THE EFFECT OF ISCHEMIC RE-PERFUSIONINJURY PLUS PARTICLE INFUSION EMBOLISM ON THE APOPTOSIS OF RATS WITH PANCREATIC CANCERA

OBJECTIVES: In an attempt to develop new method of treating the end- or mid-stage pancreatic cancer, we examined the effect of ischemic re-perfusion injury plus particle embolism on the pathology and cell apoptosis of pancreatic cancer in Sprague Dawely rats. METHODS: 9 mg dimethylbeneanthracine (DMBA) were implanted directly into the parenchyma of pancreatic tail of Sprague-Dawely rats. After establishment of tumor, the inferior splenic artery, a main supplying vessel to pancreatic tail was subjected to blockade and re-opening for 30 min separately, then embolism particles were infused via the artery. Afterwards, artery was ligated. Pathological changes and cell apoptosis indicators (AI) of pancreatic cancer were observed by light microscopy and ISEL respectively 14 days after the operation. RESULTS: The prevalence of pancreatic cancer among DMBA-implanted rats evaluated 3 months to 4 months after implantation was 59%. The volumes of the tumor in positive control group (B), pancreatic ischemic group (C), pancreatic ischemic re-perfusion injury group (D) were significantly larger than pancreatic ischemic re-perfusion injury plus particle thrombus group (E) ( P < 0.01). The volumes of the tumor in groups D, E were significantly smaller than that in group C (P < 0.01). There was a significant difference in tumor size between group B and group C ( P < 0. 01), but the difference was not significant between group D and group E ( P > 0.05). There was a significant infiltration of tumor tissue in group B rats, but strong inflammatory reaction was not noted. In groups C, D, E, a localized tumor growth was observed; infiltration of inflammatory cells and proliferation of fibroblasts and connective fiber were obvious, and some of these fibers grew into cancer nests and separate the tumor. The above findings were most conspicuous in group E. There was a significant difference in AI between group E (13.7 +/- 1.5) and other groups ( P < 0.01), with the difference being also significant between group C (4.3 +/- 2.4), D (8.5 +/- 1.1) and group B (1.2 +/- 0.8) (P < 0.01), and between group C and group D (P < 0.01) or between group D and group E ( P < 0.01). In the samples of group A, the apoptotic cells were not found. CONCLUSIONS: Pancreatic ischemic re-perfusion injury plus particle thrombus can cause significant infiltration of inflammatory cells in tumor tissues thereby limiting its growth, and inducing cell apoptosis of pancreatic cancer. This effect is superior to either pancreatic ischemia alone or pancreatic ischemia plus re-perfusion injury.     

乳腺癌气虚血瘀证小鼠模型的建立及益气活血法的干预研究

目的建立气虚血瘀证乳腺癌荷瘤4-T1小鼠模型,并予益气活血中药干预,分析在气虚血瘀状态下中药干预对肿瘤微环境的影响。方法雌性BALB/c小鼠48只,随机平均分为6组:空白对照组、单纯荷瘤组、气虚血瘀证荷瘤模型组(复合模型组)、复合模型低剂量组、复合模型中剂量组、复合模型高剂量组。单纯荷瘤组小鼠注射4T1单细胞悬液,复合模型组在单纯荷瘤组基础上注射利血平。从实验第15天起,复合模型低剂量组、中剂量组、高剂量组分别灌胃相应剂量中药(四君子汤合血府逐瘀汤),每日1次,连续28 d。分别于实验第7、14、21、28、35、42天对复合模型组及各中药干预组予症状量化评分表评估,第42天测量各组耳缘、脚趾、腹壁静脉微循环血流量,Western blot法测定肿瘤组织基质金属蛋白酶2(MMP2)、磷酸化细胞外信号调节激酶(p-ERK1/2)的表达。结果复合模型组、复合模型低剂量组、复合模型中剂量组、复合模型高剂量组均出现气虚血瘀症状,低剂量组量化评分值自第28天低于复合模型组(P0.05),中剂量组自第21天分值低于复合模型组(P0.05)。空白对照组静脉微循环血流量大于其余各组(P0.05),复合模型组小于单纯荷瘤组(P0.05),各中药干预组均比复合模型组提高(P0.01或P0.05)。复合模型组原位瘤MMP-2表达高于单纯荷瘤组(P0.05),各中药干预组均低于复合模型组(P0.05)。复合模型低剂量组、中剂量组原位瘤p-ERK1/2表达均低于复合模型组(P0.05或P0.01)。结论通过注射4T1单细胞悬液结合注射利血平方法建立的气虚血瘀证乳腺癌小鼠模型稳定可靠,符合中医证候特点。气虚血瘀证小鼠肿瘤组织内MMP-2、p-ERK1/2表达上调,益气活血中药干预后能纠正气虚血瘀状态同时下调MMP-2、p-ERK1/2的表达,抑制肿瘤生长。     

脑梗死,TIA患者外周血IL—1,IL—6及TNF水平的检测分析

探讨白细胞介素－１、白细胞介素－６及肿瘤坏死因子与脑缺缺血的关系。方法用双抗体夹心法测定３８例脑梗死患者、１６例短暂性脑缺血发作患者及４０例正常人周围血的ＩＬ－１、ＩＬ－６，ＴＮＦ水平。     

门静脉淤血对硬化肝脏缺血再灌注的损伤作用

研究肝硬化大鼠肝缺血再灌注(HepaticIscheiaReperfusion,HIR)损伤的机制。方法用60%四氯化碳(CCl4)溶液皮下注射的方法制作肝硬化大鼠模型。随机分为四组A组假手术组(6只),B组单纯肠系膜上静脉阻断(16只),C组肝门阻断+门腔转流(16只),D组肝门完全阻断40min(16只)。观察丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、透明质酸(HA)、肿瘤坏死因子(TNF)、7d存活率及肝、肺病理的变化。结果B、C、D组的7d存活率分别为5/10只、8/10只、4/10只;再灌注后4h血清TNF含量,B、C、D组分别为(0.631±0.198)u/ml、(0.596±0.223)u/ml、(0.789±0.371)u/ml,明显高于术前的(0.177±0.139)u/ml和A组的(0.315±0.182)u/ml(P＜0.01);D组再灌注4h的AST明显高于C组(P＜0.01);C组的AST、ALT显著高于B组(P＜0.05);同时的HA水平,D组明显高于B组,C组显著高于A组(P＜0.01);肝、肺组织学检查可见肝、肺的病理损害,程度以D组最重、B组次之。结论门脉淤血是导致肝硬化大鼠HIR损伤乃至死亡的主要原因。     

EFFECTS OF ACUTE HYPOGLYCEMIA ON THE OREXIN SYSTEM IN RAT

OREXIN isa kind of new neuropeptideisolated from rathypothalamus, which involvedin many physiologicalregulationprogressesvia orexin1receptor(OX R )and orexin2 receptor(OX R ).The physio- 1 2logicalfunctionoforexinisstillnotcompletelyknown ,butitisidentifiedthatorexinis involvedinmany physiologicalprogressesuchasfeeding,sleep,and stress.1However ,itisstillunknown abouttheregulationoforexinsystemby hypo-thalamus,theregulationtopancreasby orexinsystem,and the regulationbetween orexin,bloodglu…     

低剂量低分子肝素对内毒素诱发的急性肺损伤的影响

目的 探讨低剂量低分子肝素(LMWH)对内毒素(LPS)诱发的急性肺损伤(ALI)的影响.方法 选取36只雄性SD大鼠分为3组:正常对照组(A组),LPS组(B组)和LPS+ LMWH组(C组),每组12只.B、C组腹腔注射6 mg/kg LPS诱发ALI.C组腹腔注射低分子肝素100 U/kg,B组腹腔注入同等容积的生理盐水.6h后处死动物,光镜下观察各组大鼠肺组织病理改变,行动脉血气分析、检测肺湿质量/干质量(W/D)比值和支气管肺泡灌洗液(BALF)中总蛋白浓度;测定肺组织髓过氧化物酶(MPO)活性、丙二醛(MDA)水平;ELISA法测定血浆中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)及白细胞介素6(IL-6)水平.结果 B、C组PaO2、pH值低于A组,C组与B组相比明显升高(P＜0.05).B、C组大鼠肺W/D、BALF总蛋白及肺组织MDA、MPO水平明显高于A组(P＜0.01);C组与B组相比,肺W/D、BALF中蛋白及肺组织MDA、MPO水平明显下降(P＜0.05).B、C组大鼠血浆中TNF-α、IL-1β及IL-6水平较A组明显升高(P＜0.01),而C组较B组明显降低(P＜0.01).结论 LMWH处理能够减轻LPS诱发的急性肺损伤.     

VEGF—C对胰腺癌细胞凋亡影响的实验研究

目的研究VEGF—C对胰腺癌细胞凋亡的影响。方法建立人胰腺癌细胞株PANC-1裸鼠原位种植瘤模型,原代培养原发和淋巴结转移灶中胰腺癌细胞,通过VEGF—C反义寡核苷酸体外转染抑制其表达,应用RT—PCR及流式细胞术等方法研究对淋巴结转移胰腺癌细胞凋亡及bcl-2的影响。结果体外转染后,原代培养原发和淋巴结转移灶中PANC-1胰腺癌细胞VEGF—C的mRNA表达水平显著降低（P〈0．01）。体外转染VEGF—C反义寡核苷酸抑制其表达后,对照组、SODN组、ASODN组淋巴结转移胰腺癌细胞的凋亡率为（2．83±1．01）％、（4．98±2．05）％、（13．22±2．17）％,ASODN组细胞凋亡率显著提高（P〈0．01）,而原发灶胰腺癌细胞的凋亡率分别为（3．51±1．38）％、（4．79±2．16）％、（5．33±2．18）％,凋亡率无明显影响（P〉0．05）;淋巴结转移胰腺癌细胞的反义组bcl-2表达水平明显下调（P〈0．05）,而原发灶胰腺癌细胞bcl-2表达水平无明显变化（P〉0．05）。结论抑制淋巴结转移灶中胰腺癌细胞VEGF—C的高表达可促进胰腺癌细胞凋亡,这和bcl-2表达下调有关;但对原发灶胰腺癌细胞无明显影响。     

参附注射液对局灶性脑缺血再灌注损伤大鼠血脑屏障通透性及神经功能的影响

目的:观察参附注射液(Shenfu injection,SFI)对大鼠脑局灶性缺血再灌注(Ischemic-reperfusion,IR)损伤后血脑屏障(Blood brain barrier,BBB)通透性和神经功能的影响,评价参附注射液对大鼠局灶性脑缺血再灌注损伤有无保护作用,为参附注射液的临床应用提供参考.方法:清洁级的SD雄性大鼠80只.随机分为A、B、C、D4个组:A组为假手术加生理盐水处理组,B组为假手术加参附注射液处理组,C组为造模加生理盐水处理组,D组为造模加参附注射液处理组.A、C组于麻醉后5 min经尾静脉注射生理盐水10 ml/kg,B、D组在相同时点给予相同剂量的SFI.随后对C、D两组采用Zea Longa线栓法制备大鼠右侧大脑中动脉(Middle cerebral artery,MCA)缺血2 h再灌注24 h模型.A、B两组均不作造模处理,其余处理步骤与C、D两组相同.记录各组麻醉后苏醒时间,缺血2 h及再灌注24 h两时间点观察神经功能缺损情况,于再灌注24 h结束时在每组随机选择5只取右侧额顶叶大脑皮质部分电镜下观察血脑屏障超微结构的情况,其余15只大鼠静脉注射伊文思蓝(Evan's blue,EB),对比两组伊文思蓝通过血脑屏障进入脑组织的量.结果:各组间比较,体重差异无统计学意义.麻醉后苏醒时间A、B两组短于C、D两组(P＜0.05),A、B两组之间无差异,C组长于D组(P＜0.05).缺血2 h神经功能缺损评分(Longa score,Longa评分):A、B两组低于C、D两组(P＜0.01),A、B两组之间以及C、D两组之间差异无统计学意义.再灌注24 h后Longa评分:A、B两组低于C、D两组(P＜0.01),A、B两组之间差异无统计学意义,C组高于D组(P＜0.01).C组再灌注24 h Longa评分明显高于缺血2 h(P＜0.01),D组这两个时点比较,差异无统计学意义.缺血2 h,再灌注24h结束时,A、B两组大鼠脑组织内伊文思蓝含量明显低于C、D两组(P＜0.01),A、B两组之间差异无统计学意义,C组明显高于D组(P＜0.05).电镜下,A、B两组大鼠大脑皮质毛细血管周围及管腔未见明显变化.C组大鼠大脑皮质缺血再灌注区毛细血管周围重度水肿,毛细血管管腔明显受压.D组相应区域毛细血管周围轻度水肿,毛细血管轻度受压.结论:参附注射液可减轻局灶性脑缺血再灌注损伤大鼠血脑屏障通透性升高程度.减轻神经功能受损伤程度,发挥脑保护作用.     

终止香烟烟雾暴露后大鼠Th1/Tc1介导气道炎症及调节性T细胞的变化

目的 观察香烟烟雾暴露和终止香烟烟雾暴露后大鼠Th1/Tc1介导气道炎症及支气管肺泡灌洗液(BALF)中调节性T细胞(Treg)的变化.方法 将50只健康清洁级雄性Wistar大鼠随机分为5组:12周正常对照组(简称12周对照组)、24周正常对照组(简称24周对照组)、12周香烟烟雾暴露组(简称12周暴露组)、24周香烟烟雾暴露组(简称24周暴露组)、终止香烟烟雾暴露组(简称终止暴露组).烟熏法复制大鼠气道炎症的动物模型.12周后,12周对照组、12周暴露组处置取材,24周暴露组继续烟熏12周,终止暴露组终止烟雾暴露12周,将后2组及24周对照组处置取材.HE染色观察小气道病理改变,进行气道评分;收集BALF进行细胞学计数和分类计数;酶联免疫吸附法(ELISA)法测BALF中4种细胞因子:Th1/Th2型细胞因子干扰素(IFN)γ、白细胞介素(IL)4及促炎因子IL-8、肿瘤坏死因子(TNF)α的浓度;流式细胞术检测各组大鼠BALF中Treg细胞的比例,RT-PCR检测各组大鼠BALF中Foxp3 mRNA的表达.结果 (1)12周暴露组、24周暴露组、终止暴露组气道炎症评分较12周对照组、24周对照组明显高(均P＜0.01).24周暴露组、终止暴露组气道炎症评分均较12周暴露组高(均P＜0.01).(2)与12周对照组、24周对照组相比,12周暴露组、24周暴露组、终止暴露组BALF中IFN-γ、TNF-α和IL-8高,IL-4低(均P＜0.01).与12周暴露组相比,终止暴露组IFN-γ、IL-4、TNF-α差异均无统计学意义(均P＞0.05),IL-8较高(P＜0.01),24周暴露组IFN-γ、TNF-α和IL-8明显高(均P＜0.01).(3)BALF中Treg细胞比例,12周暴露组(7.4%±0.8%)、24周暴露组(7.8%±1.7%)、终止暴露组(7.0%±1.4%)较12周对照组(4.8%±1.2%)、24周对照组(4.7%±1.2%)高(均P＜0.01),前3组之间BALF中Treg细胞比例差异均无统计学意义(均P＞0.05).(4)12周暴露组(0.22±0.02)、24周暴露组(0.23±0.03)、终止暴露组(0.20±0.04)BALF中Foxp3 mRNA表达较12周暴露组(0.13±0.01)、24周暴露组(0.11±0.02)高(均P＜0.01).前3者之间BALF中Foxp3 mRNA表达差异均无统计学意义(均P＞0.05).结论 香烟暴露致大鼠气道Th1/Tc1介导炎症伴Treg细胞表达增高,终止香烟烟雾暴露后其炎症及Treg细胞高表达仍持续存在,提示该免疫失衡可能是导致终止香烟暴露后Th1/Tc1气道炎症仍持续进展的原因之一.     

金纳多对兔移植肺缺血再灌注损伤保护作用的实验研究

目的探讨金纳多对肺移植术供体肺缺血再灌注损伤的保护作用及机制。方法建立模拟的兔肺自体原位移植模型。分为单纯缺血再灌注(I/R)组(n=6),改良LPD液灌注(LPD)组(n=6)和金纳多治疗(LPD+E)组(n=6)。监测氧分压(PaO2)、血清肿瘤坏死因子(TNF-α)的变化,左肺作肺组织干/湿重比值(D/W)、丙二醛(MDA)含量、髓过氧化物酶(MPO)活力的测定,并在光镜下观察肺组织病理变化。结果(1)3组再灌注后15、60和90min的PaO2均有明显的下降,但LPD+E组明显好于I/R组(各时点分别为212.2mmHg±53.9mmHgvs122.5mmHg±20.7mmHg,240.5mmHg±52.5mmHgvs64.5mmHg±5.6mmHg,236.5mmHg±51.1mmHgvs100.0mmHg±8.6mmHg,P<0.01),LPD组与LPD+E组相比差异无统计学意义。(2)I/R组和LPD组再灌注后各时点及LPD+E组再灌注后60min、90min后TNF-α水平高于缺血前,但LPD+E组明显低于其余两组(分别为53.0ng/L±6.2ng/Lvs98.5ng/L±2.8ng/L、86.9ng/L±3.5ng/L,56.5ng/L±6.3ng/Lvs103.7ng/L±4.4ng/L、90.2ng/L±2.4ng/L,P<0.05)。(3)I/R组、LPD组和LPD+E组肺组织MDA含量分别为12.4nmol/mg±1.1nmol/mg、9.9nmol/mg±0.9nmol/mg、6.6nmol/mg±0.7nmol/mg,MPO活力分别为14.85U/g±1.40U/g、12.81U/g±1.04U/g、10.38U/g±1.07U/g,各组间比较P<0.01。(4)肺组织D/W比值:I/R组、LPD组和LPD+E组分别为0.1309±0.0122、0.1550±0.0096、0.1775±0.0073,各组间比较P<0.01。(5)病理学改变:I/R组肺组织损伤严重,肺泡间隔大量炎症细胞浸润,肺泡腔内炎症细胞聚集、炎性液体渗出,可见片状出血,LPD+E组病理改变最为轻微,炎症细胞浸润、炎性渗液不显著。结论金纳多对兔移植肺缺血再灌注损伤具有明显的保护作用,其作用机制可能通过与抗氧化、抑制中性粒细胞聚集和炎症因子TNF-α的释放有关。     

针刺百会、大椎穴对局灶性脑缺血大鼠皮层脑源性神经营养因子表达的影响

[目的]观察电针对不同时间段局灶性脑缺血大鼠缺血区皮层脑源性神经营养因子(BDNF)的影响。[方法]SD大鼠36只,随机分为6组,分别为假手术2周与5周组、缺血2周与5周模型组、电针2周与5周组;除假手术组外.其他动物均采用热凝闭大鼠大脑中动脉法复制局灶性脑缺血模型,观察缺血2周和5周后缺血区皮层BDNF的变化规律及针刺对其影响。[结果]假手术组大鼠相应区域仅见少量BDNF阳性细胞,且强度较弱;脑缺血2周及5周组大鼠缺血区BDNF的免疫阳性细胞数量比假手术组增多(P<0.01),但脑缺血2周组与5周组大鼠缺血区脑片比较,BDNF阳性细胞数量无显著性差异(P>0.05);电针2周组与5周组脑片中免疫阳性细胞数量增加,与模型组比较有显著性差异(P<0.01),但随着时间的推移,此表达呈下降趋势。[结论]电针可以通过提高BDNF在缺血区周围皮层的表达,保护缺血性脑损伤,并可能与大脑可塑性的形成有一定的关系。     

不同浓度七氟醚预处理对内毒素性 急性肺损伤大鼠肺组织的影响

目的：比较3种不同浓度(3.6%,2.4%和1.2%)七氟醚预处理对内毒素性急性肺损伤大鼠肺组织的影响。方法：健康雄性SD大鼠36只,随机分为6组（n=6）：对照组（A组）、单纯七氟醚吸入组（B组）、3.6%七氟醚预处理组（C组）、2.4%七氟醚预处理组（D组）、1.2%七氟醚预处理组（E组）、内毒素组（F组）。分别在给药后（LPS或生理盐水）6 h 处死大鼠, 观察肺组织病理学结果,测定肺组织湿干比(W/ D)、髓过氧化物酶(MPO)活性及肺组织ICAM-1mRNA的表达。结果：与对照组比较,F组和不同浓度七氟醚预处理组肺组织病理损伤加重,肺W/D、MPO活性、ICAM-1mRNA表达升高(P＜0.05)。与F组比较,C组肺组织病理损伤减轻,MPO活性和ICAM-1mRNA表达降低(P＜0.05),但肺W/D无明显降低（P＞0.05）;D组肺组织病理损伤减轻,肺W/D、MPO活性、ICAM-1mRNA表达均降低(P＜0.05),F组肺组织MPO活性降低(P＜0.05),但肺W/D和ICAM-1mRNA表达无明显降低（P＞0.05）。结论：2.4%七氟醚预处理可以减轻内毒素所致急性肺损伤,作用机制可能与其降低肺组织ICAM-1mRNA的表达上调,从而减少肺内中性粒细胞的浸润相关。     

KiSS-1基因及其蛋白在人胰腺癌组织中表达及其与侵袭和转移的关系

目的研究胰腺癌组织中KiSS-1表达及其与侵袭和转移的关系.方法应用RT-PCR和western blot法检测KiSS-1mRNA及其蛋白肽metastin在37例胰腺癌及27例转移灶和9例正常胰腺组织中的表达情况.结果在胰腺癌组织中KiSS-1基因及其蛋白表达水平显著低于正常胰腺组织(P＜0.01,P＜0.01).KiSS-1mRNA及其蛋白的表达与性别、年龄、肿瘤大小及组织学分级无关,与临床分期、有无转移和神经侵犯密切相关.在Ⅲ、Ⅳ期胰腺癌组织中KiSS-1基因及蛋白的表达分别为(0.135±0.121)和(9.23±2.74)μg/100μg总蛋白,明显低于Ⅰ、Ⅱ期(P＜0.05,P＜0.05).KiSS-1基因及其蛋白在无转移、有转移的胰腺癌原发灶和转移灶组织中表达水平呈降低趋势,转移灶中的表达水平最低.有神经侵犯的胰腺癌原发灶KiSS-1基因及蛋白表达显著低于无神经侵犯的胰腺癌原发灶(P＜0.01,P＜0.01),其表达与有无神经侵犯相关系数分别为Kendall τ-b=0.597(P=0.003)和Kendall τ-b=0.438(P=0.007).结论KiSS-1基因转录和表达降低与胰腺癌侵袭和转移密切相关,可能参与胰腺癌转移的调控.     

肢体缺血后处理和盐酸戊乙奎醚防治胃缺血再灌注损伤的实验观察

目的 探讨缺血后处理和盐酸戊乙奎醚对大鼠肢体缺血再灌注时胃损伤的防治作用及其可能的作用机制.方法 实验采用大鼠肢体缺血再灌注损伤模型,将雄性Wistar大鼠144只,体重220～250 g,随机分为4组:对照组(C);缺血再灌注组(IR组);缺血后处理组(IPO组)和盐酸戊乙奎醚组(IPHC组).C、IR、IPO和IPHC组再分为再灌注0 h(T0)、1 h(T1)、3 b(T3)、6 h(T6)、12 b(T12)和24 h(T24)6个亚组(n=6).分别于上述6个时间点测定血清中LDH和CK活性,TNF-α和IL-10含量及胃组织中MDA含量,SOD、MPO、XOD和LDH活性;光镜下观察病理学改变和HIF-1α的蛋白表达水平.结果 与C组比较,IR、IPO和IPHC组血清LDH[再灌注后3 h:(6.68±0.11)、(3.93±0.07)、(2.92±0.08)U/ml]和CK[再灌注后3 h:(4.04±0.27)、(2.31±0.11)、(2.07±0.15)U/ml]活性升高、TNF-α[(再灌注后3 h:106.86±8.31)、(87.80±7.31)、(74.21±1.04)mg/ml]和IL-10[再灌注后3 h:(135.42±8.62)、(151.71±4.06)、(173.36±4.96)mg/m1]含量升高(P＜0.05或P＜0.01),胃组织SOD活性降低、MPO和XOD活性及LDH活性和MDA含量升高(P＜0.05或P＜0.01);光镜可见部分胃黏膜肌层或胃黏膜与腺体间少量中性粒细胞浸润,或少许嗜酸性粒细胞浸润、偶见中性粒细胞浸润,间质血管扩张充血并黏膜深层少量出血或间质血管扩张充血和HIF-1α表达上调(P＜0.01).与IR组比较,IPO和IPHC组血清LDH和CK活性降低、TNF-α含量降低和IL-10含量升高(P＜0.01),胃组织SOD活性升高、MPO和XOD活性及LDH活性和MDA含量降低(P＜0.05或P＜0.01);胃组织病理损伤减轻和HIF-1α表达下调(P＜0.01).与IPO组比较,IPHC组血清LDH活性明显降低、CK活性先降低后升高、TNF-α含量明显降低和IL-10含量升高(P＜0.05或P＜0.01),胃组织SOD活性明显升高、MPO活性明显降低、MDA含量于3 h升高、XOD活性于12 h后升高和LDH活性于3 h升高且12 h后降低(P＜0.05或P＜0.01),组织病理损伤减轻和HIF-1α表达下调(P＜0.01).结论 可以通过减少氧自由基的产生、中性粒细胞的浸润、炎症因子的释放和促进抗炎因子的产生及改善微循环功能和细胞能量代谢,有效减轻肢体缺血再灌注继发性胃缺血再灌注损伤.盐酸戊乙奎醚后处理的抗缺血再灌注损伤作用好于肢体缺血后处理.

Abstract：

Objective To investigate the protective effects of ischemic postconditioningand and penehyclldine hydrochloride on gastric injury induced by ischemia-reperfusion of lower limb in rats.Methods The model of limb ischemia reperfusion injury was used to perform this experiment.One hundred and forty four male Wistar rats weighing 220-250 g were randomly divided into 4 groups:group Ⅰ Control (C),group Ⅱ Ischemic Reperfusion(IR),group Ⅲ Ischemic postconditioning(IPO)and group Ⅳpenehyclidine hydrochloride(IPHC);C,IR,IPO and IPHC groups has been followed for 0(T0),1(T1),3(T3),6(T6),12(T12),or 24(T24)perfusion,all the groups were secondary separated into six subgroups as time point and each subgroup contained six rats,respectively.Blood samples from the inferior vena cava were taken for determineation of LDH,CK activities and TNF-α,IL-10 content at every time point of reperfusion;the animals were killed at every time point respectively and the gastric were removed for determineation of SOD,MPO,XOD and LDH activities,MDA content,and histologyical examination and the expression of HIF-1α was analyzed.Results Compared with group C,IR,IPO and IPHC,in serum LDH and CK activeities were increased,TNF-α and IL-10 content were increased(P ＜0.05 or P ＜0.01);and in gastric tissue MPO,XOD and LDH activeities were increased and MDA content increased,while SOD activity decreased in group IR,IPO and IPHC(P＜0.05 or P＜0.01);and gastric tissue resulted in significant injury as evidenced by infiltrated of few neutrophils or eosinophils and rare neutrophils between the gastric mucosa or muscularis mucosa and the glands,interstitial vascular dilation hyperemia and small quantity hemorrhage from deep layers of mucosa or interstitial vascular dilation hyperemia,and the expression of HIF-1α was significantly increased(P ＜0.01).Compared with group IR,IPO and IPHC in serum LDH and CK activeities,TNF-α content decreased while IL-10 content were increased(P ＜0.01);and in gastric tissue MDA content,MPO,XOD and LDH activeities were decreased,and SOD activity increased in group IPO and IPHC(P ＜0.05 or P ＜0.01);and the histologyical injury were milder and the expression of HIF-1α was significantly decreased(P ＜0.01).Compared with group IPO,IPHC,in serum LDH activeities were markably decreased,CK activeities were first increased and then declined,TNF-α content markaly declined while IL-10 content were increased(P ＜0.05 or P ＜0.01),and the histologyical injury were milder and the expression of HIF-1α was markably decreased(P＜0.01)and in gastric tissue SOD activity were markably increased,MPO activeities significantly decreased,MDA content increased at T3,XOD activeities increased after T12,LDH activeities increased at T3 and declined after T12 in group IPHC(P＜0.05 or P＜0.01).Conclusion The combination of ischemia postconditioning and postconditioning with penehyclidine hydrochloride can protect the gstric from ischemia-reperfusion injury induced by ischemia reperfusion of the lower limbs in rats,the main mechanism may be reducing post-ischemic oxidative damage,inflammatory reaction,amelio-rating microcirculatory and cellular energy metabolism et al.Additionally,this study found that the protective effects of penehyclldine hydrochloride on gastric injuryinducedbyischemiareperfusionof thelowerlimbs,werebetterthanischemic postconditioning,and the mechanism might be related to its anti-inflammatory effect,antioxidant action and prevention of cell injury et al.     

氯胺酮对脓毒症大鼠肺组织HO-1表达的影响

目的探讨氯胺酮对脓毒症大鼠肺组织血红素加氧酶-1表达的影响及其可能的保护机制。方法选择健康雄性Vista大鼠40只,随机分为5组：假手术组（A组）、CLP组（B组）、氯胺酮40mg／kg＋CLP组（C组）、氯胺酮组40n=1g／kg＋锌原卟啉组IX5mg／k时CLP（D组）、锌原卟啉IX5mg／kg＋CLP组（E组）,药品均用生理盐水稀释到1ml,假手术组、CLP组均术前30min腹腔注射1ml生理盐水。各组均在9h腹主动脉放血处死大鼠,检测肺组织中MDA的含量、肺泡灌洗液中（BALF）蛋白含量（TP）,取肺组织行HE染色观察其病理变化,用免疫组化方法测定HO-1的表达。结果与A组相比,各损伤组MDA含量和BALF中总蛋白含量升高,HO-1表达上调（P〈0．01）;与B组相比,C组MDA含量和BALF中总蛋白含量降低,HO-1表达上调（P〈O．05或P〈0．01）;与c组相比,D组MDA含量和BALF中总蛋白含量升高,HO-1表达下调（P〈0．01）;B组、D组、E组之间比较,MDA含量和BALF中总蛋白含量差异无统计学意义。结论氯胺酮能够减轻脓毒症大鼠肺损伤,此作用可能与诱导HO-1表达增强有关。     

依达拉奉对吸入性肺损伤大鼠肺组织的保护作用

目的 探讨依达拉奉对黑火药烟雾所致吸入性肺损伤大鼠的保护作用.方法 将24只雄性Wistar大鼠随机分为正常对照组（C组）、吸入性肺损伤组（S组）和依达拉奉组（E组）,每组8只.除正常对照组外,其余各组大鼠使用自制发烟装置构建吸入性肺损伤模型.造模成功后依达拉奉组按9 mg/kg腹腔注射依达拉奉注射液;正常对照组、吸入性肺损伤组腹腔注射0.9％氯化钠注射液12 ml/kg.于第7天从大鼠腹主动脉取血,检测各组大鼠动脉血气;肺湿/干重比（lung wet-to-dry weight ratio,W/D）;留取各组大鼠腹主动脉血液样本,离心取血清,待检测肿瘤坏死因子-α （tumor necrosis factor-α,TNF-α）、白细胞介素-6（interleukin-6,IL-6）含量.留取肺组织,制备肺组织匀浆后测定髓过氧化物酶（myeloperoxidase,MPO）、丙二醛（malondialdehyde,MDA）含量.取部分右肺组织,经4％甲醛溶液固定后做病理苏木精-伊红（HE）染色切片,光镜观察.结果 与S组相比,E组大鼠动脉中PaO2水平升高（P＜0.01）,大鼠血清中IL-6、TNF-α水平降低并降低肺组织中MPO活性及MDA含量（P＜0.01）.光镜下观察,E组肺组织较S组肺组织水肿减轻,炎性细胞浸润减少.结论 依达拉奉可能通过抑制自由基的产生及减少部分炎性介质的产生和释放,减轻烟雾吸入性肺损伤大鼠体内炎性反应,对肺组织起到一定的保护作用.     

鞘内注射塞地塞米松联合螺内酯对大鼠根性神经痛行为的影响

目的 探讨鞘内注射地塞米松(Dex)联合螺内酯(Spir)对大鼠根性神经痛行为的影响.方法 选择成功鞘内置管后无运动障碍的雄性SD大鼠48只,分为假手术组(Sham组,n=12)、溶剂对照组(C组,n=12)、Dex组(D组,n=8)、Spir组(S组,n=8)和Dex联合Spir组(DS组,n=8),制备注射式根性神经痛模型.D组、S组和DS组分别于造模后第2～4天,每次鞘内给予Dex 4μg、Spir 3μg、Dex 4μg+Spir 3μg,每天2次.Sham组和C组则给予等量的10%酒精.于造模前后测量双侧足底机械缩足阈值(PWMT)和热辐射刺激潜伏期(PWTL).结果 与Sham组相比,CCD术后同侧PWMT和PWTL都明显降低(P＜0.01).与C组相比,鞘内注射Dex明显抑制疼痛行为(P＜0.01),并一直持续到术后第10天;Spir也明显改善PWMT(P＜0.01)和PWTL(P＜0.01),并持续到术后第7天;Dex联合Spir则产生明显的协同作用[PWMT:(13.52±0.72)g,(11.58±1.38)g,P＜0.0l;PWTL:(19.63±1.68)s,(14.14±1.52)s,P＜0.01],并且这一作用持续了至少10 d.结论 鞘内注射Dex和Spir对慢性压迫大鼠背根神经节诱导的根性神经痛有治疗作用,二者联合使用可以产生明显的协同作用.

Abstract：

Objective To investigate the effects of intrathecally coadministered dexamethasone and spironolactone in trathecally on radicular pain behaviors.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion (CCD) ,48 male SD rats successfully received intrathecal catheter implantation and without motor dysfunction were randomly divided into Sham-operation group (Sham group, n= 12),Control group ( C group, n = 12 ), Dexamethasone group ( D group, n = 8 ), Spironolactone group ( S group, n = 8 )and Dexamethasone plus spironolactone group (DS group, n=8).Rats in D group,S group or DS group were intrathecally treated with dexamethasone 4 μg, spironolactone 3 μg or dexamethasone 4 μg plus spironolactone 3 μg twice daily on day 2 ～4 after CCD respectively,while rats in C and Sham group received 10μl 10% alcohol.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before CCD and day 1,4,7,10,14,17 and 21 after CCD.Results Compared with Sham group, both PWMT and PWTL were significantly decreased after CCD surgery on the ipsilateral side(P＜0.01 ＝.Intrathecally administrated with dexamethasone significantly improved pain behaviors (P＜0.01 ＝ and these therapeutic effects lasted up to 10 days after CCD surgery.As with dexamethasone,intrathecal spironolactone also significantly attenuated PWMT (P＜0.01 ＝ and PWTL (P＜0.01 ＝ and the change lasted up to 7 days after CCD surgery.Coadministration spironolactone and dexamethasone exhibited significant synergies( PWMT: ( 13.52 ± 0.72) g vs ( 11.58 ± 1.38 ) g, P ＜0.01; PWTL: ( 19.63 ± 1.68) s vs ( 14.14 ± 1.52) s, P ＜ 0.01 ＝.These effects lasted up to at least 10 days.Conclusion Both dexamethasone and spironolactone intrathecally have therapeutic effects on radicular pain behaviors, combination injection of these two drugs could generate significant synergies.     

TLR9对胰腺癌裸鼠增殖生长及化疗耐药性的研究

目的 观察TLR9在不同的活化状态下对人胰腺癌裸鼠移植瘤生长及药物抗性的影响.方法 建立人胰腺癌PANC-1细胞裸鼠移植肿瘤模型,并随机分为6组:无菌生理盐水组、TLR9激动剂、TLR9抑制剂组、吉西它滨组、TLR9抑制剂+吉西它滨组、TLR9激动剂+吉西它滨组进行实验.游标卡尺记录肿瘤体积大小及生长情况,采用免疫组化方法检测肿瘤TLR9受体表达情况,核磁共振成像(MRI)观察肿瘤生长、转移及周围组织侵犯情况.结果 吉西他滨组、TLR9激动剂+吉西它滨组、TLR9抑制剂+吉西它滨组肿瘤摘除后体积及生长速度明显小于其他组(P＜0.05),TLR9激动剂+吉西它滨组生长速度及肿瘤摘除后体积明显大于TLR9抑制剂+吉西它滨组及吉西他滨组(P＜0.05),TLR9抑制剂+吉西它滨组与吉西他滨组比较,差异有统计学意义(P＜0.05),TLR9激动剂组、TLR9抑制剂组及生理盐水组差异无统计学意义(P＞0.05).种植后7周小鼠在MRI下观察,瘤成椭圆形,境界清楚,周围组织未见明显转移及对周围组织的侵犯;检测肿瘤组织中并鉴定表面明确有TLR9的表达.结论 胰腺癌裸鼠移植瘤中确有TLR9的阳性表达,TLR9的激活可以明显降低胰腺癌对吉西他滨化疗的敏感性,增加肿瘤的耐药性,相反促进肿瘤生长.     

不同液体小容量复苏对内毒素休克大鼠肺损伤的影响

【目的】 探讨不同液体小容量复苏对大鼠内毒素休克所致肺损伤的影响&#65377;【方法】 清洁级SD大鼠30只随机分为5组,正常对照组(C组)&#65380;内毒素(LPS)对照组(LPS,E组)&#65380;75 g/L高渗氯化钠(HSS)组(LPS + HSS, HSS组)&#65380;羟乙基淀粉液130/0.4(HES)组(LPS + HES,HES组)&#65380;75 g/L高渗氯化钠羟乙基淀粉40溶液(HSH)组(LPS + HSH,HSH组)&#65377;各组在静注LPS(1 mg/kg) 30 min后予小容量复苏(4 ml/kg)干预&#65377;观察各组大鼠肺组织病理学变化,测定肺组织干/湿质量比例,测定肺泡灌洗液中蛋白浓度,测定肺组织丙二醛(MDA)的浓度以及超氧化物歧化酶(SOD)的活性&#65377;【结果】 与空白对照组比较,内毒素模型各组病理改变均加重,肺泡灌洗液蛋白浓度均升高(P < 0.05),内毒素对照组病理评分升高(P < 0.01),干/湿质量比例下降(P < 0.05),SOD活性下降(P < 0.01),内毒素对照组和75 g/L高渗氯化钠组MDA浓度升高;与内毒素对照组比较,75 g/L高渗氯化钠组,羟乙基淀粉130/0.4溶液组和75 g/L高渗氯化钠羟乙基淀粉40组,病理改变减轻,肺病理评分&#65380;MDA浓度下降(P < 0.01),干/湿质量比例升高(P < 0.05),SOD活性升高(P < 0.01)&#65377;羟乙基淀粉130/0.4溶液组和75 g/L高渗氯化钠羟乙基淀粉40组肺泡灌洗液中蛋白浓度下降(P < 0.05, P < 0.01)&#65377;【结论】 使用75 g/L高渗氯化钠,羟乙基淀粉130/0.4溶液和75 g/L高渗氯化钠羟乙基淀粉40进行小容量复苏对大鼠内毒素休克所致肺损伤有明显保护作用&#65377;相比75 g/L高渗氯化钠,羟乙基淀粉130/0.4溶液和75 g/L高渗氯化钠羟乙基淀粉40在减轻肺毛细血管通透性方面作用更佳&#65377;