关于FAK与Pyk2的研究新进展

局部黏着斑激酶（focal adhesion kinase,FAK）是一种存在于细胞质中的酪氨酸激酶,对胚胎的生长,肿瘤、心血管疾病的发病机制都有重要作用.FAK分布广泛,序列在多种物种中高度保守.序列和结构分析显示FAK包含4个明显的区域：①N端的FERM区域;②起催化作用的酪氨酸激酶中心区域;③C端黏着斑定位序列（FAT）区域;④脯氨酸富集区域（PR1、PR2和PR3）,定位于催化作用点和FAT区域之间.FAK分子中含有6个可被酪氨酸激酶磷酸化的位点：Y397、Y407、Y576、Y577、Y861和Y925.     

Molecular mechanism of Syntenin promoting human glioma cell migration though upregulating FAK phosphorylation

目的 探讨Syntenin促进人脑胶质瘤细胞迁移的分子机制.方法 采用划痕实验和Western blot检测CHG-5、稳定表达人源性Syntenin的CHG-hS细胞在纤维连接蛋白(fibronectin,FN)和多聚赖氨酸(Poly-l-lysine,PL)两种基质表面迁移能力、FAK磷酸化位点及相关信号分子的变化情况;分别在P38MAPK特异性抑制剂SB239063和PI3K特异性抑制剂LY294002处理后,FN基质上CHG-5、CHG-hS细胞迁移能力和下游信号分子JNK和AKT磷酸化水平的改变.结果 细胞划痕实验结果显示,CHG-hS细胞在FN表面的运动能力显著高于CHG-5细胞(P＜0.05);而在多聚赖氨酸包被的培养板上,CHG-hS细胞的运动能力与CHG-5组细胞表面无显著性差异(P＞0.05).Western blot检测结果显示,FN作用下CHG-hS细胞中FAK Tyr397、FAK Tyr576、FAK Tyr925位点的磷酸化水平随时间延长而逐渐升高(P＜0.05),而FAK FAK Tyr861位点的磷酸化水平没有变化(P＞0.05);相关信号Src、JNK、AKT的磷酸化水平也随时间延长而升高(P＜0.05).分别采用SB239063和LY294002处理后,伴随着p-JNK和p-AKT的磷酸化水平减弱,CHG-hS迁移能力下降.结论 Syntenin通过与p-Src结合,随后触发FAK Tyr397、FAK Tyr925、FAK Tyr576位点的磷酸化作用,最大程度的激活FAK并上调JNK、AKT等下游信号分子的磷酸化水平,最终通过Syntenin-Src/FAK/MAPK和Syntenin-Src/FAK/PI3K两条通路增强FN相关胶质瘤细胞的迁移能力.     

Syntenin上调FAK磷酸化水平促进人脑胶质瘤细胞迁移的分子机制

目的探讨Syntenin促进人脑胶质瘤细胞迁移的分子机制。方法采用划痕实验和Western blot检测CHG-5、稳定表达人源性Syntenin的CHG-hS细胞在纤维连接蛋白(fibronectin,FN)和多聚赖氨酸(Poly-l-lysine,PL)两种基质表面迁移能力、FAK磷酸化位点及相关信号分子的变化情况;分别在P38MAPK特异性抑制剂SB239063和PI3K特异性抑制剂LY294002处理后,FN基质上CHG-5、CHG-hS细胞迁移能力和下游信号分子JNK和AKT磷酸化水平的改变。结果细胞划痕实验结果显示,CHG-hS细胞在FN表面的运动能力显著高于CHG-5细胞(P<0.05);而在多聚赖氨酸包被的培养板上,CHG-hS细胞的运动能力与CHG-5组细胞表面无显著性差异(P>0.05)。Western blot检测结果显示,FN作用下CHG-hS细胞中FAK Tyr397、FAK Tyr576、FAK Tyr925位点的磷酸化水平随时间延长而逐渐升高(P<0.05),而FAK FAK Tyr861位点的磷酸化水平没有变化(P>0.05);相关信号Src、JNK、AKT的磷酸化水平也随时间延长而升高(P<0.05)。分别采用SB239063和LY294002处理后,伴随着p-JNK和p-AKT的磷酸化水平减弱,CHG-hS迁移能力下降。结论 Syntenin通过与p-Src结合,随后触发FAK Tyr397、FAK Tyr925、FAK Tyr576位点的磷酸化作用,最大程度的激活FAK并上调JNK、AKT等下游信号分子的磷酸化水平,最终通过Syntenin-Src/FAK/MAPK和Syntenin-Src/FAK/PI3K两条通路增强FN相关胶质瘤细胞的迁移能力。     

配体诱导受体酪氨酸激酶活化的结构基础研究进展

受体酪氨酸激酶（RTK）家族是一类具有内源性蛋白酪氨酸激酶（PTK）活性的单次跨膜受体,它们在调控与细胞增殖、分化等相关的信号转导通路中起关键作用。它与配体结合后引起二聚化或结构重排而使胞内区的酪氨酸（Tyr）被自磷酸化。Tyr的自磷酸化一方面可以激活胞内PTK区的活性,另一方面可以为下游的信号蛋白提供结合位点从而完成活化过程。本文对RTK家族成员的晶体结构及其活性调节机制的研究进行了综述,阐述了由配体诱导RTK活化的结构基础,并简要讨论了RTK抑制剂可能的作用靶点。     

6种不同人类肿瘤细胞株pp60c-Src的活性及PP2抑制效应的差异

pp60c-Src是一个相对分子质量为60 000的、416位点磷酸化后激活的Src蛋白,是非受体型酪氨酸激酶家族中研究最多的成员.目前大量研究显示,Src或其家族活性异常与癌症的发生发展密切相关,它们参与了肿瘤的黏附、迁移、浸润及血管形成等过程.     

Src、Fyn激酶对脑缺血/再灌注大鼠海马NMDA受体NR2A亚基磷酸化水平的影响

目的观察脑缺血再灌注后海马Src家族酪氨酸蛋白激酶（Src family of protein tyrosine kinase,SrcPTKs）成员Src、Fyn激酶对NMDA受体（N-methyl-D-aspartate receptors,NMDAR,NR）NR2A亚基磷酸化水平的影响。方法采用Pulsinelli-Brierley四动脉阻断（four-vessel occlusion,4-VO）大鼠全脑缺血模型,缺血前连续3天脑室注射Src、Fyn反义寡核苷酸（antisense oligode oxynucletides,AS-ODNs）及错义寡核苷酸组（missenseoligodeoxynucletides,MS ODNs）后缺血15 min,用免疫沉淀和免疫印迹法检测NR2A亚基酪氨酸磷酸化水平的变化。结果Fyn反义寡核苷酸使脑缺血再灌注海马NR2A亚基磷酸化水平下降,而Src反义寡核苷酸对脑缺血/再灌注海马NR2A亚基磷酸化水平影响更加明显。结论脑缺血/再灌注过程中NMDA受体NR2A亚基的磷酸化主要是由Src激酶介导的。     

Src酪氨酸激酶抑制剂对表皮生长因子诱导的肺腺癌细胞生长的作用及机制

目的 探讨Src酪氨酸激酶抑制剂对表皮生长因子（EGF）诱导的肺腺癌细胞生长浸润的作用及其机制。方法 选用PC-9和A549细胞进行培养,分别给予不同浓度的Src酪氨酸激酶抑制剂和EGF,利用MTT比色法和Boyden-chamber法检测PC-9和A549细胞增殖及浸润情况,台盼蓝染色检测细胞活力,蛋白质印迹检验Src蛋白的表达和磷酸化及其下游信号磷酸化情况。结果 抑制Src酪氨酸激酶可以在不影响细胞活力的情况下抑制PC-9和A549细胞的增殖浸润,EGF可以增强EGF受体（EGFR）、Src、促丝裂原活化蛋白激酶（MAPK）及AKT的磷酸化,Src酪氨酸激酶抑制剂可以抑制这种磷酸化。结论 抑制Src酪氨酸激酶可以通过调节EGFR及其下游信号通路抑制肺腺癌细胞的增殖浸润。     

埃兹蛋白477位酪氨酸的磷酸化在神经生长因子前体促进乳腺癌细胞侵袭中起关键作用

目的研究神经生长因子前体（precursor of nerve growth factor, proNGF）促进乳腺癌细胞侵袭的作用与埃兹蛋白（ezrin） 表达水平及其567位苏氨酸（Thr567）和477位酪氨酸（Tyr477）的磷酸化的相互关系。方法用梯度浓度的proNGF（0、2.5、5和 10 ng/mL）刺激人乳腺癌细胞系MDA-MB-231 和MCF-7,用Transwell 侵袭实验检测proNGF对MDA-MB-231 和MCF-7 侵袭 的影响;用免疫印迹法检测ezrin 蛋白的表达水平,ezrin Thr567 和ezrin Tyr477 磷酸化水平的变化;在MDA- MB-231 中转染 pEnter-His-ezrinY477F质粒（ezrin显性负突变质粒）,研究ezrin Tyr477磷酸化在proNGF促进乳腺癌细胞侵袭中的作用。结果 proNGF 以浓度依赖的方式促进MDA-MB-231 和MCF-7 的侵袭（P<0.05）;proNGF 以浓度依赖和时间依赖的方式显著升高 ezrin Tyr477磷酸化,而对ezrin蛋白的表达以及其Thr567磷酸化无明显影响;Src激酶特异抑制剂SKI-606显著抑制proNGF对 ezrin Tyr477磷酸化的促进作用;转染pEnter-His-ezrinY477F抑制proNGF对MDA-MB-231细胞ezrin Tyr477磷酸化和侵袭能 力的促进作用（P<0.05）。结论ezrin Tyr477磷酸化在proNGF促进乳腺癌细胞侵袭中起关键作用;proNGF通过激活Src激酶使 ezrin Tyr477发生磷酸化。     

哇巴因诱导血液肿瘤细胞增殖的信号通路研究

目的揭示哇巴因诱导急性淋巴细胞白血病细胞株Jurkat增殖的信号传导途径.方法采用MTT、Western Blot等方法,观察哇巴因及不同激酶抑制剂对Jurkat细胞生长的影响,同时检测酪氨酸活化水平和丝裂原活化蛋白激酶MAPK（ERK1/2）的磷酸化程度.结果哇巴因促使Jurkat细胞膜酪氨酸磷酸化水平增高,并激活ERK1/2.激酶抑制剂PD98059,PP2,HerbimycinA和PP3均可阻断哇巴因诱导ERK1/2 的活化并抑制哇巴因诱导Jurkat细胞的增殖.结论哇巴因激活钠泵的信号传导功能,引起细胞膜酪氨酸磷酸化水平增高,MAPK（ERK1/2）信号途径级联样活化,从而导致Jurkat的增殖反应.Src激酶和EGFR参与了钠泵介导的信号传导.     

盘状结构域受体酪氨酸激酶1与炎症关系的研究进展

盘状结构域受体酪氨酸激酶1(DDR1)是新发现的一种受体酪氨酸激酶,可以与其配体——胶原蛋白特异性结合,而这种结合可导致位于胞内区的酪氨酸激酶缓慢而持续的磷酸化,这种磷酸化可参与细胞的增殖、分化、黏附、迁移以及炎症信号通路的活化,进而导致多种病理学的改变,如纤维化和恶性肿瘤的发生、发展。同时,DDR1还可参与多个脏器的炎症反应,如肝炎、肾炎、中枢神经系统炎症等。未来,需进一步研究DDR1是否参与其他脏器的炎症反应,力求全面反映DDR1在炎症中所发挥的作用。     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

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.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.