肿瘤坏死因子相关凋亡诱导配体联合化疗药物杀伤胰腺癌细胞的研究

目的研究肿瘤坏死因子相关凋亡诱导配体以及联合应用亚毒性剂量的化疗药物对胰腺癌的治疗作用。方法半定量 RT-PCR 检测 TRAILR mRNA 在胰腺癌细胞株 Canpan-2中的表达。DNA 琼脂糖凝胶电泳检测细胞凋亡情况。应用不同浓度的 TRAIL 及联合亚毒性剂量的氟尿嘧啶(5-fluorouracil)、吉西他滨(gemcitabin)处理胰腺癌细胞,通过 MTT 法检测细胞毒性作用,流式细胞仪分析细胞凋亡率。结果死亡受体 DR4、DR5及诱骗受体 DcR1、DcR2在胰腺癌细胞株 Canpan-2中均有表达。DNA 琼脂糖凝胶电泳可见到典型的凋亡梯形带。TRAIL100 ng/ml 作用胰腺癌细胞24h 后,细胞杀伤率为(29.5±1.2)%,且其作用存在浓度依赖性;联合应用亚毒性剂量的氟尿嘧啶、吉西化滨能够大大提高TRAIL 的细胞毒活性,杀伤率分别为 TRAIL+氟尿嘧啶(43.7±1.4)%,TRAIL+吉西化滨(49.8±1.2)%,联合用药前后差异有显著性(P<0.01)。结论 TRAIL 受体在胰腺癌细胞普遍表达,并存在受体类型的表达差异;TRAIL 与化疗药物氟尿嘧啶、吉西他滨有协同杀伤胰腺癌细胞的作用。     

胰胆管合流异常患者胆汁对人胆管癌细胞生长的影响

目的　研究胰胆管合流异常患者 (APBDU)胆汁对人胆管癌细胞QBC93 9生长的影响。方法　应用四唑氮蓝比色 (MTT)法检测QBC93 9增殖状况 ,应用流式细胞仪测定细胞周期和凋亡 ,采用酶联免疫吸附试验 (ELISA)检测前列腺素E2 (PGE2 )含量。结果　APBDU胆汁与正常对照组胆汁比较明显促进QBC93 9细胞增殖 (P <0 .0 5 ) ,这种促癌活性可被环氧合酶 2 (COX 2 )选择性抑制剂Celecoxib拮抗。用APBDU患者胆汁处理 2 4h的QBC93 9细胞增殖指数显著上升(P <0 .0 5 ) ,S期细胞比例为 ( 3 2 .2 4± 1.3 3 ) % ,比正常对照组 ( 10 .5 6± 1.47) %明显增高 (P <0 .0 5 ) ,G0 /G1期细胞比例为 ( 4 9.72± 4.13 ) % ,比正常对照组 ( 74.81± 5 .10 ) %明显降低 (P <0 .0 5 )。结论　APBDU胆汁具有潜在的促癌活性 ,这种促癌活性依赖COX 2和PGE2 途径。     

胆管结石患者胆汁对人胆管癌细胞增殖的影响

目的观察胆管结石患者胆汁对人胆管癌细胞QBC939生长的影响 ,探讨胆管结石与胆管癌发生、发展的关系。方法应用噻唑蓝比色法检测 2 0份胆管结石患者胆汁和 10份正常胆汁对QBC939增殖的影响 ,应用流式细胞仪测定细胞周期和凋亡。结果胆管结石患者胆汁与正常胆汁比较 ,明显促进QBC939细胞增殖 ,用胆管结石患者胆汁处理 4 8h的QBC939细胞增殖指数显著上升 (P <0 0 1) ,胆管结石胆汁组 (47%± 10 % )S期细胞比例比正常胆汁组 (2 3%± 3% )明显增高 (P <0 0 1) ,G0 /G1期细胞比例 (42 %± 8% )比正常胆汁组 (6 3%± 10 % )明显降低 (P <0 0 5 )。结论胆管结石患者胆汁具有潜在的促增殖活性 ,胆管结石与胆管癌的发生、发展关系密切     

肝癌合并肝硬化患者肝癌切除后机体免疫状态的变化

目的探讨肝癌合并肝硬化患者癌灶切除前后机体免疫状态的变化。方法以肝硬化患者为对照组 ,采用流式细胞技术 (FCM)及ELISA方法分析 18例中晚期肝癌合并肝硬化患者癌灶切除前后外周血T细胞亚群CD4、CD8、CD4 /CD8及Th1/Th2细胞因子IFN γ、IL2、IL10蛋白水平的变化。结果　肝癌切除术后CD4 (33± 3) %、CD4 /CD8(1 1± 0 1)、IL2 (71± 11)pg/ml、IFN γ(90± 15 )pg/ml回升 ,高于术前水平〔CD4 (2 9± 4 ) %、CD4 /CD8(0 9± 0 3)、IL2 (5 7± 15 )pg/ml、IFN γ(78± 13)pg/ml〕 ,但仍低于肝硬化组〔CD4 (37± 4 ) %、CD4 /CD8(1 3± 0 2 )、IL2 (82± 15 )pg/ml、IFN γ(10 4± 2 2 )pg/ml〕(P <0 0 1或P <0 0 5 ) ;CD8(32± 3) %、IL10 (70± 9)pg/ml下降 ,低于术前〔CD8(35± 6 ) %、IL10(81± 15 )pg/ml〕水平但高于肝硬化组〔CD8(2 9± 2 ) %、IL10 (6 1± 10 )pg/ml〕(P <0 0 5 )。结论　癌灶切除后 ,机体免疫功能有明显改善。但仍未恢复到肝硬化患者的水平。     

胰胆管合流异常患者胆汁对人胆管癌细胞生长的影响

目的　研究胰胆管合流异常患者胆汁对人胆管癌细胞生长的影响 ,探讨胰胆管合流异常与胆管癌发生和发展的关系。方法　应用四唑氮蓝 (MTT)比色法检测胰胆管合流异常患者胆汁对人胆管癌细胞 QBC939增殖的影响 ,应用流式细胞仪 (FCM)测定细胞周期和细胞凋亡 ,采用酶联免疫吸附试验 (EL ISA)检测前列腺素 E2 (PGE2 )比例。结果　胰胆管合流异常患者胆汁与正常对照组胆汁比较明显促进胆管癌细胞 QBC939的增殖 (P<0 .0 1) ,这种促癌活性可被 COX- 2选择性抑制剂 celecoxib拮抗。细胞周期分析用胰胆管合流异常患者胆汁处理 2 4 h的 QBC939细胞增殖指数显著上升 (P<0 .0 1) ,S期细胞比例为 32 .2 4± 1.33% ,比正常对照组 (10 .5 6± 1.4 7% )明显增高 (P<0 .0 5 ) ,G0 / G1期细胞比例为 4 9.72 %± 4 .13% ,比正常对照组 (74 .81± 5 .10 % )明显降低 (P<0 .0 4 2 ) ,但胰胆管合流异常患者胆汁对 QBC939细胞凋亡率无明显影响 (P=0 .882 )。结论　胰胆管合流异常患者胆汁具有潜在的促癌活性 ,这种促癌活性依赖 COX- 2和 PGE2 途径     

1,25二羟维生素D3对胆管癌细胞系QBC939增殖和凋亡的影响

目的 探讨1,25二羟维生素D3[1,25(OH)2 D3]对胆管癌细胞系QBC939的体外增殖及凋亡的影响.方法 将不同浓度的1,25(OH)2 D3与胆管癌细胞系QBC939共同培养,采用MTT法测定细胞增殖能力、显微镜观察细胞形态学的改变、流式细胞仪检测细胞周期与凋亡、免疫细胞化学观察bcl-2的表达.结果 0.1～0.5μmol/L的1,25(OH)2 D3对胆管癌细胞QBC939有抑制作用,呈剂量依赖性.经1,25(OH)2 D3作用72h后细胞G1期比例升高,S期比例下降,其中0.5μmol/L组细胞G1期由(50.3±1.0)%上升至(65.5±3.2)%,S期由(39.4±0.5)%下降至(23.6±0.7)%;并且可诱导细胞产生凋亡,0.5μmol/L组作用后细胞凋亡率由0.5%上升至24.6%;bcl-2的表达下调.结论 1,25(OH)2 D3能抑制胆管癌细胞系QBC939增殖并诱导细胞凋亡,其引起凋亡的机制可能与下调bcl-2的表达相关.     

肝癌细胞裂解物致敏DC疫苗诱导的肿瘤特异抗瘤效应

目的　研究肝癌细胞裂解物致敏的树突状细胞 (DC)疫苗诱导的体内、外抗瘤效应。方法　体外实验 :将肿瘤细胞裂解物致敏的DC与T细胞共培养 ,4d后收获致敏的T细胞 ,观察其对野生型肿瘤细胞的杀伤反应并检测培养液中的IFN γ分泌情况。体内实验 :观察肿瘤细胞裂解物致敏的DC疫苗对小鼠皮下肝癌发生的抑制作用。结果　在效 :靶比分别为 5 0 .0∶1.0、2 5 .0∶1.0、12 .5∶1.0时 ,肝癌细胞裂解物致敏的DC组与DC组诱导的肿瘤杀伤活性分别为 (4 5 .7±3 .2 ) %对 (2 6.5± 2 .5 ) % ;(3 1.0± 2 .7) %对 (14 .3± 3 .3 ) % ;(2 7.8± 1.7) %对 (9.9± 0 .6) % (在各效靶比 ,P <0 .0 1) ,混合细胞培养上清液中IFN γ浓度则为 (2 60 3 .3± 2 60 .0 )ng/L对 (5 0 1.0±5 0 .0 )ng/L。在体内则可有效抑制小鼠皮下肝癌的发生。结论　肝癌细胞裂解物致敏的DC疫苗可以诱导肿瘤特异的抗瘤效应     

bcl-2/bax mRNA在三氧化二砷体外诱导人胆管癌细胞株凋亡中的表达

目的 观察bcl-2、bax mRNA在三氧化二砷(As2O3)诱导人类胆管癌株QBC939、RBE细胞凋亡中的表达.方法 Rhodamine123染色检测线粒体膜通透性变化;逆转录-聚合酶链反应(RT-PCR)检测bcl-2、bax mRNA的表达.结果 As2O3干预,实验组Rhodamine123荧光染色强度较对照组明显减弱;凝胶成像检测bcl-2/β-actin mRNA A比值,QBC939、RBE细胞对照组分别为0.41±0.03、0.84±0.03;24 h实验组为0.01±0.01、0.43±0.02.bax/GAPDHmRNAA比值QBC939、RBE细胞对照组分别为0.21±0.01、0.42±0.04;24 h实验组为1.44±0.16、1.15±0.21,对照组与实验组比较差异均有统计学意义(P＜0.01).结论 As2O3可能增加胆管癌细胞株线粒体膜的通透性,下调bcl-2mRNA表达,上调bax mRNA的表达,可能启动了线粒体凋亡信号传导途径.     

肝细胞癌合并肝硬化患者肝脾联合切除术后免疫功能变化的研究

目的　探讨肝细胞癌合并肝硬化患者肝癌切除时联合脾切除术后免疫功能的变化。方法　将 16例肝癌合并肝硬化患者分成 2组 ,即肝癌切除联合脾切除组 ( 7例 )和单纯肝癌切除组 ( 9例 ) ,于术前、术后 2个月取外周血 7ml,采用流式细胞仪检测CD4、CD8、CD4 /CD8,ELISA法检测IL 2、IFN γ、IL 10。　结果　 2组患者术前CD4、CD8、CD4 /CD8、IL 2、IFN γ、IL 10水平差异无显著性 ;术后 2个月 ,切脾组CD4 ( 3 8 2 %± 3 7% )、CD4 /CD8( 1 7%± 0 3 % )高于保脾组CD4 ( 3 2 5 %± 4 0 % )、CD4 /CD8( 1 1%± 0 1% ) ,而CD8( 2 3 7%± 3 7% )低于保脾组CD8( 2 9 4 %± 4 0 % ) (P <0 0 5 ) ;切脾组IFN γ[( 10 4 4± 14 9)pg/ml]、IL 2 [( 98 6± 18 6)pg/ml]高于保脾组 [IFN γ( 70 5± 12 6)pg/ml、IL 2 ( 80 9± 13 5 )pg/ml],而IL 10 [( 5 5 5± 11 2 )pg/ml]低于保脾组 [IL 10 ( 89 4± 10 )pg/ml](P <0 0 5 )。　结论肝癌切除时联合脾切除不但没有降低机体T细胞亚群和Th细胞的平衡 ,反而促进其恢复平衡 ,并改善机体抗肿瘤免疫功能     

5-氮-2-脱氧胞苷调控p53-baX凋亡通路DNA甲基化对胆管癌细胞生长的影响

目的 探讨甲基转移酶抑制剂5-氮-2-脱氧胞苷(DAC或5-aza-dc)对胆管癌细胞QBC939生长的影响及其机制.方法 应用噻唑蓝(MTT)比色法检测DAC在不同浓度(0.1、0.5、1.0、5.0、25.0、100.0μmol/L)及不同时间(24、48、72 h)下对胆管癌细胞QBC939存活率的影响,透射电镜观察细胞形态学改变,流式细胞仪观察细胞生长周期及凋亡率的变化;甲基化聚合酶链反应检测DAC作用前后p53-bax凋亡通路DNA甲基化变化.结果 DAC对QBC939细胞生长的抑制有浓度和时间依赖性,其半数抑制率(IC50)为72h 5μmol/L;经DAC作用后电镜下胆管癌细胞凋亡增加;胆管癌细胞凋亡发生率由DAC处理前的4.31%上升至处理后的43.04%;DAC作用前p53-bax凋亡通路中p14、DAPK抑癌基因甲基化,DAC作用后p14、DAPK抑癌基因脱甲基化.结论 DAC对胆管癌QBC939细胞的生长具有抑制作用,这一抑制作用是由于DAC脱甲基化造成的,DAC对胆管癌QBC939细胞p53-bax线粒体凋亡通路DNA甲基化的影响使细胞凋亡功能得以恢复.     

胸腰椎骨折椎弓根内固定断裂与植骨融合方式的相关性分析

[目的]探讨胸腰椎骨折椎弓根螺钉内固定系统内固定术后,椎弓根螺钉断裂与植骨融合方式之间的关系,以探讨胸腰椎骨折植骨融合的最佳方式。[方法]回顾性研究1995年5月~2005年12月本院脊柱外科收治的胸腰椎骨折病人197例,其中A组单纯内固定(不植骨)患者14例,B组“H”形椎板植骨21例,C组横突间植骨67例,D组椎间、椎内联合横突间植骨95例。[结果]术后随访6~32个月,内固定断裂12例,其中A组4例,B组3例,C组5例,D组0例,4组中D组内固定断裂率显著低于其他3组(P<0.05)。[结论]椎间、椎体内联合横突间植骨重建脊柱三柱的稳定性,符合人体生物力学原理,能有效降低内固定断裂的发生。     

On the measurement of the functional properties of the CFTR

A number of methods are currently employed to assess the functional properties of CFTR channels and their response to pharmacological potentiators, correction of the defective CFTR trafficking, and vectorial introduction of new proteins. Here we review the most common methods used to assess CFTR channel function. The suitability of each technique to various experimental conditions is discussed.     

Preservation of the pylorus and resection of the head of the pancreas

The historical evolution of the pylorus-preservation resection of the head of the pancreas is traced from the first resections early in this century to relative standardization of the operation, to a lowering of the operative mortality, and to an interest in improving nutritional status after resection. There are many theoretical advantages for the function of the upper gastrointestinal tract after pylorus and gastric preservation, such as maintenance of gastric capacitance and equilibration of osmotic pressure in gastric digestants, foodstuff digestion and absorption, and bowel motility. After the pylorus-preserving resection, gastric emptying is normal, pyloric function to prevent duodenal reflux is often normal, and gastric acids and serum levels of duodenal hormones are at normal levels, whereas after standard pancreatoduodenectomy, all of these are often abnormal. No prospective blinded studies have been published comparing nutritional values after the two operative procedures, but evidence is presented of a satisfactory result with regard to gastric capacitance, body weight gain, and lack of postgastrectomy symptoms. An undoubted advantage of the pylorus-preserving feature is a simplification of the operation. These gains are achieved without increase in operative mortality, without increase in the incidence of jejunal ulcer, and without theoretical or actual decrease in value of the procedure as a cancer operation, except in patients with duodenal carcinoma proximal to the ampulla of Vater.     

下颌全牙弓后移量及最后界的测量

目的：研究下颌牙弓的有效后移量及找寻下颌牙弓移动的后界。方法：选取涉及拔除下颌第三磨牙或下颌第三磨牙缺失的病例18例（男6例,女12例）。采用种植支抗牵引下牙弓向远中,治疗完成时所有病例均明确到达下颌牙弓后界,即下颌第二磨牙远中到达下颌升支前缘软组织交界处。应用治疗前后的曲断片测量下颌第二磨牙远中到升支前缘的距离。结果：下颌第二磨牙后移量为（3.49±1.21）mm;治疗后磨牙后间隙的长度为（4.43±0.97）mm。结论：下颌牙弓可确定性地实现整体后移;最大后移量由磨牙后间隙的长度决定;其最后界止于下颌第二磨牙远中与下颌升支前缘软组织交界处。     

The Measurement of the Inclination Angle of the Hamate and Analysis of the Inclination Angle for the Rotation Deformity of the Little Finger in the Fixation of the Carpometacarpal Joint

ObjectiveComplex base fractures of the fifth metacarpal bone and dislocation of the fifth carpometacarpal joint are more prone to internal rotation deformity of the little finger sequence after fixation with a transarticular plate. In the past, we have neglected that there is actually a certain angle of external rotation in the hamate surface of transarticular fixation. This study measured the inclination angle of the hamate surface relative to the fifth metacarpal surface for clinical reference.MethodsIn a prospective single‐center study, we investigated the tilt angle of 60 normal hamates. The study included thin‐layer computed tomography (CT) data from 60 patients from the orthopaedic clinic and inpatient unit from January 2017 to March 2020, including 34 men and 26 women who were 15~59 years old, average 35 years old. The CT data of 60 cases in Dicom format of the hand was input into Mimics and 3‐Matics software for three‐dimensional (3D) reconstruction and measuring the angle α between hamate surface and the fifth metacarpal surface. According to the possible placement of the transarticular plate on the fifth metacarpal surface, we measured the angle β between the hamate surface 1 and the fifth metacarpal surface and the angle γ between the hamate surface 2 and the fifth metacarpal surface.ResultsThe average angle between the hamate surface and the fifth metacarpal surface was 11.66°. The hamate surfaces 1 and 2 have an external rotation angle of 7.30° and 7.51° on average with respect to the fifth metacarpal surface, respectively. There is no statistically significant difference in the angles between the two groups (P > 0.05).ConclusionsThe horizontal angle of the dorsal side of the hamate is different from the back of the fifth metacarpal surface, and the hamate has a certain external rotation angle with respect to the fifth metacarpal surface. No matter how the transarticular plate is placed, the plate always has a certain external rotation angle relative to the fifth metacarpal surface. When the fixation is across the fifth carpometacarpal joint, if the plate does not twist and shape, it will inevitably cause internal rotation of the fifth metacarpal, resulting in internal rotation deformity of the little finger sequence.     

静脉输注甘露醇开放血脑屏障对抗生素透过血脑屏障的影响

目的　通过快速静脉输注甘露醇可逆性开放血脑屏障 (BBB) ,探知此方法能否增加抗生素透过BBB的量 ,在何时达到最高峰 ,其通透量增加后临床上有无不良反应。方法　采用自身配伍设计 ,共 6个样本组。对照组仅使用抗生素 ;其余 5组分别在使用甘露醇前 60、3 0min ,同时使用甘露醇后 3 0、60min使用抗生素 ,各组皆取使用抗生素后 1h的脑脊液测其抗生素浓度。抗生素选用头孢三嗪。结果　测量值经过q检验 ,经 2 0 %甘露醇处理前后的CSF中的头孢三嗪浓度差异有非常显著性。全组患者经临床观察未出现神经系统的不良反应。结论　经静脉快速输注2 0 %甘露醇后可以使透过BBB的水溶性抗生素的量增加 ,两者使用的顺序是在抗生素使用 3 0min内即给予甘露醇快速滴注。该方法不会增加低神经毒性抗生素在中枢神经系统的不良反应。     

Preservation of the pylorus in resection of the head of the pancreas

Whipple's pancreatoduodenectomy was the standard operation for diseases of the head of the pancreas for more than 40 years, but the results were vitiated in part by poor gastrointestinal function and malnutrition. Reintroduced in 1978, pylorus-preserving proximal pancreatoduodenectomy (PPPP) has had an increasing impact on pancreatic surgery as its benefits have been recognized: improved nutritional status, decreased incidence of postgastrectomy syndromes, and a technically easier operation. Postoperative mortality rates and 5-year survival rates are comparable with those of the classic Whipple procedure. PPPP is indicated for most patients with chronic pancreatitis of the pancreatic head. It is also appropriate for patients with periampullary cancer and for those with pancreatic cancer arising from the lower part of ‘the head and the uncinate process. More than 650 patients have now undergone PPPP: 31% for chronic pancreatitis and 66% for periampullary and pancreatic cancers. We assess the indications for PPPP, outline the operation, and review the results.