丁酸钠诱导BEL7402人肝癌细胞恶性表型逆转的研究

丁酸钠诱导ＢＥＬ７４０２人肝癌细胞恶性表型逆转的研究梁若斯胡梅香黄炜＊（广州医学院组胚教研室＊广州医学院肿瘤研究所）癌细胞能在某些因子的诱导作用下，恶性表型发生逆转。本研究旨在证明天然物质丁酸钠能诱导ＢＥＬ７４０２人肝癌细胞恶性表型逆转。我们使用四种...

三仁汤联合美洛昔康对湿热型骨关节炎患者关节功能及血清BMP⁃2、COMP水平的干预作用

目的:探讨三仁汤联合美洛昔康对湿热型骨关节炎(OA)患者关节功能及血清细胞因子骨形态发生蛋白2(BMP⁃2)、软骨寡聚基质蛋白(COMP)水平的影响。方法:选取2017年3月-2019年3月广州医科大学附属第二医院123例湿热型OA患者作为研究对象,简单随机化分为3组,各41例。对照A组予美洛昔康治疗,对照B组予三仁汤治疗,观察组予三仁汤联合美洛昔康治疗。对比三组临床疗效、不良反应发生率及治疗前后中医与疾病证候积分、WOMAC指数、血清炎性因子(CRP、ESR、IL⁃18)、BMP⁃2、COMP水平。结果:(1)临床疗效:观察组总有效率为95.12%,显著高于对照A、B组(75.61%、78.05%)(P<0.05);(2)中医证候积分、疾病症候积分:观察组治疗后中医证候积分、疾病症候积分显著低于对照A、B组(P<0.05);(3)WOMAC指数:观察组治疗后关节僵直、关节疼痛、关节功能评分及总分显著低于对照A、B组(P<0.05);(4)炎性因子:观察组治疗后血清CRP、ESR、IL⁃18水平显著低于对照A、B组(P<0.05);(5)血清BMP⁃2、COMP水平:观察组治疗后血清BMP⁃2、COMP水平显著低于对照A、B组(P<0.05);(6)不良反应:3组不良反应发生率差异无统计学意义(P>0.05)。结论:三仁汤联合美洛昔康治疗湿热型OA,疗效确切,能有效缓解临床症状,降低炎症反应,调节血清BMP⁃2、COMP水平,改善关节功能,保证安全性。

自制稀土永磁夹板对骨折愈合作用的观察

目的观察稀土永磁夹板对家兔骨折愈合质量及骨形态发生蛋白(BMP-2)、血管内皮细胞生长因子(VEGF)表达情况的影响,从分子生物学水平探讨磁场促进骨折愈合的作用机制。方法健康成年家兔50只,建立双前肢桡骨中段骨折模型。左侧用稀土永磁夹板固定,作为实验侧;右侧用内镶铅片小夹板固定,作为对照侧。分别于术后1、2、3、4、6周摄双前肢X线片后取材,进行HE及BMP-2、VEGF免疫组化染色并在光镜下观察。结果X线片图像定量分析结果比较,实验侧愈合明显快于对照侧。两侧差异有显著性(P<0.05)。实验侧在1、2、3、4周VEGF的表达均比对照侧强,两侧差异有显著性(P<0.05)。实验侧在2、3、4周BMP-2的表达均比对照侧强,两侧差异有显著性(P<0.01)。结论静磁场在骨折愈合过程中可加强BMP-2、VEGF的表达。自制稀土永磁夹板对实验性骨折愈合有明显的促进作用。

丹皮酚对强直性脊柱炎模型小鼠Wnt和BMP/Smad信号转导通路的影响

目的:研究丹皮酚对强直性脊柱炎(AS)模型小鼠分泌型糖蛋白(Wnt)和骨形态发生蛋白(BMP)/细胞信号转导分子(Smad)通路的影响,探讨丹皮酚防治AS的机制。方法:将40只小鼠随机分为正常组、模型组、柳氮磺吡啶组(阳性对照,9 mg/kg)和丹皮酚组(3 mg/kg),每组10只。除正常组外的其余各组小鼠均采用完全弗氏佐剂+蛋白聚糖腹腔注射法复制AS模型。各给药组小鼠在成模后灌胃相应药物,正常组和模型组小鼠灌胃等体积蒸馏水,每天给药1次,连续20 d。末次给药后处死小鼠,透射电镜下观察各组小鼠骶髂关节滑膜细胞超微病理结构变化,采用酶联免疫吸附法检测小鼠血清中肿瘤坏死因子α(TNF-α)、Wnt信号通路病理性骨化相关因子(DKK-1)含量,实时荧光定量聚合酶链式反应法检测小鼠滑膜组织中骨形态发生蛋白2(BMP-2)、细胞内核心结合因子α1(Cbfα1)、Smad1 m RNA表达。结果:与正常组比较,模型组小鼠血清中TNF-α含量明显增加、DKK-1含量明显减少,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显升高,差异均有统计学意义(P<0.05或P<0.01);电镜下可见模型组小鼠滑膜细胞增生,排列紊乱,分泌活性细胞器分泌亢进,细胞间隙增宽。与模型组比较,柳氮磺吡啶组和丹皮酚组小鼠血清中TNF-α含量均明显减少,丹皮酚组小鼠血清中DKK-1含量明显增加,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显降低,差异均有统计学意义(P<0.05或P<0.01);电镜下可见丹皮酚组小鼠滑膜细胞线粒体、溶酶体、粗面内质网的形态明显改善。结论:丹皮酚防治AS的机制可能与降低血清中TNF-α含量、升高血清中DKK-1含量,下调滑膜细胞中BMP-2、Cbfα1和Smad1 m RNA表达,抑制Wnt和BMP/Smad骨化相关信号转导通路逆转滑膜细胞成骨分化有关。

Physical Blocking Neural Tube Closure Affects Radial Intercalation and Neural Crest Midline-directed Migration in Xenopus Dorsal Explants

Neural tube defects （NTDs） are severe congenital malformation diseases, which occur in 1 out of 1000 births in human. In Xenopus, several tissue movements are involved in the neural tube closure process. Immediately after the neural tube fusion, the neural crest cells get monopolar protrusion toward dorsal midline and migrate to form the roof of the neural tube. At the same time, radial intercalation takes place from the ventral neural tube and forces it to be single-layered. Here, we physically block the neural tube closure to test the cell movements and the following patterning in Xenopus laevis explants. The results show that the single-layered neural tube fails to form and the neural crest cells remain at the lateral regions in the explants with NTDs. However, the patterning of the neural tube is not affected as indicated by the normal expression of the preneural genes. These results indicate a requirement of the neural tube fusion for the radial intercalation and the dorsal midline directed neural crest migration, but not for the dorsal-ventral patterning of the neural tube.

Effect of BMPs on hematopoietic injury of acute radiation sickness in mice

Objective: To investigate the effect of bone morphogenetic proteins (BMPs) on hematopoietic injury of acute radiation sickness in mice. Methods: Mice were subjected to whole-body 60Co γ ray irradiation, then bpBMP was put into spatium intermusculare or rhBMP-2m, PBK/ hBMP-2 -NIH3T3 cells were injected into abdominal cavity. The effect of BMPs on hematopoiesis including some hematological parameters, the survival rate of 30 d and formation of bone marrow CFU-GM colony were detected at postradiation. Results: pbBMP (purified bovine bone morphogenetic protein) increased the formation of bone marrow CFU-GM colony (P<0. 05) on d 10 after irradiation. rhBMP-2m increased the survival rate of mice irradiated by 7. 5 Gys Mice in control group died in 30 days, while 10%, 15% and 35% mice survived when they were injected i. p. with 0. 5 mg, 1. 0 mg and 2. 0 mg of rhBMP-2m respectively. All hematological parameters of treated mice were significantly higher than those of control group (P<0. 01). PBK/ hBMP-2 -NIH3T3 cells were established and transplanted into mice irradiated by 7. 0 Gy γ ray with i. p. . The survival ratio of treated mice was higher than that of negative control group (P<0. 01), and all hematopoietic parameters were increased statistically significantly (P<0. 01). Conclusion: Results indicate that in adult mice, BMPs can recover or treat the hematopoietic injury of acute radiation sickness, the mechanism may be related with repairing of hematopoietic injury.

Shh信号参与调控BMP9诱导的间充质干细胞成骨分化

目的:观察sonic hedgehog(Shh)信号通路在骨形态发生蛋白9(BMP9)诱导的小鼠间充质干细胞(MSCs)C3H10T1/2和C2C12成骨分化中的作用,并初步探讨其作用机制。方法:Shh信号通路抑制剂Cyclopamine和激活剂Purmorphamine以及过表达Shh腺病毒分别作用于BMP9处理的C3H10T1/2和C2C12细胞,碱性磷酸酶(ALP)检测早期成骨指标ALP,茜素红S染色检测晚期成骨指标钙盐沉积,RT-PCR检测Shh信号相关基因以及成骨关键转录因子的表达,Western blot检测Shh的表达,荧光素酶报告基因检测Smad1/5/8的转录调控活性。结果:BMP9促进Shh信号相关基因的表达,激活Shh信号可增强BMP9诱导的C3H10T1/2和C2C12细胞早晚期成骨分化并促进了BMP9诱导的Smad荧光素酶活性,抑制Shh信号后作用相反。结论:激活Shh信号通路可促进BMP9诱导的小鼠MSCs成骨分化,抑制其活性后作用相反。

Odontoblast β-catenin signaling regulates fenestration of mouse Hertwig's epithelial root sheath

The interaction between Hertwig＇s epithelial root sheath （HERS） and the adjacent mesenchyme is vitally important in mouse tooth root development. We previously generated odontoblast-specific Ctnnbl （encodingβ-catenin） deletion mice, and demon- strated that odontoblast β-catenin signaling regulates odontoblast proliferation and differentiation. However, the role of odon- toblast β-catenin signaling in regulation of HERS behavior has not been fully investigated. Here, using the same odonto- blast-specific Ctnnbl deletion mice, we found that ablation of β-catenin signaling in odontoblasts led to aberrant HERS for- mation. Mechanistically, odontoblast-specific Ctnnbl deletion resulted in elevated bone morphogenetic protein 7 （Bmp7） ex- pression and reduced expression of noggin andfollistatin, both of which encode extracellular inhibitors of BMPs. Furthermore, the levels of phosphorylated Smadl/5/8 were increased in HERS cells. In vitro tissue culture confirmed that BMP7 treatment disrupted the HERS structure. Taken together, we demonstrated that odontoblast f3-catenin signaling may act through regula- tion of BMP signaling to maintain the integrity of HERS cells.

Morphogenesis of T-tubules in heart cells: the role of junctophilin-2

The T-tubule (TT) system forms the structural basis for excitation-contraction coupling in heart and muscle cells. The morphogenesis of the TT system is a key step in the maturation of heart cells because it does not exist in neonatal cardiomyocytes. In the present study, we quantified the morphological changes in TTs during heart cell maturation and investigated the role of junctophilin-2 (JP2), a protein known to anchor the sarcoplasmic reticulum (SR) to TT, in changes to TT morphological parameters. Analysis of confocal images showed that the transverse elements of TTs increased, while longitudinal elements decreased during the maturation of TTs. Fourier transform analysis showed that the power of ~2 m spatial components increased with cardiomyocytes maturation. These changes were preceded by increased expression of JP2, and were reversed by JP2 knockdown. These findings indicate that JP2 is required for the morphogenesis of TTs during heart development.

Role of adsorbed proteins on hydroxyapatite-coated titanium in osteoblast adhesion and osteogenic differentiation

The method of plasma-spray coating of hy- droxyapatite （HA） onto pure titanium has been demon- strated to be effective to enhance the osteogenic differentiation and accelerate bone regeneration. Yet it is still a big challenge to figure out the interplay among im- plant surface properties, adsorbed proteins and cell-surface interactions. In this study, the plasma-sprayed HA-coated titanium （HA-Ti） surface was compared with the titanium substrate in terms of protein adsorption, cell adhesion and differentiation. The phase composition, wettability and to- pography were characterized. Compared to the Ti substrate, the HA-Ti had a smaller water contact angle, but larger micro-scale roughness, and showed a poorer ability to ad- sorb fibronectin （Fn）, bovine serum albumin （BSA） and serum proteins. However, it could adsorb larger amount of recombinant human bone morphogenetic protein 2 （BMP- 2）. The osteoblasts and bone marrow mesenchymal stem cells （BMSCs） tended to adhere on the Ti substrate. By contrast, the BMSCs cultured on the HA-Ti showed a stronger tendency toward osteogenesis differentiation.

Treatment of osteoarthritis with mesenchymal stem cells

Osteoarthritis(OA)is one of the most prevalent joint diseases with prominent symptoms affecting the daily life of millions of middle aged and elderly people.Despite this,there are no successful medical interventions that can prevent the progressive destruction of OA joints.The onset of pathological changes in OA is associated with deviant activity of mesenchymal stem cells(MSCs),the multipotent precursors of connective tissue cells that reside in joints.Current therapies for OA have resulted in poor clinical outcomes without repairing the damaged cartilage.Intra-articular delivery of culture-expanded MSCs has opened new avenues of OA treatment.Pre-clinical and clinical trials demonstrated the feasibility,safety,and efficacy of MSC therapy.The Wnt/β-catenin,bone morphogenetic protein 2,Indian hedgehog,and Mitogen-activated protein kinase signaling pathways have been demonstrated to be involved in OA and the mechanism of action of MSC therapies.