Dopaminergic hypofunctions and prepulse inhibition deficits in mice lacking midkine

Midkine is a 13-kDa retinoic acid-induced heparin-binding growth factor involved in various biological phenomena such as cell migration, neurogenesis, and tissue repair. We previously demonstrated that midkine-deficient (Mdk(−/−)) mice exhibited a delayed hippocampal development with impaired working memory and increased anxiety only at the age of 4 weeks. To assess whether midkine gene could play important roles in development and maintenance of central nervous system, we investigated biochemical and behavioral parameters in dopamine and glutamate neurotransmission of Mdk(−/−) mice. The Mdk(−/−) mice exhibited a hypodopaminergic state (i.e., decreased levels of dopamine and its receptors in the striatum) with no alterations of glutamatergic system (i.e., normal level of glutamate, glutamine, glycine, d-serine, l-serine, and NMDA receptors in the frontal cortex and hippocampus). We also found prepulse inhibition deficits reversed by clozapine and haloperidol in the Mdk(−/−) mice. Our results suggested that midkine deficiency may be related to neurochemical and behavioral dysfunctions in dopaminergic system.     

rhG-HGF联合成纤维细胞生长因子促进下肢血管新生的实验研究

目的 探讨重组人肝细胞生长因子(rhG-HGF)联合成纤维细胞生长因子(bFGF)对下肢缺血动物模型血管新生的影响.方法 制作80只小鼠左下肢缺血模型,术后随机分为4组,每组20只:(1)生理盐水对照组;(2)bFGF组;(3)rhG-HGF组;(4)rhG-HGF+bFGF组.应用多谱勒超声血流测定、肌肉毛细血管密度测定,比较4组缺血肢体血流/正常肢体血流比值及肌肉毛细血管密度.结果 术后4周,4组缺血肢体血流/正常肢体血流比值及肌肉毛细血管密度为:rhG-HGF+bFGF组>rhG-HGF组>bFGF组>生理盐水组(均P<0.05).结论 rhGvHGF促进血管新生作用强于bFGF,两者联合应用有协同作用,可以更明显改善下肢缺血状况.     

Role of fibroblast growth factor receptor signaling in kidney development

Fibroblast growth factor receptors (Fgfrs) are expressed in the ureteric bud and metanephric mesenchyme of the developing kidney. Furthermore, in vitro and in vivo studies have shown that exogenous fibroblast growth factors (Fgfs) increase growth and maturation of the metanephric mesenchyme and ureteric bud. Deletion of fgf7, fgf10, and fgfr2IIIb (the receptor isoform that binds Fgf7 and Fgf10) in mice lead to smaller kidneys with fewer collecting ducts and nephrons. Overexpression of a dominant negative receptor isoform in transgenic mice has revealed more striking defects including renal aplasia or severe dysplasia. Moreover, deletion of many fgf ligands and receptors in mice results in early embryonic lethality, making it difficult to determine their roles in kidney development. Recently, conditional targeting approaches revealed that deletion of fgf8 from the metanephric mesenchyme interrupts nephron formation. Furthermore, deletion of fgfr2 from the ureteric bud resulted in both ureteric bud branching and stromal mesenchymal patterning defects. Deletion of both fgfr1 and fgfr2 in the metanephric mesenchyme resulted in renal aplasia, characterized by defects in metanephric mesenchyme formation and initial ureteric bud elongation and branching. Thus, Fgfr signaling is critical for growth and patterning of all renal lineages at early and later stages of kidney development.     

基质金属蛋白酶9高表达对大鼠皮肤成纤维细胞生物学行为的影响

目的 观察基质金属蛋白酶9(matrixmetalloproteinase 9,MMP 9)高表达对大鼠皮肤成纤维细胞生物学行为的影响,以阐述其在糖尿病足伤口愈合中的可能机制.方法 本研究在中山大学孙逸仙纪念医院内分泌专科实验室完成.大鼠皮肤成纤维细胞与高糖(22 mmol/L)+高同型半胱氨酸(100 μmol/L)联合孵育建立体外MMP 9高表达细胞模型,另设正糖(5.5 mmol/L)孵育对照组.Realtime PCR、ELISA和明胶酶谱法检测MMP 9 mRNA、蛋白表达量及活性,流式细胞仪检测细胞增殖功能,CCK-8检测细胞活力,ELISA法检测细胞胶原(羟脯氨酸)分泌能力、划痕实验评价细胞横向迁移能力、Transwell法评价细胞纵向迁移能力.计量数据以均数±标准差(x-±s)表示,两组间均数的比较采用成组t检验,以P ＜0.05为差异具有统计学意义.结果 与对照组比较,MMP 9高表达组大鼠皮肤成纤维细胞的MMP 9 mRNA、蛋白表达量及活性,分别升高6.05倍、4.12倍和1.58倍(P＜0.01);与此同时MMP 9高表达组大鼠皮肤成纤维细胞S期的比例、增殖指数、细胞活力、胶原(羟脯氨酸)分泌量、6h横向迁移率和纵向迁移细胞数,分别下降29.8％、18.1％、23.3％、68.7％、45.0％和21.4％ (P ＜0.01).结论 MMP 9高表达的皮肤成纤维细胞增殖减慢、活力下降、迁移和胶原分泌能力降低,提示MMP 9可能抑制成纤维细胞的生物学行为.     

Wnt/β-catenin pathway forms a negative feedback loop during TGF-β1 induced human normal skin fibroblast-to-myofibroblast transition

Background

Fibroblast-to-myofibroblast transition is a key event during wound healing and hypertrophic scar formation. Previous studies suggested Wnt/β-catenin signaling might be involved in the wound healing. However, its specific role in skin fibroblast-to-myofibroblast transition remains unclear.

Objective

To investigate the specific role of β-catenin during the transforming growth factor-β1 induced normal skin myofibroblasts transition.

Methods

By real-time quantitative polymerase chain reaction, Western-blot and immunocytochemistry, the activation of Wnt/β-catenin pathway in cultured human normal skin fibroblasts during TGF-β1 induced fibroblast-to-myofibroblast transition was investigated. The effects of β-catenin on myofibroblasts transition were also investigated when SB-216763, over-expression and siRNA of β-catenin were utilized. In addition, fibroblasts populated collagen lattices contraction assays were conducted to examine the effects of β-catenin on the contractility of the fibroblasts induced by TGF-β1. Furthermore, the effects of β-catenin on the expression of α-smooth muscle actin and collagen types I and III in hypertrophic scar derived fibroblasts were studied.

Results

The expression of Wnts mRNA and β-catenin protein was up-regulated by TGF-β1 stimulation during the myofibroblasts transition. Both of SB-216763 and β-catenin over-expression was paralleled with decreased expression of α-smooth muscle actin, collagen types I and III, while siRNA targeting β-catenin leads to up-regulation of α-smooth muscle actin, collagen types I and III. The increased contractility and α-smooth muscle actin expression of the fibroblasts in the collagen lattices induced by TGF-β1 was inhibited by SB-216763. In addition, the expression levels of α-smooth muscle actin, collagen types I and III in hypertrophic scar derived fibroblasts were also down-regulated by SB-216763.

Conclusion

Specifically in normal skin fibroblasts, β-catenin might be involved in the myofibroblasts transition and negatively regulate the TGF-β1-induced myofibroblast transition.     

复方消斑灵对创伤性瘢痕作用的实验研究

目的:观察复方消斑灵对体外培养的成纤维细胞的胶原蛋白合成的影响,以及对水浴烫伤动物模型的作用。方法:采用光镜、电镜研究用药前、后成纤维细胞的形态学变化,以3 H-脯氨酸掺入等方法检测成纤维细胞胶原蛋白的合成,通过建立水浴烫伤动物模型,观察复方消斑灵对增生性瘢痕的作用。结果:消斑灵能明显影响成纤维细胞的超微结构,抑制胶原蛋白的合成,并呈量效关系;在大鼠水浴烫伤实验中复方消斑灵可抑制瘢痕增生,而且大剂量使用未发现药物毒性。结论:复方消斑灵对创伤性瘢痕有良好的治疗作用。     

Short-term efficacy of intravitreal dobesilate in central serous chorioretinopathy

Purpose

To report the anatomic and functional outcome of intravitreal dobesilate to treat recurrent central serous chorioretinopathy (CSC).

Methods

This is an interventional case report in which dobesilate was intravitreally injected in a case of recurrent CSC. Main measures included fundoscopy, Snellen visual acuity (VA) testing, fluorescein angiography and optical coherence tomography (OCT).

Results

We present anatomical and functional evidences, obtained as early as eleven days after the treatment, of the efficacy of intravitreal dobesilate, in the treatment of chronic CSC condition. The effect after intravitreal dobesilate injection for CSC might be related to the normalization of retinal architecture.

Conclusions

Intravitreal dobesilate may be an effective treatment option for recurrent CSC.     

IL-32在病理性瘢痕基因治疗中的研究进展

白细胞介素(IL)32是新近发现的炎性细胞因子,能诱生细胞因子的产生,且在调节细胞增殖与凋亡过程中有重要作用。瘢痕组织中成纤维细胞IL-32基因的蛋白表达水平明显低下,提示在瘢痕成纤维细胞中存在IL-32基因的缺失。随着基因治疗技术的日益成熟,以IL-32为目的基因导入到瘢痕成纤维细胞中来调控过度增生的成纤维细胞及减少胶原等大量细胞外基质的过度产生和沉积。现就IL-32的生物学特性及在病理性瘢痕中的作用及应用前景等进行综述,以期为以后的实验及临床作铺垫。     

机械磨削联合微针导入重组人酸性成纤维细胞生长因子和基因重组人表皮生长因子促进创面愈合的临床观察

目的探讨机械磨削联合微针导入重组人酸性成纤维细胞生长因子(rhaFGF)和基因重组人表皮生长因子(rhEGF)促进创面愈合的临床效果。方法 2010年4月—2011年9月表浅瘢痕患者60例,随机分为试验组、对照组和空白对照组,每组20例。试验组采用机械磨削联合微针导入rhaFGF和rhEGF,对照组采用机械磨削联合外涂rhaFGF及rhEGF,空白对照组采用机械磨削。研究三组创面愈合时间。结果试验组平均创面愈合时间(9.35±0.87)d,对照组(10.02±0.96)d,空白对照组(10.66±1.02)d,三组比较差异有统计学意义(P<0.05);其中试验组平均创面愈合时间较对照组和空白对照组均明显缩短,差异有统计学意义(P<0.05);对照组平均创面愈合时间较空白对照组明显缩短,差异有统计学意义(P<0.05)。结论机械磨削联合微针导入rhaFGF和rhEGF可促进创面愈合,缩短了创面愈合时间,提高了愈合质量。     

硅酮凝胶喷雾剂局部喷涂对兔耳增生性瘢痕影响的实验研究

目的通过检测硅酮凝胶喷雾剂对兔耳瘢痕动物模型瘢痕纤维组织的影响,探讨硅酮凝胶喷雾剂对增生性瘢痕的作用及其可能机制。方法将8只新西兰大耳白兔随机分为空白对照组与硅酮凝胶喷雾剂组,并在每只兔耳腹侧面制作创面建立病理性瘢痕模型。于创口制备术后第28天始在瘢痕部位喷抹硅酮凝胶药物,于术后第56天处死动物,切除耳部瘢痕,HE染色检测瘢痕组织的增生指数（HI）及成纤维细胞数密度（NA）,VG染色检测瘢痕组织胶原纤维面积密度（AA）,同时采用免疫组织化学法检测Sinad3表达水平。结果硅酮凝胶喷雾剂组（硅酮凝胶组）的成纤维细胞胞体变小,胶原纤维较稀疏,排列有序;HI、NA和AA均明显低于空白对照组（P均〈0．01）;免疫组织化学法检测发现硅酮凝胶组瘢痕组织中Smad3蛋白的表达水平明显低于空白对照组（P〈0．01）。结论硅酮制剂可抑制Smad3的表达,抑制成纤维细胞增殖及胶原形成,避免过量纤维组织形成,进而抑制瘢痕增生。