PDTC对创伤复合内毒素所致大鼠ALI的保护作用

为观察（PDTC）对创伤复合内毒素所致大鼠急性肺损伤的保护作用并探讨其作用机制，用多功能小型生物撞击机对大鼠右上有胸壁撞击并经尾静脉注射内毒素5mg/kg进行致伤，致伤前经尾静脉注射PDTC120mg/kg，观察伤后48h死亡率、肺组织MPO、NF－κB活性变化，同时检测TNFα、IL-8 mRNA的表达及蛋白含量。结果发现，PDTC预处理能够显著降低复合损伤48h大鼠的死亡率，其MPO、NF－κB活性与TNFα、IL-8 mRNA表达被显著抑制。说明PDTC能够通过抑制NF－κB的活化，减少致炎细胞因子基因表达与合成释放，减轻炎细胞浸润，从而对复合损伤大鼠起到保护的作用。     

脂多糖对受照小鼠血清IL-10、IL-6和TNF-α水平的影响

目的 探讨6 Gy137Csγ射线一次性全身照射后,小鼠血清中白细胞介素10(IL-10)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-o)水平的变化及对脂多糖刺激反应性的远期影响.方法 将实验小鼠分为假照射组和照射组,假照射组不接受照射,照射组给予6Gyγ射线照射.照射后10周,将假照射组和照射组小鼠按组别分别提前24 h和1h腹腔注射脂多糖(20 mg/kg),对照组注射生理盐水(100μl/只).取小鼠外周血,利用酶联免疫吸附技术检测小鼠血清中IL-10、IL-6和TNF-t的水平.结果 假照射组小鼠在给予脂多糖刺激1h后,血清中IL-10、IL-6和TNF-α水平与其对照组相比均显著升高(t=7.31、2.71和15.09,P分别为＜0.01、＜0.05和＜0.01).照射组小鼠在给予脂多糖刺激1h后,血清中IL-10、IL-6和TNF-α水平与其对照组相比均显著升高(t=4.14、7.18和5.14,P均＜0.01).与假照射组相比,照射组小鼠在给予脂多糖刺激24h后,TNF-α和IL-6水平无明显变化,IL-10水平升高了19.9％(t=2.84,P＜0.05).结论 经6Gyγ射线照射后10周,小鼠免疫调节功能尚未完全恢复;在接受脂多糖刺激后,假照射组和照射组小鼠的抗感染能力也有差异.脂多糖对辐射后小鼠血清中IL-10、IL-6和TNF-α水平的影响及其意义仍需进一步研究.     

内毒素／感染性休克时循环血中TNF的动态变化及其发生机制

探讨不同休克条件下循环血中肿瘤坏死因子（ＴＮＦ）的动态变化规律及其可能的发生机制。实验发现内毒素休克时ＴＮＦ呈陡直的单峰曲线，持续时间较短（３～４小时）；绿脓杆菌败血症休克时ＴＮＦ呈平坦的单峰曲线，持续时间较长（６小时）；阑尾结扎穿孔（ＡＬＰ）感染性休克时ＴＮＦ呈前峰极低、后峰较高的双峰曲线，持续时间更长（１６小时）。结合不同剂量脂多糖（ＬＰＳ）在体外对肝Ｋｕｐｆｆｅｒ细胞释放ＴＮＦ的影响以及不同休克条件下血浆ＴＮＦ的变化特点，结果提示内毒素、绿脓杆菌败血症、ＡＬＰ感染性休克时循环血中ＴＮＦ不同动态变化特点可能主要取决于循环血中ＬＰＳ的不同浓度和不同持续时限。     

抗TNF单克隆抗体对内毒素休克大鼠的保护性作用

研究抗ＴＮＦ单抗及Ｌ－ＮＭＭＡ对内毒素休克时血流动力学的影响并探讨其作用机制。     

熊果苷对乙酰胆碱-氯化钙混合液诱导心房颤动大鼠心脏保护作用及其机制研究

目的 探讨熊果苷(AR)对乙酰胆碱-氯化钙(Ach-CaCl₂)混合液诱导的心房颤动(AF)大鼠的心脏保护作用及其可能的机制。方法 将30只健康雄性SD大鼠随机分为3组：正常对照组(Sham组)、AF模型组(AF组)与AR治疗组(AR+AF组),每组各10只。采用心电图及小动物超声心动图检测各组大鼠的心功能。采用Masson染色检测各组大鼠心房纤维化程度。通过酶联免疫吸附试验检测各组大鼠血清炎症因子。应用蛋白免疫印迹法检测各组大鼠炎症相关蛋白,并探讨AR对AF大鼠的保护作用及炎症相关作用机制。结果 AF组大鼠AF持续时间显著多于Sham组、AR+AF组,差异均有统计学意义(P<0.05)。AF组大鼠左心房内径大于Sham组、AR+AF组,差异均有统计学意义(P<0.05)。与Sham组比较,AF组、AR+AF组的白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)水平明显升高,差异均有统计学意义(P<0.05)。与AF组比较,AR+AF组的IL-6、TNF-α水平明显降低,差异均有统计学意义(P<0.05)。与Sham组比较,AF组蛋白...     

丙泊酚对内毒素刺激小鼠腹腔巨噬细胞促炎细胞因子mRNA的影响

目的：研究不同浓度丙泊酚(异丙酚)对内毒素刺激小鼠腹腔巨噬细胞TNF-α，IL-6，IL-8 mRNA表达的影响。方法：分离巯基乙酸盐诱导的小鼠腹腔巨噬细胞，随机分为7组。A组：阴性对照组；B组：阳性对照组．脂多糖(LPS)10n/ml孵育细胞；C组：单用丙泊酚组，500μg／ml孵育细胞；D，E，F，G组：丙泊酚 LPS组，不同浓度丙泊酚(0．5，5，50，500μg/ml)孵育2h后加入LPS 10ng/nl继续孵育1～4h。用RT-PCR法检测巨噬细胞TNF-α，IL-6，IL-8 mRNA的水平。结果：在内毒素诱导下，腹腔巨噬细胞TNF-α，IL-6，IL-8 mRNA的水平均显著增高；丙泊酚对LPS刺激后TNF-α，IL-6 mRNA表达水平的增高有抑制作用，并呈浓度依赖性；但对LPS刺激下IL-8 mRNA表达水平增高的影响无统计学意义。结论：丙泊酚能在转录水平抑制内毒素刺激下小鼠腹腔巨噬细胞促炎性细胞因子的产生。     

免疫吸附清除内毒素休克兔循环TNF对IL—1,IL—6水平的影响

内毒素休克并非由内毒素直接作用所致，而是藉体内过度生成和释放的多种内源性细胞因子介导产生的，其中ＴＮＦ起着关键作用。但目前有关内毒素休克发病的分子机制尚不完全清楚。本实验采用免疫吸附方法清除内毒素休克早期循环ＴＮＦ，观察对ＩＬ－１、ＩＬ－６水平的影响...     

活性炭、大孔树脂Amberlite XAD－7血液灌流对大鼠内毒素休克的防治作用

观察了活性炭、大孔树脂ＡｍｂｅｒｌｉｔｅＸＡＤ－７血液灌流对大鼠内毒素休克的防治作用。表明：活性炭可显著降低大鼠血浆内毒素（ＬＰＳ）、白细胞介素１（ＩＬ－１）水平（Ｐ＜０．０１），而对肿瘤坏死因子（ＴＮＦ）的降低作用不明显。ＡｍｂｅｒｌｉｔｅＸＡＤ－７灌流组ＬＰＳ、ＩＬ－１水平与假灌流组比较无显著差异，ＴＮＦ水平明显低于假灌流组（Ｐ＜０．０５）。在开始灌流后３ｈ，两个治疗组的收缩压均较假灌流组明显改善（Ｐ＜０．０５）。并且活性炭、树脂血液灌流组存活时间与假灌流组比有显著差异（Ｐ＜０．０５）。ＬＰＳ攻击后６ｈ，活杀大鼠进行病理检查，两个治疗组肺、肠、肾、肝等脏器损害较轻。由此提示，通过体外血液灌流清除循环中致病递质可能是治疗内毒素休克的另一有价值的新途径。     

免疫吸附抗介质治疗内毒素休克的实验研究

目的观察应用免疫吸附的方法特异性清除循环肿瘤坏死因子－α（ＴＮＦ－α）对内毒素休克的影响。方法利用新西兰白兔内毒素休克模型，研究经抗ＴＮＦ－α单克隆抗体亲和免疫吸附柱血液灌流对平均动脉压、血浆ＴＮＦ－α活性、肝肾功能及存活率的影响。结果免疫吸附治疗后，血浆ＴＮＦ－α水平迅速下降，低血压状态明显改善，延缓和显著减轻了肝肾功能损害及心肌、细胞膜和溶酶体损伤的发生，实验动物存活率明显提高。结论免疫吸附特异性清除循环ＴＮＦ－α可能是一条治疗内毒素休克的新的有效途径。     

Irradiation induces increased production of haemopoietic and proinflammatory cytokines in the mouse lung

PURPOSE: To investigate cytokine expression following irradiation of mice, predominantly in lung tissue but also in selected other tissues. MATERIALS AND METHODS: Mice of strain ICR were whole-body (unilaterally) exposed to 3-20 Gy of (60)Co gamma-rays. Colony-stimulating activity (CSA) of lung-conditioned media (LCM), and also other non-haemopoietic and haemopoietic organs, and blood serum of mice was assayed using a GM-CFC bioassay. The production of GM-CSF, IL-6 and TNF-alpha protein in LCM and sera was determined by an ELISA method. RESULTS: Greatest CSA was detected in conditioned media from the lungs and was induced in a dose- and time-dependent fashion, peaking at 3-9 days after irradiation with a lethal dose of 9 Gy. Conditioned medium prepared from lungs that had been irradiated with a dose of 9 Gy in vitro did not exhibit an increase in CSA. However, whereas the lung-conditioned medium from irradiated mice was found to produce CSA, sera from normal or irradiated mice did not lead to this effect. A significant increase in CSA in sera was observed in the presence of a suboptimal concentration of IL-3, implying that they comprise the co-stimulatory activity (CoSA). The results showed that radiation exposure increased GM-CSF and TNF-alpha protein levels but did not affect IL-6 production in LCM. In contrast, IL-6 and TNF-alpha protein levels in serum were increased after irradiation but no GM-CSF production could be detected. CONCLUSION: Whole-body irradiation enhances CSA in lungs as well as in other haemopoietic and non-haemopoietic organs. The increase of CSA correlates with increased levels of haemopoietic and proinflammatory cytokines in lung.     

Irradiation induces increased production of haemopoietic and proinflammatory cytokines in the mouse lung

Purpose : To investigate cytokine expression following irradiation of mice, predominantly in lung tissue but also in selected other tissues. Materials and methods : Mice of strain ICR were whole-body (unilaterally) exposed to 3-20 Gy of 60 Co γ-rays. Colony-stimulating activity (CSA) of lung-conditioned media (LCM), and also other non-haemopoietic and haemopoietic organs, and blood serum of mice was assayed using a GM-CFC bioassay. The production of GM-CSF, IL-6 and TNF- α protein in LCM and sera was determined by an ELISA method. Results : Greatest CSA was detected in conditioned media from the lungs and was induced in a dose- and time-dependent fashion, peaking at 3-9 days after irradiation with a lethal dose of 9 Gy. Conditioned medium prepared from lungs that had been irradiated with a dose of 9 Gy in vitro did not exhibit an increase in CSA. However, whereas the lung-conditioned medium from irradiated mice was found to produce CSA, sera from normal or irradiated mice did not lead to this effect. A significant increase in CSA in sera was observed in the presence of a suboptimal concentration of IL-3, implying that they comprise the co-stimulatory activity (CoSA). The results showed that radiation exposure increased GM-CSF and TNF- α protein levels but did not affect IL-6 production in LCM. In contrast, IL-6 and TNF- α protein levels in serum were increased after irradiation but no GM-CSF production could be detected. Conclusion : Whole-body irradiation enhances CSA in lungs as well as in other haemopoietic and non-haemopoietic organs. The increase of CSA correlates with increased levels of haemopoietic and proinflammatory cytokines in lung.     

内毒素休克小鼠肝血管靶向性结合肽的初步研究

目的研究肽CLTTWAPAC与内毒素休克BALB/c小鼠肝血管的靶向性结合作用。方法构建pET14b-SBP和pET14b-SBP-CLTTWAPAC原核表达载体,转化大肠埃希菌BL21(DE3),诱导表达纯化蛋白SBP和SBP-CLTTWAPAC。建立内毒素休克小鼠模型,将9只内毒素休克BALB/c小鼠随机均分为3组:①PBS对照组(经尾静脉先后注射灭活空噬菌体、PBS和噬菌体CLTTWAPAC,间隔5min);②SBP对照组(经尾静脉先后注射灭活空噬菌体、SBP和噬菌体CLTTWAPAC,间隔5min);③SBP-CLT-TWAPAC组(经尾静脉先后注射灭活空噬菌体、SBP-CLTTWAPAC和噬菌体CLTTWAPAC,间隔5min)。小鼠左心室灌流后取肝匀浆称重,行噬菌体计数,计算各组肝噬菌体单位重量滴度。结果 PBS对照组与SBP对照组肝噬菌体单位重量滴度差异无统计学意义(435185±57824pfu/gvs468290±74106pfu/g,P0.05),而SBP-CLTTWAPAC组肝噬菌体单位重量滴度(176059±98506pfu/g)显著降低,与两对照组比较差异均有统计学意义(P0.05)。结论 CLTTWAPAC可竞争性抑制噬菌体CLTTWAPAC与内毒素休克小鼠肝血管的结合,进一步证明CLTTWAPAC靶向性结合于内毒素休克小鼠肝血管。     

The gene expression of cytokines and chemokines induced by tourniquet shock in mice

Traumatic shock is one of the major fields in forensic pathology, but its mechanism remains elusive from the pathophysiological aspects. Tourniquet shock has been established as one of the animal models of traumatic shock, and we examined the gene expression of cytokines and chemokines in the lung and liver in tourniquet shock using mice. Tourniquet was conducted by the application of elastic bands with five turns at both the thighs as high as possible for 2 h, followed by reperfusion. In this procedure, more than 90% mice died within 48 h after reperfusion. Serum hepatic transaminase and hematocrit values significantly increased at 2 h after reperfusion, and their elevation was still evident after 10 h. Histopathologically, hemorrhages, congestion and leukocyte recruitment were observed in the lung and liver specimens after 6 h of reperfusion. Immunohistochemical analysis with anti-myeloperoxidase antibody demonstrated a massive neutrophil infiltration in the lung and liver at 2 h or more after reperfusion. RT-PCR analyses demonstrated that the gene expression of interleukin-1beta, tumor necrosis factor-alpha, monocytes chemoattractant protein-1, macrophage inflammatory protein (MIP)-1alpha, MIP-2, KC and vascular endothelial adhesion molecule-1 was most enhanced in the lung and liver at 2 h after reperfusion. Thus, the gene expression of cytokines and chemokines is presumed to be closely related with the onset of tourniquet shock. From the forensic aspects, these cytokines and chemokines are considered to be useful markers for the early diagnosis of tourniquet shock.     

创伤早期未控制性出血非手术疗法的实验研究

目的探讨延迟复苏、低容量液体复苏、脑垂体后叶素复苏对创伤早期重度未控制性失血性休克（UHS）复苏效果的影响。方法成年健康中华田园犬24只,股动脉放血,制作重度UHS模型。采用完全随机法随机分为A组（延迟复苏组）;B组（低容量液体复苏组）;C组（脑垂体后叶素组）。观察各组放血前即刻（TO）、达到目标血压时（T1）、实施三种复苏方法后1h（T2）、复苏平稳后2h（113）各时点的HR、MAP、CVP、血气分析及各组的出血量和72h存活率。结果1、2时点,A组的CVP、MAP明显低于B组和C组（P〈0．05）,B组和C组间差异无统计学意义（P〉0．05）。B组和C组在B时点的血流动力学指标接近TO;在T2时点,A组的PaO：明显低于B组和C组（P〈0．05）,A组中心静脉血乳酸（BL）和碱剩余（BE）均明显高于B组和C组（P〈0．05）,B组BE明显高于C组（P〈0．05）。T3时点,B组和C组的BL和BE均组和明显高于1、0时点（P〈0．01）,低于T1时点（P〈0．01）。三组间的Hot、出血量比较差异无统计学意义（P〉0．05）;A组72h内存活率明显低于B组和C组（P〈0．01）。结论低容量液体复苏能显著提高重度UHS时的存活率;小剂量垂体后叶素的复苏效果优于低容量液体复苏。     

抗内毒素核心区单克隆抗体及其对革兰阴性菌脓毒性休克症小鼠保护作用的研究

通过大肠杆菌Ｒｃ突变株（Ｊ５）免疫的小鼠脾细胞和Ｓｐ２／０骨髓瘤融合，建立了２２个抗内毒素单克隆抗体细胞株。对７株进行了Ｉｇ类型鉴定：５株为ＩｇＭ／Ｋ，１株为ＩｇＧ２ｂ／λ，１株为ＩｇＧ３／λ。５株ＩｇＭＪｉ体识别的表位均在内毒素的内核心区。经沉淀法和凝胶过滤法纯化腹水，抗体纯度达９０％。在Ｅ．ｃｏｌｉ临床分离物或Ｓ－ＬＰＳ所致的小鼠休克模型上，腹腔注射抗体６２．５～１２５μｇ／只，实验组存活率比对照组高３７．５％～７５．０％。体外结合和抑制试验表明，这些抗体与克雷伯杆菌、沙雷茵、鼠伤寒杆菌、痢疾杆菌、绿脓杆菌等属有很强的交叉反应，提示本组抗体可能对革兰阴性菌感染有广谱的保护作用。     

多发伤患者血清促炎介质水平与创伤预后的相关性

目的 探讨在多发伤患者发病过程中TNF-α、IL-8、C反应蛋白(CRP)、铜蓝蛋白(CER)的变化及其与创伤预后的相关性.方法 选择100例多发伤患者,根据ISS,将其分为轻伤组(ISS＜16分),重伤组(16分≤ISS＜25分),严重伤组(ISS≥25分).在入院后各时段测定其血清TNF-α、IL-8、CRP、CER的水平,比较各组的差异及其对预后的影响.结果 血清TNF-α、IL-8、CRP、CER在伤后早期即升高,4～7 d后达高峰.各组促炎介质水平若下降后再升高,则预示创伤感染或其他并发症将发生.结论 ISS的增高与致炎因子血清水平增高呈明显的正相关,通过测定TNF-α、IL-8、CRP、CER水平有助于对创伤并发症和预后的判断.     

荷电微孔滤膜去除细菌内毒素的试验研究

评价了一种由天然多糖制备的荷正电微孔滤膜，考察了该荷电微孔滤膜去除蒸馏水及葡萄糖、氯化钠等药液中细菌内毒素的能力。研究了膜孔径、细菌内毒素的来源和浓度以及使用操作程度等对脱除效果的影响，同时还试验了其它因素如药液组分及ＰＨ值的影响。结果表明，带正电的荷电微孔滤膜能有效去除水溶液中的细菌内毒素。讨论了荷电微孔滤膜在大输液及注射用水生产中的应用。