目的 从形态学上证实小鼠实验性肺纤维化上皮细胞-间充质细胞转变（EMT）及骨桥蛋白（OPN）的表达变化。方法 将60只健康雄性C57BLP6小鼠随机分为生理盐水对照组和博来霉素（BLM）模型组，采用口咽抽吸法经咽部注入气管50μl博来霉素溶液，对照组注入50μl生理盐水。分别在造模后第3d、7d、14d、21d及28d 5个时间点处死对照组及模型组小鼠。取小鼠左肺制备石蜡切片，行苏木素-伊红（HE）染色及天狼猩红染色观察肺组织形态变化及胶原增生情况;行表面活性物质相关蛋白-C（SP-C）、上皮型钙黏蛋白（E-Cadherin）、波形蛋白（vimentin）、成纤维细胞特异蛋白1（FSP1）和骨桥蛋白（OPN）免疫组织化学染色，观察肺纤维化上皮细胞-间充质细胞转变（EMT）形态与OPN表达，利用图像分析软件Image-Pro Plus 6.0（IPP6.0）测量切片积分吸光度（IA）并进行统计分析。结果 HE及天狼猩红染色显示出小鼠肺组织正常结构和纤维化形态变化，并且模型组小鼠肺组织的胶原生成量随时间推移而增加（P<0.05）。免疫组织化学结果显示,模型组小鼠肺OPN表达量随病程进展持续升高,在纤维化期达到峰值,而对照组表达量很少（P<0.05）；对照组SP-C在Ⅱ型肺泡细胞恒定表达，模型组SP-C持续增加，且胞体增大；对照组E-Cadherin表达量多，而模型组在肺实变部位几乎不表达，但肺实变部位产生波形蛋白及FSP1，这些蛋白提示肺纤维化存在EMT。结论 BLM诱导性肺纤维化中存在EMT，且与显著增加的OPN有潜在关系。

Osteoporosis is a disease of aging associated with bone loss that often occurs without symptoms until microarchitectural deterioration becomes so significant that bone fracture occurs. The effective microorganism-X (EM-X) is an antioxidant beverage derived from ferment of unpolished rice, sea weeds and papaya with effective microorganisms of lactic acid bacteria, yeast and photosynthetic bacteria (containing minerals, a-tocopherol, lycopene, ubiquinone, saponin and flavonoids). The levels of serum estradiol (E2) and the bone density of the middle and epiphysis of femurs were assessed in order to determine the effect of EM-X on osteoporosis in ovariectomized rat (an animal model of postmenopausal osteoporosis). EM-X (1ml/rat/day) was initially administrated by gavage to rats which were then allowed to consume 10% (v/v) EM-X in water freely for 3 months. There was no statistical significance of E2 level between sham operation group and control group, indicating that sham operation did not affect E2 level. However, the E2 levels in the ovariectomized rats tended to increase after treatment of EM-X for 3 months. The bone density of the middle and epiphysis of femur in both sham operation and ovariectomy group decreased with time. Rats receiving EM-X for 3 months after sham operation or ovariectomy had increased bone density of the middle of femur that was statistically significant (P<0.01 and P<0.05). The bone density of the epiphysis of femur in both sham operation and ovariectomy group were significantly increased, an outcome highly suggestive of the beneficial effects of EM-X on bone density of the middle and the epiphysis of femur in the rats with or without ovariectomy.

目的 研究丝裂原活化蛋白激酶（MAPKs）的3个亚家族成员细胞外信号调节激酶（ERK）、c-Jun N-端激酶（JNK）和p38 MAPK的活化形式在睾丸中的定位，了解MAPKs在生精过程中所起的作用。方法 免疫组织化学方法检测正常大鼠睾丸中磷酸化的p-ERK、JNK、p38MAPK的表达情况。结果 正常大鼠睾丸中p-ERK主要分布于精原细胞、细线前期到粗线期的初级精母细胞以及9～12期长形精子细胞的细胞核，p-JNK则主要位于支持细胞与支持细胞、支持细胞与生精细胞（尤其是19期精子细胞）之间，而p-p38 MAPK除了在生精小管的部分细胞胞质中有分布外,其表达最明显的部位是在间质细胞的细胞质。结论 ERK、JNK和p-38 MAPK分别定位于正常大鼠睾丸内的不同部位，提示MAPKs不同的亚家族成员分别在精子发生的不同环节中发挥主要作用。ERK可能参与生精细胞增殖、分化的信号转导,JNK则可能通过调节细胞的黏附而最终影响生精细胞的迁移与精子释放过程，而p38MAPK除了可能与JNK一起参与精子释放的调节外，最主要的作用可能是睾酮合成分泌的调节。

目的：探讨低剂量的棉酚12 mg/kg/day与甲基睾丸酮20 mg/kg/day和炔雌醇100 &micro;g/kg/day联合用药24周是否能够影响大鼠生精干细胞的功能。方法：采用生精细胞原位凋亡、生精干细胞移植和生殖能力自主恢复试验。结果:联合用药24周后在大鼠的睾丸组织切片中发现了凋亡信号阳性细胞，凋亡信号阳性细胞主要是精母细胞和精子细胞，没有发现精原细胞出现凋亡阳性信号。当把用药组大鼠的生精干细胞移植进入受体裸鼠的曲细精管内二至三个月后在受体裸鼠的曲细精管中观察到了来自供体大鼠的长型精子细胞。停止用药六周后用药组大鼠的生殖能力与对照组相比完全恢复。由恢复生殖能力的用药组大鼠产生的F1代仔鼠在经过一系列的检测后被证明为身体机能和智力水平正常。结论：通过以上的结果我们认为此联合用药在用药24周内没有影响到用药大鼠生精干细胞的生存和功能。

Background Myocardial fibrosis is a major component of ventricular remodeling after myocardial infarction (MI). The aim of the present study was to determine the outcome of transplantation into ischemic myocardium of bone marrow derived stem cell (MSC) on left ventricular (LV) function and remodeling, and to look closely at extracellular matrix gene expression. Methods and Results MI was induced by direct ligation of the left anterior descending coronary artery in rats, followed by MSC transplantation into the ischemic myocardium. Hemodynamic evaluations were performed at 7 and 28 days after coronary ligation. Changes in the mRNA expressions of collagen type I and type III, matrix metalloproteinase-1, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and transforming growth factor (TGF)-·1· were investigated using reverse transcription polymerase chain reaction and in situ hybridization after MI. Heart/body weight ratio in the MI+MSC group decreased after coronary ligation. However, LV systolic pressure and LV peak velocities of contraction and relaxation significantly increased compared with the MI group (p<0.01). There was marked up-regulation of the mRNA expressions of collagen types I and III, TIMP-1 and TGF-·1· in the MI rats, all of which were significantly attenuated by MSC transplantation. Conclusions MSC transplantation could inhibit LV remodeling, improve heart function and reduce the expression of extracellular matrix genes. (Circ J 2005; 69: 1275–1283)

目的 探索β-连环蛋白促进毛囊干细胞增殖作用的分子机制。方法 利用含有氨基端删除的β-连环蛋白的表达质粒转染毛囊细胞，观察转染细胞增殖，利用Westen blotting及原位杂交法观察其目的基因c-myc的表达。结果 转染△N90β-连环蛋白基因后细胞集落形成率增加，细胞K19阳性率亦有所增加；转染细胞c-myc的表达在蛋白质水平与mRNA水平均增加。结论 β-连环蛋白对毛囊细胞增殖分化的促进作用可能是通过促进c-myc基因的表达实现的。

目的 探讨敏乐定促进毛囊生长的机制。方法 选用无血清成人毛囊器官培养模型，体外加入不同浓度的敏乐定，测量毛囊生长速度。在不同时间点取培养毛囊，进行组织化学及免疫组织化学染色，检测I型胶原蛋白、基质金属蛋白酶-I（MMP-1）、转化生长因子区（TGFβ1）、整合素β1及K19的表达情况。结果 培养毛囊在前3d增长较快，而后期增长趋缓，1mg/L敏乐定对毛囊生长促进作用最明显。敏乐定组毛囊退化表现出现较晚，I型胶原蛋白含量较对照组多，MMP-I表达减少；TGFβ1表达较对照组减少；整合素β1和K19与对照组相比没有显著差异。结论 敏乐定能够延缓器官培养毛囊的退化期出现而延长生长期，促进毛囊的增长。敏乐定的这种作用可能是通过抑制TGFβ1的过度表达而实现的。

Objective The present study was to determine the effects of mesenchymal stem cell (MSC) transplantation into ischemic myocardium on left ventricular (LV) function and remodeling, especially the changes in extracellular matrix (ECM) and in transforming growth factor (TGF)-β1 after myocardial infarction (MI). Methods In this study, we investigated the changes in ECM involving collagen type I, collagen type III, matrix metalloproteinase (MMP)-1, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and in TGF-β1 mRNA expressions by using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization after MSC transplantation into infarcted myocardium in rats. Protein expressions were detected by immunohistochemistry and western blot. Seven and 28 days after coronary artery ligation, hemodynamic evaluations were performed. Results Heart : body weight ratio in the MI +MSC group decreased after MSC transplantation into infarcted myocardium compared with the MI group. However, LV systolic pressure (LVSP) and LV peak velocities of contraction and relaxation (+ dP/dtmax and –dP/dtmax) significantly increased compared with the MI group (P < 0.01). There was marked up-regulation of mRNA transcription for ECM and TGF-β1 proteins in MI rats. Immunohistochemical and western blot studies confirmed that the expressions of these ECM and TGF-β1 proteins were significantly increased in the MI group. The mRNA and protein expressions of collagen type I, collagen type III, TIMP-1 and TGF-b1 were all decreased by MSC transplantation in our study, although no significant difference was found for the mRNA and protein expressions of MMP-1 between the MI group and the MI+MSC group. Conclusions Our results demonstrated that MSC transplantation could inhibit LV remodeling and improve heart function. Coron Artery Dis 16: 245-255

目的 寻找一种快速简便的分离毛囊干细胞的方法，探讨β-连环蛋白（β-catenin）信号分子在毛囊干细胞增殖分化中的调节作用。方法 利用毛囊干细胞黏附性强的特点，通过将毛囊细胞悬液黏附于包被有人胶原的平板而使富毛囊干细胞分离。利用毛囊干细胞标志物Keratin 19（K19）及体外集落形成能力进行鉴定。采用免疫细胞化学及原位杂交技术检测分选后的毛囊干细胞β-catenin的表达情况。结果 毛囊干细胞悬液黏附于人胶原20min后，K19的表达增加1.6倍，集落形成能力增加3.5倍。β-catenin在细胞核中的蛋白表达增加。结论 快速黏附法可以使富集毛囊干细胞分离，β-catenin参与了毛囊干细胞的增殖及分化调控。