酪蛋白空肠灌注对急性坏死性胰腺炎大鼠胰腺外分泌的影响

目的 观察空肠内灌注酪蛋白对急性坏死性胰腺炎(ANP)大鼠胰腺外分泌的影响,并初步探讨其神经机制.方法 30只SD大鼠按随机数字法分为对照组、ANP组和肠内营养组.后2组以胰管内逆行注射牛磺胆酸钠法制作ANP模型.造模后24 h空肠插管分别灌注酪蛋白溶液(肠内营养组)和牛理盐水(埘照组和ANP组).每15 min收集胰液1次,共6次,记录胰液分泌量和检测胰液蛋白含量.取大鼠延髓孤束核,采用免疫组化法检测c-Fos蛋白表达.结果 ANP组和肠内营养组组内各时间段之间胰液分泌最无显著差别,两组相应时间段之间胰液分泌最也无显著差别,但均显著低于对照组对应时间段的胰液分泌量(P<0.05).对照组、ANP组和肠内营养组空肠灌注过程中胰液蛋白含量水平稳定,不同时间段间无明显差异,但ANP组和肠内营养组空肠灌注后0～15 min、15～30 min、30～45 min、75～90 min时间段内胰液蛋白含量均低于对照组(P<0.05).肠内营养组灌注后延髓孤束核c-Fos蛋白表达阳性,而埘照组和ANP组延髓孤束核c-Fos表达阴性.结论 空肠内酪蛋白灌注可促进延髓孤束核c-Fos表达,但不增加胰液分泌量和胰液蛋白含量.     

脂联素对大鼠心肌缺血再灌注损伤的保护作用

目的 观察脂联素对大鼠心肌缺血再灌注损伤的保护作用并探讨其机制.方法 32只8周龄雄性大鼠随机分为假手术组、缺血再灌注组、地尔硫革组和脂联素组,每组8只.(1)假手术组:只穿线,旷置90 min.(2)缺血再灌注组:先阻断血流30 min,再灌注60 min.(3)地尔硫(革)组和脂联素组:先阻断血流30 min,于再灌注开始时,从鼠尾静脉分别注射地尔硫(革)(3.5 μg·g~(-1)min~(-1))或脂联素(60 ng·g~(-1)·min~(-1)),注射2 min,再灌注60 min.各模型组于再灌注60 min后处死大鼠.测定心肌组织一氧化氮(NO),心肌组织半胱氨酸蛋白酶3(Caspase 3)活性,心肌组织腺苷酸活化蛋白激酶(AMPK)活性,过氧化物酶体增殖物激活受体γ(PPARγ)的含量,同时用透射电镜观察大鼠心肌线粒体结构.结果 (1)缺血再灌注组心肌组织中Caspase 3活性显著高于假手术组[(168.50±30.08)μmol/L比(53.25±11.41)μmol/L,P<0.01],AMPK活性、PPARγ含量均显著低于假手术组[(0.74±0.59)IU/ml比(25.63±4.61)IU/ml,P<0.01;0.1894比0.7949,P<0.01],心肌组织中NO含量显著低于假手术组[(6.359±1.355)μmol/L比(10.396±1.901)μmol.L,P<0.01].(2)脂联素组心肌组织中Caspase 3活性显著低于缺血再灌注组[(88.75±6.92)μmol/L比(168.50±30.08)μmol/L,P<0.01],AMPK活性、PPARγ含量均显著高于缺血再灌注组[(27.22 ±4.76)IU/ml比(0.74±0.59)IU/ml,P<0.01;0.8613比0.1894,P<0.01],心肌组织中NO含量显著高于缺血再灌注组[(15.755±1.045)μmol/L比(6.359±1.355)μmol/L,P<0.01].脂联素可保护急性心肌缺血再灌注过程中大鼠心肌细胞线粒体结构的完整性,上述作用优于地尔硫(革).结论 脂联素对缺血再灌注造成的心肌损伤有一定的保护作用,机制可能与其增加心肌细胞AMPK、PPARγ表达,以及抗心肌细胞凋亡作用有关.     

清胰汤对大鼠急性胰腺炎肺损伤时SP-A表达的影响

目的:探讨重症急性胰腺炎(SAP)肺损伤(ALI)时肺表面活性蛋白A(SP-A)的表达及其在ALI发病中的作用,并观察清胰汤对SP-A表达和病情转归的影响.方法:采用胆胰管内逆行注入1.5%去氧胆酸钠建立大鼠SAP时ALI模型.将SD大鼠随机分为假手术组(n=10)、模型组(n=10)和清胰汤组(n=10).假手术组仅行剖腹术,翻动胰腺.清胰汤组在建立SAP模型后30 min和12 h清胰汤ig(10 mL/kg).各组大鼠在术后24 h测PaO_2和血淀粉酶.应用逆转录聚合酶链式反应(RT- PCR)检测肺SP-A mRNA的表达强度,Western- blot观察SP-A表达,并观察胰、肺病理变化及肺泡Ⅱ型上皮细胞的电镜下变化.结果:模型组血淀粉酶(7144.19 U/L±727.91 U/L)显著高于清胰汤组(4283.51 U/L±527.52 U/L)和假手术组(1193.41 U/L±192.54 U/L,P<0.01).模型组PaO_2显著低于假手术组和清胰汤组(79.24±5.84 vs 96.78±3.81.79.24±5.84 vs 88.16±5.07,P<0.01).清胰汤组肺SP-A mRNA表达显著高于模型组(P<0.01),肺SP-A蛋白的表达显著高于模型组,SP-A mRNA的表达与肺损伤的程度呈负相关.清胰汤组胰、肺病理及电镜改变较模型组减轻.结论:SAP时肺泡Ⅱ型上皮细胞功能受损,SP-A mRNA表达降低导致急性肺损伤的机制之一.清胰汤能保护肺泡Ⅱ型上皮细胞功能,恢复SP-A mRNA正常表达,维持肺泡功能,从而对肺组织起保护作用.     

不同浓度雌二醇对去卵巢小鼠T淋巴细胞凋亡的影响及作用机制

目的 研究不同浓度雌二醇(E2)对去卵巢小鼠脾脏T淋巴细胞凋亡的影响并探讨其作用机制.方法 分离出小鼠脾脏T淋巴细胞,分为10组,分别为青年组、假手术组、去卵巢对照组、去卵巢+E2(10-11mol/L)组、去卵巢+E2(10-10mol/L)组、去卵巢+E2(10-8mol/L)组、去卵巢+E2(10-6mol/L)组、去卵巢+E2(10-10mol/L)+雌激素受体拮抗剂IC1182 780(10-7mol/L)组、去卵巢+E2(10-10mol/L)+1-甲基-4-苯基-四氢吡啶(MPP,10-6mol/L)组、去卵巢+E2(10-10mol/L)+四氢化吡咯二硫代氨基甲酸酯(PDTC,10-6mol/L)组.流式细胞仪检测各组细胞凋亡率,Westernblot方法检测各组细胞雌激素受体(ER)α、ERβ、Bax、Bcl-2蛋白表达及细胞核中P65蛋白水平.结果 去卵巢对照组细胞凋亡率为(19.4±2.5)%,高于青年组(14.6±2.4)%和假手术组(14.5±2.3)%,Bcl-2、核内P65蛋白水平分别为0.25±0.05、0.09±0.01,低于青年组(0.40±0.07、0.15±0.02)和假手术组(0.38±0.05、0.15±0.02),均P<0.01;去卵巢+E2(10-10mol/L)组凋亡率为(16.6±1.8)%,低于去卵巢对照组(P<0.05),Bel-2及核内P65蛋白水平高于去卵巢对照组(P<0.01),而去卵巢+E2(10-8mol/L,10-6mol/L)组凋亡率高于去卵巢对照组(分别为P<0.05、P<0.01).去卵巢对照组ERα、ERβ蛋白水平分别为0.23±0.01、0.22±0.03,低于青年组(0.27±0.02、0.29±0.04)和假手术组(0.28±0.03、0.29±0.02),去卵巢+E2(10-11mol/L、10-10mol/L、10-8mol/L)组ERα、ERβ蛋白水平均高于去卵巢对照组,而去卵巢+E2(10-mol/L)组(0.09±0.01、0.14±0.02)低于去卵巢对照组(均P<0.01).去卵巢+E2(10-10mol/L)+IC1182 780组及去卵巢+E2(10-10mol/L)+PDTC组凋亡率与去卵巢对照组差异无统计学意义,去卵巢+E2(10-10mol/L)+MPP组凋亡率与去卵巢+E2(10-10mol/L)组比较,差异无统计学意义.结论 去卵巢小鼠T淋巴细胞凋亡增多,E2对去卵巢小鼠T淋巴细胞的凋亡呈剂量依赖的双相调节作用,生理范围内可部分逆转去卵巢小鼠T淋巴细胞凋亡,而超过生理范围则增加其凋亡.生理剂量E2可能通过ERβ及核因子-kB途径抑制去卵巢小鼠的T淋巴细胞凋亡.     

罗格列酮对脂多糖诱导人脐静脉内皮细胞高迁移族蛋白B1分泌的影响

目的探讨1mg/L脂多糖诱导体外培养的人脐静脉内皮细胞(HUVEC)表达高迁移族蛋白B1(HMGB1)及罗格列酮的干预作用。方法取体外培养的第3⁷代HUVEC进行试验。实验分组:(1)空白对照组。(2)脂多糖组,用1mg/L脂多糖分别作用HUVEC 6、12、24h。(3)罗格列酮+脂多糖组(罗格列酮组):分别以罗格列酮0μmol/L、5μmol/L、10μmol/L、15μmol/L 4个浓度预先作用2h,然后再分别以罗格列酮加1mg/L脂多糖共作用24h。采用四唑盐比色法(MTT法)检测96孔板细胞的吸光度值。ELISA测定细胞培养液中HMGB1蛋白含量以检测HUVEC分泌HMGB1含量。结果与空白对照组比较,脂多糖组和0μmol/L罗格列酮组6、12、24h HMGB1分泌明显升高(P<0.01)。与脂多糖组比较,5μmol/L罗格列酮组6、12、24hHMGB1分泌明显降低(P<0.01)。5、10、15μmol/L罗格列酮组24hHMGB1分泌较脂多糖组明显降低[(165.77±20.29)ng/ml,(136.63±15.90)ng/ml,(112.25±12.23)ng/ml vs(338.74±18.22)ng/ml,P<0.05]。10、15μmol/L罗格列酮组24hHMGB1分泌较5μmol/L罗格列酮组明显降低(P<0.05,P<0.01)。结论脂多糖诱导HUVEC分泌HMGB1,罗格列酮可呈剂量依赖性地抑制脂多糖诱导的HMGB1分泌。     

糖尿病大鼠胃化学感觉异常及其机制研究

目的研究糖尿病大鼠给予盐酸刺激后脑孤束核c-fos蛋白表达的改变,探讨其是否存在胃化学感觉异常,并初步探讨其发生机制。方法雄性SD大鼠共24只,随机分为正常对照组、糖尿病组、糖尿病+5-HT4激动剂组。后两组用链脲霉素腹腔注射建立糖尿病大鼠模型。予0.1 mmol/L盐酸灌胃刺激后,应用免疫组化染色法检测各实验组大鼠脑孤束核c-fos蛋白表达水平。糖尿病+5-HT4激动剂组予盐酸灌胃前给予替加色罗灌胃3天。结果糖尿病组大鼠脑孤束核c-fos蛋白表达水平显著低于正常对照组大鼠(P<0.05);糖尿病+5-HT4激动剂组大鼠脑孤束核c-fos蛋白表达水平显著高于糖尿病组(P<0.05),与对照组比较差异无统计学意义。结论5-HT4受体激动剂能增加糖尿病大鼠胃对化学刺激的敏感性,提示该感觉减退可能与胃5-HT4受体有关。     

心力衰竭大鼠解偶联蛋白2表达及其对线粒体功能的影响

目的观察心力衰竭大鼠心肌线粒体解偶联蛋白2(uncoupling protein 2,UCP2)表达及其对线粒体功能的影响。方法45只雄性SD大鼠随机分为腹主动脉缩窄组(实验组)、假手术组和正常对照组,实验组行腹主动脉缩窄术。术后20周心脏超声检测心功能,测定血清游离脂肪酸(free fatty acid,FFA)浓度,免疫印迹法分析心肌线粒体UCP2蛋白表达,密度梯度法提取大鼠心肌线粒体,氧电极法测定线粒体的呼吸活性,罗丹明123法测定线粒体膜电位。结果实验组术后20周左心室射血分数比假手术组和正常对照组降低[(41.0±6.7)%vs.(84.0±2.5)%,P<0.01;(41.0±6.7)%vs.(84.0±2.3)%,P<0.01]、舒张末期内径比假手术组和正常对照组增加[(6.80±0.58)mm vs.(5.20±0.78)mm,P<0.01;(6.80±0.58)mm vs.(5.00±0.67)mm,P<0.01],大鼠心力衰竭建模成功。与正常对照组和假手术组比较,实验组血清FFA浓度明显增高[(372±14)μmol/L vs.(218±16)μmol l/L,P<0.01;(372±14...     

Interleukin-1 beta up-regulates tissue inhibitor of matrix metalloproteinase-1 mRNA and phosphorylation of c-jun N-terminal kinase and p38 in hepatic stellate cells

AIM: To study the relationship between interleu-kin-1beta (IL-1β) up-regulating tissue inhibitor of matrix metalloproteinase-1 (TIMMP-1) mRNA expression and phosphorylation of both c-jun N-terminal kinase (JNK) and p38 in rat hepatic stellate cells (HSC). METHODS: RT-PCR was performed to measure the expression of TIMMP-1 mRNA in rat HSC. Western blot was performed to measure IL-1β-induced JNK and p38 activities in rat HSC. RESULTS: TIMMP-1 mRNA expression (1.191±0.079) was much higher after treatment with IL-1β(10 ng/mL) for 24 h than in control group (0.545±0.091) (P<0.01). IL-1βactivated JNK and p38 in a time-dependent manner. After stimulation with IL-1βfor 0, 5, 15, 30, 60 and 120 min, the JNK activity was 0.982±0.299, 1.501±0.720, 2.133±0.882, 3.360±0.452, 2.181±0.789, and 1.385±0.368, respectively. There was a significant difference in JNK activity at 15 min (P<0.01), 30 min (P<0.01) and 60 min (P<0.01) in comparison to that at 0 min. The p38 activity was 1.061±0.310, 2.050±0.863, 2.380±0.573, 2.973±0.953, 2.421±0.793, and 1.755±0.433 at the 6 time points (0, 5, 15, 30, 60 and 120 min) respectively. There was a significant difference in p38 activity at 5 min (P<0.05), 15 min (P<0.01), 30 min (P<0.01) and 60 min (P<0.01) compared to that at 0 min. TIMMP-1 mRNA expression trended to decrease in 3 groups pretreated with different concentrations of SP600125 (10μmol/L, 1.022±0.113; 20μmol/L, 0.869±0.070; 40μmol/L, 0.666±0.123). Their decreases were all significant (P<0.05, P<0.01,P<0.01) in comparison to control group (without SP600125 treatment, 1.163±0.107). In the other 3 groups pretreated with different concentrations of SB203580 (10μmol/L, 1.507±0.099; 20μmol/L, 1.698±0.107; 40μmol/L, 1.857±0.054), the expression of TIMMP-1 mRNA increased. Their levels were higher than those in the control group (without SB203580 treatment, 1.027±0.061) with a significant statistical significance CONCLUSION: IL-1βhas a direct action on hepatic fi-brosis by up-regulating TIMMP-1 mRNA expression in rat HSC. JNK and p38 mitogen-activated protein kinases (MAPKs) are involved in IL-1β-induced TIMMP-1 gene expression, and play a distinct role in this process, indicating that p38 and JNK pathways cooperatively mediate TIMP-1 mRNA expression in rat HSC.     

氢化可的松对人NK细胞增殖及其对胰腺癌SW1990细胞杀伤效率的影响

目的 探讨氢化可的松对人外周血NK细胞增殖及其对胰腺癌SW1990细胞杀伤力的影响.方法 分离健康人外周血单个核细胞加入到含IL-15细胞因子的NK细胞培养基中诱导培养NK细胞.当NK细胞纯度达到70％以上时,加入10-6、10-5、10-4、10-3 μmol/L氢化可的松继续培养7d.以未加氢化可的松的NK细胞作为对照组.采用锥虫蓝染色计数细胞;采用流式细胞术检测CD3-CD56+ NK细胞含量及其穿孔素、颗粒酶B和IFN-γ的表达;以20∶1的效靶比将NK细胞与SW1990细胞共培养,用细胞增殖-毒性检验法(CCK-8)检测NK细胞对SW1990细胞的杀伤效应.结果 用氢化可的松处理7d后NK细胞量平均达到70.72％ ～ 76.39％,与对照组的(72.61±3.76)％差异无统计学意义;10-6、10-5、10-4、10-3 μmol/L氢化可的松处理组NK细胞的增殖倍数分别为(9.13±0.94)、(9.67±1.51)、(10.33± 1.07)、(8.40±1.47)倍,均显著高于对照组的(4.23±0.82)倍(P值均＜0.01);NK细胞对SW1990细胞的杀伤效率分别为(58.58±4.89)％、(62.27±5.63)％、(64.02±5.79)％、(63.88±3.61)％,均较对照组的(57.46±5.11)％增强,其中10-4μmol/L组的差异具有统计学意义(P ＜0.05);10-4、10-3μmol/L氢化可的松处理组NK细胞穿孔素表达分别为(96.71±3.04)％、(97.56±2.18)％,显著高于对照组的(92.40±3.53)％(P ＜0.05或0.01);10-5 μmol/L氢化可的松处理组NK细胞的颗粒酶B表达为(78.23±2.94)％,显著高于对照组(73.68±3.52)％(P＜0.05);10-5、10-4、10-3 μmol/L氢化可的松处理组NK细胞IFN-γ表达分别为(96.61±2.04)％、(97.58±2.17)％、(98.00±1.77)％,均显著高于对照组的(92.44±2.74)％(P值均＜0.01).结论 氢化可的松可促进IL-15活化的NK细胞增殖,适当浓度条件下增强对SW1990细胞杀伤活性,其机制可能与其穿孔素、颗粒酶B和IFN-γ表达上调有关.     

生长抑素及质子泵抑制剂对大鼠胰腺腺泡细胞慢活化电压依赖性钾离子通道的影响

目的 通过检测慢活化电压依赖性外向钾离子通道电流(Iks)的变化,探讨生长抑素及质子泵抑制剂(PPI)对胰腺腺泡细胞慢活化电压依赖性钾离子通道及胰腺外分泌的影响.方法 分离SD大鼠胰腺组织,取得胰腺腺泡细胞悬液,用EPC10膜片钳放大器记录电压钳制下的不同组别胰腺腺泡细胞Iks的电流值并比较其间差异.分组如下:冲洗液处理为对照组,5μmol/L和20μmol/L 293B组,5 nmol/L促胰液素组,5 nmol/L促胰液素+100 nmol/L或10 nmol/L或1 nmol/L生长抑素组,0.3 mmol/L 8溴—环单磷酸腺苷(8Br-cAMP)组,0.3 mmol/L 8Br-cAMP+100 nmol/L生长抑素组,1 μmol/L乙酰胆碱(Ach)组,1μmol/L Ach+100 nmol/L生长抑素组,5 nmol/L促胰液素+10-5mol/L或10-6mol/L或10-7mol/L奥美拉唑组,1μmol/L Ach+10-5 mol/L或10-6 mol/L或10-7 mol/L奥美拉唑组.组间比较采用单因素方差分析,P＜0.05为差异有统计学意义.结果 大鼠胰腺腺泡细胞静息膜电位为(-40±0.8)mV,当去极化至+10 mV时,对照组的Iks为(420.0±3.2) pA.经5 μmol/L和20 μmol/L 293B作用后,大鼠胰腺腺泡细胞Iks减少为对照组的(60.4±4.2)％和(30.2±3.1)％(F=6.87,P＜0.05).5 nmol/L促胰液素刺激后,Iks增加为(823.0±2.2)pA,分别加入100 nmol/L、10 nmol/L和1 nmol/L生长抑素,Iks降低至(510.0±3.2)pA,(584.0±2.8)pA和(789.0±6.9)pA(F=5.67,P＜0.05);分别加入10-5 mol/L、10-6 mol/L和10-7mol/L奥美拉唑后,Iks峰值未见明显变化,分别为(806.5±3.6)pA,(814.8±3.2) pA和(816.3±2.9) pA (P＞0.05).1 μmol/L Ach刺激后,Iks的峰值为(966.0±3.2) pA;加入100 nmol/L生长抑素后,Iks峰值为(942.0±6.3)pA;分别加入10-5 mol/L,10-6 mol/L和10-7 mol/L奥美拉唑后,其Iks峰值未见明显变化,分别为(956.3±10.3) pA,(957.5±8.6)pA和(960.0±8.4)pA (P＞0.05).结论 生长抑素能抑制胰腺腺泡细胞慢活化电压依赖性钾离子通道的开放,PPI对该通道没有明显抑制作用.     

青少年高血压的研究进展

随着人们生活和行为方式的改变,高血压发病明显呈年轻化趋势。在青少年时期识别高血压病高危人群有助于早期进行有效干预和治疗,降低未来高血压的发生率及其严重性。现试从青少年高血压的诊断、发病因素、特点、治疗策略等方面的研究进展作一综述。     

Morbidity in cardiovascular diseases in immigrants in Sweden

INTRODUCTION: Although immigration to Sweden has increased in the last few decades, the incidence rates of cardiovascular disease and coronary heart disease in immigrants are unknown. The aim of the present study is to estimate whether place of birth affects the incidence rates of cardiovascular disease and coronary heart disease. MATERIAL AND METHODS: The study was designed as a follow-up study on morbidity in cardiovascular disease and coronary heart disease between 1 January 1997 and 31 December 1998, including three and a half million persons with age range 35-64 years, of whom 550 000 were born abroad, from the database MigMed consisting of the whole Swedish population. Incidence rates and relative risks were estimated by indirect standardization and a proportional hazard model. RESULTS: The age-adjusted risk of coronary heart disease was higher in most foreign-born groups than in Swedes. For example, in nine of 12 male groups, the relative risks varied between 1.1 and 2.2, and in seven of 12 female groups, the relative risks varied between 1.4 and 2.5. When also adjusting for level of education and employment status, the risks were still high, but on a lower level. CONCLUSIONS: Foreign-born people possess an over-risk of cardiovascular or coronary heart disease(CVD/CHD) compared with Swedish-born persons, also when level of education and employment status are taken into account.     

C·肉毒杀鼠素(冻干剂)应用研究

目的为研究C·肉毒杀鼠索对杀灭达乌尔黄鼠(简称黄鼠)的大面积应用情况和对家畜、家禽的毒害作用,进行了C·肉毒杀鼠素的应用研究.方法大面积投毒采用ES-2药饵撒播机[1],间隔约80m进行条投.羊、鸡采用直接灌胃.结果大面积应用的灭鼠率为83.72%.对羊、鸡最高剂量分别为500万MLD、150万MLD,均未出现中毒现象.结论 C·肉毒杀鼠素是较为理想的草原大面积杀灭黄鼠的理想、首选药物.     

Pulmonary sporotrichosis in Oklahoma in susceptibilities in vitro

Six cases of pulmonary sporotichosis were observed in 2 institutions in Oklahoma City, Okla. Three of the patients were treated with iodides with or without surgery. Although one patient required a second course of iodides, the patients have remained well after at least 34 months of follow-up. Three patients treated with amphotericin B, single course as well as multiple courses, and other antifungal agents (hydroxystilbamidine and miconazole) have all relapsed. These cases and a reviewed of more than 40 cases of pulmonary sporotrichosis susceptibilities of Sporothrix schenckii that we observed in vitro suggest that amphotericin B is not an effective agent for the treatment of pulmonary sporotrichosis. It is our opinion that the treatment of choice for pulmonary sporotrichosis is a supersaturated solution of potassium iodide. If the patient is allergic to the medication or fails to respond, then a combination of amphotericin B plus flucytosine may be tried.