创伤性脑损伤患者血清中sFas、FasL、NSE的变化与临床观察

目的　观察创伤性脑损伤患者不同伤情、不同时期血清中可溶性 Fas(Soluble Fas,s Fas)、Fas配体(Fas ligand,Fas L)、神经元特异性烯醇化酶 (neuron- specific enolase,NSE)的动态变化及其内在联系 ,借此间接了解创伤性脑损伤后神经细胞损伤的规律 ,为探索新的神经保护性治疗措施提供理论依据。方法　选择 35例单纯创伤性颅脑损伤患者 ,依据入院时 GCS(Glasgow coma score)评分将其分为轻型组、中型组和重型组。入选患者分别于伤后2 4 h、72 h、7d、1 4 d抽取外周静脉血 ,运用双抗体夹心酶联免疫分析法 (EL ISA)测定血清中 Fas L、s Fas、NSE含量。结果　 (1 )伤后 2 4 h Fas L、s Fas均升高 ,并于伤后 72 h左右达到高峰 ,后逐渐下降 ,至 2周左右接近正常。 (2 ) NSE于伤后 2 4 h即开始升高 ,且损伤越重升高越显著。 (3) s Fas、Fas L与 NSE呈正相关。结论　 (1 )创伤性脑损伤后神经细胞的损伤存在原发和继发两种机制。 (2 ) Fas/ Fas L系统在继发性脑损伤中起重要作用 ,s Fas的表达增强在一定程度上对神经组织起到保护作用。 (3)创伤性脑损伤后测定血清中 Fas L、s Fas可间接了解继发性脑损伤的程度 ,借此可以为临床探索新的神经保护措施提供理论依据     

Dysregulation of apoptosis in scorpion envenomed children: its reflection on their outcome.

In the present study, 46 children in Upper Egypt (less than 13 years old) were admitted to the Pediatric Intensive Care Unit for scorpion envenomation. They were compared with 20 apparently healthy children of matching age and sex as controls. Out of the studied victims, 25 children (54%) showed signs of severe envenomation and multiple organ dysfunction (MOD), while 21 victims (46%) showed signs of mild envenomation. The serum levels of apoptotic markers, soluble Fas (sFas), soluble Fas ligand (sFasL) and Bcl-2, were determined for both victims and controls. In addition, the serum levels of nitric oxide (NO) and lipid peroxides (LPO) were also measured. Scoring of MOD was evaluated using Logistic Organ Dysfunction System Score (LODS) for the severely envenomed victims. All victims (both severe and mild cases) showed significantly higher mean levels of sFas, LPO and NO and significantly lower serum levels of Bcl-2 in comparison to the controls. The level of sFasL was not detectable in the sera of the healthy control group. The case fatality rate was 15%. The severely envenomed children with MOD as well as the non-survivors showed significantly higher serum levels of sFas, sFasL, LPO and NO and significantly lower serum levels of Bcl-2 in comparison to the mild envenomed victims and the surviving victims of severe cases, respectively. The LODS score of the severely envenomed victims showed significant positive correlations with sFas and LPO and significant negative correlation with Bcl-2. In all victims, a significant positive correlation was detected between sFas and NO. On the other hand, Bcl-2 was significantly negatively correlated with both sFas and LPO. In conclusion, our study revealed that scorpion envenomation can increase apoptosis as shown by up-regulation of sFas/sFasL system and down-regulation of Bcl-2 that was associated by elevation of LPO and NO. This dysregulation of apoptosis was increased with the severity of scorpion envenomation and its extent increased as MOD score and outcome increased. Therefore, sFas and Bcl-2 may be of value in predicting the outcome of these cases. The increase of the extent of apoptosis detected in this study seems to play a role in the outcome of scorpion envenomation, and hence, should be taken into consideration for strategies of therapeutic regimen.     

低剂量毒死蜱重复染毒诱发雄性大鼠生殖毒性及其机制

目的探讨毒死蜱(CPF)低剂量重复染毒对雄性大鼠生殖功能的影响及其可能作用机制。方法健康雄性Wistar大鼠ig给予CPF 1,5和10 mg.kg-1,每日1次。连续染毒12周后取大鼠附睾和睾丸称重计算脏器系数,进行组织病理学、精子数量以及活力及畸形率等检查,测定睾丸碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、γ-谷氨酰转肽酶(γ-GT)和乳酸脱氢酶(LDH)的活性;TUNEL法检测细胞凋亡,免疫组化及Western印迹法检测胱天蛋白酶和Fas,FasL蛋白的表达。结果与正常对照组相比,CPF染毒大鼠睾丸脏器系数显著升高,精子数及精子活力降低,精子畸形率显著升高(P<0.05),睾丸AKP和γ-GT活性抑制明显。组织病理检查显示,睾丸曲细精管疏松和生精细胞减少。CPF染毒组生精细胞凋亡增加(P<0.05)。Fas/FasL蛋白表达量升高。结论 CPF低剂量重复染毒能够诱导雄性大鼠生殖毒性,其作用机制可能与激活Fas/FasL受体途径而引起睾丸生精细胞凋亡有关。     

p,p′-DDE induces testicular apoptosis in prepubertal rats via the Fas/FasL pathway

1,1-Dichloro-2,2 bis(p-chlorophenyl) ethylene (p,p′-DDE), the major metabolite of 2,2-bis(4-chlorophenyl)-1,1,1-trichloroethane (DDT), is a known persistent organic pollutant and male reproductive toxicant. It has antiandrogenic effect. However, the mechanism by which p,p′-DDE exposure causes male reproductive toxicity remains unknown. To elucidate the mechanism underpinning the testicular effects of p,p′-DDE, we sought to investigate Fas/FasL apoptotic pathway in the testis of prepubertal rats, including Fas, FasL, caspase-8, -3, and NF-κB. Animals were administered with different doses of p,p′-DDE (0, 20, 60, 100 mg/kg b.wt) every other day by intraperitoneal injection for 10 days. The results indicated that p,p′-DDE exposure at over 20 mg/kg b.wt showed the induction of apoptotic cell death. p,p′-DDE could induce increase in the MDA level, and decrease in SOD and GSH-Px activity. Significant elevations in the mRNA levels of Fas along with an increase in FasL, caspase-3, -8 were observed in 100 mg/kg b.wt group. In protein level, p,p′-DDE could induce increase of FasL and reduction of procaspase-8. NF-κB p65 was activated by p,p′-DDE treatment in rat testis. In addition, the activities of caspase-3, -8 were increased in 100 mg/kg b.wt group. Taken together, these results lead us to speculate that in vivo exposure to p,p′-DDE might induce testicular apoptosis in prepubertal rats through the Fas/FasL pathway.     

Role of the Fas/Fas Ligand death receptor pathway in ginseng saponin metabolite-induced apoptosis in hepg2 cells

This research team found in previous studies, that the ginseng saponin metabolite IH901 induces apoptosis in HepG2 cells via a mitochondrial-mediated pathway, which resulted in the activation of caspase-9 and subsequently of caspase-3 and -8. Based on these results, the involvement of the Fas/Fas ligand (FasL) death-receptor pathway, in IH901-induced apoptosis in HepG2 cells, was investigated. Levels of Fas and the Fas ligand (FasL) mRNA or protein were not increased by IH901, rather they were decreased significantly at 18 h post treatment. Soluble FasL (sFasL) was detectable by immunoprecipitation analysis in the medium of HepG2 cells treated with IH901. Increased levels of sFasL were inversely correlated with the levels of FasL. Preincubation of HepG2 cells with antagonistic anti-Fas antibody showed little protective effect, if any, on IH901-induced cell death. At a 30 microM (24 and 48 h) and 40 microM (24 h) concentration of IH901, the cytotoxic effect of IH901 was less then 50%, anti-Fas antibody prevented IH901-induced cell death. However, at a 60 microM (24 and 48 h) and 40 microM (48 h) concentration of IH901, cell death rates were about 80% or more and most of the chemopreventive and chemotherapeutic effects of IH901 were manifested. Blocking the Fas receptor did not influence IH901-induced cell death. These results indicate that the Fas/FasL system is engaged, but not required for IH901-induced cell death, at pharmacologically significant concentrations.     

Involvement of Fas/FasL system in apoptotic signaling in testicular germ cells of male Wistar rats injected i.v. with microcystins

Previous studies have shown that gonads were the second target organ of microcystins (MCs), and that MCs exposure exerted obvious toxic effects on male reproductive system of mammals. However, relevant molecular evidences are still lacking. Fas-signaling pathway plays a key role in toxicant-induced germ cell apoptosis. This study was to evaluate the responses of Fas/FasL system related genes and proteins in testes of rats injected intravenously with MCs. Enhanced apoptosis of germ cells in the testes of MCs-treated rats was detected by the terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling (TUNEL) associated with up-regulation of the Fas/FasL system. Both Fas and FasL protein expression were induced evidently from 1 h post-injection, and this high expression level maintained throughout the experiment. In addition, the activation of caspase-8 and caspase-3 protein was also observed, which were indicators of apoptosis. These results suggested the likely involvement of Fas/FasL system in the MCs-induced germ cell apoptosis. It is also suggested that MCs can cause damage to Sertoli cells directly.     

急性心肌梗死患者血清Hcy、sFas和sFasL的水平及相关分析

目的探讨急性心肌梗死患者血清Hcy水平的变化和可溶性Fas(sFas)及可溶性Fas配体(sFasL)水平的相关性。方法应用酶联免疫吸附法对42例急性心肌梗死患者进行了血清Hcy和血清sFas、sFasL检测并与40例正常健康人做比较。结果急性心肌梗死患者血清Hcy与sFas、sFasL水平非常显著地高于正常对照组(P<0.01),血清Hcy水平与sFas、sFasL水平互呈明显正相关(P<0.01)。结论急性心肌梗死患者血清Hcy与sFas、sFasL水平密切相关。     

p,p'-DDE induces apoptosis and mRNA expression of apoptosis-associated genes in testes of pubertal rats

One,1‐dichloro‐2,2 bis(p‐chlorophenyl) ethylene (p,p'‐DDE), the major metabolite of 2,2‐bis(4‐chlorophenyl)‐1,1,1‐trichloroethane (DDT), is a known persistent organic pollutant and male reproductive toxicant. It has antiandrogenic effect. However, the mechanism by which p,p'‐DDE exposure causes male reproductive toxicity remains unknown. To elucidate the mechanism underpinning the testicular effects of p,p'‐DDE, we sought to investigate apoptotic effects and mRNA expression of apoptosis‐associated genes in the testis of pubertal rats, including Fas, FasL, calpain‐1, cytochrome c, Bax, Bcl‐w, Bak, and caspase‐3, ‐8, ‐9, ‐12. Animals were administered with different doses of p,p'‐DDE (0, 20, 60, 100 mg/kg body weight) every other day by intraperitoneal injection for 10 days. The results indicated that p,p'‐DDE exposure at over 20 mg/kg body weight showed the induction of apoptotic cell death. p,p'‐DDE could induce decrease in SOD and GSH‐Px activity of serum in 60 mg/kg body weight group. Significant elevations in the mRNA levels of Fas, FasL, calpain‐1, cytochrome c, Bax, Bak, and caspase‐3, ‐8, ‐9, ‐12 were observed in testis of rat treated with p,p'‐DDE. Taken together, these results lead us to speculate that in vivo exposure to p,p'‐DDE might induce testicular apoptosis in pubertal rats through the involvement of Fas/FasL, mitochondria and endoplasmic reticulum‐mediated pathways. © 2011 Wiley Periodicals, Inc. Environ Toxicol 2013.     

急性心肌梗死合并糖尿病患者血清细胞凋亡因子sFas和sFasL的变化及意义

目的了解糖尿病对急性心肌梗死患者血清细胞凋亡因子sFas和sFasL的影响。方法急性心肌梗死患者根据是否合并糖尿病将其分为非糖尿病组和糖尿病组。应用ELISA方法测定两组患者血清sFas和sFasL的含量。结果AMI合并DM患者血清sFas和sFasL的浓度明显高于非糖尿病患者,P〈0.01。心功能killip3/4级患者血清sFas和sFasL的浓度也高于killip1、2级患者,P〈0.05。结论急性心肌梗死合并糖尿病患者血清细胞凋亡因子sFas和sFasL的浓度高于非糖尿病患者。糖尿病可能通过促进Fas/FasL系统的激活而加速急性心肌梗死时心肌细胞凋亡的发生。     

血脂康预处理对大鼠在体心脏急性缺血-再灌注心肌Fas/FasL mRNA表达的影响

目的探讨血脂康对大鼠在体心脏急性缺血-再灌注心肌Fas/FasLmRNA表达的影响。方法40只雄性SD大鼠随机分成5组,每组8只。假手术组,双蒸水灌胃1周;模型对照组,双蒸水2ml灌胃1周;甲羟戊酸组,甲羟戊酸150mg/kg·d溶于2ml双蒸水灌胃1周;血脂康组,血脂康原粉2.8g/kg·d溶于2ml双蒸水配成悬液灌胃1周;血脂康 甲羟戊酸组,血脂康原粉2.8/kg·d联合甲羟戊酸150mg/kg·d溶于2ml双蒸水灌胃1周。1周后应用0000无创丝线结扎左冠脉前降支,结扎30min后松开恢复冠脉血流复制在体心脏急性缺血-再灌注模型,假手术组仅穿线而不结扎前降支,其他手术步骤相同。再灌注120min后用RT-PCR检测各组心肌Fas和FasLmRNA的表达。结果与假手术组大鼠心肌Fas/FasLmRNA表达相比较,其他4组大鼠心肌Fas/FasLmRNA表达明显升高(P<0.01);与模型对照组比较,血脂康组大鼠心肌Fas/FasLmRNA表达显著减少(P<0.01),血脂康 甲羟戊酸组和甲羟戊酸组大鼠心肌Fas/FasLmRNA表达无差异(P>0.05)。结论血脂康能减少大鼠心脏急性缺血-再灌注心肌Fas/FasLmRNA的表达。     

雷公藤内酯醇对雄性大鼠生殖毒性的机制研究(Ⅱ)

目的：考察雷公藤内酯醇对大鼠睾丸细胞相关凋亡基因mRNA表达的影响,研究雷公藤内酯醇生殖毒性的分子机制。方法：以低（0.025 mg·kg^-1）、中（0.05 mg·kg^-1）、高（0.1 mg·kg^-1）剂量的雷公藤内酯醇对健康雄性Wistar大鼠连续灌胃染毒30 d,每天一次,于末次染毒24 h后处死大鼠,取睾丸组织进行病理学检查,并通过实时荧光定量PCR测定Bcl-2、Bax、Fas、FasL、CREM和Caspase-3基因mRNA的表达情况。结果：与阴性对照组相比,雷公藤内酯醇染毒组睾丸组织生精细胞明显减少,精索内几乎无精子;高剂量组Bcl-2、CREM mRNA表达降低;而Bax mRNA表达水平在中、高剂量组时呈显著地高表达;Fas和FasL mRNA表达水平在高剂量组显著上升;Caspase-3 mRNA表达水平呈现依赖剂量的高表达,中、高剂量时呈现显著性差异。结论：提示在本实验染毒剂量范围内,特别是高剂量的雷公藤内酯醇能够使生殖细胞相关凋亡基因Bcl-2、Bax、Fas、FasL、CREM和Caspase-3不同程度的表达异常,这很可能是雷公藤内酯醇诱导大鼠生殖细胞凋亡的重要原因,为进一步深入阐述雷公藤内酯醇雄性生殖毒性的分子机制提供了依据。     

联检慢性心力衰竭患者血清sCD40L、可溶性Fas和sFasL的临床意义

目的 探讨慢性心力衰竭患者治疗前后血清可溶性CD40L(sCD40L)、sFas和sFasL水平的变化及意义.方法 应用放免法对40例慢性心力衰竭患者进行了治疗前后血清sCD40L、sFas和sFasL的检测,并与40例正常健康人作比较.结果 在慢性心力衰竭治疗前,患者血清sCD40L、sFas和sFasL水平非常显著地高于正常人组(P＜0.01),治疗后2周,则与正常健康人组比较差异无统计学意义(P＜0.05).结论 检测慢性心力衰竭患者血清sCD40L、sFas和sFasL水平的变化对了解病情、观察预后均有重要的临床价值.     

Isoliquiritigenin inhibits the proliferation and induces the apoptosis of human non-small cell lung cancer a549 cells

1. Isoliquiritigenin (ISL) is a natural pigment with the simple chalcone structure 4,2',4'-trihydroxychalcone. In the present study, we report, for the first time, ISL-induced inhibition of the proliferation of the human non-small cell lung cancer A549 cell line. 2. The results showed that ISL not only inhibited A549 cell proliferation, but also induced apoptosis and blocked cell cycle progression in the G1 phase. An ELISA assay demonstrated that ISL significantly increased the expression of p53 and p21/WAF1 protein, which caused cell cycle arrest. 3. An enhancement in Fas and its two ligands, namely membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), may be responsible for the apoptotic effect induced by ISL. 4. Taken together, the results indicate that the p53 and Fas/FasL apoptotic system may participate in the antiproliferative activity of ISL in A549 cells.     

海洛因依赖患者外周血清可溶性Fas及Fas配体的变化

目的:探讨海洛因依赖患者血清可溶性Fa(ssFas)和Fas配体(FasL)水平的变化及其意义。方法:采用双抗体夹心酶联免疫吸附法(ELISA),检测46例海洛因依赖患者血清中sFas和FasL水平,与正常人群作对照研究,并比较不同方式依赖者的sFas和FasL变化。结果:海洛因依赖者血清中sFas水平与对照组比较无统计学差异,而FasL水平明显高于对照组,差异具有统计学意义(P<0.01)。24例注射依赖者血清FasL水平较对照组高,差异具有统计学意义(P<0.05)。结论:海洛因依赖者血中FasL水平异常,提示海洛因滥用增加了Fas介导的细胞凋亡,且与用毒的方式有一定关系。这可能是海洛因依赖者免疫功能低下的重要原因之一。     

昆布提取物J201对博莱霉素诱导的大鼠肺纤维化的保护作用研究

目的：研究昆布提取物J201对大鼠肺纤维化的治疗作用及其可能的作用机制。方法：Wistar大鼠64只，随机分为4组。气管内灌注博莱霉素（BLM）诱导肺纤维化模型，观察BLM诱导的大鼠肺纤维化经不同剂量的J201治疗后，肺组织形态学、肺系数、丙二醛（MDA）和羟脯氨酸（HYP）含量的变化以及Fas、Fas配体（FasL）、基质金属蛋白酶-9（MMP-9）、组织金属蛋白酶抑制剂-2（TIMP-2）表达的变化。结果：J201能明显改善BLM诱导的大鼠肺纤维化肺组织形态学改变，并呈现一定的量效关系；能显著降低肺系数、肺组织MDA和HYP的含量，对肺组织Fas、FasL、MMP-9、TIMP-2的表达有明显的抑制作用，对肺系数、MDA、HYP、Fas、FasL、MMP-9 TIMP-2的影响呈现一定的量效关系。结论：J201对实验性大鼠肺纤维化有一定的保护作用，抗氧化作用及抑制Fas、FasL、MMP-9、TIMP-2的表达是其可能的机制之一。     

GABA tea prevents cardiac fibrosis by attenuating TNF-alpha and Fas/FasL-mediated apoptosis in streptozotocin-induced diabetic rats

GABA tea is a tea product that contains a high level of gamma-aminobutyric acid (GABA). This study investigated the effects of GABA tea on the heart in a diabetic rat model. Male Wistar rats were injected with 55 mg/kg streptozotocin (STZ) to induce diabetes for 2 weeks and then orally given dosages of 4.55 and 45.5 mg/kg/day GABA tea extract for 6 weeks. The results revealed that fasting blood glucose levels returned to normal levels in GABA tea-treated diabetic rats, but not in the untreated diabetic rats. Additionally, GABA tea effectively inhibited cardiac fibrosis induced by STZ. Further experiments showed that the STZ-induced protein levels of tumor necrosis factor-alpha (TNF-alpha), Fas, activated caspase-8 and caspase-3 were significantly inhibited by the GABA tea treatment. Therefore, our data suggest that the inhibiting effect of GABA tea on STZ-induced cardiac fibrosis in diabetic rats may be mediated by reducing blood glucose and further attenuating TNF-alpha expression and/or Fas/Fas ligand (FasL)-mediated apoptosis. These findings will provide implications for the potential anti-diabetic properties of GABA tea.     

Mitochondrial signaling pathway is also involved in bisphenol A induced germ cell apoptosis in testes

Bisphenol A (BPA) is a potential endocrine disruptor and testicular toxicant. An earlier study showed that BPA-induced germ cell apoptosis through the Fas/FasL apoptotic pathway. In the present study, we aimed to investigate whether the mitochondrial pathway is also involved in the process of BPA-mediated germ cell apoptosis in testes. Male mice were administered with BPA (160 or 480 mg/kg) by gavage daily from postnatal day 35 (PND35) to PND49. Germ cell apoptosis in testes was determined by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL). As expected, the number of TUNEL+ germ cells per tubule and the percentage of tubules with TUNEL+ germ cells were significantly increased in testes of mice treated with BPA during puberty. TUNEL+ germ cells were observed mainly in stages VII–VIII seminiferous tubules in testes. An increase in the level of Fas and FasL was observed in testes of mice exposed to BPA during puberty. In addition, pubertal BPA exposure evoked the activation of caspase-8 and caspase-3 in testes. Interestingly, pubertal BPA exposure also caused the translocation of cytochrome c from mitochondria into cytosol. In addition, pubertal BPA exposure upregulated the level of Bax and active caspase-9 in testes. Taken together, these results suggest that pubertal BPA exposure induces germ cell apoptosis in testes through not only the Fas/FasL signaling pathway but also the mitochondrial apoptotic pathway.     

泻肺汤对肺纤维化模型鼠肺组织病理形态变化及Fas/FasL蛋白表达的影响

目的 观察泻肺汤对博莱霉素致肺纤维化模型鼠肺组织病理形态变化及Fas/FasL蛋白表达的影响。方法 SPF级SD大鼠84只随机分为空白对照组，博来霉素模型对照组，泻肺汤低、中、高剂量组，醋酸泼尼松组，每组14只。除空白对照组外，其余各组采用气管滴注博莱霉素法制备肺纤维化大鼠模型，空白对照组气管内滴注等量生理盐水，造模24 h后，泻肺汤低、中、高剂量组灌胃泻肺汤（10，20，40 g·kg^-1·d^-1），醋酸泼尼松组灌胃醋酸泼尼松（1.8 mg·kg^-1·d^-1），空白对照组和博来霉素模型对照组用等体积生理盐水灌胃，第29天处死动物，HE染色法观察肺组织的病理形态变化，ELISA法测定各组大鼠细胞间黏附因子-1（intercellular cell adhesion molecule-1，ICAM-1）和血清结缔组织生长因子（connective tissue growth factor，CTGF）的含量，实时荧光定量PCR法测定各组大鼠肺组织Fas和FasL的基因表达，Western blotting检测各组大鼠肺组织Fas和FasL的蛋白表达。结果 与博来霉素模型对照组相比，泻肺汤各剂量组血清ICAM-1和CTGF均显著降低，肺组织Fas和FasL的基因表达也均显著减弱（P<0.05或P<0.01）；同时，泻肺汤中、高剂量组肺组织Fas和FasL的蛋白表达均显著减弱（P<0.05或P<0.01）。结论 泻肺汤可降低ICAM-1、CTGF含量和Fas/FasL表达来显著改善博来霉素诱导的肺纤维化，表明泻肺汤对博来霉素诱导的大鼠纤维化改善与Fas/FasL凋亡通路有关。     

基因转移FasL诱导人脑胶质瘤细胞凋亡的体外实验研究

目的：探讨体外基因转移Fas配体（Fas-Ligand,FasL)对恶性人脑胶质瘤细胞凋亡的影响，方法：用携带人FasL cDNA的缺陷型重组腺病毒载体（Ad-FL)，在体外转导5株人脑胶质瘤细胞，并使其表达，通过流式细胞仪，RT－PCR，TUNEL法及荧光显微镜分别进行Fas/FasL表达检测和相关凋亡检测，分析，结果：RT－PCR和流式细胞仪检测5株胶质瘤细胞表达均表达Fas，不表达FasL,而Ad-FL转导的5株胶质瘤细胞均能表达FasL,Fas表达水平与抗Fas抗体诱导胶质瘤细胞凋亡敏感性无相关性，Ad-FL能显著诱导5株胶质细胞瘤在体外凋亡或抑制其生长，结论：重组腺病毒FasL在体外诱导人脑胶质瘤凋亡效果显著。