Effects of activated protein C on coagulation and fibrinolysis in rabbits with endotoxin induced acute lung injury

Background Sepsis induced acute lung injury （ALI） as a common syndrome in clinical practice has a high mortality. Recombinant human activated protein C （APC） can significantly reduce the mortality of patients with severe sepsis. Several studies have implicated that APC may be protective in ALI. Methods Twenty-one rabbits were operatively prepared and randomly divided into sham, control, or APC groups （n=7 in each group）. After a tracheotomy had been performed, ALI was produced in the control and APC groups by infusion of Escherichia coil endotoxin 100 μg/kg per hour intravenously for 1 hour. The sham group received only the vehicle, infusion of 20 ml of 0.9% saline. The rabbits were studied under anesthesia for 6 hours and were ventilated with 40% oxygen. Bovine APC （25 μg·kg^-1·h^-1） was intravenously administered. The infusion was initiated half an hour post-injury and lasted for 4 hours. The animals were resuscitated with Ringer＇s lactate solution. Results In comparison with nontreatment in the control group, the infusion of APC significantly reduced the increase of thrombomodulin level （TM; control group was （0.68±0.06） ng/ml, vs APC group of （0.62±0.07） ng/ml at 6 hours, P 〈0.05）, and significantly attenuated the fall in protein S （PS; control group was （2.32±0.03） μg/ml at 2 hours, （2.24±0.06） μg/ml at 4 hours and （2.21±0.09）μg/ml at 6 hours, vs APC group （2.46±0.04） μg/ml at 2 hours, （2.40±0.05） μg/ml at 4 hours and （2.39±0.07） μg/ml at 6 hours, P 〈0.01）. In addition, APC limited the increase in plasminogen activator inhibitor-1 （PAI-1） both in plasma （control group was （0.68±0.12） ng/ml at 1 hour, （0.84±0.06） ng/ml at 2 hours, （0.87±0.08） ng/ml at 4 hours and （0.91±0.05） ng/ml at 6 hours, vs APC group （0.42±0.16） ng/ml at 1 hour, （0.43±0.04） ng/ml at 2 hours, （0.45±0.09） ng/ml at 4 hours and （0.45±0.14） ng/ml at 6 hours, P 〈0.01） and in bronchoalveolar lavage fluid （at 6 hour     

Protective effects of α1-antitrypsin on acute lung injury in rabbits induced by endotoxin

Objective To investigate whether pretreatment with α1-antitrypsin (AAT) can attenuate acute lung injury (ALI) in rabbits induced with endotoxin.Methods Thirty-two healthy adult New Zealand rabbits were anaesthetized, tracheotomized and mechanically ventilated. They were then randomly divided into four groups (n=8): ① Infusion of Escherichia coli endotoxin ［Lipopolysaccharide (LPS) 500 μg/kg］ without AAT (Group LPS). ② Infusion of AAT 120 mg/kg at 15 minutes after LPS (Group LAV). ③ Infusion of AAT 120 mg/kg without endotoxin (Group AAT). ④ Infusion of saline 4 ml/kg as control (Group NS). Arterial blood gases, peripheral leukocyte counts and airway pressure were recorded every hour for eight hours. Physiologic intrapulmonary shunting (Qs/Qt) was measured every four hours. After eight hours, blood samples were collected for measurement of plasma concentration and activity of AAT. Then, the animals were sacrificed, and bronchoalveolar lavage fluid (BALF) was collected for measurement of concentrations of total protein (TP), interleukin-8 (IL-8), tumor necrosis factor (TNFα, the activities of NE and AAT, total phospholipids (TPL) and disaturated phosphatidylcholine (DSPC). In addition, the wet-to-dry lung weight ratio (W/D) was measured.Results The infusion of endotoxin induced decreases in arterial oxygen pressure (PaO2), peripheral leukocyte counts, total respiratory compliance (TLC) and the increases in peak pressure (Ppeak), Qs/Qt compared with the baseline values (P<0.05). The increased plasma concentration but reduced activity of AAT was also found in contrast to that in Group NS (P<0.05). In the BALF, the activity of AAT, TPL, DSPC/TPL were lower than those in Group NS (P<0.05), but the concentrations of albumin, IL-8, TNFα, the activity of NE and the ratio of W/D were higher than those in Group NS (P<0.05). The pretreatment of AAT attenuated the deterioration of oxygenation, the reduction of compliance and the deterioration of other physiological and biochemical parameters mentioned above.Conclusion Pretreatment with AAT could attenuate endotoxin-induced lung injury in rabbits. Those beneficial effects of AAT might be due, in part, to reduction in the levels of mediators that could activate neutrophils, in addition to the direct inhibitory effect on neutrophil elastase.     

Relationship between contents of plasma atrial natriuretic peptide and serum lipids of atherosclerosis in rabbits

Changes in plasma atrial natriuretic peptide(ANP)and serum lipids were observed us-ing dietetic atherosclerosis(AS)models.The results showed that plasma ANP level of the ASgroup was significantly higher than that of the control group(14.33±3.58μg/L vs 9.43±3.14μg/L).There was also a marked increase in serum Tch,TG,LDL-ch and VLDL-ch comparedwith the control group(P<0.01),suggesting that release of ANP increased with disturbance ofthe serum lipids and during AS formation,and that change in ANP was closely related to Tchand LDL-ch(P<0.05).Possible mechanisms causing these changes are also discussed.     

Protective effects of Wulingwan on liver injury in rabbits, mice and rats

ＰｒｏｔｅｃｔｉｖｅｅｆｆｅｃｔｓｏｆＷｕｌｉｎｇｗａｎｏｎｌｉｖｅｒｉｎｊｕｒｙｉｎｒａｂｂｉｔｓ，ｍｉｃｅａｎｄｒａｔｓＷａｎｇＳｈｅｎｇｃｈｕｎ（王胜春）；ＦａｎｇＫｕｎｑｕａｎ（方坤泉）；ＪｉａｎｇＹｏｎｇｐｅｉ（蒋永培）；ＺｈａｏＤｅｈｕａ...     

Expression of BMP-7 and its receptors in renal tubolo-interstitial fibrosis induced with unilateral ureteral obstruction in rats

Objective:To study the changes of the expression of bone morphogenetic protein-7(BMP-7) and its receptors(BMPR- Ⅰ ,ALK2,ALK3 and ALK6) in the renal tubulointerstitial fibrosis induced with unilateral ureteral obstruction in rats. Methods: Sixty Wistar male rats were divided randomly into the normal control,sham-operation and unilateral ureteral obstruction(UUO) groups and the rats were killedon the 1^st , 3^rd, 7^th and 14^th postoperative days respectively. The mRNA level of BMP-7, BMPR- Ⅰ , ALK2, ALK3 and ALK6 was determined with RT-PCR. The site and level of protein expression of BMP-7 were observed with immunohistochemical staining. Results: The mRNA level of BMP-7, BMPR- Ⅰ ,ALK2 and ALK3 was significantly decreased in the rats of UUO group than in those of the sham-operation group but the mRNA level of ALK6 showed no obvious changes in all the rats. Immunohistochemical staining revealed that the protein of BMP-7 was mainly expressed in the renal tubules and interstitial tissue of the kidneys in normal rats but it was decreased gradually along with the unilateral ureteral obstruction. Conclusion: It is found that the loss of BMP-7 and its receptors including BMPR- Ⅰ ,ALK2 and ALK3 occursin the early phase of renal tubulointerstitial fibrosis before the appearance of other pathological changes in the kidney and may play an important role in the occurrence and progress of renal tubulointerstitial fibrosis.     

Functional changes of pulmonary surfactant in rats with lung injury induced by endotoxin

We studied the functional changes of pulmonary surfactant (PS) in acutelung injury models produced by endotoxin injection (E.coli O_(55)B_5) in rats.The sur-face properties of the lung lavage liquid and the total phospholipids (TPL) ex-tracted from it were assessed on a modified Wilhelmy film balance.γ-A isothermof the lavage liquid revealed an increase in minimum surface tension and a de-crease in hysteresis area,recruitment index and stability index,whereas that ofTPL extracted from it did not show any change except for hysteresis area.Thesurface activity correlates positively with the TPL content but negatively with thetotal protein content in the lavage liquid.The findings indicated that there was adysfunction of PS in rats with the lung injury induced by endotoxin,suggestingthat the function deficiency of PS might be caused by decreased phospholipidsand increased proteins in the alveoli.     

Influence of therapy of resolving accumulation and dissipating binds on expression of integrin-linked kinase induced by TGF-β1 in human renal tubular epithelial cells

目的 研究消癥散结法对转化生长因子β1(TGF-β1)刺激人肾小管上皮细胞表达整合素连接激酶(ILK)的影响,探讨其在肾间质纤维化中的干预作用.方法 将消癥散结法复方总方拆为补方、散方,运用血清药理学方法分别制备总方、补方、散方、盐酸贝那普利含药血清.体外培养人肾小管上皮细胞,分为6组:空白组、TGF-β1组、贝纳普利组、总方组、补方组、散方组.将5％各含药血清培养基加入体外培养的肾小管上皮细胞,除空白组外均加入TGF-β1 10 μg/L,48 h后,提取细胞总蛋白及总RNA,分别采用免疫印迹法及逆转录-聚合酶链反应方法检测各组ILK蛋白及其mR-NA的表达.结果 空白组肾小管上皮细胞胞浆内ILK及其mRNA有少量表达,TGF-β1刺激后能够诱导肾小管上皮细胞ILK及其mRNA表达显著增加(与空白组相比P＜0.01);与TGF-β1组相比,贝纳普利组和总方组、散方组肾小管上皮细胞ILK及其mRNA含量明显降低(P＜0.01);补方组与TGF-β1组相比差异不明显(P＞0.05).结论 消癥散结法复方总方含药血清能抑制细胞ILK及其mRNA的表达,干预TGF-β1/ILK信号通路可能是其防治肾间质纤维化的机制之一.     

Role of connective growth factor in plasminogen activator inhibitor-1 and fibronectin expression induced by transforming growth factor β1 in renal tubular cells

Background Connective tissue growth factor (CTGF) contributes greatly to renal tubulointerstitial fibrosis, which is the final event leading to end-stage renal failure. This study was designed to investigate the effects of CTGF antisense oligodeoxynucleotides (ODNs) on the expressions of plasminogen activator inhibitor-1 (PAI-1) and fibronectin in renal tubular cells induced by transforming growth factor β1 (TGF-β) in addition to the role of CTGF in the accumulation and degradation of renal extracellular matrix (ECM). Methods A human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipidmediated CTGF antisense ODNs were transfected into HKC cells. After HKC cells were stimulated with TGF-β1 (5 μg/L), the mRNA levels of PAI-1 and fibronectin were measured by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 and fibronectin in the medium were determined by Western blot and ELISA, respectively. Results TGF-β was found to induce tubular CTGF, PAI-1, and fibronectin mRNA expression. PAI-1 and fibronectin mRNA expression induced by TGF-β was significantly inhibited by CTGF antisenes. ODNs CTGF antisense ODNs also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 and fibronectin protein secreted into the medium. Conclusions CTGF may play a crucial role in the accumulation and degradation of excessive ECM during tubulointerstitial fibrosis, and transfecting CTGF antisense ODNs may be an effective way to prevent renal fibrosis.     

Influence of vasectomy on cholesterol level and activities of CPK, GPT, AKP in serum of rabbits

采用动物模型对输精管结扎引起的血清胆固醇水平及一些血清酶活性的改变进行了研究，结果表明：①增龄可使血清胆固醇含量增高；②血清中肌酸磷酸激酶（CPK）、乳酸脱氢酶（LDH）、谷草转氨酶（GOT）、α-羟丁酸脱氢酶（HBDH）水平随年龄增长略有增高，但结扎组（VG）和假手术组（SOG）比较无差异；③血清酸性磷酸酶（ACP）水平，25月组明显高于6月组（P＜0．05），VG与SOG比较无明显差异（P＞0．05）；血清碱性磷酸酶（AKP），25月组明显低于6月组，VG与SOG无明显差异；血清谷丙转氨酶（GPT）与γ-谷氨酰转肽酶（γ-GD）的活性各组间无显著差异。提示：血清酶水平与衰老有关，输精管结扎对其无影响。     

镉诱发兔肾损害时血清和尿液中胍基乙酸的变化

本文采用给兔皮下注射氯化镉的方法造成肾损害模型,对照组皮下注射生理盐水,研究了血清和尿液中胍基乙酸的变化,并探讨了导致这些变化的原因及胍基乙酸作为肾小管受损的早期诊断指标的可能性。结果表明,镉处理组血清中胍基乙酸随实验时间延长有逐渐上升的趋势,4周后呈明显升高,尿液中胍基乙酸于第1周时有轻微升高,第2周后却呈非常明显的下降。     

尿酶谱对镉诱导兔肾损害的早期诊断价值研究

本文给兔皮下注射氯化镉生理盐水诱导肾损害，探讨了尿酶作为发现镉诱导肾损害的早期指标的可能性。结果显示尿ALP、GGT、AAP及TREH排出量在镉处理一周后显著升高，早于尿NAG、LDH、LYS的升高，更早于蛋白尿的出现，尿AST在镉处理6周后才显著升高。血中以上各酶活性无明显变化，镉处理组肾小管刷状缘膜上ALP、GCP、AAP及TREH活性显著降低。结果提示除AST外，其它几种尿酶可用于早期发现镉诱导肾小管损害，尿中来源于肾小管刷状缘膜的ALP、GGT、AAP、TREH优于尿中NAG、LYS及LDH的测定。     

亚慢性镉中毒大鼠肾近曲小管细胞凋亡的初步研究

本试验对雄性ＳＤ大鼠进行了亚慢性镉染毒，经ＨＥ染色、甲基缘一派若宁双重染色和过碘酸－雪夫反应（ＰＡＳ）染片后，在光学显微镜下观察了大鼠肾脏的改变。结果显示，大鼠肾脏未见明显的肾小管坏死、间质纤维化或炎细胞浸润，但肾近曲小管上皮细胞发生凋亡。镉处理组的肾近曲小管的凋亡细胞数（５２．２５±５．１９个／ｍｍ２）明显高于对照组（１２．５０±３．５４个／ｍｍ２），两者之间差异有显著性意义（Ｐ＜０．０１）。     

急性镉中毒重要器官功能与镉含量测定

目的：探讨急性镉中毒时,心、肝、肺、肾等重要器官的功能改变。方法：同步测定急性镉染毒大鼠心、肝、肺、肾等重要器官的功能及血液、脑和上述器官中的镉含量。结果：通过呼吸道和静脉染毒,所引起的肝、肾功能的损害均低于心功能的损害;静脉急性镉染毒致死时,心、肝、肾、脑。血的镉含量与呼吸道染毒致死时基本一致。结论：急性镉中毒时,心、肝、肾、脑、血有一稳定的镉浓度,同时揭示,急性镉中毒不是由于肝、肾、肺功能衰竭所致死。     

急性肾损伤患者尿液与血液中肾脏损伤分子-1的表达及其对肾衰竭的早期诊断价值

目的:探讨急性肾损伤患者尿液与血液中肾脏损伤分子-1的表达及其对肾衰竭的早期诊断价值.方法:回顾性分析在本院肾内科接受诊治的84例急性肾损伤患者的临床资料.根据其是否发生肾衰竭分为RF组和非RF组,ELISA检测两组入院时尿液与血液中肾损伤分子-1(Kim-1)的水平,并绘制受试者工作特征曲线(ROC曲线),评价Kim-1的表达及其对急性肾损伤患者肾衰竭的早期诊断价值.结果:RF组尿液与血液中Kim-1水平均明显高于非RF组,差异具有统计学意义(P＜0.05);尿液Kim-1 ROC曲线下面积为0.855(P=0.000),血液中Kim-1 ROC曲线下面积为0.640(P=0.028).结论:尿液与血液中肾脏损伤分子-1的水平对急性肾损伤患者肾衰竭的发生具有预测性,可作为急性肾损伤患者肾衰竭早期诊断标志物.     

慢性镉中毒肾功能损伤的研究

本实验用0.8％CdCl_2生理盐水按1 mgCd/kg体重,隔日1次皮下注射3个月,建立兔中毒模型。测定了血尿素氮、肌酐含量及肾皮质的Na~+/K~+-ATPase,Ca~(2+)-ATPase,γ-GT的活性。结果表明,在肾中毒早期,尿素氮、肌酐无明显变化,几种酶活性明显下降,P值分别<0.05及<0.001。提示,这些酶活性下降,可能是镉中毒所致肾功能紊乱和病理改变的启动环节之一和进一步导致肾衰的机理之一。     

血清胱抑素C和肾小管损伤标志物对急性肾损伤诊断价值的研究

目的评估血清胱抑素C（sCysC）、尿N-乙酰-β-D-氨基葡萄糖苷酶（uNAG）和尿白蛋白/肌酐比值（uACR）在急性肾损伤（AKI）检测和预后预测的诊断价值及最优组合。方法收集ICU135例危重患者,采用肾脏经疾病改善整体预后肌酐标准来确定AKI患者,包括需行肾脏替代治疗（RRT）的患者。由急性生理学和慢性健康评估（APACHE）II分,器官衰竭评估（SOFA）评分衡量病情严重程度。使用Spearman相关分析各生物标志物之间的相关性,采用回归分析评估收治EICU后sCysC、uNAG和uACR的浓度与AKI最高分期和RRT治疗之间的相关性,并且运用ROC曲线比较sCysC、uNAG和uACR以及不同组合联合检测对于AKI的诊断效能。结果符合入组标准的患者135例,其中58例发生AKI,其中合并有肾功能不全和脓毒症的比例以及ICU停留时间和使用RRT疗法明显高于非AKI组（均P＜0.05）,两组间删失率无统计学差异（P＞0.05）,收治ICU后24h再次评估,之前无AKI的18例（31%）患者均发生不同程度的肾损伤（P＜0.01）。需RRT治疗的患者26例（44.8%）较入院时明显增多（P＜0.01）。两组相比3种生物标志物均有统计学差异（均P＜0.01）,sCysC与uNAG（r=0.731,P＜0.01）,sCysC与uACR（r=0.70,P＜0.01）,uNAG与uACR（r=0.651,P＜0.01）之间均呈正相关。其中sCysC与AKI各个期和需要RRT治疗存在相关性（P＜0.05）。ROC曲线结果显示sCysC的AUC高于uNAG和uACR,分别为0.972、0.842、0.869（P＜0.01）。sCysC+uNAG组的AUC-ROC值为0.973,均高于单个生物标志物和其他两个联合组（P＜0.01）。sCysC+uNAG组比单个生物标志物或其他两个组显示出更高的特异度和灵敏度（分别为0.99和0.82）。结论sCysC与AKI的最坏分期和需要RRT治疗之间具有最强的独立相关性。sCysC和uNAG的组合可以显著增加AKI的诊断效能。     

血清胱抑素C、超敏CRP和尿微量白蛋白联合诊断痛风患者早期肾功能损伤

[摘要]目的探讨血清胱抑素C、超敏C反应蛋白(hs-CRP)和尿微量白蛋白对痛风患者早期肾功能损伤的诊断价值。方法将100例痛风患者和50例健康志愿者分成3组：急性发作期组50例、间歇期组50例、健康对照组50例;采用全自动生化仪检测各组研究对象血清超敏C反应蛋白(hs-CRP)、胱抑素C(Cys-C) 和尿肌酐(Cr)水平,用免疫荧光分析仪测定尿微量白蛋白(mALB),分析各组检测结果。结果急性发作期组、间歇期组、健康对照组的mALB/Cr\[mg/(g·Cr)\]、hs-CRP(mg/L)和Cys-C(mg/L)分别为：11.45±3.55、5.15±1.98、2.62±0.83;8.96±1.78、3.28±1.23、0.93±0.26;3.89±0.76、1.91±0.57、0.78±0.22;急性发作期组、间歇期组mALB/Cr、hs-CRP和Cys-C均高于健康对照组,组间比较差异有统计学意义(P<0.05),且发作期组各指标均高于间歇期组(P<0.05);87%的痛风患者存在hs-CRP+Cys-C+mALB/Cr异常。结论联合测定mALB/Cr、hs-CRP、Cys-C可能有助于评判痛风患者早期肾功能损伤。     

镉致肾脏损伤与锌代谢及其功能的关系

目的探讨锌代谢的变化与镉肾损伤之间的关系。方法将12只清洁级大鼠随机分成染镉组和对照组,分别测定各组大鼠血、尿和肾组织中镉和锌含量,测定尿液中NAG活性、β2微球蛋白含量,测定肌酐以校正尿量。结果染镉组血、尿、肝组织和肾组织中镉含量升高;肾组织和血清锌含量有下降趋势,肝组织锌含量有升高趋势;尿液中NAG活性和β2微球蛋白含量升高;且染镉组血、尿、肝肾组织中镉含量均与尿液中NAG活性和β2微球蛋白含量呈正相关关系;而与肾组织锌含量呈负相关关系;肾组织锌含量与尿液中NAG活性、β2微球蛋白含量呈负相关关系;结论镉能影响大鼠体内锌分布改变,而锌分布改变与镉引起的肾脏损伤有关。     

青春期至成年早期铅、镉联合暴露对雄性小鼠肝脏与肾脏功能的影响

目的:观察青春期至成年早期小鼠经饮水暴露铅和镉对肝脏和肾脏功能的影响。方法:健康4周龄雄性ICR小鼠60只,随机分为对照组、铅组、镉组、铅+镉组,各15只。4组小鼠分别经口饮用纯净水、醋酸铅水溶液、氯化镉水溶液和醋酸铅+氯化镉混合水溶液,暴露40 d后摘眼球取血,处死小鼠后取肝脏和肾脏,称脏器质量,计算脏器系数,检测血清丙氨酸氨基转移酶（ALT）与碱性磷酸酶（ALP）活力以及血清肌酐（Cr）和尿素氮（BUN）含量,用HE染色观察肝脏和肾脏组织形态学改变。结果:对照组、镉组和铅+镉组小鼠的每日饮水量均少于铅组（P<0.05~P<0.01）,铅组和镉组小鼠每日进食量均少于对照组和铅+镉组（P<0.05~P<0.01）,而4组小鼠4~10周龄体质量差异均无统计学意义（P>0.05）。连续暴露40 d后,均未引起铅、镉组和铅+镉组小鼠体质量、肝脏质量、肝脏系数、血清ALT和ALP活力以及肝脏组织形态学改变。镉+铅组小鼠肾脏质量低于单纯镉组（P<0.05）,但铅组、镉组和铅+镉组小鼠肾脏质量、肾脏组织形态学、血清Cr和BUN含量与对照组差异均无统计学意义（P>0.05）。结论:青春期和成年早期经饮水暴露一定剂量铅与镉均未引起雄性小鼠肝脏和肾脏功能损害。