促进细胞分裂与增殖作用 ,IL - 1β ,TNF -α,作用软骨细胞 4 8h也有促进细胞增殖作用。     

骨髓间质干细胞移植梗死心肌后细胞因子分泌及对血管新生的影响

目的研究骨髓间质干细胞(MSCs)移植梗死心肌后细胞因子的分泌及其对血管新生的作用.方法贵州香猪24只,按照计算器随机数法随机分为对照组、实验组.抽取骨髓液3 ml,按照Wakitani的方法培养出骨髓间质干细胞,经5-氮胞苷(5-aza)诱导后,5-溴脱氧尿苷(BrdU)标记备用.开胸结扎左冠状动脉前降支,自体骨髓间质干细胞分别经局部注射和结扎的左冠状动脉前降支远端灌注移植入自体急性心肌梗死区域,对照组以DMEM作对照.术后3周、6周取标本,免疫组织化学法、计算机图象分析检测组织血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)和Ⅷ因子表达.结果实验组梗死区3周、6周bFGF、VEGF灰度值明显高于对照组(P<0.01),微血管计数明显增多(P<0.01).结论自体骨髓间质干细胞移植梗死心肌后能分泌VEGF、bFGF,诱导血管新生.     

骨髓细胞移植对梗死区心肌基本结构及左心室功能的影响

目的 探讨骨髓细胞移植对梗死区心肌基本结构及左心室功能的影响。方法 骨髓细胞注射人心肌梗死区域，1、2、4、8周后取心肌标本，组织切片染色观察组织结构；术后1、2、4、8周通过微机生物信号记录分析系统测定左心室功能指标。结果 移植组梗死区心肌观察到带荧光的骨髓细胞，部分已分化成肌源性细胞，电子显微镜观察到不同分化阶段的新生心肌细胞，并与成熟心肌细胞以闰盘相联接。HE染色显示对照组梗死区结构紊乱，而移植组梗死区内细胞呈有序排列；VG染色显示移植组胶原纤维融合较少，排列基本处于有序状态。移植组左心室功能的恢复明显优于对照组，且其恢复随时间的延长而增加。结论 骨髓细胞移植后对梗死心肌的基本结构起到了保护作用，并可促进左心室功能的恢复。     

自体内皮祖细胞移植对急性心肌梗死近、中期心肌收缩力的影响

目的比较自体内皮祖细胞（EPCs）移植急性心肌梗死区域后近、中期心功能恢复的程度。方法将SD大鼠120只随机分为实验组和对照组（每组60只），抽取自体外周血，体外分离出内皮祖细胞。开胸结扎左冠状动脉前降支，实验组用自体内皮祖细胞经局部注射移植入急性心肌梗死区域；对照组以IMDM（Iscove＇s modified Dulbecco＇s Medium）作对照。术后3、6周、6、8和12个月取标本，体外药物刺激离体肌条检测心肌收缩情况。免疫组织化学法、计算机图象分析检测组织血管内皮生长因子（VEGF），碱性成纤维生长因子（bFGF），血管内皮第八因子（Ⅷ因子）的表达。结果实验组梗死区3、6周、6个月bFGF、VEGF灰度值明显高于对照组（P〈0．01），微血管计数较对照组明显增多（P〈0．01），心肌收缩力明显较对照组增强（P〈0．01），且随时间推移恢复程度增加（P〈0．05）。但从8个月开始，实验组以上指标不再明显增加。结论自体内皮祖细胞移植可能通过血管发生、血管新生等方式增加心肌细胞的收缩力，但中期疗效并不能随着时间推移而持续增加，在较长时间内保持稳定。     

碱性成纤维细胞生长因子对成骨细胞中转化生长因子-β1和碱性成纤维细胞生长因子mRNA表达的影响

目的　探讨外源性碱性成纤维细胞生长因子 (bFGF)对体外成骨细胞中的生长因子 :转化生长因子 β1 (TGF β1 )和bFGFmRNA表达的影响。 方法　将体外培养的新生SD大鼠颅骨成骨细胞 ,用不同浓度的bFGF(5～ 50 μg/L)进行处理 ,利用核酸原位分子杂交 ,检测两种生长因子在细胞中mRNA的表达。结果　依bFGF浓度的增加成骨细胞内TGF β1mRNA表达分别是对照组的 1 .5、1 .6、1 .9和 2 .0倍 ;bFGFmRNA表达则分别是对照组的 1 .2 8、1 .37、1 .40和 1 .51倍。bFGF组中 ,TGF β1和bFGFmRNA的表达量均明显高于对照组 (P <0 .0 1 )。结论　外源性bFGF可以对成骨细胞自分泌bGFG产生影响 ,还能够促进TGF β1的合成     

急性胰腺炎血中抗炎症细胞因子的意义

目的　探讨急性胰腺炎时血中抗炎症细胞因子白介素 10 (IL 10 )和转化细胞生长因子 β(TGF β)的变化及其意义。方法　实验用SD大鼠 ,分为正常对照组 (n=6 )和胰腺炎组 (n=2 0 ) ,后者采用开腹胰管注射 5 %牛磺胆酸钠(1.0ml/kg)诱导急性胰腺炎。分别于术后 2小时 (n=6 )、6小时 ((n=6 )和 2 4小时 (n=8)处死动物 ,抽血用酶联免疫吸附法 (ELISA)检测血中IL 10和TGF β及胰淀粉酶和胰腺湿重。 结果　对照组动物血IL 10为 32 .0 5± 14 .87pg/ml;TGF β为6 6 .40± 13 .2 0 pg/ml。胰腺炎后血IL 10和TGF β均升高 ,IL 10在 2、6、2 4小时时段分别为 36 .2 5± 9.76pg/ml、37.75± 6 .5 4pg/ml和 6 8.13± 19.90 pg/ml;TGF β在 2、6、2 4小时分别为 6 4.5 8± 10 .5 6pg/ml、72 .87± 18.34pg/ml和 10 3 .77± 2 8.95pg/ml。胰腺炎后 2 4小时血IL 10和TGF β与正常组相比差异有显著性意义 (P <0 .0 5 )。 结论　急性胰腺炎时血中抗炎症细胞因子升高 ,并可能和胰腺炎后期的免疫抑制效应有关。     

Antitumor Killer Lymphocytes in the Peripheral Blood of a Patient with Transitional Cell Carcinoma of the Bladder

Background: Peripheral blood lymphocytes (PBL) from patients with bladder cancer also contain cells possessing cytotoxic activity against autologous tumor cells. These cells are phenotypically heterogenous and include natural killer (NK) and cytotoxic T cells. This study investigated the role of cytotoxic lymphocytes directed against autologous bladder cancer cells.
Methods: PBL were obtained at intervals before and after surgery and analyzed for cytotoxic activity against autologous bladder cancer cells in 4-hour⁵¹ Cr release assay. PBL stimulated with autologous tumor cells were also transformed with human T-lymphotropic virus type-1, establishing a cell line (KB31) which was analyzed for phenotype and cytotoxic activity against the autologous tumor cells.
Results: PBL preoperative cytotoxic activity was low, but increased after surgery. Cytotoxic activity was found not only against autologous bladder cancer cells, but also against heterologous bladder cancer (KK-47) and myeloid leukemia (K562) cells, with the highest activity against the heterologous cell lines. The cytotoxic activity of KB31 was 40|X% against autologous tumor cells 6 weeks after initiation of the cell line, but decreased to 5|X% by 6 months. This activity was lower than that against the other cell lines, and was similar to that of PBL in short-term culture. Fluorescence-activated cell sorter (FACS) analysis demonstrated that in KB31 cells at 6 weeks, CD8+ cells were dominant, but CD56+ cells predominated at 6 months.
Conclusion: These results suggest that the presence of cytotoxic activity in the peripheral blood of the patient was due to both cytotoxic T cells and NK cells. The cytotoxic activity was lowest prior to surgery and increased postoperatively.     

干细胞向雄性配子诱导分化研究进展

人类雄性配子的重要功能之一是完整地将父代遗传信息传递给子代。配子发生,尤其雄性配子发生是个极其复杂的细胞分化过程,它包括有丝分裂与减数分裂两大过程。然而由于缺乏可重复、高效率研究雄性配子发生的体外培养体系,生殖细胞发生、发育机制研究进展缓慢。干细胞经体外诱导向雄性生殖细胞分化的研究,将推进生殖细胞发育研究,甚至生殖生物学的发展。本文综述近年原始生殖细胞体外培养及干细胞向雄性配子诱导分化的研究进展。     

睾丸Leydig细胞干细胞研究进展

Leyd ig细胞是机体合成和分泌雄激素的主要细胞,利用Leyd ig细胞干细胞的体外定向诱导分化扩增至具有雄激素合成功能的成熟细胞再植入机体中,对雄激素缺乏的治疗具有非常广阔的临床应用前景。     

肝干细胞来源的研究进展

从广义上讲,肝干细胞并非特指某一类细胞,而是与肝脏胚胎发育及再生有关的各类具有干细胞特性细胞类型的总称,是一种具有分化为成熟肝细胞和胆管细胞能力和自我更新能力的多潜能干细胞.根据其起源的不同,通常可分为肝源性肝干细胞和非肝源性肝干细胞.     

Involvement of apoptosis in the control of Sertoli and pre-meiotic germ cell numbers in the developing rabbit testis

The numbers of Sertoli and pre-meiotic germ cells in the developing rabbit testis were investigated as an initial step in determining the physiological meaning of the control of cell number in the testis by apoptosis. Sections were stained immunohistochemically for the detection of apoptotic cells and counterstained with haematoxylin. The number resulting from subtraction of the number of apoptotic cells from the total cell number was defined as the viable cell number. The number of viable premeiotic germ cells in the adluminal compartment of seminiferous tubules decreased during the pre-natal period, although neither apoptotic nor necrotic figures were detected. After birth, the numbers of total and apoptotic Sertoli and pre-meiotic germ cells increased, maintaining a stable ratio of their viable cell populations until the induction of meiosis. During induction of meiosis, the increase in the number of viable Sertoli cells was significantly accelerated because of the rapid decrease in the number of apoptotic Sertoli cells. Just after spermatids were formed the number of viable spermatogonia increased, reflecting an active supply of differentiated sper matogonia entering meiosis. In conclusion, apoptosis of Sertoli and pre-meiotic germ cells plays an important role in the acquisition of a suitable ratio of both cell types, and in providing intratubular environments for further progression of spermatogenesis, by controlling numbers of both cell types during the post-natal period.     

睾丸支持细胞骨架的研究

本文就睾丸支持细胞的骨架构造、功能以及研究进展进行了阐述。特别对近年来骨架中波形蛋白作用与功能进行了重点介绍。理化因素对细胞骨架的影响,特别是模拟微重力和空间真实微重力下细胞微丝骨架结构的变化进行介绍。首次提出了精液中可以检出支持细胞骨架,以抛砖引玉的愿望,引起同道进行探讨。     

大鼠肝窦内皮细胞分离、培养及其生物学特性的初步研究

目的　建立大鼠肝窦内皮细胞 (SEC)的原代分离、培养方法 ,并研究其生物学特性。方法　胶原酶灌注结合Percoll密度梯度离心加选择性贴壁的方法分离SEC ;用光镜、扫描电镜观察培养SEC的形态学变化及超微结构 ;用Ⅷ因子和CD14免疫细胞化学染色及RECA 1单抗间接免疫荧光法观察SEC表面分子的表达。结果　分离培养的SEC得率为 (2 .0 6± 0 .35 )× 10 7/只大鼠 ,活力≥ 92 % ,纯度达 90 % ;光镜下细胞形态典型 ,SEM下可观察到特征性的窗孔结构 ;Ⅷ因子染色阴性而CD14染色阳性 ,RECA 1单抗间接免疫荧光染色阳性。结论　分离培养的SEC细胞得率、活力及纯度较高 ,形态典型且具有一般细胞所没有的窗孔结构及表面标志 ,为其功能和作用机制的深入研究奠定了基础。     

Development of primordial germ cells in the mouse.

Primordial germ cells in the mouse are known to be derived from the epiblast. They can be identified histochemically, by their high alkaline phosphatase activity. At 8 d post coitum they have been observed within the embryonic part of the egg cylinder, at the posterior end of the primitive streak. Earlier, at 7.25 days post coitum, we have observed them embedded in the extra-embryonic mesoderm, as a tight clump. Germ cell counts over the 7-8 d period of gastrulation have been made. They are consistent with either of two models: (1) derivation of the germ cell lineage from a very small stem cell pool, followed by a constant rate of proliferation, and (2) derivation from a larger initial stem cell pool, followed by a period when germ cells are differentiating but not dividing. From their initial extra-embryonic location, germ cells spread into the mesoderm of the primitive streak, and the endoderm of the yolk sac and hind gut. Active locomotion is probably required for their passage up the dorsal mesentery and into the genital ridges. Mutant alleles at two loci, W (White-spotting) and Sl (Steel), drastically reduce the number of germ cells reaching the ridges. Since those that succeed in reaching the ridges suffer little if any delay, the defect is unlikely to be due to reduced powers of locomotion, but rather to a failure of proliferation or survival. W acts cell-autonomously: its gene product is the c-kit polypeptide, a transmembrane tyrosine kinase receptor.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of fatty acids and eicosanoid synthesis inhibitors on the growth of two human prostate cancer cell lines

Dietary fatty acids (FAs) may be involved in the carcinogenic process within the prostate gland and progression to clinically manifest disease. We have shown that growth of the androgen-unresponsive PC-3 human prostate cancer cell line is stimulated in vitro by the presence of linoleic acid (LA), an omega-6 polyunsaturated FA. The response was positively related to the FA concentration over the entire range examined (5-750 ng/ml). Conversely, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), two omega-3 FAs present in fish oils, inhibited PC-3 cell growth in a dose-dependent manner; both were equally effective, with an approximately 65% reduction in growth occurring at a concentration of 2.0 micrograms/ml (P less than 0.001). The DU 145 human prostate cancer cell line, which is also androgen-unresponsive, showed no growth response to LA and was less susceptible to growth inhibition when cultured in the presence of omega-3 FAs. Growth experiments with indomethacin, esculetin, and piroxicam, pharmacological inhibitors of eicosanoid biosynthesis with differing sites of action, indicated that human prostate cancer cell growth requires intact metabolic pathways for both leukotriene and prostaglandin production.