缺血预处理对大鼠肝脏缺血/再灌注早期核因子κB活性及细胞凋亡的影响

目的 观察缺血预处理对大鼠肝脏缺血/再灌注早期核因子KB活性、促凋亡基因Fas和FasL蛋白表达以及肝细胞凋亡的影响,以进一步阐明肝脏缺血预处理的抗凋亡作用机制.方法 建立SD大鼠肝缺血(40min)再灌注(120 min)损伤及肝缺血预处理的模型.24只大鼠随机分成3组(n=8).①假手术对照组(C组),仅分离肝十二指肠韧带,不阻断肝门,不进行其他干预处理.②缺血再灌注组(IR组),在第一肝门用小血管夹阻断尾状叶及左肝叶血流40 min松开血管夹肝脏再灌注2 h,再灌注开始后关腹.③缺血预处理组(IP组),先行3个循环的缺血预处理,阻断第一肝门10 min,开放再灌注10 min为1个循环,随后操作同缺血/再灌注组.实验结束后全自动生化分析仪检测血清谷草转氨酶和血清谷丙转氨酶活性:TUNEL法检测肝组织的细胞凋亡指数(AI):EMSA法测定肝组织核因子κB的结合活性:Western blotting免疫印迹法检测Fas及FasL蛋白的表达水平.结果 IR组和IP组的ALT、AST及细胞凋亡指数(AI)均明显高于S组(P<0.01),但与IR相比,IP组则明显较低(P<0.01).与C组比较,IR组的核因子κB活性、促凋亡基因Fas和FasL蛋白表达水平明显增高,而IP组仅轻度增高.结论 缺血预处理可通过降低肝脏缺血/再灌注早期核因子κB的转录活性,并下调促凋亡基因Fas及FasL的表达,从而发挥抗细胞凋亡和损伤保护效应.     

缺血预处理对大鼠肝细胞凋亡及HIF-1α和Survivin蛋白表达的影响

目的:观察缺血预处理(IP)对缺血再灌注(IR)损伤引起的肝细胞凋亡及对调控基因Survivin、HIF-1α蛋白表达的影响,探讨IP对鼠肝IR损伤保护作用的机制。方法:将90只SD大鼠分为假手术(SO)组10只、IR组和IP组各40只。SO组术中只牵拉分离肝十二指肠韧带,操作完成后关腹;IR组阻断十二指肠韧带,造成缺血30min;IP组缺血30min前持续5min缺血及5min再灌注。IR组及IP组分别于再灌注2、6、12和24h后各处死10只动物,取肝脏标本;SO组于术后2h取肝组织标本。检测细胞凋亡指数(AI)及Survivin、HIF-1α蛋白表达水平。结果:IR组、IP组AI较SO组明显增加(P<0.05),相同时相点IP组较IR组细胞AI明显降低(P<0.05)。IR组、IP组Survivin、HIF-1α蛋白表达先升后降,6h左右达高峰;HIF-1α(除24h时)和Survivin蛋白表达在相同时相点IP组较IR组高(P<0.05)。结论:IP对肝组织IR损伤的保护作用机制之一可能是通过上调HIF-1α、Survivin蛋白的表达抑制细胞凋亡,从而发挥肝脏保护作用。     

缺血预处理和Caspase抑制剂对缺血再灌注损伤保护作用的比较

目的　比较缺血预处理和Caspase抑制剂治疗对大鼠肝缺血再灌注的保护作用及其机制。方法　SD大鼠随机分为 6组 :( 1)缺血再灌注1 (IR1 )组 ;( 2 )IR2 组 ;( 3 )缺血预处理1 (IP1 )组 ;( 4 )IP2 组 ;( 5 )Caspase抑制剂治疗1 (T1 )组 ;( 6)T2 组。比较各组大鼠 70 %肝脏 60min或 12 0min缺血 ,在再灌注 3h时的肝组织Caspase 3活性 ,肝细胞凋亡率和血清AST和ALT水平 ,及实验动物 7d存活率。结果　在 60min缺血及 3h再灌注时间 ,IP1 组和T1 组的保护作用相同 ,在 12 0min缺血及 3h再灌注时 ,T2 组对Caspase活性和肝细胞凋亡的抑制优于IP2 组 (P <0 .0 1) ,但两者的AST和ALT水平及动物 7d存活率均无显著性差异。结论　缺血预处理和Caspase抑制剂治疗对鼠肝缺血再灌注损伤都有保护作用 ,两者的保护效果无显著性差异。缺血预处理对缺血再灌注损伤的保护更简便、经济、安全 ,临床应用前景十分广阔。     

缺血后处理对大鼠肝缺血再灌注时肝细胞线粒体损伤的影响

目的 评价缺血后处理对大鼠肝缺血再灌注时线粒体损伤的影响.方法 雄性SD大鼠30只,体重180～230 g,随机分为3组(n=10):假手术组(S组)、肝缺血再灌注组(IR组)和缺血后处理组(Ipo组).IR组和Ipo组采用阻断肝门60 min再灌注6 h的方法 制备肝缺血再灌注模型,Ipo组缺血60 min时再灌注10 s、缺血10 s,反复6次,进行缺血后处理.于再灌注6 h时取静脉血样,测定血清谷丙转氨酶(ALT)及天门冬氨酸氨基转移酶(AST)活性,然后取肝组织,制备病理切片及分离肝细胞,电镜下观察线粒体超微结构,测定线粒体膜电位及线粒体Na+-K+-ATP酶活性.结果 与S组比较,IR组和Ipo组血清ALT和AST活性升高,线粒体Na+-K+-ATP酶活性及线粒体膜电位降低(P<0.01);与IR组比较,Ipo组血清ALT和AST活性降低,线粒体Na+-K+-ATP酶活性及线粒体膜电位升高(P<0.05或0.01).Ipo组线粒体损伤程度轻于IR组.结论 缺血后处理可减轻大鼠肝缺血再灌注时肝细胞线粒体损伤.     

常温下缺血预处理对肝细胞凋亡调节基因表达影响的临床研究

目的探讨常温下肝脏缺血预处理后细胞凋亡调节基因C-jun和bcl-XL的表达及其临床意义.方法16例肝癌患者随机分成缺血再灌注组(IR组)和缺血预处理组(IP组),每组8例.在肝门阻断前和再灌30min时抽血查肝损害标记酶,同时取肝组织切片行肝细胞凋亡、C-jun和bcl-XL基因及PCNA表达检测,并行光镜和电镜检查.结果IR组和IP组ALT,AST,LDH及凋亡指数(AI值)在再灌注30min时均较阻断前明显升高(P<0.05或P<0.01),IR组升高更显著(P<0.01),且IR组再灌注30min时有肝细胞坏死和超微结构不可逆性改变;与IP组比较,IR组再灌注30min时凋亡诱导基因C-jun和PCNA表达明显增强(P>0.05或P<0.01);与IR组比较,IP组再灌注30min时凋亡抑制基因bcl-XL表达明显增强(P<0.01).结论(1)常温下缺血预处理对肝脏缺血再灌注损伤的保护作用可能是通过调节肝细胞凋亡调节基因C-jun和bcl-XL表达而实现的;(2)肝脏IR损伤可能与激活C-jun基因表达促发肝细胞凋亡发生有关;(3)IP可能通过激活bcl-XL基因表达而抑制肝细胞凋亡发生.     

缺血预处理减轻大鼠肝缺血再灌注损伤的免疫机制研究

目的探讨大鼠肝缺血再灌注损伤(HIRI)的免疫机制和缺血预处理(IPC)的保护作用。 方法80只大鼠被随机分为假手术组(A组)、肝门阻断20 min组(B组)、30 min组(C组)、40 min组(D组)以及肝门阻断30 min前预处理组(E组),每组再分为再灌注2 h亚组和24 h亚组,各8只。检测再灌注后2 h、24 h的血丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、白细胞介素10、12(IL-10、IL-12)以及外周血T淋巴细胞亚群的水平,观察再灌注后2 h、24 h时的存活率及肝脏病理情况。 结果随着肝门阻断时间的延长,ALT、AST显著升高,肝内炎症细胞浸润增加,24 h存活率逐渐降低。D组再灌注2 h时,CD8+ T淋巴细胞显著升高,CD4+/CD8+比值下降,调节性T淋巴细胞显著减少,血清IL-10显著降低,而IL-12水平显著升高。再灌注后24 h,B组大鼠各项指标逐步恢复至假手术组水平,而D组大鼠CD4+T淋巴细胞、CD4+/CD8+比值尤其是Treg显著升高,且IL-10水平显著升高,IL-12水平明显降低。与C组相比,E组阻断30 min后无一例死亡,再灌注2 h的ALT、AST水平显著降低,Treg和IL-10水平显著升高,IL-12水平明显降低,再灌注24 h后各项指标恢复并接近假手术组水平,肝脏病理损伤较轻。 结论肝缺血再灌注引起肝脏损伤甚至死亡,可能与诱导T淋巴细胞尤其是Treg和细胞因子的紊乱有关。缺血预处理可以增加再灌注早期的Treg细胞,有效纠正免疫紊乱,减轻损伤。     

N-乙酰半胱氨酸预处理对肝脏缺血再灌注损伤的保护作用

目的 探讨N-乙酰半胱氨酸(N-acetylcysteine,NAC)预处理对肝脏缺血再灌注损伤的保护作用.方法 18 例肝血管瘤手术患者随机分为3 组(n=6):对照组(C0)、NAC 预处理组(PR)和NAC 后处理组(PO),其中PR和PO 组分别于肝门阻断前和开放后给予NAC 120 mg/kg.三组患者均于切皮前(T0)、肝门阻断开放后1 h(T1)及6 h(T2)抽血检测谷丙转氨酶活性(ALT)和谷草转氨酶(AST)水平,并于肝门阻断开放后1 h(T1)取肝组织测定丙二醛(MDA)含量及核因子κB(NF-κB)活性.结果 肝门阻断开放后1 h(T1)和6 h(T6),各组的ALT 和AST 水平均明显高于肝门阻断前(T0)水平(P<0.01);与对照组相比,PR 和PO 组的MDA 含量在肝门阻断开放后1 h(T1)明显减少(P<0.01),但NF-κB 的活性仅在PR 组显示有明显的降低(P<0.01).结论 NAC 预处理可有效防治肝脏缺血再灌注损伤,其机制与抑制再灌注后NF-κB 的启动激活有关.     

缺血后处理减轻肠缺血再灌注引起的肝损伤的机制研究

目的：探讨缺血后处理减轻肠缺血再灌注引起的肝损伤的作用机制,为外科防治缺血再灌注损伤提供策略。方法将36只SD大鼠随机分为假手术组（SO组,仅手术显露肠系膜上动脉）、缺血再灌注组（IR组,阻断肠系膜上动脉60min,再灌注120min）、缺血后处理组（IP组,阻断肠系膜上动脉60min后行3个循环的灌注30s/阻断30s,再持续灌注117min）,每组12只。建立模型2h后采集各组大鼠动、静脉血及部分小肠、肝组织,检测血肿瘤坏死因子α（TNF-α）、白细胞介素10（IL-10）、丙氨酸氨基转氨酶（ALT）、天门冬氨酸氨基转移酶（AST）水平,测定血清及肝组织内丙二醛（MDA）、髓过氧化酶（MPO）水平,光学显微镜下观察小肠及肝脏病理学改变,免疫组织化学法检测肝脏组织中核因子κBp65（NF-κBp65）和缺氧诱导因子1α（HIF-1α）的表达变化。结果与SO组比较,IR组小肠、肝脏病理损伤加重,肝组织NF-κBp65和HIF-1α的表达显著升高,血清和肝组织中MDA、MPO水平及血清TNF-α、IL-10、ALT和AST水平升高;与IR组比较,IP组小肠、肝脏损伤减轻,肝组织NF-κBp65表达下降而HIF-1α的表达显著升高,血清和肝组织中MDA、MPO水平及血清TNF-α、ALT和AST水平均显著下降,血清IL-10水平增加,差异均有统计学意义（P＜0.05）。结论缺血后处理可以促进抗炎因子的激活,抑制NF-κB信号通路调控的炎症级联反应,上调HIF-1α的表达,减轻小肠缺血再灌注引起的肝损伤。     

氯胺酮后处理及其联合缺血后处理对大鼠肝脏再灌注损伤的作用

目的 探讨氯胺酮后处理及其联合缺血后处理对大鼠肝脏再灌注损伤的作用.方法 雄性SD大鼠24只,体重200～230 g,随机均分为缺血损伤组(A组)、氯胺酮后处理组(B组)、氯胺酮后处理联合缺血后处理组(C组)、假手术组(D组).按照Nauta介绍的方法建立缺血模型,在缺血1 h后恢复灌注时,B组由尾静脉推注氯胺酮10 mg/kg,C组由尾静脉推注等量氯胺酮的同时对肝脏进行缺血后处理,D组仅分离肝十二指肠韧带.再灌注4 h后采集下腔静脉血测丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST);取肝组织测超氧化物歧化酶(SOD)、丙二醛(MDA)含量,检测Bcl-2和Bax的表达和通过电镜观察肝脏的损伤程度.结果 B、C组ALT、AST、MDA均低于A组、SOD高于A组(P<0.05),Bcl-2表达高于A组,Bax表达低于A组(P<0.05).透射电镜下观察B、C组细胞损伤程度轻于A组.结论 氯胺酮后处理可以减轻大鼠肝脏的再灌注损伤,但是联合缺血后处理后其保护效应并没有加强.     

异丙酚后处理联合缺血后处理对大鼠肝脏缺血再灌注损伤的影响

目的 探讨异丙酚后处理联合缺血后处理对大鼠肝脏缺血再灌注损伤的影响.方法 健康雄性sD大鼠30只,体重200～250 g,随机分为5组(n=6),假手术组(Ⅰ组)仅开腹;缺血再灌注组(11组)肝脏缺血1 h再灌注4 h;缺血后处理组(Ⅲ组)肝脏缺血1 h后,再灌注10 8,缺血10 8,重复6次进行缺血后处理;异丙酚后处理组(Ⅳ组)肝脏缺血1 h后经尾静脉注射异丙酚10 mg/kg,随后静脉输注异丙酚40 mg·kg~(-1)·h~(-1) h;异丙酚后处理+缺血后处理组(V组)肝脏缺血1 h后进行异丙酚后处理及缺血后处理.于再灌注4 h时测定血清ALT活性、肝组织MDA含量、SOD活性、Bcl-2及Bax的蛋白表达水平,电镜下观察肝细胞超微结构.结果 与Ⅰ组比较,Ⅱ组～Ⅴ组血清ALT活性及肝组织MDA含量升高,肝组织Bcl-2蛋白表达上调,Ⅲ组-Ⅴ组肝组织SOD活性升高,Ⅱ组、Ⅲ组及Ⅴ组肝组织Bax蛋白表达上调(P<0.05或0.01);与Ⅱ组比较,Ⅲ组-Ⅴ组血清ALT活性及肝组织MDA含量降低,肝组织SOD活性升高,Bcl-2蛋白表达上调,Bax蛋白表达下调(P<0.05或0.01);与Ⅲ组比较,Ⅳ组血清ALT活性及肝组织MDA含量降低(P<0.05或0.01).Ⅲ组-Ⅴ组肝组织病理学损伤较Ⅱ组明显减轻.结论 异丙酚后处理联合缺血后处理可减轻大鼠肝脏缺血再灌注损伤,与异丙酚后处理单独应用时效果相同,其机制可能与抑制肝组织脂质过氧化反应及细胞凋亡有关.     

Hyperbaric Oxygen and N-Acetylcysteine Treatment in L-Arginine-Induced Acute Pancreatitis in Rats

ABSTRACT

Background: This study was designed to evaluate the combined effects of hyperbaric oxygen (HBO) and N-acetylcysteine (NAC) on acute necrotizing pancreatitis in rats. Methods: Experiments were performed in 50 male Wistar rats, which were divided into five groups (N = 10 for each group). The first group received normal saline (0.9% NaCl) intraperitoneal and served as the control group. In the second group, acute pancreatitis was induced by 3.2-g/kg body weight L-arginine intraperitoneal twice at an interval of 1 hr, which has been shown previously to produce severe necrotizing acute pancreatitis. In the third group, NAC treatment (1000 mg/kg) was given after 1 hr of the induction of acute pancreatitis twice 24 hr apart. In the fourth group, animals received HBO, 6 hr after the induction of pancreatitis twice 12 hr apart. In the fifth group, animals received together NAC as in Group 3 and HBO treatment as in Group 4. Groups 1, 2, and 3 were left under normal atmospheric pressures. Twelve hours after last treatment, the animals were killed by exsanguinations. Blood samples were studied for amylase, calcium, and lactate dehydrogenase (LDH), pancreatic histology, pancreatic tissue malondialdehyde, superoxide dismutase, and glutathione levels. Results: Acute pancreatitis is reduced by the treatment of NAC, HBO, NAC + HBO. HBO + NAC groups performed statistically the best in preventing L-arginine-induced acute necrotising pancreatitis. Conclusions: NAC especially combined with HBO, decreases oxidative stress parameters, serum amylase, calcium, and LDH levels, as well as histopathologic score.     

硝普钠阴茎海绵体内注射治疗阳萎的临床研究

本研究选择４２例阳萎患者，采用硝普钠进行阴茎海绵体注射（ＩＣＩ），并选择罂粟碱／酚妥拉明进行对照，结果表明，硝普钠ＩＣＩ后：（１）阴茎外形性状（长度、周径等）明显改变。（２）Ｖｉｒａｇ硬度计点表明硝普钠与罂粟碱／酚妥拉明效果之间无明显差别。（３）所有测试患者无一例出现低血压或局部不适等副反应，与罂粟碱／酚妥拉明相比各有优劣，但总体差异不大，这充分表明，硝普钠作为一种ＮＯ供体可导致阴茎平滑肌松弛，血窦充盈阴茎勃起，其副反应较小，有其临床应用之价值。     

糖皮质激素对肿瘤坏死因子引起的大鼠中性粒细胞粘附的影响

采用中性粒细胞（ＰＭＮ）与玻璃珠粘附和ＰＭＮ与血管内皮细胞（ＥＣ）粘附两种模型，以肿瘤坏死因子（ＴＮＦ），作为ＰＭＮ的刺激因子，研究糖皮质激素（ＧＣ）对ＴＮＦ引起的大鼠ＰＭＮ粘附的影响，同时给予糖皮质激素受体（ＧＲ）阻断剂ＲＵ３８４８６观察ＧＲ在粘附中的作用。结果发现，ＴＮＦ能明显增强大鼠ＰＭＮ的粘附（Ｐ＜０．０１）；Ｄｅｘ不能抑制经ＴＮＦ预处理的ＰＭＮ的粘附（Ｐ＞０．０５），但有一定的预防作用；经ＴＮＦ预处理再同时给予Ｄｅｘ和ＲＵ３８４８６的ＰＭＮ粘附同样明显增强（Ｐ＜０．０１）。     

Kelfiprim,a new sulpha-trimethoprim combination,versus cotrimoxazole,in the treatment of urinary tract infections: A multicentre,double-blind trial

Summary A new combination of trimethoprim with a sulphonamide, named Kelfiprim, differs from cotrimoxazole in that: a) the sulpha drug is sulphamethopyrazine instead of sulphamethoxazole; b) the trimethoprim to sulpha ratio is 5:4 instead of 1:5;c) the presence of a long-acting sulphonamide allows the administration of a daily dose of one capsule, following an initial loading dose of two capsules; d) a reduced amount of trimethoprim is given, as compared to cotrimoxazole, without any decrease of efficacy. Kelfiprim [KP] was compared to contrimoxazole [Co] in a multicentre double blind trial. Sixty four patients suffering from acute and chronic infections of the upper and lower urinary tract entered the study. Urine sterilisation and clinical improvement without relapses showed no differences from the two treatment groups. Tolerance was excellent except in two patients, one treated with KP and the other treated with Co, who showed a transient exanthema.     

Time-dependent hexaminolaevulinate induced protoporphyrin IX distribution after topical application in patients with cervical intraepithelial neoplasia: A fluorescence microscopy study

BACKGROUND AND OBJECTIVES: Compared to the conventional management of cervical intraepithelial neoplasia (CIN) the potential advantage of photodynamic therapy (PDT) for the treatment of cervical human papilloma virus (HPV)-related disease encompasses a minimal invasive procedure with reduced risk of profuse bleeding as a consequence of conization, and possibly more favorable long-term results avoiding cervical stenosis. At present little is known about the precise time-dependent distribution and histological localization of hexaminolaevulinate (HAL) induced protoporphyrin IX (PPIX) fluorescence in healthy tissue and in CIN. The aim of this study was to use ex vivo fluorescence microscopy to determine whether PPIX is selectively induced by neoplastic cells of the cervical epithelium at various times after topical application. STUDY DESIGN/MATERIALS AND METHODS: Cold cream containing 0.5% HAL was applied by means of cervical cap over various periods of time. We analyzed 52 healthy cervical mucosa and 84 CINs. RESULTS: At time delay 100 (+/-10) minutes, high epithelial fluorescence and a significant selectivity between epithelium and underlying lamina propria was found. By contrast, no significant difference between healthy and neoplastic tissues, or between low and high-grade epithelial dysplasia (P > or = 0.05), was observed at any time point. CONCLUSIONS: Application of HAL 0.5% cream to the cervix induced selective fluorescence in epithelial cells. The optimal ratio with a homogeneous PPIX distribution was obtained after 100 ( +/- 10) minutes cream application, which should be evaluated further for PDT.     

An updated review on the principles of intraoperative neurophysiological monitoring and the anaesthetic considerations

It is known that any surgery to the nervous system poses risks to neural structures and their surrounding structures. These mechanisms of injury are the result of mechanical manipulations, haemodynamic alterations, chemical or thermal injuries. Intraoperative neurophysiological monitoring (IONM), using various modalities, is employed to facilitate the assessment of the functional integrity of neural structures, and it is used to provide a real-time alerting system when changes caused by surgically induced insults are detected. The primary goal of IONM is reducing the risk of postoperative neurological deficits during these surgical procedures. It is used to provide information that allows the surgeon to correct any surgical interventions that may have compromised these systems and this also in turn provides guidance on what neurological deficits to anticipate postoperatively. Apart from being utilized as an alerting system to avoid catastrophic outcomes, IONM also assists as a guidance system using stimulation techniques to map out eloquent areas within the cortex, allowing identification of specific neuronal structures, particularly when landmarks cannot be easily recognized due to infiltration by pathological lesions.In this article, we focus on updating our previous paper published in 2019 and again, to provide attention to the various neurophysiological modalities that are employed in IONM. We will look at the basic underlying physiological principles and their individual indications for use clinically. We will explain the information that each modality provides. Importantly, and the primary reason for this article, we look at the various anaesthetic agents, their effects on each neurophysiological modality and other anaesthetic considerations such as haemodynamic and temperature effects. We will also recommend the use of an alert checklist for the multidisciplinary team should an intraoperative alert be issued during surgical procedures.