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1.
目的研究单甲基亚砷酸(MMAIII)联合隐丹参酮(CPT)对多发性骨髓瘤U266细胞的协同抑制效应,阐明其诱导多发性骨髓瘤凋亡的作用机制。方法多发性骨髓瘤U266细胞用含10%gibco胎牛血清的RPMI1640培养液培养,按隐丹参酮单药组、MMAIII单药组和联合组分别加药处理。cell-counting-kit-8(CCK8)法检测细胞存活率;AnnexinⅤ-FITC/PI流式细胞术检测细胞凋亡;Western Blot检测凋亡标记蛋白PARP的总蛋白和剪切表达水平;酶标仪检测酸性鞘磷脂酶活性;免疫荧光技术检测细胞膜神经酰胺和脂筏标记物。结果 CCK8法检测结果显示,隐丹参酮浓度15μM时细胞存活率为(80.9±5.4)%(24h)、(60.0±8.4)%(48h),单甲基亚砷酸浓度1μM时细胞存活率为(82.5±5.8)%(24h)、(67.7%+9.7)%(48h),隐丹参酮(15μM)联合单甲基亚砷酸(1μM)作用U266细胞24h、48h后细胞存活率为(29.1±7.0)%(24h)、(18.0±2.7)%(48h);流式细胞术结果显示,联合用药组细胞凋亡率为(41.7±4.4)%、单甲基亚砷酸组为(10.6±4.0)%、隐丹参酮组为(10.5±3.0)%;Western Blot显示,联合作用组凋亡蛋白PARP剪切活化水平较单甲基亚砷酸单药组及隐丹参酮单药组有显著升高;酸性鞘磷脂酶活性测定结果显示,联合用药组上调多发性骨髓瘤U266细胞酸性鞘磷脂的活性;免疫荧光技术检测结果显示,联合用药后多发性骨髓瘤U266细胞膜上荧光强度增加,加入Filipin后荧光强度较联合用药减少。结论 MMAIII与CPT协同作用能诱导人多发性骨髓瘤U266细胞凋亡发生,其作用机制主要与促进细胞膜鞘磷脂水解生成神经酰胺、诱导脂筏聚集促进凋亡相关蛋白剪切活化密切相关。  相似文献   

2.
摘要 目的 验证丹参二萜醌类活性成分对胰腺癌和多发性骨髓瘤的抑制效应,阐明其诱导胰腺癌和多发性骨髓瘤凋亡的作用机制。方法 胰腺癌细胞AsPC-1、BxPC-3用含10% gibco胎牛血清的RPMI1640培养液培养,按隐丹参酮组(30μM)、丹参新酮组(15μM)、去氢丹参新酮组(15μM)分别加药处理。cell-counting-kit-8(CCK8)法检测细胞存活率;AnnexinⅤ-FITC/PI流式细胞术检测细胞凋亡;Western Blot检测相关PKC同工酶磷酸化水平;对AsPC-1和BxPC-3细胞进行siRNA转染,Western Blot检测相关PKC同工酶磷酸化水平。结果 隐丹参酮组AsPC-1、BxPC-3细胞存活率分别为40.1%±5.0%、36.2%±5.4%;丹参新酮组AsPC-1、BxPC-3细胞存活率分别为52.1%±5.1%、47.2%±5.7%;去氢丹参新组AsPC-1、BxPC-3细胞存活率分别为46.1%±5.0%、42.2%±5.4%(P<0.01)。流式细胞术结果显示:AsPC-1组内空白对照组、隐丹参酮组、丹参新酮组、去氢丹参新酮组细胞的凋亡率分别为4.71%、30.10%、52.26%、42.30%;BxPC-3组内空白对照组、隐丹参酮组、丹参新酮组、去氢丹参新酮组细胞的凋亡率分别为5.10%、30.66%、33.76%、51.76%(P<0.01)。Western Blot检测显示隐丹参酮组、丹参新酮组、去氢丹参新酮组较空白对照组,胰腺癌AsPC-1、BxPC-3细胞p-PKD/PKCμ ser916、p-PKCδ thr505、p-PKD/PKCμ ser744/748的水平降低。Western Blot检测显示,siRNA沉默胰腺癌AsPC-1、BxPC-3细胞PKCδ,胰腺癌AsPC-1、BxPC-3细胞PKD/PKCμ ser744/748的磷酸化水平下调。结论 隐丹参酮、丹参新酮、去氢丹参新酮通过抑制PKCδthr505的磷酸化水平,继而PKD1μser744/748磷酸化水平下调,从而显著促进胰腺癌AsPC-1和BxPC-3细胞凋亡发生。  相似文献   

3.
目的:阐明苦参碱对多发性骨髓瘤(MM)细胞的作用和内在机理。方法:MTT法检测苦参碱对多发性骨髓瘤细胞株RPMI8226和多发性骨髓瘤原代细胞增殖的影响;根据中效原理分析苦参碱联合亚砷酸对RPMI8226细胞株的联合作用;应用DAPI染色检测细胞凋亡。结果:M1TT比色法检测结果显示,苦参碱对RPMI8226细胞及MM原代细胞均有生长抑制作用,与空白对照组比较,差异有统计学意义(P<0.05)。体外联合亚砷酸作用RPMI8226细胞抑制率>50%时联合指数(CI)<1。DAPI荧光染色,荧光显微镜下观察苦参碱以终浓度2.0g/L作用于RPMI8226细胞12h后可见明显的凋亡小体出现。结论:苦参碱对RPMI8226细胞和MM原代细胞均有生长抑制作用,体外联合亚砷酸作用MM细胞株RPMI8226细胞,抑制率>50%时呈协同作用。苦参碱通过诱导细胞凋亡抑制RPMI8226细胞的增殖。  相似文献   

4.
目的 研究去甲斑蝥素(norcantharidin,NCTD)对多发性骨髓瘤细胞株的增殖及其凋亡的影响及其可能的机制.方法 四甲基偶氮唑盐[3-(4,5-dimethyl-2 thiahiazoyi)-3,5-di-phenyl-tetrazolium bromide,MTT]检测5、20、40、80、160μmol/L的NCTD对U266细胞增殖的影响;用流式细胞术检测不同浓度NCTD对细胞周期及细胞凋亡的影响,并应用半定量反转录聚合酶链反应检测survivin基因表达的变化.结果 在20~160μmol/L的浓度内,NCTD对U266细胞的增殖有抑制,抑制与NCTD的浓度呈剂量依赖关系(P<0.05).不同浓度的NCTD能诱导多发性骨髓瘤细胞株U266凋亡,使细胞明显阻滞于G2 /M 期;在NCTD作用下,U266 细胞survivin 及其机制可能与诱导凋亡、细胞周期阻滞有关.结论 NCTD能抑制U266细胞的增殖,并诱导其凋亡的机制可能与抑制survivin的表达及G2 /M 期阻滞有关.  相似文献   

5.
目的 探讨硼替佐米联合WEE1激酶抑制剂AZD-1775对人多发性骨髓瘤(MM)细胞凋亡的影响。方法 培养RPMI8226和U266细胞,逆转录PCR(RT-PCR)检测WEE1 mRNA表达。采用随机数字表法将两种细胞分为对照组、AZD-1775组、硼替佐米组和联合处理组。AZD-1775组细胞加入AZD-1775(10μmol/L)孵育24 h;硼替佐米组细胞加入硼替佐米(20 nmol/L)孵育24 h;联合处理组细胞先加入硼替佐米(20 nmol/L)孵育24 h,再加入AZD-1775(10μmol/L)孵育24 h。采用肿瘤细胞成球实验检测不同处理对RPMI8226和U266细胞成球率影响;采用流式细胞术检测不同处理对RPMI8226和U266细胞凋亡率影响;采用Westen blot检测不同处理对RPMI8226细胞Bcl-2和Caspase-3蛋白表达影响。结果 RT-PCR检测显示,RPMI822和U266细胞均存在WEE1 mRNA表达。与对照组相比,AZD-1775组、硼替佐米组和联合处理组细胞球形成率均降低(P<0.05),而联合处理组细胞球形成率低于AZ...  相似文献   

6.
目的:研究力达霉素(LDM)联合硼替佐米(BZM)的抗骨髓瘤作用,并探讨两药联合
协同抗骨髓瘤的作用机制。方法:选取处于对数生长期人多发性骨髓瘤细胞SKO-007随机分
为对照组、LDM组、BZM组和LDM+BZM联合组,采用MTS法和流式细胞术检测各组细胞存活率
、细胞周期分布和凋亡率;采用Western blotting 法检测各组SKO-007细胞凋亡相关蛋白
和内质网应激(ER
S)相关蛋白的表达量。结果:细胞培养48 h后,LDM+BZM联合组细胞存活率显著低于LDM组
、BZM组和对照组(P<0.05);LDM+BZM联合组
SKO-007细胞
G2/M期的细胞比例、细胞凋亡率、caspase-3和poly ADP-ribose polymerase (PAR
P)的
切割片段蛋白表达量、GRP78/Bip蛋白表达量、CHOP/GADDl53蛋白表达量和c-Jun氨基末端  相似文献   

7.
目的 观察白凤菜总黄酮(TFG)对多发性骨髓瘤U266细胞的增殖和凋亡的影响。方法 MTT实验检测U266细胞增殖;倒置显微镜及荧光显微镜分别观察U266细胞经TFG处理24 h后细胞形态及凋亡的变化;流式细胞术检测细胞凋亡及细胞周期的变化。结果 TFG明显抑制U266细胞的增殖,P<0.05,且呈浓度和时间依赖性;100 μg /mLTFG作用组的U266细胞凋亡明显增加;TFG使U266细胞周期阻滞于S期。结论 TFG可抑制U266细胞增殖,可能同细胞周期的捕获和细胞凋亡的诱导相关。  相似文献   

8.
目的 探讨三萜类化合物熊果酸(ursoIic acid,UA)对多发性骨髓瘤(multiple myeloma,MM)细胞株U266的凋亡诱导作用及分子机制.方法 用熊果酸处理U266细胞,Cell Counting Kit-8法检测细胞增殖抑制情况;用20μmol/L熊果酸处理U266细胞24 h,瑞氏法染色后在光学显微镜下观察细胞凋亡的形态学变化;Annexin-V标记,在流式细胞仪上检测细胞凋亡率变化;用实时荧光定量PCR方法及Western blot方法检测FGFR3、Bcl-2、CCND1、MYC及p53的表达变化.结果 经熊果酸处理后,U266细胞的增殖受到抑制,光学显微镜下可观察到细胞形态不规则样改变,胞膜小泡状形成,细胞质内空泡增多,细胞核染色质浓缩、固缩;流式细胞术检测Annexin-V阳性细胞比例随用药浓度增加及用药时间延长而增加;细胞内FGFR3、Bcl-2、CCND1、MYC的mRNA和蛋白质表达随用药浓度增加及用药时间延长呈显著下降趋势,而p53的mRNA及蛋白表达逐渐增高.结论 熊果酸可抑制U266细胞的增殖并诱导其凋亡,体外有抗多发性骨髓瘤细胞作用,为多发性骨髓瘤选择新的靶向治疗方案提供实验依据.  相似文献   

9.
目的探讨胃泌素受体拮抗剂丙谷胺与特异性环氧合酶-2(COX-2)抑制剂NS-398对人胃腺癌细胞株MKN-45增殖、凋亡的调控作用。方法MKN-45细胞常规培养于RPMI-1640培养液中,细胞长至亚单层后加丙谷胺(终浓度5.0mmol/L)和/或NS-398(10.0μmol/L),连续培养48h。四甲基偶氮唑蓝(MTT)比色分析检查细胞增殖,流式细胞仪检测细胞凋亡,逆转录聚合酶链反应(RT-PCR)和Western印迹法检测凋亡抑制基因bcl-2mRNA及蛋白表达。结果丙谷胺和NS-398协同抑制MKN-45细胞增殖;丙谷胺组、NS-398组和联合用药组的细胞凋亡率分别为24.7%±3.2%,26.7%±3.4%和36.1%±4.6%,显著高于对照组的1.6%±0.6%(均P<0.01);联合用药组的细胞凋亡率明显高于单一用药组(P<0.05)。丙谷胺和NS-398显著下调bcl-2mRNA及蛋白表达(均P<0.05)。结论丙谷胺、NS-398抑制MKN-45细胞增殖,通过下调bcl-2基因表达而诱导细胞凋亡,两者联合具有协同抗癌作用。  相似文献   

10.
目的观察丹参二萜醌类活性成分对胰腺癌的抑制效应,探讨其诱导胰腺癌凋亡的作用机制。方法胰腺癌细胞AsPC-1、BxPC-3用含10%gibco胎牛血清的RPMI1640培养液培养,按隐丹参酮组(30μM)、丹参新酮组(15μM)、去氢丹参新酮组(15μM)分别加药处理。Cell-countingkit-8(CCK8)法检测细胞存活率;AnnexinⅤ-FITC/PI流式细胞术检测细胞凋亡;Western blot检测相关PKC同工酶磷酸化水平;对AsPC-1和BxPC-3细胞进行siRNA转染,Western blot检测相关PKC同工酶磷酸化水平。结果隐丹参酮组AsPC-1、BxPC-3细胞存活率分别为(40.1±5.0)%、(36.2±5.4)%;丹参新酮组AsPC-1、BxPC-3细胞存活率分别为(52.1±5.1)%、(47.2±5.7)%;去氢丹参新组AsPC-1、BxPC-3细胞存活率分别为(46.1±5.0)%、(42.2±5.4)%(P<0.01)。流式细胞术结果显示,AsPC-1组内空白对照组、隐丹参酮组、丹参新酮组、去氢丹参新酮组细胞的凋亡率分别为4.71%、30.10%、52.26%、42.30%;BxPC-3组内空白对照组、隐丹参酮组、丹参新酮组、去氢丹参新酮组细胞的凋亡率分别为5.10%、30.66%、33.76%、51.76%,组间比较,差异有统计学意义(P<0.01)。Western blot检测显示,隐丹参酮组、丹参新酮组、去氢丹参新酮组较空白对照组,胰腺癌AsPC-1、BxPC-3细胞p-PKD/PKCμser916、p-PKCδthr505、p-PKD/PKCμser744/748的水平降低。Western blot检测显示,siRNA沉默胰腺癌AsPC-1、BxPC-3细胞PKCδ,胰腺癌AsPC-1、BxPC-3细胞PKD/PKCμser744/748磷酸化水平下调。结论隐丹参酮、丹参新酮、去氢丹参新酮可能通过抑制PKCδthr505磷酸化水平,下调PKD1μser744/748磷酸化水平,促进胰腺癌AsPC-1和BxPC-3细胞凋亡发生。  相似文献   

11.
肱骨远端全骺分离   总被引:1,自引:0,他引:1  
肱骨远端全骺分离在5岁以下儿童中是较常见的肘部损伤,且极易误诊为肱骨髁上骨折或肘关节脱位。本组25例肱骨远端全骺分离均为向内侧移位,年龄11个月至14岁,平均5.6岁,属Salter-HarrisⅠ型或Ⅱ型。对本症骨骺损伤的解剖学和组织学,诸骨(和骨化中心)之间的对应关系以及骨骺损伤的诊治进行了简短的讨论。  相似文献   

12.
通过对右手虎口挛缩以及左拇指皮肤剥脱损伤各一例应用直指背侧皮瓣修复的成功经验,提出该手术的适应症和优点,并重点介绍其应用解剖学和手术方法。  相似文献   

13.

Objective

To investigate the hypoglycemic, hypolipidemic and antioxidant activities of aqueous extract of Terminalia paniculata bark (AETPB) in streptozotocin (STZ)-induced diabetic rats.

Methods

Acute toxicity was studied in rats after the oral administration of AETPB to determine the dose to assess hypoglycemic activity. In rats, diabetes was induced by injection of STZ (60 mg/kg, i.p.) and diabetes was confirmed 72 h after induction, and then allowed for 14 days to stabilize blood glucose level. In diabetic rats, AETPB was orally given for 28 days and its effect on blood glucose and body weight was determined on a weekly basis. At the end of the experimental day, fasting blood sample was collected to estimate the haemoglobin (Hb), glycosylated haemoglobin (HbA1c), serum creatinine, urea, serum glutamate-pyruvate transaminase (SGPT), serum glutamate-oxaloacetate transaminase (SGOT) and insulin levels. The liver and kidney were collected to determine antioxidants levels in diabetic rats.

Results

Oral administration of AETPB did not exhibit toxicity and death at a dose of 2 000 mg/kg. AETPB treated diabetic rats significantly (P<0.001, P<0.01 and P<0.05) reduced elevated blood glucose, HbA1c, creatinine, urea, SGPT and SGOT levels when compared with diabetic control rats. The body weight, Hb, insulin and total protein levels were significantly (P<0.001, P<0.01 and P<0.05) increased in diabetic rats treated with AETPB compared to diabetic control rats. In diabetic rats, AETPB treatment significantly reversed abnormal status of antioxidants and lipid profile levels towards near normal levels compared to diabetic control rats.

Conclusions

Present study results confirm that AETPB possesses significant hypoglycemic, hypolipidemic and antioxidant activities in diabetic condition.  相似文献   

14.
15.

Objective

To determine the association of smoking, alcohol and nonsteroidal anti-inflammatory drugs (NSAIDs) use with presence and virulence of Helicobacter pylori (H. pylori) infection in a representative sample of a random adult population of asymptomatic subjects.

Methods

Non virulent 16S rRNA and virulent cag A and T genes from salivary samples of 854 asymptomatic subjects were determined using polymerase chain reaction. The presence and absence of virulent and non virulent infection was statistically compared with consumption of smoking, alcohol and NSAIDs.

Results

The prevalence of infection in male and female subjects was found to be 69.25% and 66.90%, respectively. The prevalence of infection in the population of asymptomatic subjects with respect to consumption of alcohol was as follows: current (31.22%), former (52.20%) and never (43.58%). The prevalence of infection in the population of asymptomatic subjects with respect to smoking of cigarettes was as follows: current (88.80%), former (57.14%) and never (33.33%). The prevalence of infection in the subject population consuming NSAIDs and not consuming NSAIDs frequently was found to be 82.75% and 21.16%, respectively. Virulence in male and female subjects was found to be 60.00% and 50.00%, respectively. The presence of virulent infection in the population of asymptomatic subjects with respect to consumption of alcohol was as follows: current (28.57%), former (40.15%) and never (50.00%). The prevalence of virulent infection in the population of asymptomatic subjects with respect to smoking of cigarettes was as follows: current (79.32%), former (75.00%) and never (50.00%). The prevalence of virulent infection in the subject population consuming NSAIDs and not consuming NSAIDs frequently was found to be 88.23% and 66.66%, respectively.

Conclusions

It can be concluded that smoking and NSAIDs consumption are aggravating factors for virulence of H. pylori and alcohol can inhibit H. pylori infection in asymptomatic subjects.  相似文献   

16.
综述分析1954~84年国内外文献,探讨了大肠多发原发性癌的诊断标准、发病情况、临床表现、发病危险因素、外科处理以及预防和诊断检测技术等问题,强调了大肠腺瘤,遗传基础、免疫缺陷、慢性溃疡性结肠炎等在发病上的作用,及高危险组患者应按方案进行常规检测监护之重要性。述介了大肠多发原发性癌系一种原发性大肠癌多中心发生的形式,在诊治上前者与后者有一致性,亦各有其独特性。报告宁夏医学院附属医院肿瘤科1977~84年收治的8例住院大肠多发原发性癌病例的临床资料,对诊治经过进行了初步总结。  相似文献   

17.
目的:研究开发一种涂覆(HA/CHI-Van)5多层膜结构的骨科内植入物以用于预防骨内植入物感染。方法:将透明质酸(HA)和壳聚糖-万古霉素(CHI-Van)溶液进行层层自组装,制备得到(HA/CHI-Van)5多层膜结构;通过zeta电位检测组装过程,通过活细菌染色实验评价多层膜的抗细菌黏附效果,通过越狱实验和FE-SEM实验检测体外抗菌效果。SD大鼠随机分为空白组、(HA/CHI-Van)5组和假手术组,并以X线、Micro-CT、骨组织切片等方法检测该材料的体内动物实验效果。结果:每次组装不同的单层,zeta电位交替变化,证明多层膜组装成功。活细菌染色实验显示多层膜结构中的CHI具有一定的抗细菌黏附作用。越狱实验和FE-SEM实验证明(HA/CHI-Van)5多层膜结构在体外具有良好的抗菌作用。在体外实验中,(HA/CHI-Van)5组的X线评分为0.7,而空白组的评分为12.3,(HA/CHI-Van)5组在骨密度、骨小梁数量、骨体积分数、骨组织切片方面均优于空白组(均P<0.05),且非常接近假手术组(均P>0.05)。结论:通过在骨科内植入物表面涂覆(HA/CHI-Van)5多层膜结构能够获得一定的抗细菌黏附功能和良好的抗菌作用,可以预防骨内植入物感染。  相似文献   

18.

Objective

To evaluate the efficacy and side effects of praziquantel (PZQ) in the treatment of schistosomiasis in Ethiopia.

Methods

In a cross-sectional study, stool specimens were collected from randomly selected 299 school children in Shesha Kekele Elementary School, Wondo Genet, Southern Ethiopia, in April 2010. Stool specimens were examined using a single Kato-Katz thick smear for Schistosoma mansoni (S. mansoni) ova. Children who were found positive for S. mansoni were treated with a single oral dose of PZQ at 40 mg/kg bw and interviewed for treatment-related symptoms 24 hours after drug administration. Four weeks post-treatment, stool specimens were collected from the same children and examined following the same procedure as in the pre-treatment. Drug efficacy was determined based on cure and egg reduction rates.

Results

Pre-treatment prevalence of S. mansoni infection was 74.9% with geometric mean egg count of 268. The evaluated generic PZQ produced an overall cure rate of 73.6% (P<0.000 1, OR: 8.33, CI: 5.3–13.1) and egg reduction rate of 68.2% (P=0.03, F=0.64). The cure rate showed significant association with age (χ2=11, P=0.004), the highest rate being observed in the 15–22 age group. 83% of S. mansoni infected children showed various treatment-related symptoms, the most frequent being headache, nausea, and abdominal pain. These symptoms were associated with age (P<0.001) and pre-treatment intensity of infection (P<0.05).

Conclusions

The present observations revealed relatively lower cure and egg reduction rates of the PZQ evaluated as compared to previous reports for other PZQ brands in Ethiopia. Hence, in depth studies are recommended to clarify whether the present relatively lower cure rate is the actual cure rate of the praziquantel evaluated, treatment failure, or reduced susceptibility of the parasite. Treatment-related side effects observed were transient and tolerable.  相似文献   

19.
目的:制备具有肿瘤血管靶向的NGR修饰唑来膦酸长循环脂质体,并对其进行表征研究。方法:采用薄膜分散法制备NGR修饰唑来膦酸长循环脂质体,,并对其进行表征研究,并测定包封率。结果:制备得到的NGR-PEG-ZOL-LPs平均粒径为(271.43±9.23)nm,Zeta电位为-(36.20±0.20)mv,多分散系数(PDI)为(0.15±0.03),理化性质相对稳定;NGR-PEG-ZOL-LPs包封率为8.41%;电镜下显示NGR-PEG-ZOL-LPs呈类圆形,大小分布均匀,无明显聚集。结论:采用薄膜分散法制备的NGR修饰唑来膦酸长循环脂质体具有较高的稳定性,从而为体内药效学研究奠定基础。  相似文献   

20.

Objective

To evaluate the acaricidal activity of extracts of three essential oils of chamomile, marjoram and Eucalyptus against Tetranychus urticae (T. urticae) Koch.

Methods

Extracts of three essential oils of chamomile, marjoram and Eucalyptus with different concentrations (0.5%, 1.0%, 2.0%, 3.0% and 4.0%) were used to control T. urticae Koch.

Results

The results showed that chamomile (Chamomilla recutita) represented the most potent efficient acaricidal agent against Tetranychus followed by marjoram (Marjorana hortensis) and Eucalyptus. The LC50 values of chamomile, marjoram and Eucalyptus for adults were 0.65, 1.84 and 2.18, respectively and for eggs 1.17, 6.26 and 7.33, respectively. Activities of enzymes including glutathione-S-transferase, esterase (α-esterase and β-esterase) and alkaline phosphatase in susceptible mites were determined and activities of enzymes involved in the resistance of acaricides were proved. Protease enzyme was significantly decreased at LC50 of both chamomile and marjoram compared with positive control. Gas chromatography-mass spectrometer (GC-MS) proved that the major compositions of Chamomilla recutita are α-bisabolol oxide A (35.251%), and trans-β-farersene (7.758%), while the main components of Marjorana hortensis are terpinene-4-ol (23.860%), p-cymene (23.404%) and sabinene (10.904%).

Conclusions

It can be concluded that extracts of three essential oils of chamomile, marjoram and Eucalyptus possess acaricidal activity against T. urticae.  相似文献   

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