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目的探讨转化生长因子-β1（TGF-β1）抗体对体外培养的翼状胬肉成纤维细胞增殖的抑制作用及对TGF-β1-Smad4信号转导通路的影响,评估其在翼状胬肉发病机制中的作用。方法人翼状胬肉切除后,体外培养其成纤维细胞并传至4~6代时,分别用6.25、12.5、25、50mg/mL TGF-β1抗体处理24、48、72h,采用MTT法检测成纤维细胞的生长抑制率,RT-PCR检测TGF-β1mRNA表达,Western blot检测Smad4蛋白的表达。所得数据采用SPSS19.0统计软件处理。结果 TGF-β1抗体可不同程度抑制成纤维细胞的增殖;TGF-β1抗体处理组TGF-β1mRNA的表达明显低于对照组,且不同浓度组间差异有统计学意义（P〈0.05）,随着浓度增大和作用时间延长,TGF-β1抗体处理组TGF-β1 mRNA的表达量呈逐渐降低趋势。不同浓度的TGF-β1抗体对Smad4的蛋白表达具有明显增强作用,且各组间差异有统计学意义（P〈0.05）。结论 TGF-β1抗体对人翼状胬肉体外培养的成纤维细胞生长具有明显抑制作用,其可能通过调节TGF-β1和Smad4的表达而实现。

目的 对人头皮游离毛囊（HF）切片制作及凋亡检测方法进行比较。方法 分别将石蜡切片法和冰冻切片法应用于游离毛囊；利用原位缺口末端标记法（TUNEL）进行荧光染色与3，3＇-二氨基联苯胺（DAB）染色，检测未经培养的游离毛囊及培养5d的游离毛囊的不同部位凋亡细胞情况。结果 在对游离毛囊形态检测中，各组毛囊在外根鞘（ORS），毛球（HB）下部，真皮鞘（DS）及毛乳头（DP）中的凋亡细胞数并无显著性差异（P〉0．05）。结论 冰冻切片法应用于游离毛囊简单有效，TUNEL原位荧光结合DAB染色适合于检测毛囊细胞凋亡情况。

目的 观察神经生长因子(nerve growth factor，NGF)对离体培养条件下的人头皮毛囊生长的影响。方法 在离体人头皮毛囊培养模型中，加入100μg·L^-1的NGF，测量毛囊长度的变化以及24h DNA合成率。结果 目镜测微器观测100μg·L^-1的NGF与125mg·L^-1的米诺地尔一样能明显地促进游离的人头皮毛囊生长(P<0．05)，尤以前8d较为明显；^3H—TdR掺入检测的DNA合成率亦明显增高。结论 100μg·L^-1的NGF对游离的人头皮毛囊的生长有促进作用。

目的 了解丹参酮对人肺癌细胞（SPC-A-1）的凋亡诱导作用，并探讨其可能的分子机理，方法 体外培养的SPC-A-1细胞经无毒剂量的丹参酮ⅡA（0.5mg/L)处理5天，光镜，电镜观察细胞凋亡情况，流式细胞仪检测凋亡指数及凋亡相关基因的蛋白表达，并以全反式维甲酸及顺铂作为对照。结果 SPC-A-1细胞经丹参酮ⅡA处理后，电镜观察可见多量凋亡细胞，流式细胞仪检测；丹参酮组凋亡指数显著高于顺铂组及对照组，而与维甲酸组比较无显著差异。丹参酮组促凋亡基因p53,Fas,Bax表达水平明显升高，而凋亡抑制基因Bcl-2的表达水平则显著降低。结论 丹参酮ⅡA具有诱导肺癌SPC-A-1细胞凋亡的作用。其分子机理可能是上调p53,Bax,Fas及下调Bcl-2的表达。

目的 通过研究氟对牙胚发育早期骨形成蛋白(BMP)表达的影响，初步探讨氟在早期牙胚发育中的可能作用机制。方法 4个月的人胚胎，取乳牙胚用RPMI 1640培养液进行器官培养8d。免疫组化法研究25mg·L-1和50mg·L-1的氟对分泌前期牙胚的影响，观察培养第2、4、6、8天时BMP表达的变化。采用图像分析仪对免疫组化染色结果进行灰度分析。结果 MBP的表达主要在造釉器。成釉细胞，中间层细胞和星网状层细胞均有BMP表达，牙乳头细胞或成牙本质细胞不表达BMP。25mg·L-1氟时从培养第6天开始BMP表达增强；50mg·L-1氟时从培养第2天到第6天BMP的表达增强，培养第8天时BMP的表达降低。结论 氟可能通过促进BMP某一亚型或某几个亚型的表达而在牙胚发育中起作用。

Muscle stem cells （MuSCs, satellite cells） are the major contributor to muscle regeneration. Like most adult stem cells, long-term expansion of MuSCs in vitro is difficult. The in vivo muscle regeneration abilities of MuSCs are quickly lost after culturing in vitro, which prevents the potential applications of MuSCs in cell-based therapies. Here, we establish a system to serially expand MuSCs in vitro for over 20 passages by mimicking the endogenous microen- vironment. We identified that the combination of four pro-inflammatory cytoklnes, IL-1α, IL-13, TNF-α, and IFN-γ, secreted by T cells was able to stimulate MuSC proliferation in vivo upon injury and promote serial expansion of MuSCs in vitro. The expanded MuSCs can replenish the endogenous stem cell pool and are capable of repairing mul- tiple rounds of muscle injuries in vivo after a single transplantation. The establishment of the in vitro system provides us a powerful method to expand functional MuSCs to repair muscle injuries.

Various liver diseases result in terminal hepatic failure, and liver transplantation, cell transplantation and artificial liver support systems are emerging as effective therapies for severe hepatic disease. However, all of these treatments are limited by organ or cell resources, so developing a sufficient number of functional hepatocytes for liver regeneration is a priority. Liver regeneration is a complex process regulated by growth factors （GFs）, cytokines, transcription factors （TFs）, hormones, oxidative stress products, metabolic net- works, and microRNA. It is well-known that the function of isolated primary hepatocytes is hard to maintain; when cultured in vitro, these cells readily undergo dedifferentia- tion, causing them to lose hepatocyte function. For this mason, most studies focus on inducing stem cells, such as embryonic stem cells （ESCs）, induced pluripotent stem cells （iPSCs）, hepatic progenitor cells （HPCs）, and mesenchymal stem cells （MSCs）, to differentiate into hepatocyte-like cells （HLCs） in vitro. In this review, we mainly focus on the nature of the liver regeneration process and discuss how to main- tain and enhance in vitro hepatic function of isolated primary hepatocytes or stem cell-derived HLCs for liver regenera- tion. In this way, hepatocytes or HLCs may be applied for clinical use for the treatment of terminal liver diseases and may prolong the survival time of patients in the near future.

The potential of spermatozoa to become motile during post-testicular maturation, and the relationship between the cytoplasmic droplet and fertilizing capacity are reviewed. Post-testicular maturation of spermatozoa involves the autonomous induction of motility, which can occur in vivo in testes with occluded excurrent ducts and in vitro in testicular explants, and artefactual changes in morphology that appear to occur in the testis in vitro. Both modifications may reflect time-dependent oxidation of disulphide bonds of head and tail proteins. Regulatory volume decrease （RVD）, which counters sperm swelling at ejaculation, is discussed in relation to loss of cytoplasmic droplets and consequences for fertility. It is postulated that： （i） fertile males possess spermatozoa with sufficient osmolytes to drive RVD at ejaculation, permitting the droplet to round up and pinch off without membrane rupture; and （ii） infertile males possess spermatozoa with insufficient osmolytes so that RVD is inadequate, the droplet swells and the resulting flagellar angulation prevents droplet loss. Droplet retention at ejaculation is a harbinger of infertility caused by failure of the spermatozoon to negotiate the uterotubal junction or mucous and reach the egg. In this hypothesis, the epididymis regulates fertility indirectly by the extent of osmolyte provision to spermatozoa, which influences RVD and therefore droplet loss. Man is an exception, because ejaculated human spermatozoa retain their droplets. This may reflect their short midpiece, approximating head length, permitting a swollen droplet to extend along the entire midpiece; this not only obviates droplet migration and flagellar angulation but also hampers droplet loss.

Aim： Sulforaphane （SFN）, a natural dietary isothiocyanate, is found to exert beneficial effects for cardiovascular diseases. This study aimed to investigate the mechanisms underlying the protective effects of SFN in a model of myocardial hypoxia/reoxygenation （H/R） inj u ry in vitro. Methods： Cultured neonatal rat cardiomyocytes pretreated with SFN were subjected to 3-h hypoxia followed by 3-h reoxygenation. Cell viability and apoptosis were detected. Caspase-3 activity and mitochondrial membrane potential （△ψm） was measured. The expression of ER stress-related apoptotic proteins were analyzed with Western blot analyses. Silent information regulator 1 （SIRT1） activity was determined with SIRT1 deacetylase fluorometric assay kit. Results： SFN （0.1-5 pmol/L） dose-dependently improved the viability of cardiomyocytes, diminished apoptotic cells and suppressed caspase-3 activity. Meanwhile, SFN significantly alleviated the damage of △ψm and decreased the expression of ER stress-related apoptosis proteins （GRP78, CHOP and caspase-12）, elevating the expression of SIRT1 and Bcl-2/Bax ratio in the cardiomyocytes. Co-treatment of the cardiomyocytes with the SIRTl-specific inhibitor Ex-527 （1 μmol/L） blocked the SFN-Jnduced cardioprotective effects.Conclusion： SFN prevents cardiomyocytes from H/R injury in vitro most likely via activating SIRT1 pathway and subsequently inhibiting the ER stress-dependent apoptosis.

Aim： Honokiol （HNK） is a natural compound isolated from the magnolia plant with numerous pharmacological activities, including inhibiting epithelial-mesenchymal transition （EMT）, which has been proposed as an attractive target for anti-tumor drugs to prevent tumor migration. In this study we investigated the effects of HNK on EMT in human NSCLC cells in vitro and the related signaling mechanisms. Methods： TNF-α （25 ng/mL） in combination with TGF-β4 （5 ng/mL） was used to stimulate EMT of human NSCLC A549 and H460 cells. Cell proliferation was analyzed using a sulforhodamine B assay. A wound-healing assay and a transwell assay were performed to examine cell motility. Western blotting was used to detect the expression levels of relevant proteins, siRNAs were used to knock down the gene expression of c-FLIP and N-cadherin. Stable overexpression of c-FLIP L （H157-FLIP L） or Lac Z （H157-Lac Z） was also performed.Results： Treatment with TNF-α＋TGF-β1 significantly enhanced the migration of A549 and H460 ceils, increased c-FLIP, N-cadherin （a mesenchymal marker）, snail （a transcriptional modulator） and p-Smad2/3 expression, and decreased IKB levels in the cells; these changes were abrogated by co-treatment with HNK （30 pmol/L）. Further studies demonstrated that expression level of c-FLIP was highly correlated with the movement and migration of NSCLC cells, and the downstream effectors of c-FLIP signaling were NF-κB signaling and N-cadherin/snail signaling, while Smad signaling might lie upstream of c-FLIP. Conclusion： HNK inhibits EMT-mediated motility and migration of human NSCLC cells in vitro by targeting c-FLIP, which can be utilized as a promising target for cancer therapy, while HNK may become a potential anti-metastasis drug or lead compound.