ATRA联合Genistein对A549人肺腺癌细胞株MUC1表达及转移潜能的影响

目的 MUC1是一种跨膜粘蛋白,在肿瘤进展过程中起重要作用,MUC1可作为多种肿瘤预后判断的有用标记,本实验探讨了全反式维甲酸(all-trans retinoic acid,ATRA)联合三羟异黄酮(Genistein)对肺腺癌细胞MUC1蛋白、mRNA的表达及转移潜能的影响.方法 ATRA、Genistein及两者联合处理人肺腺癌细胞株A549,免疫组化法检测MUC1蛋白的表达情况,荧光定量PCR法检测MUC1 mRNA的表达,根据细胞穿过铺有Matrigel胶多孔膜的数量检测细胞侵袭力.结果 ATRA和Genistein联合处理A549肺癌细胞后,具有协同下调细胞MUC1蛋白、mRNA的表达,抑制A549细胞体外侵袭能力的作用.结论 ATRA联合Genistein可能通过协同下调MUC1蛋白及mRNA的表达,从而抑制肺癌A549细胞的转移潜能.     

c-Met基因调控肺腺癌A549细胞放射敏感性

目的探讨上调和下调肺腺癌细胞株A549细胞中c-Met基因表达水平后,该细胞鼠移植瘤放射敏感性的变化情况。方法利用电穿孔法将c-Met siRNA和重组表达载体pc DNA3.1-c-Met分别转染入肺癌A549细胞后采用G418筛选得到稳定转染的肺癌细胞系。转染48 h后用RT-PCR法和Western blot法分别检测细胞中c-Met基因和蛋白表达水平,采用CCK-8法检测细胞增殖活性;克隆形成实验研究分别上调和下调c-Met基因表达水平对肺腺癌细胞株A549放射敏感性的影响;TUNEL法检测细胞凋亡影响,移植瘤实验检测基因沉默和激进c-Met基因表达水平并联合放射射线照射对肺腺癌细胞生长的抑制作用。结果 c-Met下调组中肺癌A549细胞内c-Met基因和蛋白表达水平明显降低,而上调组中的表达水平则得到了明显升高。c-Met下调组肺癌A549细胞放射敏感性升高,c-Met上调组肺癌A549细胞放射敏感性降低(P0.05)。下调组中肺癌A549细胞的增殖活性受到抑制而细胞凋亡率显著增加(P0.05),上调组肺癌细胞的增殖能力明显增强而细胞凋亡率明显降低(P0.05);下调组裸鼠移植瘤的平均体积明显小于对照组,而上调组裸鼠移植瘤的瘤体平均体积则显著大于对照组(P0.05)。结论基因沉默c-Met的基因表达水平能显著降低肺癌细胞的增殖活性,抑制人肺癌A549细胞裸鼠移植瘤的生长并明显提高移植瘤瘤体的放射敏感性。     

GANT61 阻断Hedgehog 信号通道调控肺腺癌A549 / DDP 细胞耐药的机制研究

目的:探讨GANT61阻断Hedgehog信号通道调控肺腺癌A549/DDP细胞增殖、细胞周期及耐药的机制研究。方法:应用CCK8法检测顺铂(DDP)对肺腺癌细胞株A549和A549/DDP的半数抑制浓度(IC50),从而计算A549/DDP细胞的耐药指数;同时采用Western blot检测A549、A549/DDP细胞中Gli1、Ptch1、Shh、XRCC1蛋白的表达水平。用不同浓度的GANT61干预A549/DDP细胞后,CCK8法检测增殖活力。流式细胞术检测GANT61对A549/DDP细胞凋亡率和细胞周期的影响。Western blot检测GANT61对A549/DDP细胞周期和凋亡相关蛋白表达的影响。采用20μmol/L GANT61联合不同浓度的顺铂干预A549/DDP细胞,CCK8法检测各组细胞增殖抑制率变化及Western blot检测各组细胞中XRCC1蛋白的表达水平。结果:A549/DDP细胞的耐药指数为5. 34。A549/DDP细胞与A549细胞相比,Gli1、Ptch1、Shh、XRCC1在蛋白表达水平明显升高,差异有统计学意义(P 0. 01或P 0. 05)。使用GANT61处理A549/DDP细胞,当其浓度分别≥20μmol/L、≥10μmol/L时,分别培养24 h和72 h,与对照组相比肿瘤细胞增殖活性下降(P0. 01),且呈现剂量依赖性。GANT61可通过下调c-myc、cyclin D1表达(P0. 01),阻滞细胞周期于G1期(P0. 01)。GANT61能够明显诱导A549/DDP细胞发生凋亡(P0. 05),其分子机制可能与促进caspase-3的剪切活化相关(P0. 05)。GANT61联合顺铂用药后使A549/DDP细胞抑制率显著增加,呈协同效应,且伴随XRCC1蛋白表达下降(P0. 01)。结论:Gli1在人肺腺癌耐药细胞A549/DDP中的表达明显高于顺铂敏感株A549,提示Gli1可能与肺腺癌顺铂耐药相关,且XRCC1蛋白在A549/DDP细胞高表达。故采用Gli1抑制剂GANT61能够明显抑制A549/DDP细胞增殖,阻滞细胞周期,并诱导细胞凋亡。体外试验中,GANT61与顺铂联合用药可以协同抑制增殖作用,其机制可能与下调XRCC1蛋白表达有关。     

PLCE1通过调控p53抑制肺腺癌A549细胞凋亡

目的:探讨磷脂酶Cε1(PLCE1)抑制肺腺癌A549细胞凋亡的作用机制。方法:选用人肺腺癌细胞株A549作为研究对象。采用real-time PCR和Western blotting法分别检测PLCE1抑制剂U-73122处理前、后肺腺癌细胞株A549中PLCE1和p53 mRNA和蛋白水平的表达;流式细胞术检测细胞凋亡。结果:肺腺癌细胞株A549高表达PLCE1,低表达p53;抑制PLCE1表达后A549细胞中p53表达上调,细胞凋亡明显增加。结论:PLCE1通过抑制肺腺癌A549细胞株中p53的表达,从而抑制A549细胞凋亡。     

MADD在肺腺癌组织中的表达及对肺腺癌A549细胞增殖和凋亡的影响

目的:探讨MADD在肺正常组织及肺腺癌组织中的表达及其对肺腺癌细胞增殖和凋亡的影响。方法:收集肺临床病理组织标本,免疫组化法检测肺正常组织和肺癌组织MADD表达;培养人肺腺癌A549细胞,逆转录PCR检测其IG20基因表达;用携带MADD基因的质粒和能够沉默MADD表达的慢病毒载体分别转染A549细胞,Western blot、MTT分析和流式细胞术检测其MADD表达、增殖及凋亡。结果:肺腺癌组织MADD表达水平明显高于肺正常组织和肺鳞癌组织;A549细胞能够表达MADD;高表达MADD能抑制A549细胞凋亡,提高其增殖活力,而沉默MADD表达则能促进A549细胞凋亡,降低其增殖活力。结论:肺腺癌组织MADD表达明显增高;MADD可通过抑制凋亡来促进肺腺癌细胞生存。     

Polo-like激酶l反义RNA对肺癌细胞A549生长的实验研究

目的： 探讨Polo-like激酶1（Plk1）基因表达下调对肺癌细胞周期分布及其生长的影响。方法： 培养肺腺癌细胞株A549，构建表达Plk1反义RNA的质粒pcDNA3-Plk1，通过脂质体介导转染A549细胞，采用RT-PCR和Western blotting的方法检测Plk1基因的表达，细胞计数、BrdU脉冲标记检测细胞增殖，流式细胞仪分析细胞周期变化和凋亡，MTT法检测长春瑞宾（NVB）对各组细胞的生长抑制率。结果： A549细胞转染pcDNA3-Plk1后24 h，Plk1 mRNA及蛋白表达均下降；细胞变圆、漂浮、增殖减慢；S期细胞百分数（BrdU标记指数）显著低于对照组（P<0.05）；转染后48 h A549细胞出现G₂/M期阻滞（P<0.05）并发生凋亡；等浓度化疗药物诺维本对转染pcDNA3-Plk1细胞的抑制率明显高于各对照组（P<0.05），转染pcDNA3与未转染的对照细胞差异无显著（P>0.05）。结论： pcDNA3-Plk1的转染能下调Plk1基因的表达，抑制A549细胞增殖，诱导凋亡，并能增加A549细胞对化疗药物的敏感性。     

雷帕霉素对顺铂作用下人肺腺癌A549及耐药A549/DDP细胞增殖、侵袭、黏附及自噬凋亡的影响

 目的:研究雷帕霉素（Rap）对顺铂（DDP）作用下人肺腺癌A549及耐药A549/DDP 细胞增殖、迁移、黏附及其自噬凋亡的影响。方法: 培养人肺腺癌A549及耐药A549/DDP细胞株,利用MTT方法分别检测Rap和DDP单独与联合作用对A549及耐药A549/DDP细胞增殖抑制率的影响;Transwell方法检测Rap和DDP单独与联合作用对A549及耐药A549/DDP细胞体外侵袭能力的影响;黏附实验检测Rap和DDP单独与联合作用对A549及耐药A549/DDP细胞体外侵袭能力的影响;流式细胞术检测Rap和DDP单独与联合作用对A549及耐药A549/DDP细胞凋亡的影响;Western blotting检测Rap和DDP单独与联合作用对A549及耐药A549/DDP细胞自噬标志蛋白beclin-1和LC3表达的影响。 结果: 与Rap或DDP单独作用组相比,Rap和DDP联合作用能够同时显著抑制人肺腺癌A549及耐药A549/DDP细胞增殖、体外侵袭能力及细胞黏附能力,并能够促进细胞凋亡和自噬标志蛋白beclin-1和LC3的表达（均P＜0.05）。结论: Rap能够通过促进细胞自噬而增强DDP的作用,进而抑制人肺腺癌A549及耐药A549/DDP细胞的增殖、侵袭、黏附并促进细胞凋亡作用,具有协同作用。     

RNA干扰抑制乙酰肝素酶对肺腺癌细胞A549增殖、侵袭和凋亡的影响

目的 探讨抑制乙酰肝素酶表达对人肺腺癌细胞A549增殖、侵袭和凋亡的影响.方法 构建靶向人乙酰肝素酶基因短发夹状双链RNA(shRNA)真核表达质粒(pshRNA-Hpa),用脂质体将其转入A549细胞,建立稳定转染细胞株系,经逆转录.PCR和Western blot法检测转染效率后,分别用四甲基偶氮唑蓝(MTT)、Transwell侵袭实验和流式细胞术(AnnexinV-FITC/PI双染法)检测其对A549细胞增殖、侵袭和凋亡的影响.采用Western blot法检测转染后磷酸化蛋白激酶B(PKB/Akt-Ser473)的表达.结果 转染pshRNA-Hpa的A549细胞乙酰肝素酶mRNA及蛋白表达明显低于未处理A549细胞,抑制率分别为54.09%和48.26%,细胞增殖速度减缓,侵袭能力降低,流式细胞术显示早期细胞凋亡率达(12.53±0.34)%,较空白对照组、空载体转染细胞组和阴性对照组明显增加(P＜0.01).同阴性对照质粒相比,转染pshRNA-Hpa的A549细胞Akt-Ser473的磷酸化水平降低52.15%.结论 靶向乙酰肝素酶重组shRNA质粒pshRNA-Hpa能有效抑制A549细胞乙酰肝素酶的表达,并可能通过下调磷酸化蛋白激酶B途径抑制细胞的增殖侵袭、促进细胞的凋亡.     

Polo-like激酶1反义RNA对肺癌细胞A549生长的实验研究

目的:探讨Polo-like激酶1(Plk1)基因表达下调对肺癌细胞周期分布及其生长的影响。方法:培养肺腺癌细胞株A549,构建表达Plk1反义RNA的质粒pcDNA3-Plk1,通过脂质体介导转染A549细胞,采用RT-PCR和Western blotting的方法检测Plk1基因的表达,细胞计数、BrdU脉冲标记检测细胞增殖,流式细胞仪分析细胞周期变化和凋亡,MTT法检测长春瑞宾(NVB)对各组细胞的生长抑制率。结果:A549细胞转染pcDNA3-Plk1后24h,Plk1mRNA及蛋白表达均下降;细胞变圆、漂浮、增殖减慢;S期细胞百分数(BrdU标记指数)显著低于对照组(P<0.05);转染后48hA549细胞出现G2/M期阻滞(P<0.05)并发生凋亡;等浓度化疗药物诺维本对转染pcDNA3-Plk1细胞的抑制率明显高于各对照组(P<0.05),转染pcDNA3与未转染的对照细胞差异无显著(P>0.05)。结论:pcDNA3-Plk1的转染能下调Plk1基因的表达,抑制A549细胞增殖,诱导凋亡,并能增加A549细胞对化疗药物的敏感性。     

3-溴丙酮酸通过抑制糖酵解阻止A549肺腺癌细胞增殖并诱导其凋亡

目的探究3-溴丙酮酸(3-BP)对人肺腺癌A549细胞的增殖和凋亡的影响及机制。方法采用噻唑蓝(MTT)法检测(10、 20、 40、 80、 160)μmol/L3-BP处理24、 48 h,对肺腺癌A549细胞存活率的影响;采用(0、 50、 100、 150)μmol/L 3-BP处理肺腺癌A549细胞24 h,异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双染法结合流式细胞术检测A549细胞的凋亡率;试剂盒检测A549细胞ATP水平、 2′, 7′-二氯二氢荧光黄二乙酸酯(DCFH-DA)染色结合流式细胞术检测细胞活性氧(ROS)水平变化,Western blot法检测糖酵解及凋亡相关蛋白己糖激酶2(HK-2)、单羧酸转运蛋白1(MCT1)、丙酮酸脱氢酶激酶1(PDK1)和凋亡相关B细胞淋巴瘤分子2(Bcl2)、 Bcl2相关X蛋白(BAX)的表达。近红外荧光蛋白(iRFP)和荧光素酶双标记的A549细胞接种至裸鼠皮下,当皮下肿瘤体积达到100 mm~3时,将所有裸鼠随机分成PBS组、 5 mg/kg顺铂(DDP)组、 7 mg/kg 3-BP组;每组5只,采用腹腔注射方式进行给药。HE染色检测肿瘤病变情况,免疫组织化学染色检测KI-67抗原(ki67)表达确定对肿瘤细胞增殖的影响,原位末端转移酶标记技术(TUNEL)检测肿瘤细胞凋亡情况,评估3-BP的体内抗肿瘤效果并进行肝肾功能检测确定3-BP肝肾毒性。结果 3-BP可以抑制A549细胞增殖,诱导其凋亡;与对照组相比,3-BP处理组ATP水平明显下降,ROS水平明显升高,HK-2、 MCT1、 PDK1、 Bcl2蛋白表达明显降低,BAX表达明显增加。动物模型结果显示,与PBS组相比,3-BP可以抑制裸鼠肺腺癌移植瘤的生长,3-BP可以抑制肿瘤细胞的增殖并诱导其凋亡,3-BP对裸鼠肝肾毒性较为轻微,安全性较高。结论 3-BP通过抑制糖酵解抑制A549人肺腺癌细胞增殖并诱导其凋亡,对荷肺癌裸鼠的抗肿瘤效果较好且肝肾毒性较低。     

Photoperiodicity and the function of the hypophysis and the adrenal cortex

Summary The ACTH content of the hypophysis of rats rises from a minimum in the morning to a maximum in the evening hours. These fluctuations are associated with the diurnal rhythm of light and darkness, and are abolished under conditions of continual light or darkness for 30 days. Rhythmic fluctuations of the ascorbic acid content of the adrenals are also found, with maximum values in the evening, and these are likewise abolished by maintenance under conditions of constant illumination.Presented by Active Member AMN SSSR V. V. Parin     

Characteristics of the topographic interrelation of the head of the pancreas and the duodenum and pathology of the organs of the pancreatoduodenal area

Seventy pancreatoduodenal complexes of 55 patients with chronic pancreatitis and tumours of this zone and 15 patients died from other diseases are studies histotopographically . The pieces of the pancreatic head tissue in the medial wall of the duodenum were found in 12 cases of the first group and in 4 control cases. The pancreatic tissue consisted either of all elements of this organ or cystically dilated ducts and seemed to infiltrate different layers of the duodenum wall. Three variants of the pancreatic head structure are suggested on the basis of anatomo-topographical interrelationships of the pancreatic head and duodenum. In 12 out of 14 cases chronic pancreatitis and carcinoma of organs of this zone were combined with the variants of the pancreatic head structure, in 2 cases there was a true heterotopy . Pathogenetic significance of these variants for the development of chronic pancreatitis is discussed.     

指背腱膜滑动与近侧指间关节屈曲关系及临床意义

目的:探讨指背腱膜滑动距离与近侧指间关节(PIP)屈曲关系,为临床修复提供解剖学基础。方法:男性成人新鲜尸体标本10侧30指(示、中、环指各10指),切除手指皮肤,不破坏腱鞘、屈肌支持带、伸肌支持带、内在肌及外在肌,使肌腱保持正常的生理状态,分别测量各指中央束(CS)、侧束(LB)在PIP屈曲45°和90°时的滑动距离。结果:当PIP屈曲45°时,CS滑动距离为(2.7±0.4)mm,LB滑动距离为(2.8±0.6)mm;当PIP屈曲90°时,CS滑动距离为(4.3±0.7)mm,LB滑动距离为(4.8±0.6)mm。结论:指背腱膜滑动距离减少,严重影响手指的屈曲功能。对于指背腱膜的新鲜性损伤应予以精确修复;对于陈旧性损伤的修复应确保指背腱膜的正常滑动范围。     

产后妇女全血钙和骨源性碱性磷酸酶（ABAP）活性的检测与分析

目的结合我院特点,测定来我院进行产后检查妇女的骨源性碱性磷酸酶（ABAP）活性以及全血钙。方法采用金标法测定产妇3031人骨源性碱性磷酸酶（ABAP）的活性,并从中随机选择855人采用BH-5100型多通道原子吸收光谱仪检测全血钙。结果经测定,3031名产妇中679人骨源性碱性磷酸酶（ABAP）的活性异常,异常者检出率为22.4%,其中83.5%的产妇轻度缺钙,而16.5%的产妇严重缺钙;而全血钙异常检出率为17.0%。结论ABAP活性检测方法比全血钙检测方法测定体内钙营养水平更具敏感性,可作为检测评价产妇钙营养状况的一种简易可靠的方法,为产后骨质疏松提供了一种可靠、便捷、科学合理的指标。     

Cycloversion and cyclovergence: the effects of the area and position of the visual display

Rotation of a display in the frontal plane evokes a conjugate nystagmic rotation of the eyes (cycloversion) about the visual axes, with slow phases in the direction of stimulus motion — a response known as torsional optokinetic nystagmus (TOKN). Antiphase rotation of large dichoptic displays evokes a disconjugate rotation of the eyes about the visual axes, a response known as cyclovergence. Using the scleral-coil technique for monitoring eye movements we recorded TOKN evoked by black-and-white sectored displays rotating about the visual axis at an angular velocity of 30°/s. The display was confined to central areas with diameters ranging from 5° to full field or with the central 5° to 75° occluded. A 5° central display evoked TOKN with 40% of the gain for the full-field display and gain increased as a function of the size of the display. The gain of TOKN decreased with increasing size of a central occluder. These characteristics of TOKN are similar to those of horizontal OKN. Cyclovergence was virtually absent with a 5° display but was immune to occlusion of the central 40°. Cyclovergence therefore differs from cycloversion in showing no preference for centrally placed stimuli. These effects are free from the influence of stationary edges, since these were concentric with the stimulus motion. The effects are also free from the influence of voluntary pursuit, since humans do not normally have voluntary control over torsional eye movements.     

Organization and distribution of the upper and lower esophageal motoneurons in the medulla and the spinal cord of the rat

The upper cervical esophagus is exerted on swallowing and peristalsis by somatic and visceral motoneurons, whereas the lower esophagus is exerted on only peristalsis by visceral motoneurons. We examined the origin of the esophageal motoneurons and whether there were any differences between the distributions of the upper and the lower esophageal motoneurons in the medulla and the spinal cord using cholera toxin subunit b (CTb) as the retrograde tracer. Following injection of CTb into the cervical esophagus resulted in heavy labeling of the neurons in the nucleus ambiguus including the compact (AmC), semicompact (AmS) and loose (AmL) formations, and the medial column of lamina IX at the C1-C5 levels of the cervical spinal cord corresponding to the spinal accessory nucleus. A few labeled neurons were found in the inferior salivatory nucleus, the rostral division of the dorsal motor nucleus of the vagus (DMX), the accessory facial nucleus and the lateral column of lamina IX at the C2 and C3 levels. All these labeled neurons showed ChAT immunoreactivity. When CTb was injected into the cut end of the unilateral recurrent laryngeal nerve, many labeled neurons were found in the ipsilateral AmC, the AmL, and the bilateral medial column at the C1 and C2 levels. Following injection of CTb into the subdiaphragmatic esophagus resulted in heavy labeling of the neurons only in the AmC and the DMX. When CTb was injected into the sternomastoid muscle, many labeled neurons were found in the medullary reticular formation, the facial nucleus, the medial column at the C1-C3, C5 and C6 levels, and the lateral column at the C2, C3, C5 and C6 levels. Injections of a Fluoro-Gold into the cervical esophagus and a CTb into the sternomastoid muscle or the subdiaphragmatic esophagus in the same animal showed many double labeled neurons in the medial column of the accessory nucleus at the C1 and C2 levels, but no double labeled neurons in the AmC. These results indicated that the upper cervical esophagus is innervated by the visceral medullary vagal motoneurons as well as the somatic spinal accessory motoneurons. The lower esophagus is innervated only by the visceral medullary vagal motoneurons.