重组人促红细胞生成素纯化工艺

目的:建立适合大规模生产重组人促红细胞生成素(rhEPO)的纯化工艺.方法:采用无血清培养分泌rhEPO工程细胞株,收集培养基上清,经过滤、染料亲和层析,离子交换层析、分子筛层析纯化.结果:所纯化的rhEPO纯度达99%以上,比活(包括体内、体外)为2.0×105IU/mg以上,活性回收率为40%,相对分子量为38000左右,等电点为3.7～4.2.结论:该纯化工艺简便、重复性好、产品质量高,适合大规模生产.     

重组人促红细胞生成素治疗肿瘤相关性贫血

综述了肿瘤相关性贫血的病因及重组人促红细胞生成素在治疗中的意义。     

重组人促红细胞生成素鼻腔给药对大鼠慢性脑缺血的保护作用

目的：观察重组人促红细胞生成素（rhE-PO）鼻腔给药对大鼠慢性缺血性脑损伤的保护作用。方法：以大脑中动脉栓塞诱导大鼠局灶性脑缺血,手术当日术前30 min和术后2 h各给药一次,第2天起隔天一次,持续到术后14 d。缺血后1、3、7、14、21、283、5 d进行神经症状评分和斜板试验,缺血后35 d计算脑梗死体积和缺血周边区神经元密度。结果：rhEPO鼻腔给药（24、48 U）、依达拉奉和rhEPO腹腔给药（5000 U/kg）能改善缺血后神经症状评分,提高斜板角度,降低脑梗死体积,增加缺血侧周边区存活神经元密度。结论：rhEPO鼻腔给药对大鼠慢性缺血性脑损伤具有保护作用,其有效剂量范围较腹腔给药低。     

重组人促红细胞生成素生产工艺研究

采用Bellco公司转瓶机,用无血清培养基培养分泌RhEPO的工程细胞株CHOEPO-2.所收集的上清液预处理后经染料亲和层析-离子交换层析-分子筛层析纯化后,所得EPO纯度达99%以上,比活性大于1.5×105IU/mg,整个纯化全过程的EPO体内活性回收率为45%.所纯化的EPO分子量为37kD.分析表明,所纯化的EPO可与抗EPO的单克隆抗体特异性结合.该工艺纯化路线简单,重复性好,生产成本低,产品活性高,适合大规模生产重组人促红细胞生成素.     

重组人促红细胞生成素对尿毒症血清瘦素影响

目的观察重组人促红细胞生成素(rhuEPO)对维持性血液透析(MHD)的尿毒症患者血清瘦素水平的影响及其意义。方法38例行MHD的慢性肾衰竭(CRF)患者分为不用rhuEPO治疗组和使用rhuEPO治疗组;使用rhuEPO治疗的又分三组(2周组,4周组,8周组),采用放射免疫分析法测定血清瘦素水平。结果使用rhuEPO治疗8周后,血清瘦素水平明显低于治疗前及未用rhuEPO治疗组(P<0.05)。结论MHD患者的血清瘦素水平显著高于正常人群,注射rhuEPO后能显著降低患者血清瘦素水平,但不能降至正常人群的水平。     

重组人促红细胞生成素防治早产儿贫血的疗效观察

目的:探讨重组人促红细胞生成素(rHu-Epo)防治早产儿贫血的疗效。方法:将60例早产儿随机分为治疗组和对照组各30例,治疗组出生第三天即rHu-Epo250IU/(kg·d),隔日1次,每周3次皮下注射,共6周;对照组未用rHu-Epo。两组早产儿自出生第三天起口服铁剂[元素铁5mg/(kg·d)]。结果:治疗组网织红细胞及血红蛋白明显高于对照组(P<0.01);治疗组输血率与对照组比较明显减少;治疗组体重增长速率明显高于对照组。结论:早期应用大剂量rHu-Epo能有效地防治早产儿贫血,减少输血,体重增长快,且用药安全,无明显不良反应。     

重组人促红细胞生成素引起纯红细胞再生障碍性贫血的研究综述

随着重组人促红细胞生成素在临床的广泛应用，由其引起的不良反应一纯红细胞再生障碍性贫血也越来越受到人们重视。本文重点阐述重组人促红细胞生成素引起的纯红细胞再生障碍性贫血的发病情况，影响重组人促红细胞生成素免疫原性的可能原因及危险因子，以及诊断治疗措施.     

重组人促红细胞生成素联合铁剂治疗早产儿贫血效果观察

目的观察重组人促红细胞生成素（rhEPO）联合铁剂治疗早产儿贫血的效果。方法148例贫血早产儿按入院时间单双号分为对照组68例与观察组80例。对照组给予口服蛋白琥珀酸亚铁剂治疗,观察组在铁剂治疗基础上加用rhEPO治疗;治疗期间如果Hb≤80g／L或Hb≤100g／L伴贫血表现,予以输血治疗。两组均于治疗前后复查RBC、Hb和HCT。结果两组治疗前Hb、RBC、HCT水平接近,治疗后各项指标均有升高;但观察组高于对照组,差异有统计学意义。对照组接受输血治疗的百分率高于观察组,差异有统计学意义。结论在一定剂量下rhEPO联合铁剂治疗早产儿贫血,用药安全,疗效显著,可减少输血。     

重组人促红细胞生成素的临床应用

促红细胞生成素(EPO)是由肾脏分泌的一种活性糖蛋白,能刺激骨髓红系造血母细胞的增殖和分化。临床研究显示,EPO除可用于慢性肾性贫血治疗外,对多种疾病如获得性免疫缺陷综合征、肿瘤、充血性心力衰竭、危重疾病以及丙型肝炎病毒感染等疾病所致贫血亦有较好的疗效。本文就该药的作用机制、临床研究、不良反应及注意事项进行简要阐述。     

我院重组人促红细胞生成素合理性应用评价分析

目的:了解我院重组人促红细胞生成素(rhEPO)的使用情况,为临床合理用药提供参考。方法:采用回顾性的分析方法,收集我院2019年7–12月使用rhEPO的住院患者信息,共计428例,对患者使用rhEPO的适应证、用法用量、治疗前后血红蛋白水平、合并用药和不良反应等进行统计分析。结果:428例患者中肾性贫血202例(47.20%)、肿瘤化疗引起的贫血132例(30.84%)和外科围手术期的红细胞动员23例(5.37%),合并使用铁剂的194例(45.33%)。存在的主要问题有适应证不适宜、用药时机不适宜和未根据治疗后血红蛋白水平调整剂量等。结论:rhEPO在我院的应用仍存在一定问题,需要进一步规范用药,确保患者用药安全有效。     

汉防己甲素对大鼠门静脉高压性胃病的保护作用

目的观察汉防己甲素(Tet)对大鼠门静脉高压性胃病(PHG)的保护作用。方法采用四氯化碳制备肝硬化门静脉高压性胃病大鼠模型。分别用Tet 15、30 mg/kg,连续给药14 d。测定生化指标[丙氨酸转氨酸(ALT)、Ⅲ型前胶原(PCⅢ)、NO、前列环素代谢产物(6-keto-PGF1α)]同时测定胃黏膜下血管面积并进行图像分析及进行组织学观察。结果Tet可降低血中ALT、PCⅢ、NO、6-keto-PGF1α含量,使黏膜下血流量减少、黏膜血流量相对增加,尤以Tet 30 mg/kg作用显著(P<0.05)。结论Tet对门静脉高压性胃病有保护作用。     

Early detection of renal injury using urinary vanin‐1 in rats with experimental colitis

Renal complications are often detected in patients with inflammatory bowel disease (IBD). Because conventional markers such as serum creatinine and β2‐microglobulin are not sensitive and/or specific, a new renal biomarker is needed. We have recently identified urinary vanin‐1 as an early biomarker for the detection of nephrotoxicant‐induced renal injury. In this study, we compared the usefulness of urinary vanin‐1 with other newly developed biomarkers [urinary monocyte chemoattractant protein‐1 (MCP‐1), kidney injury molecule‐1 (Kim‐1) and N‐acetyl‐beta‐D‐glucosaminidase (NAG)] for the detection of renal complications in rats with experimental colitis. On day 2 after intracolonic injection of 2,4,6‐trinitrobenzene sulfonic acid (TNBS), male Wistar rats developed not only colitis, but histologically evident renal injury. Urinary vanin‐1 started to elevate on day 1, whereas serum creatinine and urinary excretions of glucose, total protein, albumin, Kim‐1, MCP‐1 and NAG significantly increased only on day 2. The mRNA expressions of vanin‐1 and Kim‐1 significantly increased in the kidney, but not in the colon. In addition, vanin‐1 did not appear in the blood. On the other hand, colonic mRNA expression and the serum concentration of MCP‐1 were significantly higher in the TNBS‐treated rats than in the control animals. These results suggest that, in contrast to MCP‐1, urinary vanin‐1 and Kim‐1 mainly originated from the kidney rather than the colon in this model. Compared with Kim‐1 and MCP‐1, vanin‐1 might be an earlier biomarker for the detection of renal injury in rats with experimental colitis. Copyright © 2013 John Wiley & Sons, Ltd.     

Susceptibility of newborn rats to hepatotoxicity of 1,3-dibromopropane and 1,1,2,2-tetrabromoethane, compared with young rats

Newborn rat studies were conducted with oral administration of 1,3-dibromopropane (DBP) and 1,1,2,2-tetrabromoethane (TBE) from postnatal Days 4 to 21 to allow comparison of NOAELs and unequivocally toxic levels with those from 28-day young rat studies starting at 5-6 weeks of age. The unequivocally toxic level was estimated by our specified criteria, requiring simultaneous change of organ weights, histopathology, some biochemical parameters and body weights, because in this study only hypertrophy of hepatocytes was observed as a major histopathological change. DBP caused centrilobular hypertrophy of hepatocytes with alteration in biochemical parameters, as well as lowering of body weights, regardless of sex, in both newborn and young rats. NOAELs and unequivocally toxic levels were considered to be 50 and 150 mg/kg/day in newborn rats and 10 and 250 mg/kg/day in young rats, respectively. In the newborn rat study of TBE, some hepatic effects observed at the top dose of 50 mg/kg were not considered adverse because of the lack of histopathological changes. Significant lowering of body weight was noted at 200 mg/kg in the dose-finding study but histopathological data were not available. In the young rat study, there was no definite toxicity at 6 mg/kg and hypertrophic changes in liver and thyroids without body weight change occurred at 200 mg/kg. There were no clear sex differences in both the newborn and young rat studies. NOAELs were considered to be 50 and 6 mg/kg/day in newborn and young rats, respectively, but unequivocally toxic levels for both rats could not be estimated. Abnormalities of external and sexual development and reflex ontogeny in the newborn were not observed with either chemical. Based on these results, it can be concluded that the target organ of DBP and TBE is the liver in both newborn and young rats, and that while the doses at which toxic signs began to appear are higher in newborn rats, those causing clear toxicity may be paradoxically lower in the newborn case.     

实验性溃疡性结肠炎大鼠模型的建立

目的采用三硝基苯磺酸（TNBS）诱导大鼠溃疡性结肠炎动物模型,探讨与人溃疡性结肠炎（UC）的相似程度。方法 TNBS/乙醇法制作溃疡性结肠炎动物模型,分别于造模后1、3、7周3个时间段动态观察模型鼠的一般状态、结肠黏膜大体和病理及电镜变化。同时检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素-2（IL-2）、白细胞介素-4（IL-4）含量。结果大鼠的一般情况、大体病理及组织学病理及电镜变化均与人类溃疡性结肠炎类似,并且血清TNF-α、IL-2、IL-4含量变化亦与人类溃疡性结肠炎的变化基本一致。结论 TNBS可以成功诱导大鼠溃疡性结肠炎动物模型,一般情况、大体与病理表现与人UC的相似度高,有急、慢性转变过程等,而且是免疫学模型,可用于UC免疫学方面的研究。     

己酮可可碱对新生大鼠缺血缺氧性脑病的干预作用

目的 :观察己酮可可碱 (pentoxifylline,PTX )对新生大鼠缺血缺氧性脑病 (hypoxic- ischemicencephalopathy,HIE)模型的干预作用。方法 :建立新生大鼠 HIE模型 ,72只出生 7d的 Wistar大鼠随机分为假手术组、HIE模型组和 PTX组 ,于缺血缺氧后 72 h取左脑 ,测定脑组织水含量、大脑皮层神经细胞内游离钙离子浓度、超氧化物歧化酶 (SOD)、丙二醛 (MDA)、一氧化氮合酶 (NOS)及一氧化氮 (NO)。结果 :与 HIE模型组相比 ,PTX组脑组织含水量、游离钙离子浓度和 MDA、NOS以及 NO水平显著降低 (P <0 .0 5、0 .0 1、0 .0 5、0 .0 5、0 .0 5 ) ,SOD水平显著升高 (P <0 .0 5 )。结论 :PTX对新生大鼠缺血缺氧后脑组织有保护作用 ,其机制可能与 PTX改善能量代谢及减少局部细胞毒性物质有关。     

Higher susceptibility of newborn than young rats to 3-methylphenol

To determine susceptibility of infants to 3-methylphenol, a repeated dose toxicity study was conducted with oral administration to newborn and young rats. In an 18-day newborn study from postnatal days 4 to 21 at doses of 30, 100 and 300 mg/kg/day, various clinical signs including deep respiration, hypersensitivity on handling and tremors under contact stimulus, and depressed body weight gain were observed at 300 mg/kg. At 100 mg/kg, hypersensitivity and tremors were also noted in a small number of males only on single days during the dosing period. No adverse effects were observed in the 30 mg/kg group. There were no abnormalities of physical development, sexual maturation and reflex ontogeny. The no observed adverse effect level (NOAEL) for newborn rats was considered to be 30 mg/kg/day and the unequivocally toxic level 300 mg/kg/day. In a 28-day study starting at 5 weeks of age, clinical signs and depression of body weight gain, as observed in the newborn rats, appeared in both sexes at 1000 mg/kg but not 300 mg/kg. The NOAEL and the unequivocally toxic level were 300 mg/kg/day and 1,000 mg/kg/day, respectively. From these results, newborn rats were concluded to be 3 to 10 times more susceptible to 3-methylphenol than young rats. However, the realistic no adverse effect dose for the newborn must be slightly lower than 100 mg/kg/day, at which the toxicity incidence was very low, rather than 30 mg/kg/day. Based on this speculation and the equal toxicity at unequivocally toxic levels, the differences in the susceptibility to 3-methylphenol could be concluded to be 3 to 4 times. This is consistent with the results of our previous comparative studies on 4-nitrophenol, 2,4-dinitrophenol and 3-aminophenol, which showed 2 to 4 times differences in the susceptibility between newborn and young rats.     

Molecular and toxicologic research in newborn hypospadiac male rats following in utero exposure to di-n-butyl phthalate (DBP)

The objective of this study was to first evaluate the developmental abnormalities and carry out the molecular analysis of external genitalia in newborn hypospadiac male rats induced by maternal exposure to di-n-butyl phthalate (DBP). Timed-pregnant rats were given DBP by gastric intubation at dose of 750 mg/kg body weight (bw)/day from gestation day (GD) 14 to GD18 to establish a hypospadiac rat model. The incidence of hypospadias was 46.67% in male offsprings. On postnatal day (PND) 7, at the newborn stage, decreased body weight and anogenital distance (AGD)/body weight ratio were observed in newborn hypospadiac male rats. The general image and transverse serial histological analysis of genitalia of newborn hypospadiac male rats confirmed the malformation. Autopsy analysis revealed development of reproductive organs (testes, genital tubercle (GT)), hollow organs (stomach, bladder), and solid organs (brain, heart, liver, spleen, lung, kidney, pancreas) in newborn hypospadiac male rats affected by DBP. Moreover, significantly decreased gene expression of important signaling molecules necessary for GT formation including sonic hedgehog signaling molecules (Shh and Ptched 1), bone morphogenetic proteins signaling molecules (Bmp4 and Bmp7), fibroblast growth factor signaling molecules (Fgf8, Fgf10 and Fgfr2), and the transforming growth factor-β superfamily signaling molecules (TGF-β1 and TGF-β receptor III) were observed, for the first time, in the GT of newborn hypospadias induced by DBP. These results showed that the reproductive system and development conditions of newborn hypospadiac rats were damaged by DBP. These disturbed signaling pathways which orchestrating genital development might play an important role in the toxic process of DBP induced hypospadias.     

川芎嗪对大鼠急性坏死性胰腺炎肺损伤的保护作用

目的探讨微循环障碍在急性坏死性胰腺炎肺损伤中的作用,同时观察川芎嗪对胰腺炎肺损伤的干预效应。方法SD大鼠192只,随机均分为对照组(C)、胰腺炎组(P)、川芎嗪治疗组(T)。大鼠胰腺被膜下均匀注射5%牛磺胆酸钠4ml·kg-1,制作急性坏死性胰腺炎模型,C组胰腺被膜下注射等量生理盐水,T组经股静脉注射0.6%川芎嗪10ml·kg-1。每组中8只SD大鼠采用放射性生物微球技术,在制模后0.5、2.0、6.0、12.0h分别测定肺的血流量,另8只用于胰腺、肺的病理评分,以肺组织的过氧化酶(MPO)活性、肺湿/干重比作为衡量肺损伤的指标。结果P、C组相比,各时相肺脏的血流量均明显降低(P<0.05);各时相病理改变明显加重,时间越长,损害越重,其程度与血流量相关;肺干/湿明显加重,组织MPO活性明显增加(P<0.05或P<0.01)。T、P组相比,各时相肺的血流量较P组均明显降低(P<0.01);各时相病理改变明显减轻,肺干/湿无明显变化(P>0.05),组织MPO活性明显下降(P<0.05)。急性坏死性胰腺炎(ANP)时,肺组织血流量与肺湿/干重比率、肺组织MPO活性、肺组织损伤程度呈显著负相关。结论微循环障碍在急性坏死性胰腺炎的肺组织损伤中起着重要的作用。川芎嗪可以改善微循环,减轻胰腺及肺损害。