Hedgehog信号通路与肿瘤的关系

Hedgehog信号通路是调节动物胚胎正常发育的经典信号通路之一,主要由信号分子HH、膜受体Ptch、Smo以及转录因子Gli等组成.近年来,有关Hedgehog信号通路的研究取得了较大进展,对其在肿瘤发生、发展中的作用有了新的认识,并为肿瘤治疗提供了新的靶点.本文就Hedgehog信号通路的异常激活与肿瘤的关系作一综述.     

Hedgehog信号通路分子在食道鳞状细胞癌中的表达及临床意义

目的:探讨Hedgehog信号通路分子Gli1、Hip、PDGFRα、Smo和Su(Fu)基因在食道鳞状细胞癌组织中的表达及临床意义。方法:应用原位杂交方法检测内蒙古医科大学附属医院胸外科2013年3月-2014年5月经手术切除、病理科确诊的12例食道鳞状细胞癌组织及距肿瘤边缘2 cm处经病理证实为正常组织的12例食管黏膜(阳性对照)中Hedgehog信号通路分子Gli1、Hip、PDGFRα、Smo和Su(Fu)基因的表达。结果:12例癌组织中,10例癌组织中均检测到Hedgehog信号通路分子Gli1、Hip、PDGFRα、Smo基因的表达,8例癌组织中检测到Hedgehog信号通路分子Su(Fu)基因的表达,而其在对照组中均未表达,两组比较差异均有统计学意义(P0.05)。Gli1、Hip、PDGFRα、Smo及Su(Fu)基因的表达与患者的年龄、性别、组织学分级、临床分期无关,差异无统计学意义(P0.05)。结论:Hedgehog信号通路在食道鳞状细胞癌组织中存在高表达,其可能作为肿瘤治疗的新靶点。     

白藜芦醇调控Hedgehog信号通路抑制肝星状细胞活化的研究

【目的】观察白藜芦醇对HSC-T6细胞内Hedgehog信号通路及其下游靶基因表达的影响,探讨白藜芦醇抗肝纤维化的作用机制,为白藜芦醇的临床应用提供理论依据。【方法】体外培养大鼠肝星状细胞系HSC-T6,用含体积分数10%胎牛血清的DMEM高糖培养基培养种板,细胞贴壁后加入白藜芦醇(4、8、16μmol/L)或环耙明(100μmol/L)预处理30 min,再加入瘦素(100 ng/mL)孵育24 h诱导细胞活化。采用四甲基偶氮唑盐(MTT)法检测细胞增殖水平,蛋白免疫印迹(Western Blot)法检测细胞α平滑肌肌动蛋白(α-SMA)的表达,实时聚合酶链反应(PCR)检测细胞内Hedgehog信号通路成员Shh、Smo、Patched和Gli1 mRNA及Gli1下游靶基因Cyclin D1、Cyclin D2及Bcl-2的表达。【结果】与空白组比较,模型组细胞增殖率及α-SMA蛋白表达均呈显著增高趋势,Hedgehog信号通路成员Shh、Smo、Patched、Gli1 mRNA及Gli1下游靶基因Cyclin D1、Cyclin D2及Bcl-2的表达亦显著上升(P 0.05或P 0.01)。与模型组比较,白藜芦醇低、中、高剂量组及环耙明组均可显著抑制HSC-T6细胞增殖率及α-SMA蛋白表达(P 0.01),亦可明显抑制Hedgehog信号通路成员Shh、Smo、Patched、Gli1及靶基因Cyclin D1、Cyclin D2及Bcl-2 m RNA的表达(P 0.05或P 0.01)。【结论】白藜芦醇可通过调控Hedgehog信号通路抑制HSCT6细胞的增殖与活化,达到抗肝纤维化的作用。     

Sonic hedgehog信号通路Smo蛋白及其下游转录因子Glil蛋白在胃癌组织中的表达及其意义

目的 探讨Sonic hedgehog信号通路Smo蛋白及其下游转录因子Gli1蛋白在胃癌组织中的表达及其意义.方法 收集85例胃癌组织及25例正常胃粘膜组织构建成组织芯片,应用免疫组化S-P法检测胃癌组织中Smo蛋白及G1il蛋白的表达,并以正常胃粘膜组织作对照,分析两者的相关性.结果 Smo和Gli1蛋白均在正常胃粘膜中不表达或弱表达;在胃乳头状腺癌、管状腺癌、低分化腺癌中,两者的表达强度和阳性表达率显著高于正常胃粘膜(P＜0.05,并随着胃腺癌分化程度下降而上升;相关分析显示Smo和Gli1蛋白在胃癌组织中的表达呈正相关,相关系数为0.989,P＜0.001.结论 胃癌发生过程中的Sonic hedgehog信号通路异常激活可能通过Smo蛋白高表达上调下游转录因子Gli1蛋白的表达参与部分胃腺癌的发生.     

Sonic hedgehog信号通路Smo蛋白及其下游转录因子Glil蛋白在胃癌组织中的表达及其意义

目的探讨Sonic hedgehog信号通路Smo蛋白及其下游转录因子Gli1蛋白在胃癌组织中的表达及其意义。方法收集85例胃癌组织及25例正常胃粘膜组织构建成组织芯片，应用免疫组化S—P法检测胃癌组织中Smo蛋白及Gli1蛋白的表达，并以正常胃粘膜组织作对照，分析两者的相关性。结果Smo和Gli1蛋白均在正常胃粘膜中不表达或弱表达；在胃乳头状腺癌、管状腺癌、低分化腺癌中，两者的表达强度和阳性表达率显著高于正常胃粘膜（P〈0．05，并随着胃腺癌分化程度下降而上升；相关分析显示Smo和Gli1蛋白在胃癌组织中的表达呈正相关，相关系数为0．989，P〈0．001。结论胃癌发生过程中的Sonic hedgehog信号通路异常激活可能通过Smo蛋白高表达上调下游转录因子Gli1蛋白的表达参与部分胃腺癌的发生。     

三氧化二砷对急性早幼粒细胞白血病细胞株Hedgehog信号通路的影响

［摘要］目的: 探讨三氧化二砷（As2O3）对急性早幼粒细胞白血病细胞Hedgehog信号转导通路相关分子的作用,为将其扩大应用于其他具有Hedgehog异常活化的肿瘤提供理论基础。方法： 将不同浓度的As2O3作用于急性早幼粒细胞白血病NB4细胞株48 h,采用实时定量PCR（qRT PCR）和免疫印迹技术检测Hedgehog信号转导通路关键分子Smoothened（Smo）、Ptch、Gli1和Gli2在mRNA和蛋白水平的变化。结果： As2O3处理后,NB4细胞的Smo、Ptch、Gli2在蛋白和mRNA水平均受到明显抑制,差异有统计学意义。结论： As2O3可能是通过抑制Gli2蛋白和mRNA发挥抗Hedgehog信号转导通路的作用,而Hedgehog信号转导通路可能是As2O3发挥抗急性早幼粒细胞白血病的机制之一。     

Smo蛋白及Gli1蛋白在结直肠癌组织中的表达及其意义

目的探讨Sonic hedgehog信号通路中Smo和Gli1蛋白在结直肠癌组织中的表达及其意义。方法应用免疫组化S—P法检测125例结直肠癌组织及30例正常结直肠黏膜组织中Sonic hedgehog信号通路中Smo和Gli1蛋白的表达情况。结果Smo和Gli1蛋白在结直肠癌组织的表达明显高于其正常结直肠黏膜（P〈0．001）,与肿瘤部位、分化程度、Dukes分期以及有无淋巴结转移无关（P〉0．05）,而与肿瘤组织学类型有关（P〈0．001）,相关分析显示Smo及GIi1蛋白在结直肠癌组织中的表达呈正相关,相关系数为0．690（P〈0．001）。结论结直肠癌的发生与Smo和Gli1蛋白的高表达有关,可能与Smo蛋白高表达上调其下游转录因子Gli1蛋白的表达有关。     

Hedgehog信号通路调节与胰腺癌研究

胰腺癌的发生发展与许多信号通路如:Hedgehog(Hh)、Wnt/Notch、K-ras、TGF-β等途径都有关系,Hh信号通路是其核心之一,主要由配体蛋白(Hh蛋白)、膜受体(Ptch和Smo)、核转录因子Gli蛋白家族、HSC(Hh信号复合体)和调节因子等组成,在胰腺癌中Hh信号通路100%的相关分子存在基因突变。该文主要从Hedgehog信号通路的组成、关键位点、调节作用以及与胰腺癌的关系进行总结,为寻求中医药防治恶性肿瘤的新途径和作用靶点提供实验理论依据。     

白藜芦醇诱导大鼠骨髓基质细胞的神经元样细胞分化伴Shh信号激活

目的 研究Sonic Hedgehog(Shh)信号在白藜芦醇诱导大鼠骨髓基质细胞(marrow stromal cells,MSCs)分化为神经元样细胞中的作用.方法 采用全骨髓贴壁法分离培养MSCs.MSCs分为3组,白藜芦醇诱导组:用含白藜芦醇的无血清DMEM/F12诱导MSCs分化;对照组:用无血清DMEM/F12诱导MSCs分化;正常组:不做诱导处理.倒置显微镜下观察细胞形态,免疫荧光法、免疫印迹法检测NSE、MAP-2、GFAP、Smo和Gli1蛋白的表达和移位.结果 白藜芦醇诱导后细胞胞体收缩,伸出长突起,类似神经元.免疫荧光显示正常组及对照组MSCs轻度表达神经元NSE蛋白,白藜芦醇诱导组MSCs NSE、MAP-2阳性,但诱导前后细胞始终未表达胶质细胞GFAP蛋白.免疫荧光显示,MSCs具有初级纤毛,并表达Smo和Gli1.白藜芦醇诱导组Smo从细胞质进入初级纤毛,Glil从细胞质进入细胞核,同时,Smo、Glil蛋白的表达增加(P＜0.01).对照组则无明显变化.结论 白藜芦醇能诱导MSCs分化为神经元样细胞;此过程中Shh信号通路被激活,推测Shh信号在MSCs分化过程中可能发挥着重要作用.     

调控Smo活性对成年大鼠神经干细胞增殖和迁移的影响

目的 探讨Shh信号通路中G蛋白偶联受体Smoothened（Smo）对成年大鼠神经干细胞（ANSCs）增殖、迁移的影响并阐明其作用机制。方法 培养ANSCs,分别添加Smo的激动剂Purmorphamine及其抑制剂Cyclopamine,细胞增殖-毒性实验（CCK8）检测其对ANSCs增殖的影响;细胞划痕实验检测其对ANSCs迁移能力的影响;实时荧光定量PCR（RT-PCR）技术检测Sonic Hedgehog（Shh）信号通路的细胞表面Patched蛋白（Ptch1）、G蛋白偶联受体Smo、转录因子（Gli1）以及神经导向因子（Slit1）和脑源性神经营养因子（BDNF）表达量的变化。结果 Smo激动剂PM对ANSCs的增殖有明显促进作用,48 h后PM组ANSCs的吸光度明显升高（P<0.01）;PM可以促进ANSCs的迁移,48 h后的PM组ANSCs划痕面积显著减小（P<0.01）;Smo抑制剂CPM对ANSCs的增殖和迁移均有显著的抑制作用;RT-PCR实验显示PM组细胞中的Ptch1、Smo、Gli1及Slit1、BDNF表达水平均明显增加（P<0.05）。结论 调控Smo的活性可以促进或抑制成年神经干细胞的增殖和迁移,其作用机制可能与调节BDNF和Slit1的表达有关。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.