弥猴肠缺血再灌注后生长抑素含量与中性粒细胞寿命的相关性探讨

目的探讨多器官功能障碍综合征时中性粒细胞凋亡延迟的潜在原因,以增加对全身炎症反应内源性保护机制的认识。方法对照组、实验组各5只弥猴用放射免疫分析法测定其外周血、肠黏膜生长抑素浓度;生长抑素与猕猴外周血中性粒细胞共同孵育后,观察猕猴外周血中性粒细胞的形态变化,流式细胞仪测定细胞凋亡率,电泳分析中性粒细胞DNA断裂片段,生物分子相互作用系统检测中性粒细胞上生长抑素受体(SSTR-1、SSTR-2)。结果猕猴肠缺血再灌注后,猕猴外周血及小肠黏膜中生长抑素含量均明显下降;中性粒细胞凋亡率由阻断前的15.4%±1.4%,显著降低至3.5%±0.5%(P<0.05);腹腔巨噬细胞凋亡率由阻断前的14.1%±1.7%增加至20.2%±1.8%(P<0.05);体外实验表明,生长抑素可使中性粒细胞体积缩小,细胞质密度增加,染色质固缩,细胞核变小;中性粒细胞凋亡率(50.2%±1.8%)明显高于对照组(20.0%±2.2%,P<0.01);中性粒细胞的DNA片段断裂分析电泳可见Ladder带。中性粒细胞上生长抑素受体能与生长抑素受体SSTR-1抗体、SSTR-2抗体发生结合反应,其特异结合量分别为(548±20)RU/m l及(28±21)RU/m l。结论生理状态下,生长抑素通过猕猴外周血中性粒细胞上生长抑素受体介导,诱导其凋亡;肠缺血再灌注后猕猴外周血及小肠黏膜中生长抑素含量明显下降,这使中性粒细胞凋亡延迟,从而推动了全身炎症反应综合征和多器官功能障碍综合征的发生。     

结核杆菌抗原对中性粒细胞自发性凋亡的抑制作用

目的: 探讨结核杆菌抗原(Mtb-Ag)对体外培养中性粒细胞自发性凋亡的影响。方法: 取健康成人外周血分离中性粒细胞,加Mtb-Ag培养24 h,或用Mek抑制剂PD98059预处理30 min后,Wright-Giemsa染色形态学方法和Annexin-V染色流式细胞术观察细胞凋亡。结果: 与中性粒细胞体外培养24 h后自发凋亡(55±10)%相比,加入Mtb-Ag (1.125 mg/ml)后,中性粒细胞自发凋亡率(31±3)%明显降低(P<0.01);Mek抑制剂PD98059(50μmol/L)可阻断Mtb-Ag的抑制凋亡作用(P<0.01)。结论: Mtb-Ag对中性粒细胞自发性凋亡有抑制作用,这一作用可能涉及Mek-Erk途径。     

睾丸复位固定术对大鼠隐睾Bcl-2表达及生精细胞凋亡的影响

目的研究实验性大鼠隐睾复位固定术对Bcl-2表达及生精细胞凋亡的影响。方法24只雄性SD大鼠建立左侧隐睾模型,右侧睾丸切断引带后还纳入阴囊作为假手术组;模型建立7天后实施左侧隐睾复位固定术,隐睾复位固定术前、术后第24天及48天分别处死大鼠8只,采用免疫组织化学检测各组睾丸Bcl-2表达的变化,运用TUNEL法检测各组睾丸生精细胞凋亡。 结果隐睾内生精细胞凋亡增加,复位固定术能降低生精细胞凋亡,增强睾丸内Bcl-2表达。假手术组凋亡指数(4.86%±0.27%)与复位固定术后48天组(5.59%±0.67%）比较差异无显著性（P＞0.05),但均低于复位固定术后24天组(14.48%±0.70%）及复位固定术前的隐睾组(29.45%±1.19%）（P＜0.05);Bcl-2表达在假手术组、复位固定术后24天组及复位固定术后48天组之间比较差异无显著性（P＞0.05),但均明显高于隐睾组（P＜0.05)。结论睾丸复位固定术能通过促进隐睾内Bcl-2表达来抑制生精细胞凋亡。     

视黄酸对肿瘤坏死因子α诱导肺泡上皮细胞损伤的保护作用

目的探讨肿瘤坏死因子α(TNF-α)对肺泡Ⅱ型上皮细胞株A549增殖凋亡的影响与视黄酸(RA)对其调控作用。方法用不同浓度的TNF-α、1μmol/LRA和10μg/LTNF-α+1μmol/LRA处理肺泡Ⅱ型上皮细胞株A549细胞24h和48h后MTT法测定细胞增殖能力,流式细胞仪检测细胞凋亡。用10μg/LTNF-α处理24h或48h后继续培养,用台盼蓝染色计算24、48和72h活细胞数。结果分别用0、0.1、1、5、10μg/L浓度的TNF-α培养A549细胞24h和48h测定细胞增殖数(%):24h分别为95.0%、90.0%、79.6%、72.4%和59.6%,方差分析结果F值为18.04,P<0.01;48h分别为93.2%、82.7%、61.5%、50.3%和44.7%,F值为40.61,P<0.01。用10μg/LTNF-α处理A549细胞24h,其对细胞的增殖抑制作用可逆转,但处理48h后的A549细胞增殖抑制作用不能逆转。用TNF-α和视黄酸培养A549细胞12、24与48h后,凋亡的细胞数(%)TNF-α组分别为14.3%±3.2%、18.6%±2.9%和43.4%±3.5%,与对照组(6.3%±1.2%,8.2%±1.3%和26.1%±2.5%)比较差异有统计学意义;视黄酸加TNF-α组为4.8%±1.1%,5.2%±1.3%和16.4%±2.3%,明显少于TNF-α组。结论视黄酸通过抑制TNF-α对肺泡Ⅱ型上皮细胞的破坏,可以达到减轻肺泡上皮细胞损伤,保护肺表面活性物质的作用。     

凉膈散对内毒素致急性肺损伤大鼠中性粒细胞凋亡的影响*

目的：观察凉膈散对内毒素（LPS）致急性肺损伤（ALI）大鼠肺组织病理学及血中性粒细胞（PMN）凋亡的影响。方法：选取健康雄性SD大鼠72只，随机分为6组，每组12 只，分别为正常对照组、ALI模型组、凉膈散大剂量组、凉膈散中剂量组、凉膈散小剂量组和强的松治疗组。给予正常对照组大鼠气管内滴注生理盐水，其余各组大鼠均气管内滴注LPS，每天一次，连续2天，气道内滴注1小时后分别以生理盐水、凉膈散不同剂量、强的松灌胃，每天2次。分别于第二次气管滴注后12小时、24小时每组处死6只。取外周血中性粒细胞，流式细胞仪检测PMN的凋亡改变。结果 ：病理学积分：12h模型组（2.08±0.12）高于正常对照组（0.33±0.03）（P<0.01）, 12h凉膈散大剂量组（0.83±0.09）、中剂量组（1.00±0.09）、小剂量组（1.17±0.08）均低于模型组（2.08±0.12）(P<0.01或P<0.05)，24h模型组（2.25±0.11）高于正常对照组（0.41±0.04）（P<0.01），凉膈散大剂量组（0.67±0.05）、中剂量组（1.17±0.07）、小剂量组（1.33±0.06）均低于模型组（2.25±0.11）(P<0.01或P<0.05)。中性粒细胞凋亡指数：12h模型组（51.36±3.99）低于正常对照组（71.71±4.13）（P<0.01）, 12h凉膈散大剂量组（66.37±4.25）、中剂量组（61.6±5.02）高于模型组（51.36±3.99）(P<0.05), 24h模型组（63.04±4.89）低于正常对照组（80.82±5.19）（P<0.01）, 凉膈散大剂量组（71.32±5.32）高于模型组（63.04±4.89） (P<0.05)。结论：凉膈散可诱导急性肺损伤时的中性粒细胞凋亡，抑制中性粒细胞发挥活性作用， 从而减轻急性肺损伤肺部炎症反应，推测其机制可能是通过抑制NF- κB表达。     

手术应激对儿童外周血淋巴细胞凋亡的影响

目的　探讨手术应激后儿童外周血淋巴细胞的凋亡。方法　选择择期手术患儿 16例 ,分别于术前 2 4h、术后 2 4h、术后 72h采集外周血 3.5～ 5ml。用梯度离心法分离出淋巴细胞 ,置于RPMI 16 4 0培养基中 ,培养 15h。然后用 7 AAD染色 ,通过流式细胞仪检测CD4 +、CD8+细胞凋亡。结果　术后 2 4hCD4 +、CD8+淋巴细胞凋亡率 ( % )明显增加 (术前 2 4h分别为 2 .4 4± 0 .4 5和 2 .96± 0 .6 0 ,术后 2 4h为 6 .5 3± 1.96和 5 .13± 1.19) (P <0 .0 5 ) ,术后 72h又恢复至术前水平 ( 3.6 0± 1.0 8和 4 .0 1± 0 .93,P >0 .0 5 )。结论　手术创伤可使儿童外周血CD4 +、CD8+淋巴细胞凋亡增加。     

卡维地洛对大鼠急性心肌梗死后心肌细胞凋亡和bcl-2/bax基因表达的干预作用

目的评价新一代β受体阻断剂卡维地洛对大鼠急性心肌梗死(AM I)后心肌细胞凋亡及其相关基因表达的干预作用。方法将冠状动脉结扎术后24 h存活的雌性SD大鼠(83只)分为AM I对照(M I组,43只)和卡维地洛(10 mg.kg-1.d-1)治疗组(C组,40只),另设假手术组(S组,27只)。各组再按观察时点分为48 h和4周两个亚组。C组于术后24 h直接灌胃法给药。用TUNEL法和DNA凝胶电泳检测心肌细胞凋亡。用免疫组化方法和W estern印迹法检测“抑制凋亡复合基因”bc l-2/bax的表达。结果(1)M I组梗死/瘢痕区、梗死边缘区和非梗死区的心肌细胞凋亡指数(48 h 21.2%±15.6%、19.8%±12.0%和6.3%±4.5%;4周23.5%±13.0%、8.0%±4.4%和3.2%±1.6%)高于假手术组(48 h 1.3%±1.1%,4周1.2%±0.3%,均P<0.05);凋亡促进基因bax的表达M I48 h组明显高于假手术组,但“凋亡抑制基因”bc l-2仅在M I48 h组梗死区心肌细胞中表达增加;bc l-2/bax的比值M I48 h组(1.06)低于假手术组(1.87)。(2)C48 h组3个区域的心肌细胞凋亡指数与M I组比较,差异无统计学意义(均P>0.05),但梗死区及边缘区bc l-2的表达均增加;C4周组梗死/瘢痕区及边缘区的心肌细胞凋亡指数(4.0%±2.0%,2.9%±1.6%)均低于M I组(23.5%±13.0%,8.0%±4.4%,均P<0.05),且瘢痕区心肌细胞中bax的表达亦明显降低,但bc l-2的表达均无明显变化;C组大鼠48 h和4周时的心肌细胞bc l-2/bax的比值(1.72,2.23)高于M I组(1.06,1.8),与假手术组(1.87,2.25)相当。结论大鼠出现AM I后,长时间(4周)用卡维地洛治疗能有效防止梗死/瘢痕区及其边缘区心肌细胞凋亡的发生,并使bc l-2/bax的比值增加。     

单甲基亚砷酸联合隐丹参酮抑制多发性骨髓瘤U266增殖的作用机制研究

摘要 目的：研究单甲基亚砷酸(MMAIII)联合隐丹参酮(CPT)对多发性骨髓瘤U266的协同抑制效应，阐明其诱导多发性骨髓瘤凋亡的作用机制。方法：cell-counting-kit-8(CCK8)法检测细胞存活率；AnnexinⅤ-FITC/PI流式细胞术检测细胞凋亡；Western Blot检测凋亡标记蛋白PARP的总蛋白和剪切表达水平；酶标仪检测酸性鞘磷脂酶活性；免疫荧光技术检测细胞膜神经酰胺和脂筏标记物。结果：CCK8结果显示隐丹参酮浓度为15μM时细胞存活率为80.9%±5.4%(24h)、60%±8.4%(48h)，单甲基亚砷酸浓度为1μM时细胞存活率为82.5%±5.8%(24h)、67.7%+9.7%(48h)，隐丹参酮（15μM）联合单甲基亚砷酸（1μM）作用U266细胞24h、48h后细胞存活率为29.1%±7.0%(24h)、18%±2.7%(48h)；流式细胞术结果显示联合用药组细胞凋亡率为41.7%±4.4%、单甲基亚砷酸组为10.6%±4.0%、隐丹参酮组为10.5%±3.0%；Western Blot显示联合作用组凋亡蛋白PARP剪切活化水平较单甲基亚砷酸单药组及隐丹参酮单药组有显著升高；酸性鞘磷脂酶活性测定结果显示联合用药组上调多发性骨髓瘤U266细胞酸性鞘磷脂的活性；免疫荧光技术检测结果显示联合用药后U266细胞膜上荧光强度增加，加入Filipin后荧光强度较联合用药减少。结论：MMAIII与CPT协同作用能诱导人多发性骨髓瘤U266细胞凋亡发生，其作用机制主要与促进细胞膜鞘磷脂水解生成神经酰胺、诱导脂筏聚集促进凋亡相关蛋白剪切活化密切相关。     

TGF-β_1和TL1A在高糖所致血管内皮细胞凋亡中的作用

目的:观察高糖条件下,内皮细胞分泌的TGF-β1和TL1A对细胞凋亡的影响。方法:以22.4mmol/L葡萄糖培养细胞不同时间(0h,6h,12h,24h,48h),RT-PCR和Westernblot检测内皮细胞TGF-β1和TL1A表达,Annexin V-FITC/PI双染色、流式细胞仪测定细胞凋亡率。结果:高糖条件下HUVECTGF-β1和TL1A表达增加(均P<0.01),22.4mmol/L葡萄糖培养条件下,TGF-β1的表达高峰在12h,TL1A高表达峰值在48h。高糖条件下细胞凋亡率明显增加(21.06%±3.05%vs3.09%±0.32%,P<0.01)。在正常糖培养的HUVEC中加入外源性的TGF-β1和TL1A,均导致细胞凋亡率显著增高(15.26%±2.93%,55.70%±8.91%vs3.09%±0.32%,均P<0.01),在高糖培养液中加入抗TL1A抗体能明显抑制细胞凋亡发生(4.28%±0.48%vs21.06%±3.05,P<0.01),但加入抗TGF-β1抗体却不能逆转高糖诱导的细胞凋亡(20.93%±3.21vs21.06%±3.05%,P>0.05)。结论:高糖上调内皮细胞TGF-β1和TL1A的表达,TL1A介导高糖引起内皮细胞凋亡的作用,高糖引起内皮细胞凋亡与TGF-β1无关。     

外周血T细胞亚群及NK细胞比例在转移性结肠癌患者预后评估中的临床意义

目的 探讨反复化疗的转移性结肠癌患者(mCRC)T淋巴细胞亚群和自然杀伤(NK)细胞比例及其临床意义.方法 将我院30例转移性结肠癌患者作为观察组,回顾性分析外周血T淋巴细胞亚群及NK细胞的检测结果,与30例健康受检者(对照组)进行比较分析,并于化疗4周期后复查结肠癌患者的外周血T淋巴细胞亚群及NK细胞,对比有效与无效患者的检测结果.结果 mCRC患者中化疗有效组化疗后外周血中B细胞、CD4+细胞、NK细胞及CD4+/CD8+比值均明显高于同组化疗前水平(11.2%±0.98%vs 6.7%±1.05%,P<0.05;30%±2.01%vs 26%±2.02,P<0.05;10%±1.02%vs 8.9%±0.72%,P<0.05;1.2±0.03 vs 0.9±0.06,P<0.05),而化疗无效组化疗前后差异均无统计学意义(P>0.05).化疗无效组外周血中B细胞、CD4+细胞、NK细胞及CD4+/CD8+比值均明显低于正常对照组(2.4%±0.01%vs 15%±1.10%,P<0.05;8%±0.03%vs 37%±2.10%,P<0.05;4.4%±0.12%vs 25%±1.80%,P<0.05;0.44±0.01 vs 1.23±0.02,P<0.05).结论 NK细胞与外周血T淋巴细胞亚群对评价结肠癌化疗的疗效、判断预后具有重要价值.     

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.     

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…     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

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.