NOTCH配体Jagged2和Delta4对32D细胞的分化抑制研究

目的探讨Notch信号传导系统中相关配体Jagged2和Delta4对髓系前体细胞32D的分化抑制作用.方法构建Delta4高表达载体pTracer.CMV.Delta4.FLAG,制备Delta4-CHO细胞;将Notch1-32D细胞加入含G-CSF培养液的CHO、Jagged2-CHO及Delta4-CHO细胞中,观察Notch1-32D细胞分化及分化抑制情况. 结果构建成功Delta4高表达载体pTracer.CMV.Delta4.FLAG并稳定转染CHO细胞.在G-CSF存在下,Notch1-32D细胞可分化为成熟的粒细胞; Jagged2能抑制G-CSF引起的Notch1-32D细胞分化,但Delta4不能抑制G-CSF引起的Notch1-32D 细胞分化.结论分化抑制实验发现NOTCH 配体中,Delta4不能抑制G-CSF引起的Notch1-32D细胞分化,但Jagged2能抑制G-CSF引起的Notch1-32D细胞分化,为进一步揭示Notch信号传导系统的作用机制及其对造血系统的影响提供重要理论依据.     

高表达Delta4增加Notch基因的表达

Notch信号传导系统在介导造血细胞增殖与分化过程中具有重要作用。通过分子克隆技术成功构建含标记基因FLAG的Notch配基Delta4的高表达载体pTracer．CMV．Delta4．FLAG,瞬时转染COS7细胞,48h后收获细胞并制备融合蛋白,SDS-PAGE凝胶电泳和Western-blot证实后,将其稳定转染CHO细胞,Western-blot鉴定并筛选高表达Delta4的Delta4-CHO1-4及Delta4-CHO1-5细胞株,利用Luciferase分析方法进行Delta4功能性研究。研究结果发现Delta4对Notchl及Notch2均具有信号功能活性,且对后者的活性功能水平大于前者,说明Delta4为Notchl及Notch2的配基。与其他配基功能比较：对Notchl,Delta4的功能活性高于Deltal及Jaggedl,但略低于Jagged2;对Notch2,Delta4的功能活性低于Deltal、Jaggedl及Jagged2,但亦具有较强的活性水平。     

Delta 1与Jagged 1在诱导T细胞分化中的不同作用

Delta 1（又称delta—like 1或DⅡ 1）与Jagged 1（又称Serrate 1）是脊椎动物Notch的两个配体，它们与相同或不同的Notch受体结合激活Notch信号通路，决定细胞分化的命运，参与调控许多组织的生长发育。单独敲除Delta1或Jagged 1小鼠呈现不同的表型，提示这两个配体具有非重叠作用。Delta 1与Jagged 1在T细胞、抗原提呈细胞（antigen presenting cells，APC）如树突状细胞、B细胞和巨噬细胞等表面均有表达，在诱导T细胞分化中起着重要作用。近来发现，Delta 1可以诱导外周T细胞向1型辅助性T细胞（helper T cell 1，TH 1）分化，而Jagged 1诱导其向TH 2分化；     

Delta 1与Jagged 1在诱导胸腺细胞分化中的不同作用（文献综述）

Delta 1与Jagged 1是脊椎动物Notch的两个配体，它们与相同或不同的Notch受体结合激活Notch信号通路，决定细胞分化的命运，参与调控许多组织的生长发育。单独敲除Delta 1或Jagged 1小鼠呈现不同的表型，提示这两个配体具有非重叠作用。Delta 1与Jagged 1在造血细胞、骨髓间质细胞、淋巴细胞、抗原提呈细胞（antigen presenting cells，APC）等表面均有表达，诱导淋巴细胞的分化。近来发现，Delta 1诱导胸腺细胞其向T细胞分化，而Jagged 1则诱导胸腺细胞向NK细胞分化。     

与Sertoli细胞共培养大鼠大脑皮质神经前体细胞分化中Notch信号相关基因表达分析

目的:检测与Sertoli细胞共培养条件下大鼠大脑皮质神经前体细胞分化过程中Notch信号相关分子的表达变化.方法:分离大鼠Sertoli细胞和大脑皮质神经前体细胞.神经前体细胞与Dertoli细胞共培养条件下,用Real-time PCR相对定量法,分别检测Notch受体Notch1、Notch2、Notch3、Notch4及其配体Delta1、Delta3、Jagged1、Jagged2,以及ngn1、hes1基因的表达变化.结果:培养细胞汇片后第5天,Sertoli细胞Notch1～4基本表达维持不变;其配体Jagged2表达较高,但不表达Delta1;同时也不表达ngn1、hes1.神经前体细胞Notch表达水平较高,尤以Notch1和Notch2为高;而其配体以Jagged1、Jagged2表达较高,前神经因子Hes1有较高表达,而Ngn1表达较低.细胞共培养条件下,Notch受体、配体以及Hes1与神经前体细胞组相比都有明显的下调,而Ngn1却相对有所提高.结论:Sertoli细胞可通过Notch配体的作用,调节神经前体细胞Notch信号分子的表达.在共培养条件下,Sertoli细胞对神经前体细胞Hes1表达有抑制作用,但对Ngn1有正向调节作用,从而影响神经前体细胞的分化.     

Notch配体在诱导淋巴细胞分化中的不同作用

脊椎动物Notch配体为一组单次跨膜蛋白,包括Delta1, Delta3, Delta4, Jagged1 和 Jagged2,它们与Notch受体结合激活Notch信号通路,决定细胞分化的命运.这些Notch配体具有不同的生物学特性,在诱导造血干/祖细胞和胸腺细胞向T、B和NK细胞分化中起着不同作用,可能为免疫性疾病的治疗提供新的药物靶点.     

Notch1在前列腺癌细胞PC3中对其配体Jagged1表达的调控

目的探讨在前列腺癌细胞PC3中,Notch1和Jagged1对细胞生长的影响,及Notch1对其配体Jagged1表达的调控作用。方法通过siRNA干扰的方法分别抑制Notch1和Jagged1蛋白的表达,用MTT法检测PC3细胞的生长;分别通过siRNA干扰和转染质粒的方法,抑制和促进PC3细胞中Notch1蛋白的表达,用Western blot检测Jagged1蛋白水平,用Real-timePCR检测Jagged1 mRNA水平。结果抑制Notch1和Jagged1蛋白的表达后,PC3细胞的生长减慢;抑制Notch1的表达引起Jagged1的蛋白水平下降而过表达Notch1引起Jagged1的蛋白水平上升,同时,Jagged1蛋白水平与mRNA水平的变化不一致。结论Notch1和Jagged1对前列腺癌细胞PC3的生长有重要影响。Notch1可以调控其配体Jagged1的表达。     

天花粉蛋白及其肽段诱导的免疫抑制依赖APC表面Notch配体的表达

新近发现,Notch信号途径参与调节外周成熟T细胞及其亚群的分化和功能发挥。本研究应用天花粉蛋白及其衍生肽处理骨髓来源的小鼠树突状细胞(DC),检测Notch配体家族分子的表达及DC对CD8+T细胞分泌细胞因子的影响。结果表明,天花粉蛋白或其衍生肽PB处理DC可使Notch配体Jagged1、Delta1分子表达明显增加,并改变CD8+T细胞细胞因子分泌格局,明显抑制Th1相关细胞因子IFN-γ的分泌,而Th2相关细胞因子IL-4和IL-10分泌明显增加。Notch信号的阻断剂可以部分逆转Tk及肽段的抑制作用。表明天花粉蛋白及其衍生肽可诱导一群具有抑制能力的CD8+T细胞,该作用依赖于DC表面Notch配体的表达。     

类风湿性关节炎患者外周血单个核细胞MALAT1表达降低、 hsa-miR155-3p表达增加并参与Notch信号通路调节

目的观察类风湿性关节炎(RA)患者长链非编码RNA(lncRNA)转移相关肺腺癌转录本1(MALAT1)、 hsa-miR155-3p表达及其与Notch信号通路关系,探讨RA可能发病机制。方法采取RA患者(RA组)、健康对照者(NC组)外周血,采用高通量基因测序筛选差异表达的lncRNA和mRNA。利用反转录PCR检测筛选的lncRNA MALAT1和hsa-miR155-3p及Notch信号通路受体配体表达。结果通过lncRNA测序分析发现,共有9158条差异表达的lncRNA。基因本体(GO)功能分类注释得出差异表达的mRNA参与免疫炎症反应、细胞转录调节等。通过通路(pathway)分析显示差异表达的mRNA与免疫炎症、应激、 Notch通路等功能基因变化有关。经顺式(Cis)和反式(Trans)预测, Jagged1/2、 Delta1/2、 Notch1/2可能与RA发生密切相关。与NC组相比, RA组MALAT1降低, hsa-miR155-3p明显升高; Notch通路配体Delta1、 Delta2、 Jagged1、 Jagged2及受体Notch1、 Notch2表达升高。相关性分析显示, hsa-miR155-3p与MALAT1呈负相关, hsa-miR155-3p与Notch通路Delta1、 Jagged1呈正相关, MALAT1与Jagged2呈负相关。结论 RA患者lncRNA MALAT1降低、 hsa-miR155-3p增加,共同调节Notch信号通路变化。     

Jagged1蛋白抑制神经干细胞向神经元分化的实验

目的：研究Jagged1蛋白对神经干细胞分化的影响。方法：分离小鼠胚胎脑神经干细胞,用Jagged1蛋白、Jagged1蛋白 γ泌肽酶体外诱导神经干细胞分化,观察分化后神经元所占的比例。结果：分离的细胞能持续增殖,并能分化为神经元、星形胶质细胞和少突胶质细胞;在Jagged1蛋白的影响下,分化后神经元数量明显减少;γ泌肽酶抑制剂能阻断Jagged1蛋白的诱导作用。结论：培养的细胞为神经干细胞,并表达Notch受体;Jagged1蛋白能抑制干细胞向神经元分化,这种分化作用是通过Notch受体实现的。     

Level of functioning of siblings and parents of probands of varying degrees of retardation

A further analysis of already published data supports the position that retardates of low ability level less frequently have retarded siblings, retarded parents, and parents low in occupational level than do retardates higher in ability level. The analysis supports the position that there are two types of retarded individuals, persons retarded as a result of gene or chromosomal anomalies, brain injury, etc., who more frequently occur in the lower-level retardate group, and persons whose retardation represents polygenic segregation, who more frequently occur in the higher-level group.     

局部注射辛伐他汀对骨质疏松大鼠股骨髁骨小梁的改建效应

背景：局部注射具有成骨作用的辛伐他汀,可显著增加骨质疏松大鼠股骨颈及股骨髁部的骨密度及力学强度,分析局部注射辛伐他汀对股骨髁骨小梁的影响。 目的：进一步研究骨质疏松大鼠股骨内局部注射辛伐他汀对股骨髁骨小梁的影响。为将辛伐他汀应用于临床骨质疏松局部治疗提供实验基础。 方法：18只雌性SD大鼠双侧卵巢切除后3个月,制备大鼠骨质疏松模型。实验大鼠随机数字表法均分为3组,分别在实验大鼠的右侧股骨髓腔内单次注射辛伐他汀溶液5 mg、10 mg,对照组单纯注射空白载体。分别在注射后1个月处死大鼠并取材。Micro-CT扫描并定量分析骨组织形态变化。 结果与结论：给药后1个月,Micro-CT扫描结果显示,辛伐他汀治疗组的骨微结构参数如骨皮质厚度、骨小梁密度及连接率明显优于对照组。说明疏松骨骼单次注射小剂量辛伐他汀可显著促进股骨髁部骨小梁改建,改善骨骼微结构,可为强化局部、防治骨质疏松骨折的新选择进一步提供实验基础。     

石斛属民族药用植物的分类及生药学研究

中药石斛早在《神农本草经》中就被列为上品,其应用历史悠久,具有养阴生津、补肾益气、润喉护嗓、活血明目、抗癌防老等功效。本文对国内外石斛属民族药用植物的分类及生药学研究进行了综述。     

膝关节骨性关节炎的关节镜治疗体会

目的我科自2001～2005年4月对59例67膝的骨性关节炎（0A），进行关节镜检查及镜下清理术。方法镜检：膝关节骨性关节炎伴不同程度滑膜炎，滑膜皱壁粘连纤维片，关节软骨不同程度破坏。镜下清除增生滑膜，松解粘连，去除剥脱软骨、修复关节面。结果随访2月～4年，平均1.5年，术后综合评估2月～1年，优良率94％；1～2年，优良率79.2％；2年以上优良率54.8％。结论关节镜对膝关节骨性关节炎诊断能提供了比较全面的资料，并对骨性关节炎早期有良好疗效，具有创伤小、恢复快、并发症少和重复治疗等优点。     

Anatomical study of distribution of valves of the cutaneous veins of adult''s limbs

Since the cutaneous veins of the four limbs have been used as autogenous grafts in the treatment of cardiovascular disorders, knowledge of distribution of the valves is increasingly required depending upon the use. In the gross anatomical study of distribution of valves of the trunci venae, there has been argument about locational relationships between the venous roots and the valves in the vicinity of the roots and the inter-valvular distance. However, there have been only few reports discussing detailed information about valves of the cutaneous veins of the four limbs. The authors observed patterns of distribution of the cutaneous venous valves of the four limbs of cadavers prepared for practice in anatomy. The following parts were excised from each cadaver: the cephalic, basilic, and the great saphenous veins, which originate from the acral venous network and flow into the proximal deep veins, and the venous roots communicating with these veins. An incision was made on each excised vein in the direction of the long axis under observation with a stereoscopic microscope, and the inter-valvular distance and the distance between the valve and the orifice of venous root in the vicinity of the valve were measured. The inter-valvular distance varied with type of the truncus venae, and it varied according to area even in the same truncus venae.     

Pathways of the realization of immunomodulating effects of neurotransmitters

Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 115, N ^o 4, pp. 401–404, April, 1993     

Characterization of sterols of three digenetic trematodes of buffalo

Sterols of three digenetic trematodes were isolated and characterized by infrared and 1H nuclear magnetic resonance spectroscopies, and gas-liquid chromatography and gas chromatography-mass spectrometry. Sterols identified were cholesterol, cholestanol, 24-methylcholesterol, 24-methylcholestanol, 24-ethyl-22-dehydrocholesterol, 24-ethyl-22-dehydrocholestanol, 24-ethylcholesterol and 24-ethylcholestanol.     

Mechanisms of generation of slow components of bioelectric activity in diagnostics of functional state of the gastrointestinal tract

New electrophysiologic devices were used for the diagnostics of the state of excitable structures in the gastrointestinal tract and correction of their motor functions. Bioelectrical and biomechanical activities form the basis of functioning of internal organs. The mechanisms of generation of slow bioelectrical activity that are important for clinical and physiological studies are described. One of these mechanisms is a capacitance parametric transducer converting the energy of contractions into specific electric signals reflecting muscle functions. Another mechanism results from slow oscillations of resting potentials of interrelated excitable cells in large neuromuscular structures of internal organs. The elaborated procedure is efficient for preventing early postoperative paresis of the gastrointestinal tract. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 10, pp. 448–452, October, 1999     

胃小弯侧血管弓的MDCT解剖

目的：研究正常胃小弯侧血管弓及其属支血管的多排螺旋CT表现。方法：回顾性分析51例正常成人的上腹部CT资料,观察胃小弯侧血管弓及其属支血管的显示情况、血管弓形态及其与胃壁的关系。结果：51例均显示胃小弯侧血管弓,血管弓贲门部属支、胃体部属支和幽门部属支分别显示46例、15例和10例;血管弓与胃壁紧邻41例。结论：MDCT检查可良好显示正常胃小弯侧血管弓及其属支血管。     

Assessment of the efficacy of treatment of hemangiomas

Assessment of the aftereffects of cryoexposure and ultrahigh-frequency cryoexposure on hemangioma tissue of various types, cavernous and squamous, showed a higher cryogenic effect in hemangiomatous tissue preexposed to ultrahigh-frequency waves. A quantitative criterion is proposed for assessing the efficacy of the studied methods of exposure. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 6, pp. 669–672, June, 1995