染色体平衡易位46,XY,t（4;7）（q21;p15）致生长激素缺乏症的临床综合分析

目的探讨平衡易位携带者对子代的影响。方法患者及家系成员外周血淋巴细胞常规培养制备染色体,进行核型分析。对患者进行复合刺激试验,采用化学发光法检测生长激素（GH）、促卵泡生成素（FSH）和促黄体生成激素（LH）水平,同时检测其他代谢相关生化指标及相关影象检查。结果患者核型为46,XY,t（4;7）（q21;p15）,患者母亲核型为46,XX,t（4;7）（q21;p15）,其他成员核型正常。患者复合刺激试验示GH分泌反应低下,FSH、LH反应正常,血清胰岛素样生长因子-I（IGF-1）、胰岛素样生长因子结合蛋白-3（IGFBP-3）、促肾上腺皮质激素（ACTH）、硫酸去氢表雄酮（DHEA-S）、总睾酮（T）低于参考范围的下限。骨龄11岁,垂体MRI排除脑组织异常。结论患者的异常表型是由于生长激素缺乏所致,常染色体平衡易位可致子代生长激素缺乏症。     

Injectable biodegradable hydrogel composites for rabbit marrow mesenchymal stem cell and growth factor delivery for cartilage tissue engineering

We investigated the development of an injectable, biodegradable hydrogel composite of oligo(poly(ethylene glycol) fumarate) (OPF) with encapsulated rabbit marrow mesenchymal stem cells (MSCs) and gelatin microparticles (MPs) loaded with transforming growth factor-beta1 (TGF-beta1) for cartilage tissue engineering applications. Rabbit MSCs and TGF-beta1-loaded MPs were mixed with OPF, a poly(ethylene glycol)-diacrylate crosslinker and the radical initiators ammonium persulfate and N,N,N',N'-tetramethylethylenediamine, and then crosslinked at 37 degrees C for 8 min to form hydrogel composites. Three studies were conducted over 14 days in order to examine the effects of: (1) the composite formulation, (2) the MSC seeding density, and (3) the TGF-beta1 concentration on the chondrogenic differentiation of encapsulated rabbit MSCs. Bioassay results showed no significant difference in DNA amount between groups, however, groups with MPs had a significant increase in glycosaminoglycan content per DNA starting at day 7 as compared to controls at day 0. Chondrocyte-specific gene expression of type II collagen and aggrecan were only evident in groups containing TGF-beta1-loaded MPs and varied with TGF-beta1 concentration in a dose-dependent manner. Specifically, type II collagen gene expression exhibited a 161+/-49-fold increase and aggrecan gene expression a 221+/-151-fold increase after 14 days with the highest dose of TGF-beta1 (16 ng/ml). These results indicate that encapsulated rabbit MSCs remained viable over the culture period and differentiated into chondrocyte-like cells, thus suggesting the potential of OPF composite hydrogels as part of a novel strategy for localized delivery of stem cells and bioactive molecules.     

Oxidized alginate hydrogels for bone morphogenetic protein-2 delivery in long bone defects

Autograft treatment of large bone defects and fracture non-unions is complicated by limited tissue availability and donor site morbidity. Polymeric biomaterials such as alginate hydrogels provide an attractive tissue engineering alternative due to their biocompatibility, injectability, and tunable degradation rates. Irradiated RGD-alginate hydrogels have been used to deliver proteins such as bone morphogenetic protein-2 (BMP-2), to promote bone regeneration and restoration of function in a critically sized rat femoral defect model. However, slow degradation of irradiated alginate hydrogels may impede integration and remodeling of the regenerated bone to its native architecture. Oxidation of alginate has been used to promote degradation of alginate matrices. The objective of this study was to evaluate the effects of alginate oxidation on BMP-2 release and bone regeneration. We hypothesized that oxidized-irradiated alginate hydrogels would elicit an accelerated release of BMP-2, but degrade faster in vivo, facilitating the formation of higher quality, more mature bone compared to irradiated alginate. Indeed, oxidation of irradiated alginate did accelerate in vitro BMP-2 release. Notably, the BMP-2 retained within both constructs was bioactive at 26 days, as observed by induction of alkaline phosphatase activity and positive Alizarin Red S staining of MC3T3-E1 cells. From the in vivo study, robust bone regeneration was observed in both groups through 12 weeks by radiography, micro-computed tomography analyses, and biomechanical testing. Bone mineral density was significantly greater for the oxidized-irradiated alginate group at 8 weeks. Histological analyses of bone defects revealed enhanced degradation of oxidized-irradiated alginate and suggested the presence of more mature bone after 12 weeks of healing.     

Differential effects of growth differentiation factor-5 on porcine dental papilla- and follicle-derived cells

In this study, the effect of growth differentiation factor-5 (GDF-5) on the growth and differentiation of porcine dental papilla- and follicle-derived cells was investigated. Furthermore, the effect was compared with that of BMP-2. Recombinant mouse GDF-5 (rmGDF-5) enhanced alkaline phosphatase (ALP) activity in dental papilla-derived cells in a dose-dependent manner, while ALP activity in dental follicle-derived cells was reduced. In rmGDF-5 stimulated dental papilla-derived cells, the expressions of odontoblast-marker genes were up-regulated. Conversely, recombinant human BMP-2 (rhBMP-2) enhanced ALP activity dose-dependently in both dental papilla- and follicle-derived cells. When combined, GDF-5 did not further enhance BMP-2-induced ALP activities. Rather, GDF-5 reduced BMP-2-induced ALP activities in both dental papilla- and follicle-derived cells. This suggests that affinity of GDF-5 to the shared receptors may be higher than that of BMP-2 in both cell types. These observations indicate that GDF-5 regulates differentiation of both dental papilla and follicle during odontogenesis, co-operatively with other growth factors such as BMP-2.     

Effects of insulin-like growth factor 2 and its receptor expressions on corneal repair

Limbal stem cell (LSC) on the basal layer of cornea plays an important role in the epithelial repair after corneal injury as it can proliferate, differentiate and migrate into injury sites under the direction of cytokines. This study explored the signaling pathway and cellular mechanism between corneal epithelial cells LSC, on a mouse model with mechanic corneal injury. Ipsilateral corneal mechanic injury model was prepared on mice using the contralateral eye as the control. Tissues from both central and peripheral regions of cornea were collected, cultured and quantified for expression of various cytokines including epidermal growth factor (EGF), fibroblast growth factor-β (FGF-β), heparin-like growth factor (HGF), keratinocyte growth factor (KGF), transforming growth factor-β1 (TGF-β1), IGF-1 and IGF-2. The effects of these factors on the differentiation of LSC and fibroblasts were also studied. Most of those cytokines had elevated gene expressions after the corneal injury. Among those IGF-2 had significantly increased expression, along with the high expression of IGF-2 receptor in corneal peripheral cells. IGF-2 also induced the differentiation of LSC into keratin-12-positive cells. Further studies showed the prominent expression of α-actin in injured tissues, suggesting the potential transformation of fibroblasts into myofibroblasts. Both IGF-2 and its receptor had elevated expressions after corneal injury. They may facilitate the transformation of LSC into epithelial cells, in addition to the role in transformation from fibroblasts to myofibroblasts.     

转化生长因子β和骨形成蛋白2联合诱导小鼠成骨细胞系MC3T3-E1细胞的增殖与分化

背景:生长因子是骨组织形成、改建、修复的关键要素之一,多种生长因子联合诱导骨干细胞成骨分化相较于单一生长因子具有一定的优势,但是何种类型的联合、什么样的浓度组合才能发挥最佳的诱导作用?目前该方向研究较少.目的:探索转化生长因子β和骨形成蛋白2联合诱导对小鼠MC3T3-E1细胞增殖和分化的影响.方法:将不同质量浓度的转化...     

Immunohistochemical detection of bone morphogenetic protein-2 and transforming growth factor beta-1 in tracheopathia osteochondroplastica

 Tracheopathia osteochondroplastica (TO) is an unusual condition characterized by cartilaginous or bony submucosal nodules in the tracheobronchial tree. Bone morphogenetic protein-2 (BMP-2) and transforming growth factor beta-1 (TGF-β1) are potent inducers for new bone formation. We studied the precise localization of BMP-2 and TGF-β1 in two autopsied cases of TO, using immunohistochemical methods. Positive BMP-2 immunoreactivity was detected in numerous mesenchymal cells and chondroblasts lining the nodules in the tracheal submucosa. BMP-2 was not found in mature lamellar bony nodules. TGF-β1 was not seen in mesenchymal cells, though it did appear in chondrocytes and osteocytes in the nodules. These results suggest that BMP-2 plays an important role in nodule formation and acts synergistically with TGF-β1 to promote the nodules inductive cascade in the tracheal submucosa. Received: 9 December 1996/Accepted: 10 April 1997     

Controllable mineral coatings on PCL scaffolds as carriers for growth factor release

In this study, we have developed mineral coatings on polycaprolactone scaffolds to serve as templates for growth factor binding and release. Mineral coatings were formed using a biomimetic approach that consisted in the incubation of scaffolds in modified simulated body fluids (mSBF). To modulate the properties of the mineral coating, which we hypothesized would dictate growth factor release, we used carbonate (HCO₃) concentration in mSBF of 4.2 mm, 25 mm, and 100 mm. Analysis of the mineral coatings formed using scanning electron microscopy indicated growth of a continuous layer of mineral with different morphologies. X-ray diffraction analysis showed peaks associated with hydroxyapatite, the major inorganic constituent of human bone tissue in coatings formed in all HCO₃ concentrations. Mineral coatings with increased HCO₃ substitution showed more rapid dissolution kinetics in an environment deficient in calcium and phosphate but showed re-precipitation in an environment with the aforementioned ions. The mineral coating provided an effective mechanism for growth factor binding and release. Peptide versions of vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP2) were bound with efficiencies up to 90% to mineral mineral-coated PCL scaffolds. We also demonstrated sustained release of all growth factors with release kinetics that were strongly dependent in the solubility of the mineral coating.     

全身应用神经生长因子对大鼠胫骨干骨折早期愈合作用及骨形态发生蛋白2和血管内皮生长因子表达的影响

文题释义：神经生长因子：神经生长因子家族包括神经生长因子、脑源性神经营养因子、神经营养素3、神经营养素4/5、神经营养素6和神经营养素7等,主要为前4种。神经生长因子为神经生长因子家族中最主要的一类,在组织中主要以前体的形式存在,在颌下腺中加工形成成熟的神经生长因子,能促进中枢和外周神经元的生长、发育、分化、成熟,维持神经系统的正常功能,加快神经系统损伤后的修复。神经生长因子广泛分布于机体各组织器官中(包括脑),在靶组织中的浓度与交感神经和感觉神经在靶区分支的密度和mRNA的含量有关。骨折愈合：是一个复杂而连续的过程,从组织学和细胞学的变化,通常将其分为3个阶段,但三者之间又不可截然分开,而是互相交织逐渐演进,包括血肿炎症机化期、原始骨痂形成期、骨板形成塑形期。背景：骨折愈合是一种十分复杂的多阶段生理过程,由多种细胞协调参与。有关神经因素对于骨折愈合过程的影响逐渐被重视和深入研究,神经生长因子是一种能够促进细胞生长的生物活性复合蛋白,其促进骨折愈合过程中所发挥的作用机制尚不明确。 目的：观察神经生长因子对大鼠胫骨骨折愈合的促进作用及愈合过程中机制。 方法：将72只雄性SD大鼠随机分为模型组和神经生长因子组,构建胫骨干闭合骨折模型并应用髓内固定。神经生长因子组给予肌肉注射,注射用鼠神经生长因子;模型组给予肌肉注射生理盐水。实验于2018-05-30经福建医科大学实验动物伦理委员会批准,批准号：FJYKDX-2018-035。结果与结论：①干预2周和4周时,神经生长因子组骨痂体积显著高于模型组,干预6周时,神经生长因子组骨折线消失,髓腔再通,骨折早于模型组达到完全愈合;②苏木精-伊红染色显示干预2周和4周时,神经生长因子组骨小梁生长致密,可见大量成骨细胞生长;③干预2周和4周时,神经生长因子组血清碱性磷酸酶含量明显高于模型组;④免疫组化结果显示,干预2周和4周时,神经生长因子组骨痂内骨形态发生蛋白2和血管内皮生长因子阳性表达均显著多于模型组;⑤定量PCR结果显示,干预2周和4周时,神经生长因子组骨形态发生蛋白2和血管内皮生长因子 mRNA表达水平均高于模型组;⑥提示全身应用神经生长因子可增强大鼠胫骨骨折端成骨能力,促进骨折早期愈合;同时神经生长因子可促进骨形态发生蛋白2和血管内皮生长因子的表达,并在不同时期这种促进作用表现出一定的规律,进而诱导成骨细胞分化增殖及细胞外基质的形成和钙化,促进骨形成及骨愈合。ORCID: 0000-0002-8369-5711(刘国铭) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Immunolocalization of transforming growth factor-beta1, connective tissue growth factor,phosphorylated-SMAD2/3, and phosphorylated-ERK1/2 during mouse incisor development

Background/aims: Connective tissue growth factor (CTGF) is a downstream mediator of transforming growth factor-beta 1 (TGF-β₁) and TGF-β₁-induced CTGF expression is regulated through SMAD and mitogen-activated protein kinase (MAPK) signaling pathways. However, little is known about the localization of CTGF and TGF-β₁ signaling cascades during incisor development. Therefore, we aimed to investigate the distribution pattern of TGF-β₁, CTGF, phosphorylated-SMAD2/3 (p-SMAD2/3), and phosphorylated-ERK1/2 (p-ERK1/2) in the developing mouse incisors.

Materials and methods: ICR mice heads of embryonic (E) day 16.5, postnatal (PN) day 0.5 and PN3.5 were processed for immunohistochemistry.

Results: From E16.5 to PN3.5, moderate to strong staining for TGF-β₁ and CTGF was localized in stellate reticulum (SR), transit amplifying (TA) cells, outer enamel epithelium (OEE), preameloblasts (PA), preodontoblasts (PO), and dental papilla (DP). p-SMAD2/3 was weakly positive in SR and OEE at E16.5 and PN0.5 but was strongly positive in SR and OEE at PN3.5. Particularly, in the stem cell niche, p-SMAD2/3 was only localized in SR cells adjacent to OEE. There was no staining for p-SMAD2/3 in TA cells, PA and PO, although weak to moderate staining for p-SMAD2/3 was seen in DP. From E16.5 to PN3.5, p-ERK1/2 was negative in TA cells, OEE, PA and PO, whereas weak to moderate staining for p-ERK1/2 was observed in SR. DP was moderately stained for p-ERK1/2.

Conclusions: TGF-β₁ and CTGF show a similar expression, while p-SMAD2/3 and p-ERK1/2 exhibit differential distribution pattern, which indicates that CTGF and TGF-β₁ signaling cascades might play a regulatory role in incisor development.     

胰岛素样生长因子-1通过细胞外信号调节激酶1/2通路下调转录因子基本转录元件结合蛋白表达抗心肌细胞凋亡

目的 探讨胰岛素样生长因子-1(IGF-1)对大鼠心肌细胞凋亡保护作用的基因调控机制。方法 体外培养新生大鼠心肌细胞,10nmol/L IGF-1刺激的同时,分别加入磷脂酰肌醇-3激酶（PI3K）、细胞外信号调节激酶（ERK）1/2和Raf-1 3条通路抑制剂（20μmol/L）,通过RT-PCR及Western blotting方法观察IGF-1调节基本转录元件结合蛋白（BTEB）的基因表达及其通路调控。100μmol/L H2O2处理诱导心肌细胞凋亡,通过DNA梯度分析、Annexin V-FITC/PI双染色法、Caspase-3活性测定、Hoechest33258染色法观察用BTEB特异性siRNA人为下调BTEB基因表达后对心肌细胞凋亡的影响。结果 大鼠心肌细胞经IGF-1刺激60min后,BTEB mRNA和蛋白表达均明显下降;与对照组相比,加入ERK1/2通路抑制剂PD98059组BTEB的mRNA和蛋白表达均明显增高（P＜0.01）;H2O2诱导的大鼠心肌细胞于下调BTEB表达后,DNA片段化改善,心肌细胞凋亡率下降（P＜0.05）,Caspase-3活性降低（P＜0.05）,凋亡小体减少,与IGF-1的抗心肌细胞凋亡效果相似。
结论 IGF-1可以通过ERK1/2通路下调转录因子BTEB基因表达而发挥抗心肌细胞凋亡的作用。     

不同组织部位诱导膜血管化及成骨因子表达的比较

文题释义 诱导膜技术：又称膜诱导技术、Masquelet技术,1986年由Masquelet等最先提出并使用,是一种重建超临界尺寸骨缺损的有效方法。该技术在清创后所形成的骨缺损处植入骨水泥占位器,以形成一层生物膜,并在所形成的膜内进行植骨对骨缺损进行二次重建。该技术修复大段骨缺损具有重建时间短、骨愈合率高、并发症发生率较低等优势,其疗效已经被广泛证实。 血管内皮生长因子：是一种生长因子,在血管生成中发挥重要作用。血管内皮生长因子与血管内皮表面的特异性受体结合,可以促进内皮细胞的增殖、迁移,并可增加局部毛细血管的通透性,促使纤维蛋白原渗出,为血管内皮细胞的迁移和血管形成提供基质。在促进骨愈合方面,血管内皮生长因子通过促进血管增生,协调软骨细胞的消长、细胞外基质的改建,加快软骨内成骨。血管内皮生长因子还可以趋化并促进成骨细胞的分化,使碱性磷酸酶活性增强,局部钙盐沉积增加,促进骨愈合。 背景：研究发现在皮下、肌肉等部位植入聚甲基丙烯酸甲酯骨水泥也可形成诱导膜,膜内同样具有微血管的形成并分泌多种成骨因子。 目的：比较皮下、肌肉、股骨骨缺损处诱导膜内血管化程度和成骨因子表达的差异。 方法：将36只雄性SD大鼠(购自广州中医药大学实验动物中心)随机分为3组,分别在后肢皮下、肌肉内、股骨骨缺损处植入聚甲基丙烯酸甲酯抗生素骨水泥占位器,每组12只。植入6周后,取出骨水泥周围的诱导膜,苏木精-伊红染色观察诱导膜组织形态结构变化,Western Blot印迹方法、RT-qPCR法、免疫组织化学染色法检测诱导膜中骨形态发生蛋白2、转化生长因子β1、血管内皮生长因子的表达情况。实验获得广州中医药大学动物实验伦理委员会批准,批准号：20181101006。 结果与结论：①苏木精-伊红染色显示3组均可形成诱导膜,但骨缺损组诱导膜组织切片外层血管数量较肌肉组、皮下组多,靠近骨水泥侧的内层成纤维细胞和肌纤维细胞数量较肌肉组、皮下组多;②免疫组织化学染色显示,骨缺损组骨形态发生蛋白2、转化生长因子β1、血管内皮生长因子阳性表达最多,皮下组最少;③Western Blot与RT-qPCR检测显示,骨缺损组骨形态发生蛋白2、转化生长因子β1、血管内皮生长因子表达均多于肌肉组、皮下组(P < 0.001);④结果表明,不同的周围组织条件对诱导膜的组织结构和成骨因子表达有重要影响,在骨缺损处植入聚甲基丙烯酸甲酯骨水泥可提高诱导膜的形成质量,膜内新生血管更丰富,骨生长因子的表达量更多。ORCID: 0000-0003-1405-0765(李树源) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

重组人骨形态发生蛋白2诱导骨修复的应用及前景

背景：重组人骨形态发生蛋白2具备诱导成骨时间更早、成骨量较天然骨形态发生蛋白2多、生物学活性好、生物相容性好、成本低等特点,已成为近年来临床骨科创伤疾病防治研究的热点。 目的：总结重组人骨形态发生蛋白2在骨组织工程及骨修复领域应用中的优势、不足及目前国内外的研究进展。 方法：经第一作者检索CNKI数据库及SPRINGERLINK数据库2005至2011年与重组人骨形态发生蛋白2在诱导骨再生、骨组织修复有关研究进展方面的文献,英文检索词为“rhBMP-2,bone tissue engineering,bone repair materials”,中文检索词为“重组人骨形态发生蛋白2,骨组织工程,骨修复”。共检索出98篇,最终保留30篇进行归纳总结。 结果与结论：骨骼内天然骨形态发生蛋白2含量稀少、提取成本高昂,临床应用严重受限。重组人骨形态发生蛋白2有显著的成骨诱导能力,在骨组织工程及骨修复领域展现了巨大的潜在应用价值。体外试验无细胞毒性具有良好的生物相容性可供临床应用,其中重组人骨形态发生蛋白2和重组人骨形态发生蛋白7现已被应用于外科整形手术诱导骨再生,但由于重组人骨形态发生蛋白2是外源性细胞生长因子,临床应用多为超生理剂量,故潜在有软组织水肿,皮肤红疹、局部炎症反应、异位骨化和免疫反应等不良后果的危险,所以重组人骨形态发生蛋白2用于人体后的安全性研究还须长期密切关注。找到理想的载体,有效控制其在体内缓释是重组人骨形态发生蛋白2应用研究的关键问题。     

骨水泥间隔器表面微观形貌改变对诱导膜内成骨因子表达的影响

文题释义： 聚甲基丙烯酸甲酯骨水泥：为临床上常用的骨科植入材料,具有很大的抗压能力,抗压缩能力为97 MPa,也具有良好的生物相容性。1987年Galibert等首次报道用聚甲基丙烯酸甲酯骨水泥来治疗椎体血管瘤,并将此技术称之为经皮椎体成形。1970年Buchholz首次应用载抗生素骨水泥控制关节感染,目前以聚甲基丙烯酸甲酯骨水泥作为抗生素的缓释载体被广泛应用于人工全髋关节置换及骨髓炎的治疗。 转化生长因子β1：是具有多种功能的蛋白多肽,是“转化生长因子β超家族”成员之一,大量存在于骨组织与血小板中,可以刺激间充质干细胞的增殖、分化,并抑制间充质干细胞向脂肪细胞分化,也促进成骨细胞、成软骨细胞的增殖及细胞外基质的合成,诱导膜内成骨和软骨内成骨。 背景：目前国内外学者试图通过改变植入材料的种类和形貌、改良诱导膜厚度、光滑程度等机械化学性能来促进植骨生长。 目的：比较大鼠股骨骨缺损处不同表面粗糙程度聚甲基丙烯酸甲酯骨水泥形成的诱导膜在膜内血管化程度和部分成骨因子表达的差异。 方法：取48只雄性SD大鼠(购自广州中医药大学实验动物中心)建立大鼠临界尺寸股骨缺损模型,按随机数字表法分为A、B、C、D组,分别在股骨骨缺损处植入表面粗糙度<1.5 µm、1.5-2.0 µm、5.0-7.0 µm、14.0-20.0 µm的聚甲基丙烯酸甲酯骨水泥占位器。植入6周大鼠体内诱导膜形成后取出骨水泥周围的诱导膜,苏木精-伊红染色观察诱导膜病理组织形态结构变化,采用Western Blot印迹方法和免疫组织化学染色法对诱导膜中骨形态发生蛋白2、转化生长因子β1、血管内皮生长因子蛋白进行定量和定性分析。实验获得广州中医药大学动物实验伦理委员会批准,批准号：20181101006。 结果与结论：①苏木精-伊红染色显示,4种表面粗糙程度不同的骨水泥均可以形成较为规则的诱导膜,4组诱导膜之间血管化程度和细胞的数量大体相似;②Western Blot印迹检测显示,各组诱导膜内骨形态发生蛋2、转化生长因子β1、血管内皮生长因子蛋白平均含量基本相似(P > 0.05);③免疫组织化学染色显示,各组诱导膜内骨形态发生蛋2、转化生长因子β1、血管内皮生长因子蛋白阳性表达基本相似(P > 0.05);④结果表明,骨水泥表面粗糙程度改变对诱导膜的组织形态结构和骨形态发生蛋2、转化生长因子β1、血管内皮生长因子表达在6周时无明显影响。 ORCID: 0000-0003-1405-0765(李树源) 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

Bone morphogenetic protein is required for fibroblast growth factor 2-dependent later-stage osteoblastic differentiation in cranial suture cells

Background: Understanding the pathophysiological process of calvarial bones development is important for the treatments on relative diseases such as craniosynostosis. While, the role of fibroblast growth factor (FGF) and bone morphogenetic protein (BMP) and how they interacted in osteoblast differentiation remain unclear. Methods: we digested bone fragments around the coronal and sagittal sutures from newborn rats to harvest suture cells. Markers expression at different osteoblast differentiation stage was analyzed by increasing FGF2 concentration and BMP2 blocking in these cells. Results: BMP2 expression could be stimulated by FGF2 in a dose and time dependent manner. FGF2 stimulation may decrease early marker of osteoblast differentiation (collagen type-1, COL-1) and increase the expression of continuously-expressed or late markers (alkaline phosphatase, ALP; osteocalcin, OC and bone sialoprotein, BSP) to accelerate mineralization. Inhibition of BMP2 signaling by Noggin weakens the effect of FGF2 on induction of later-stage osteoblastic differentiation of cranial suture cells. Conclusion: Our data suggest that BMP2 signaling is required for FGF2-dependent induction of later-stage of cranial suture cell osteoblastic differentiation.     

Interleukin-1β, interleukin-6, epidermal growth factor and transforming growth factor-α are elevated in the brain from parkinsonian patients

Interleukin (IL)-1β, IL-6, epidermal growth factor (EGF), and transforming growth factor-α (TGF-α) were measured for the first time in the brain (caudate nucleus, putamen and cerebral cortex) from control and parkinsonian patients by highly sensitive sandwich enzyme immunoassays. The concentrations of IL-1β, IL-6, EGF, and TGF-α in the dopaminergic, striatal regions were significantly higher in parkinsonian patients than those in controls, whereas those in the cerebral cortex did not show significant differences between parkinsonian and control subjects. Since these cytokines and growth factors may play important roles as neurotrophic factors in the brain, the present results suggest that they may be produced as compensatory responses in the nigrostriatal dopaminergic regions in Parkinson's disease, and may be related, at least in part, to the process of neurodegeneration in Parkinson's disease.     

Human epidermal growth factor receptor 2, Na+/H+ exchanger regulatory factor 1, and breast cancer susceptibility gene-1 as new biomarkers for familial breast cancers

The aim of this study is to evaluate the analysis of markers related with progression, to further characterize familial breast cancers. Here, we investigated the expression of breast cancer susceptibility gene-1, hypoxia-inducible factor-1α, vascular endothelial growth factor receptor 1, and Na+/H+ exchanger regulatory factor 1 in 187 microarrayed breast carcinomas from 94 familial and 93 sporadic breast cancer patients by immunohistochemical staining. Furthermore, the expression levels of these biomarkers were compared with triple-negative phenotype. Familiarity was significantly associated with younger age (P < .000), higher tumor grade (P = .038), negative estrogen receptor hormonal status (P = .036), and high proliferative activity (P = .029). The familial cancers were immunonegative for membranous Na+/H+ exchanger regulatory factor 1 expression compared with sporadic cancers (P = .001); notably, vascular endothelial growth factor receptor 1 staining correlated with cytoplasmic Na+/H+ exchanger regulatory factor 1 expression in familial tumors (P = .009). In multivariate analysis, the "new biomarkers," including negative human epidermal growth factor receptor 2 status (odds ratio, 4.538; 95% confidence interval, 1.756-11.728), negative membranous Na+/H+ exchanger regulatory factor 1 expression (odds ratio, 7.686; 95% confidence interval, 1.876-31.483) and positive nuclear breast cancer susceptibility gene-1 (odds ratio, 0.3982; 95% confidence interval, 0.169-0.936), significantly correlated with family history of breast cancer. We hypothesize that the evaluation of human epidermal growth factor receptor 2, Na+/H+ exchanger regulatory factor 1, and breast cancer susceptibility gene-1 could be clinically useful to identify familial breast tumors and to select patients candidate to breast cancer susceptibility genes 1/2 gene sequencing.     

重组人骨形态发生蛋白2调节人脂肪间充质干细胞表达血管内皮生长因子

背景：重组人骨形态发生蛋白2可以促进组织工程骨血管化,但是对于其作用于人体细胞时的生物学规律不明确。目前对于重组人骨形态发生蛋白2调节人体细胞血管内皮生长因子表达的规律国内还未见相关报道。 目的：从基因和蛋白水平观察比较不同时间点重组人骨形态发生蛋白2诱导下人脂肪间充质干细胞血管内皮生长因子的表达。 方法：从成人脂肪组织中分离培养脂肪间充质干细胞,取第3代细胞用于实验,分为诱导组和对照组。诱导组采用终浓度为100 μg/L重组人骨形态发生蛋白2诱导人脂肪间充质干细胞,分别诱导3,6,12,18,24,36,48 h后收集样本,用RT-PCR和ELISA分别从基因水平和蛋白水平检测血管内皮生长因子的表达,并与空白对照组比较。 结果与结论：重组人骨形态发生蛋白2调节人脂肪间充质干细胞表达血管内皮生长因子具有时间依赖性,在不同时间点血管内皮生长因子表达量不同。与空白对照组相比,3-6 h时间段重组人骨形态发生蛋白2抑制血管内皮生长因子表达(P < 0.05),18-24 h时间段重组人骨形态发生蛋白2促进血管内皮生长因子表达(P < 0.05),当利用重组人骨形态发生蛋白2促进组织工程骨血管化时这两个时间段应当引起特别关注。     

Anti-fibrotic effects of bone morphogenetic protein-7-modified bone marrow mesenchymal stem cells on silica-induced pulmonary fibrosis

Silicosis is an occupational lung disease caused by exposure to small particles of crystalline silica, which ultimately results in diffuse pulmonary fibrosis. Evidence indicates an anti-fibrotic role of bone morphogenetic protein-7 (BMP-7) and bone marrow mesenchymal stem cells (BMSCs) in lung diseases. Therefore, strategies incorporating genetic engineering and stem cell biology might have a tremendous potential to treat critical injuries and diseases. Therefore, we modified BMSCs to overexpress the BMP-7 gene (BMP-7-BMSCs) by lentivirus transduction, and then evaluated whether fibrotic processes were inhibited by these cells in vivo. Wistar rats were divided into four groups: control, silica, BMSCs, and BMP-7-BMSCs. The control group received saline, the silica group received silica and saline, the BMSCs group received silica and BMSCs, and the BMP-7-BMSCs group received silica and BMP-7-BMSCs. Rats were sacrificed on days 15 or 30 after silica instillation. Hematoxylin and eosin, and Masson's trichrome staining were performed for histological examination. The severity of fibrosis was evaluated by the levels of hydroxyproline, fibronectin (FN), and transforming growth factor (TGF)-β1. Restoration of the alveolar epithelium was detected by the epithelial marker surfactant protein (SP)-C and aquaporin (AQP)-5. Histopathological results showed that BMP-7-BMSCs could remarkably block the progression of silica-induced fibrosis. Hydroxyproline, FN, and TGF-β1 contents in the BMP-7-BMSCs-treated group were significantly lower than those in the BMSCs group (P < 0.05). Furthermore, the expression of SP-C and AQP-5 in the BMP-7-BMSCs-treated group was significantly higher than those in the BMSCs group (P < 0.05). In conclusion, the pulmonary fibrosis induced by silica in rats was significantly reduced by treatment with BMP-7-BMSCs and BMSCs. The anti-fibrotic effect of BMSCs can be strengthened by BMP-7. Treatment with BMP-7-BMSCs might be a potential therapeutic intervention for silicosis.