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1.
应用α-糜蛋白酶和盐酸胍作为抑制剂选择性测定待测样品中乳酸脱氢酶同工酶Ⅰ(LD-1).采用蛋白水解酶和抑制剂来破坏乳酸脱氢酶同工酶2~5(LD2~5)的活性,因此不需预处理待测样品,可直接用于自动生化分析仪.使用日立7150分析仪建立了LD-1分析仪建立了LD-1测定参数,方法变异系数(CV)2.7%~4.4%;测得结果(y)与LD-1电泳方法(x)相关良好,y=0.967x-1.957,r=0.992(n=37);检测了197名健康男女血清样品,确定LD-1参考值范围为29.78~59.26U/L,x±s=(44.58±7.39)U/L.  相似文献   

2.
建立了人肌型特异烯醇化酶(hMSE)的放免分析法,抗血清的亲和常数为5.1×109L/mol,采用改良的BHR法制备了 125I-hMSE,后者非特异结合率为3%,与抗血清(1∶103)结合率达50.16%;批内和批间CV分别为8.6%和13%.回收率为95%-105%.标准曲线范围为5-320μg/L.最小检出率为5μg/L.最佳反应条件:0.1mol/LpH7.4PBS(含5mmol/L MgSO4,0.1%吐温-20,0.1%NaN3);反应温度和时间:37℃反应0.5h和4℃,0.5h,作为快速测定;或选用4℃反应24h.65例健康人血清hMSE浓度为23.9±10.9μg/L(x±s).hMSE超过57μg/L(x+3s).为阳性值.测定了AMI和肌病患者,血清hMSE明显升高.  相似文献   

3.
应用免疫沉淀法测定了乳酸脱氢酶同工酶1(LD1).血清与抗血清用量为10:1,加入抗血清5min后再加入与血清量相等的饱和硫酸铵溶液.离心后测定上清液中的LD.总酶活性在618U/L内线性关系良好.二份标本批内精度CV值分别为3.76%和4.70%,批间CV值为7.00%,免疫沉淀法与电泳法高度相关(n=22,r=0.976).72例正常人LD参考值为102.31±16.4U/L,LD1为23.65±4.39U/L,LD1/LD为23.12±3.85%.免疫沉淀法的优点是特异性高,操作简单,可用于自动生化分析仪.精确度高,线性关系好,测定范围宽,是测定LD1的理想方法.  相似文献   

4.
最近的研究表明,蛋白质内含子(intein)介导的B区缺失型凝血因子Ⅷ (BDD-FⅧ)的轻链和重链剪接可顺式促进后者的分泌,而且剪接反应在细胞内、外均可发生.为进一步提高基于蛋白质内含子的双载体转BDD-FⅧ基因的功效,将具有促进重链分泌作用的位于Pro1640~Ser1690的酸性区3(acidic region-3,AR-3)引入重链,检验对蛋白质内含子剪接的BDD-FⅧ蛋白分泌和活性的影响.用融合蛋白内含子的附加ar-3重链(HCAR3IntN)基因和轻链(IntCLC)基因共转染培养的HEK293细胞,分别用ELISA和Coatest法定量分析分泌至培养上清中剪接BDD-FⅧ蛋白量和生物活性,并用免疫印迹观察了细胞内的BDD-FⅧ剪接.结果显示,共转HCAR3IntNIntCLC基因细胞,分泌至上清的剪接BDD-FⅧ蛋白量和活性分别为(173±26) μg/L和(1.31±0.15) U/ml,明显高于未添加ar-3的蛋白质内含子融合重链(HCIntN)与轻链(IntCLC)基因共转染细胞[(102±12) μg/L和(0.79±0.09) U/ml],提示AR-3对蛋白质内含子剪接的BDD-FⅧ蛋白分泌和活性有明显改善作用.而且,分别转HCAR3IntNIntCLC基因细胞混合培养后的上清中,亦检测到剪接的BDD-FⅧ蛋白和活性[(35±7) μg/L和(0.28±0.08) U/ml],表明蛋白质内含子可进行不依赖细胞机制的蛋白质剪接.另外,转基因细胞总蛋白呈现明显的可与FⅧ多克隆抗体进行反应的剪接BDD-FⅧ蛋白条带,直观地反映细胞内BDD-FⅧ的剪接.为动物模体内运用蛋白质反式剪接技术的双腺相关病毒载体(AAV)转BDD-FⅧ基因实验提供了依据.  相似文献   

5.
研究外源GA3对盐胁迫下黄瓜种子萌发和幼苗生长的影响。结果表明,添加不同质量浓度GA3的各处理,其发芽率、发芽势和发芽指数均显著高于NaCl胁迫处理,其中以100 mg/L GA3处理的发芽势、发芽率和发芽指数最高,幼苗的叶面积、根长、根冠比也最大,同时叶片中叶绿素含量最高,幼苗叶片的光合速率(Pn)、气孔导度(Gs)、胞间CO2摩尔分数(Ci)及蒸腾速率(Tr)等均达到最大;而当赤霉素的质量浓度为50 mg/L时,叶片中的POD活性为2 005 U/(g·min),达最大值。  相似文献   

6.
用琼脂糖凝胶电泳法同时测定肌酸激酶(CK)同工酶及其亚型(CK-MB亚型、CK-MM亚型),其特点是以不连续缓冲液体系为基础,在较低电压条件下,30min完成分离过程,能同时报告CK-MB同工酶,CK-MB1,CK-MB2以及CK-MM1,CK-MM2,CK-MM3的百分含量.CK-MB和CK-MM同工酶的亚型最低检出限分别为2.2U/L,3.2U/L.该法具有快速、灵敏度高、重复性好、无需特殊仪器、操作简单等优点.  相似文献   

7.
The relationships between dark respiration rate (R D) and net photosynthetic rate (P N) in Quercus ilex L. shrubs growing at the Botanical Garden in Rome were analysed. Correlation analysis of the data sets collected in the year 2006 confirmed the dependence among the considered leaf traits, in particular, R D was significantly (p<0.05) correlated with P N (r = 0.40). R D and P N increased from March to May [1.40±0.10 and 10.1±1.8 μmol(CO2) m−2 s−1 mean values of the period, respectively], when air temperature was in the range 14.8–25.2 °C, underlining the highest metabolic activity in the period of the maximum vegetative activity that favoured biomass accumulation. On the contrary, the highest R D [1.60±0.02 μmol(CO2) m−2 s−1], associated to the lowest P N rates (44 % of the maximum) and carbon use efficiency (CUE) in July underlined the mobilization of stored material during drought stress by a higher air temperature (32.7 °C).  相似文献   

8.
以2年生苹果矮化砧木M9 T337为试材,采用盆栽试验法,设置浇灌清水(CK)和盐碱胁迫(0.1 mol/L NaCl+NaHCO3溶液)+ 喷施5种浓度的H2O2 [0(T1)、0.2 mmol/L(T2)、0.4 mmol/L(T3)、0.6 mmol/L(T4)、0.8 mmol/L(T5)] 处理,测定各处理叶片叶绿素含量、光合气体交换参数、渗透调节物质含量、抗氧化酶活性和细胞膜透性,并利用相关性与主成分分析进行综合评价,以探讨外源过氧化氢(H2O2)增强其盐碱耐性的生理机制。结果表明:(1)随着盐碱胁迫(T1)的时间延长,M9 T337幼苗叶片叶绿素a(Chl a)含量、叶绿素b (Chl b)含量、叶绿素总量(Chl t)、净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)、可溶性蛋白(SP)含量均呈逐渐下降趋势;胞间CO2浓度(Ci)、可溶性总糖(TSS)含量、脯氨酸(Pro)含量、过氧化氢酶(CAT)活性、抗坏血酸过氧化物酶(APX)活性、相对电导率(REC)、丙二醛(MDA)含量均呈上升趋势;超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性均呈先升后降趋势。(2)与CK相比,盐碱胁迫+外源H2O2(T2- T5)处理后M9 T337幼苗叶片各指标均呈现不同幅度变化,且存在明显浓度效应,并以T3(0.4 mmol/L H2O2)处理叶片的Chl a、Chl b、Chl t、SP和Gs降幅最小,Ci、REC、MDA升幅最小,TSS、Pro、APX升幅最大。(3)M9 T337幼苗叶片PnTrGs、Chl a、Chl b、Chl t、SP、SOD、POD呈显著正相关,与Ci、MDA、CAT、APX、REC呈显著负相关。(4)综合评价表明,各处理对M9 T337幼苗叶片生理特性的效应依次为:CK>T3>T4>T2>T5>T1。研究发现,叶面喷施适宜浓度H2O2可有效改善盐碱胁迫下M9 T337幼苗光合能力,显著提高抗氧化酶活性和渗透调节物质的含量,降低细胞膜透性,从而达到缓解盐碱胁迫的作用,并以0.4 mmol/L H2O2处理效果最佳。  相似文献   

9.
模拟氮沉降对石栎和苦槠幼苗土壤呼吸的影响   总被引:3,自引:0,他引:3  
李凯  江洪  由美娜  曾波 《生态学报》2011,31(1):82-89
用LI-8100开路式土壤碳通量测量系统测定模拟氮沉降4种不同处理水平(0、60、120\,240 kg · hm-2 · a-1)下石栎(Lithocarpus glabra)和苦槠(Castanopsis sclerophylla)幼苗的土壤呼吸速率及土壤温度、含水量对其土壤呼吸的影响。结果表明,氮沉降对土壤呼吸的影响根据施氮水平和幼苗的种类不同而异。低氮(60 kg · hm-2 · a-1)处理下石栎和苦槠的土壤呼吸速率平均值分别为(4.014±0.812)μmol · m-2 · s-1和(5.170±0.689)μmol · m-2 · s-1,比对照组(0 kg · hm-2 · a-1)土壤呼吸速率平均值(3.802±0.948)μmol · m-2 · s-1和(3.557±0.906)μmol · m-2 · s-1分别高5%和45%;两树种在中、高氮处理下均出现对土壤呼吸明显的抑制。其中石栎中、高氮实验组的土壤呼吸速率分别为(2.653±0.681)μmol · m-2 · s-1、(2.592±0.736)μmol · m-2 · s-1, 比对照组低27%和29%。苦槠中、高氮实验组的土壤呼吸速率为(3.563±0.402)μmol · m-2 · s-1、(3.466±0.994)μmol · m-2 · s-1, 比对照组低7%和8%;石栎在高氮(240 kg · hm-2 · a-1)处理水平下,其土壤呼吸速率同10cm土壤温度之间呈现显著的指数关系(R2=0.811,P=0.001),而在低、中氮实验均未发现有明显指数关系。苦槠各处理水平下其土壤呼吸与土壤温度之间均未发现有明显的指数关系;在土壤呼吸与5cm土壤含水量的相关性方面,仅有苦槠高氮实验组表现出明显的二次方程关系(R2=0.722),而其低、中氮实验组及石栎各实验组均未有明显的相关性;与单因素(温度、含水量)拟合它们与土壤呼吸速率的方程相比,多元回归分析得到的土壤呼吸速率同土壤温度和含水量之间的拟合方程在P=0.05水平上能更好地解释土壤呼吸的变化情况。石栎和苦槠在氮沉降处理下的土壤呼吸温度系数Q10值分别为2.29、1.95、1.59和1.46、1.41、1.76,同对照组2.64和1.78相比,均有明显降低,且两者Q10值的变化分别呈递减和先减小后增大的趋势,表明氮沉降是影响石栎和苦槠土壤CO2通量的一个重要因素。  相似文献   

10.
反应体系中存在的纤维蛋白(fibrin)对尿激酶(UK)、scu-PA以及组织型纤溶酶原激活剂(t-PA)激活纤溶酶原(plasminogen)的反应有不同的作用:UK、t-PA激活plasminogen的反应可被反应体系中存在的fibrin所加强;fibrin对scu-PA激活plasminogen反应的动力学常数无明显影响;但对小分子质量scu-PA与单链抗体的嵌合分子激活plasminogen的反应起明显的抑制作用.为确定反应体系中存在的fibrin对scu-PA的K区插入突变体-InB激活plasminogen反应的影响,测定了在反应体系中存在fibrin的情况下的InB激活plasminogen反应的Kmfibrin以及kcatfibrin.Kmfibrin=4.2 μmol·L-1,远远大于无fibrin时的Km=0.379 μmol·L-1,说明有fibrin存在时突变体InB与天然底物plasminogen的亲和性降低了.kcatfibrin=0.107 s-1,也远远大于无fibrin时kcat=0.0165 s-1,说明有fibrin存在时突变体InB对plasminogen的反应活性增强了.原因可能是:与fibrin结合的plasminogen的构象发生了有利于被纤溶酶原激活剂水解的变化.  相似文献   

11.
蛇毒清胶囊对眼镜蛇咬伤患者血清CK、LDH、AST活性的影响   总被引:2,自引:1,他引:1  
黄彬  曾仲意  曲敬来  林申 《蛇志》2004,16(1):1-3
目的观察眼镜蛇咬伤后血清CK、LDH、AST活性的变化和蛇毒清胶囊对其的影响. 方法眼镜蛇咬伤2h内的患者120例,随机分为2个组,均给予常规治疗,治疗组加服蛇毒清胶囊,均以7天为1个疗程.分别于就诊时、咬伤后24h测定其血清酶学三项指标:CK、LDH、AST的活性,比较2组酶学的变化和临床疗效.结果2组患者在就诊时(伤后2h内)的酶学三项指标尚未出现明显异常,伤后24h三项指标均明显升高,对照组显著高于治疗组,治疗组疗效优于对照组(P<0.05).结论蛇毒清胶囊能抑制眼镜蛇咬伤患者血清CK、LDH、AST的升高,对眼镜蛇咬伤患者组织损伤有防治作用.  相似文献   

12.
Clinical observations have implicated the antiretroviral drug tenofovir with bone density loss during the management of HIV infection. The goal of this study was to investigate the in vitro effects of tenofovir exposure of primary osteoclasts in order to gain insights into the potential mechanisms for the drug-induced bone density loss. We hypothesized that tenofovir may alter the expression of key genes involved in osteoclast function. To test this, primary osteoclasts were exposed to physiologically relevant concentrations of the prodrug tenofovir disoproxil fumarate (TDF), then intensive microarray analysis was done to compare tenofovir-treated versus untreated cells. Specific downregulation of Gnas, Got2 and Snord32a were observed in the TDF-treated cells. The functions of these genes help to explain the basis for tenofovir-associated bone density loss. Our studies represent the first analysis of the effects of tenofovir on osteoclast gene expression and help to explain the basis of tenofovir-associated bone density loss in HIV-infected individuals.  相似文献   

13.
【背景】东湖假单胞菌HYS是本实验室从武汉东湖水域中分离并鉴定的一株高产铁载体细菌,HYS菌株对秀丽隐杆线虫具有较强毒性。前期研究发现HYS中编码精氨酸琥珀酰转移酶基因(argS)的插入突变可导致其对线虫毒性明显减弱。【目的】探究argS基因功能及其如何参与细菌毒性,为后续深入研究HYS菌株的毒性机制提供理论依据。【方法】采用生物信息学比对、遗传分析和生理生化实验确认argS基因的生物学功能及其参与的精氨酸琥珀酰转移酶(arginine succinyltransferase,AST)途径与细菌毒性的关系。【结果】生物信息学比对结果显示argS编码精氨酸琥珀酰转移酶,其蛋白序列与铜绿假单胞菌中精氨酸琥珀酰转移酶β亚基具有高达88%的相似度;缺失argS导致菌株不能利用精氨酸作为唯一碳源进行生长;精氨酸脱羧酶(arginine decarboxylase,ADC)、精氨酸脱氢酶(argininedehydrogenase,ADH)以及精氨酸脱亚胺酶(argininedeiminase,ADI)途径中关键基因缺失菌株均可正常利用精氨酸作为唯一碳源,且对线虫并无明显毒性减弱现象;添加外源精氨酸导致菌株对线虫的减毒效果更加明显,且菌株产铁载体能力显著下降。【结论】东湖假单胞菌HYS中AST途径可以通过影响菌株铁载体合成来影响其对秀丽隐杆线虫的毒性,本研究为深入了解假单胞菌精氨酸代谢和致病性机制提供了新依据。  相似文献   

14.
Sulfotransferase catalyzed sulfation is important in the regulation of different hormones and the metabolism of hydroxyl containing xenobiotics. In the present investigation, we examined the effects of hyperoxia on aryl sulfotransferase IV in rat lungs in vivo. The enzyme activity of aryl sulfotransferase IV increased 3- to 8-fold in >95% O2 treated rat lungs. However, hyperoxic exposure did not change the mRNA and protein levels of aryl sulfotransferase IV in lungs as revealed by Western blot and RT-PCR. This suggests that oxidative regulation occurs at the level of protein modification. The increase of nonprotein soluble thiol and reduced glutathione (GSH)/oxidized glutathione (GSSG) ratios in treated lung cytosols correlated well with the aryl sulfotransferase IV activity increase. In vitro, rat liver cytosol 2-naphthol sulfation activity was activated by GSH and inactivated by GSSG. Our results suggest that Cys residue chemical modification is responsible for the in vivo and in vitro oxidative regulation. The molecular modeling structure of aryl sulfotransferase IV supports this conclusion. Our gel filtration chromatography results demonstrated that neither GSH nor GSSG treatment changed the existing aryl sulfotransferase IV dimer status in cytosol, suggesting that oxidative regulation of aryl sulfotransferase IV is not caused by dimer-monomer status change.  相似文献   

15.
Erythroleukemia disease is caused by over production of malignant blood and immature large number of blood cells enters into peripheral compartment. Biophysical and biochemical changes in plasma and erythrocyte membrane in erythroleukemia treated rats were identified. Our study, leukemia is experimentally exposed in rats were injecting erythroleukemia cells (FLC) (H-2d) intravenously in adult rats and normal control rats were maintained. Significant increase in the activity of blood glucose, proteins levels, aspartate transaminase (AST) and alanine transaminase (ALT) values and significant decrease in haemoglobin (Hb), albumin levels in erythroleukemia treated rats were observed when compared with control rats. Cholesterol and low density liproprotein (LDL) levels increased significantly in erythroleukemia treated rats but triglycerides, high density lipoprotein (HDL) and very low density lipoprotein (VLDL) levels decreased significantly. Levels of red cell membrane cholesterol decreased in erythroleukemia treated rats in comparison with control while levels of phospholipids and proteins increased in erythrocytes of erythroleukemia treated rats. Red blood cell (RBC) and white blood cell (WBC) counts increased significantly and platelet count decreased. C/P (cholesterol/phospholipid) ratio decreased significantly in erythroleukemia treated rats. This study has been undertaken for the first time to investigate the effect of (FLC) (H-2d) erythroleukemia cells (treated) in intravenously in adult rats and normal control rats. Results indicate biophysical and biochemical alterations at molecular level in plasma and erythrocyte membrane.  相似文献   

16.
Frankincense (Gum Olibanum), made from resins of Burseraceae family, grows in Somalia, India and Yemen. Many years ago the oldest doctors used this plant for treatment of many diseases. This study identifies frankincense impact by biochemical analysis and histological examination on rats. In this study, forty male Wister Albino rats weighing 70–100 g were maintained in clean cages. The rats were divided into 2 groups, each group contained 20 rats. Frankincense extract was prepared by heating distilled water (400 ml) to 80 °C and soaking 20 g of herbs for about 60 min. After cooking at room temperature the dose was given orally through special drinking bottles daily. The first group acted as control drinking water. The second group served as treated group and was given frankincense in the drinking water during the whole duration of the experiment. After 15 and 30 days of treatment, the rats were anesthetized with ether, and blood was collected from the livers and kidneys; some biochemical analyses were performed including aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and non-bilirubin, urea, uric acid, and creatinine. Rats were killed by cervical decapitation of livers and kidneys. Each group was divided into 2 parts. The first part was used for the determination of glutathione (GSH), glucose 6 phosphate dehydrogenase (G6PDH), xanthine oxidase (XO), malonyldealdehyde (MDA), nitric oxide (NO), and xanthine oxidase (XO). The second part of livers and kidneys was kept in formalin solution (10%) and stained by Hematoxylin and Eosin (H & E), to be used for histological examination. I demonstrated in the biochemical analysis in the serum, tissue and histological examination, different impact between group (B) and group (A), and that frankincense is not absolutely safe and that precautions must be taken during it’s us as a traditional medicine and that increase the awareness with safety and health hazards of many other traditional medicine is critically needed.  相似文献   

17.
BackgroundNon-alcoholic fatty liver disease (NAFLD) is one of the hepatic disorders which is characterized by increasing fat deposits in the liver. This disorder may lead to elevation the activity of liver enzymes and is associated with obesity, dyslipidemia, hypertension, and type II diabetes. The aim of present study was to investigate the effect of Berberis integerrima extract on NAFLD patients compare to placebo.MethodsThe present clinical trial was performed on 42 NAFLD patients who were randomly divided into two groups. The case group received a capsule (750 mg) containing hydro-alcoholic extract of Berberis integerrima extract every 12 h for 2 months, while the control (placebo) group received a capsule containing cellulose. Baseline characteristics, biochemical factors, antioxidant parameter, functional liver and renal test were evaluated before and after the treatment.ResultsComparison rate of different parameters in case group before and after treatment demonstrated that BMI, cholesterol, triglyceride, LDL-C, fasting blood glucose, liver enzymes and renal parameters such as aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase were significantly decreased while HDL-C, glutathione peroxidase enzyme, and total antioxidant capacity were significantly elevated. The comparison of the mean parameters difference in two groups indicated that cholesterol, triglyceride, LDL-C, fasting blood glucose, liver enzymes and renal factors were significantly decreased, however HDL-C, glutathione peroxidase enzyme, and total antioxidant capacity were significantly increased in case group compared to control group.ConclusionThe study findings revealed that the Berberis integerrima extract could reduce biochemical factors of blood, except HDL-C and increases total antioxidant capacity and glutathione peroxidase enzyme. Therefore, hydro-alcoholic extract of Berberis integerrima may be used as a great supplementary medicine in treating NAFLD.  相似文献   

18.
D-aspartate was used to demonstrate possible sources of excitatory input to the suprachiasmatic nucleus (SCN) in rats. Aspartate (50 mg/kg bodyweight) was orally administrated chronically for 60 days to Wistar rats and 24 h rhythmic patterns of glucose, cholesterol, total protein and aspartate transaminase (AST) were studied under light - dark (LD 12:12 h) cycle. Our results showed acrophase advances in glucose and delays in cholesterol and AST rhythms. Increased mesor and altered amplitude values were found in all rhythms; aspartate levels in the brain were found to be significantly increased in aspartate treated animals. We hypothesised that the altered biochemical rhythms in aspartate treated rats could be due to (1) modulation of neurotransmission in SCN, (2) behavioural rhythms and (3) feeding rhythms.  相似文献   

19.
<正>共轭亚油酸(Conjugated linoleic acid,CLA)是十八碳二烯酸(亚油酸)的同分异构体,其共同特征为两个双键直接通过一个碳-碳单键连接,没有被亚甲基隔开。共轭亚油酸的双  相似文献   

20.
Polychlorinated biphenyls (PCBs) are persistent environmental pollutants. This study compared effects of two PCB mixtures, Aroclors 1221 (A1221) and 1254 (A1254) on serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), urea, creatinine and uric acid in female rats. Histopathological changes in the liver and kidney were also examined. A group of adult Wistar rats served as controls. Groups II and III were subcutaneously injected with A1221 and A1254 at 10 mg/kg every other day for 6 weeks. At the end of this period, all animals were decapitated and blood samples were collected. Serum urea, creatinine, uric acid, ALT, AST and ALP levels were determined. Liver and kidney were collected for histopathological examination. They were fixed in formaldehyde and processed for light microscopy. Both A1221 and 1254 significantly elevated serum ALT (p < 0.05) and AST (p < 0.01) levels compared to the control group. Serum ALP values were significantly increased by A1221 (p < 0.05), but they were unaffected in the A1254 group. Treatment with both A1221 and A1254 significantly increased serum levels of urea (p < 0.05), creatinine (p < 0.01) and uric acid (except in the A1221 group; p < 0.005). Distinct histopathological changes including renal corpuscular atrophy, peritubular vascular congestion and dilated cortical tubules, sinusoidal dilatation, congestion and mononuclear cell infiltration were observed. These findings suggest that PCBs may cause nephrotoxicity and hepatotoxicity in female rats.  相似文献   

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