应用Westgard多规则理论评估万古霉素血药浓度监测的室内质控

目的:探讨自制万古霉素质控血浆在血药浓度监测中的稳定性,应用Westgard多规则理论评估万古霉素血药浓度监测(TDM)的室内质量控制。方法:取万古霉素标准品12.20mg,50%甲醇溶解后加入空白血浆基质液,制备质量浓度分别为4.88,24.40,40.02μg·mL-1的万古霉素质控血浆,在连续25个工作日内对75个低、中、高质控浓度进行监测,绘制LeveyJennings质控图和Z-分数质控图,采用Westgard多规则进行质量评估。结果:万古霉素低、中、高室内质控的日内和日间RSD分别为0.59%～0.86%、3.90%～5.67%,回收率分别为98.90%～102.11%、102.93%～106.91%,符合《中国药典》2015年版(四部)生物样品测定的要求;绘制的质控图显示低、中、高浓度质控血浆均未出现失控。结论:自制万古霉素质控血浆操作方法简单、成本低、结果可靠,可以作为实验室日常质量控制;利用室内质控评价体系对TDM监测数据进行评估,可以保证血药浓度测定的准确性、可靠性,从而为临床合理化给药提供准确信息。     

均相酶扩大免疫分析法监测地高辛血浓度的质量控制与评价

目的：评价和控制均相酶扩大免疫分析法（Emit）监测患者地高辛血浓度的质量。方法：以标准质控为样本,进行预防性质量控制和室内质量控制研究。对2008年血药浓度监测中随行质控样本的测定值进行统计学分析,做回顾性的质量控制研究,同时建立新的质控规则。结果：Emit法在本单位实验条件下,地高辛低（L）、中（M）、高（H）三种浓度质控日内、日间RSD在2．19％～6．30％之间,平均回收率在97．92％-104．19％之间,符合《中国药典》生物样品检测规定。2008年随行质控的RSD,L、M、H分别为14．45％、14．11％和11．30％,适合本单位的质控规则为125／135／225／415／7tr。结论：Emit法监测地高辛血药浓度是一种较理想的测定方法,但是该法的影响因素较多,特别是温度,必须做好质量控制。     

用酶增强免疫分析法监测他克莫司血药浓度的质控评估

目的：评价酶增强免疫分析法（EMIT）监测患者全血中他克莫司浓度的质量,建立并改进他克莫司血药浓度监测的质量控制方法。方法：以标准质控为样本,进行预防性质量控制和室内质量控制研究。对2008年血药浓度监测中随行质控样本的测定值做回顾性研究并进行统计学分析,建立新的质控规则。结果：他克莫司低、中、高浓度（4.3、8.9、18.0 ng/ml）2009年质控样本的日内、日间RSD为1.8%-11.2%,平均回收率为90.5%-114.0%,符合《中华人民共和国药典》生物样品测定的要求。2008年质控样本的低、中、高浓度（3.8、7.5、15.0 ng/ml）的随行质控RSD分别为21.8%、14.2%和15.5%,适合本单位的质控规则为12S/13S/22S/41S/7tr。结论：EMIT法准确度和精密度良好,是一种较理想的他克莫司血药浓度测定方法,但影响因素较多,特别是温度对其影响较大,因此必须做好质量控制。     

荧光偏振免疫法监测环孢素A血药浓度观察与分析

吴璐刘晓慧目的观察荧光偏振免疫（FPIA）法测定环孢素A（CsA）血药浓度的可行性以及CsA监测质量控制的必要性。方法对20例环孢素A服用患者的血液样本使用荧光偏振免疫法检测血药浓度,观察日内差、日间差、回收率及相对百分偏差等数据。结果测得的低、中、高质控样本的回收率均高于100%,所测定质控样本RSD〈6%,符合中国药典要求。结论 FPIA法在测定环孢素A血药浓度具有良好的准确度和精密度,符合临床检测要求,适合在建立合理质控体系下开展治疗药物监测。     

酶放大免疫分析法监测万古霉素血药浓度的质控评估

目的：评估酶放大免疫分析法（Emit）监测患者全血中万古霉素浓度的质量,建立并改进本单位实验室条件下万古霉素药物浓度监测的质量控制方法.方法：以标准质控为样本,进行预防性质量控制和室内质量控制研究.对2012年9月至2013年4月万古霉素血药浓度监测中随行质控样本的测定值进行统计学分析,做回顾性的质量控制研究,同时建立新的质控规则.结果：Emit法在本单位实验条件下,万古霉素低（L）、中（M）、高（H）三种浓度质控日内差、日间差的相对标准偏差（RSD）在0.6％～2.1％,平均回收率在105.8％～114.3％,符合《中国药典》生物样品监测规定.随行质控L、M、H的RSD分别为0.7％、2.2％和2.5％,适合本单位的质控规则为12S/13S/32S/51s/7tr.结论：Emit法监测万古霉素血药浓度具有较好的精密度、准确度,是一种操作简便、可行的测定方法,但是必须做好质量控制,制订规范,确保测定结果准确、可靠.     

微粒子酶免疫分析法测他可莫司血药浓度的稳定性

目的:通过对微粒子酶免疫分析法(MEIA)测他可莫司(FK506)血药浓度的稳定性进行评价,为临床合理用药提供依据。方法:采用MEIA法测移植患者术后全血FK506浓度,运用统计学方法对其稳定性进行评价。结果:MEIA法测定FK506血药浓度日内差、日间差和两年中随行质控的变异系数分别为小于11.09%,8.3%,12.22%,回收率分别为101.64%~104.55%,101.45%~109.6%,103.18%~108.20%,均符合中国药典生物样品检测规定。结论:MEIA法在长期的临床应用中稳定、准确和可靠。     

HPLC法测定天元克痛方主要成分的研究

目的:建立天元克痛方的主要成分的含量测定方法.方法:应用HPLC法测定对方中的主要成分延胡索乙素和东莨菪碱进行含量测定,考察方法线形范围、精密度及回收率等.结果:采用HPLC法对延胡索乙素进行测定,标准曲线为y=0.0774x+1.1044;r=0.9999,高、中、低三个浓度的日间精密度分别为:0.15%、0.3%、0.4%;日内精密度分别为:0.43%、0.69%、0.58%;回收率分别为:100.8%、101.2%、104.5%.HPLC法对东莨菪碱进行含量测定,标准曲线为y=0.0667x-0.6352;r=0.9999,高、中、低三个浓度日内精密度分别为:3.04%、1.66%、0.46%;回收率分别为:104.34%、95.05%、95.79%.结论:采用HPLC法对天元克痛方中主要成分延胡索乙素和东莨菪碱进行含量测定,方法灵敏可靠、重现性好.     

精液常规检查850例结果分析及研究

目的通过对本地区男性精液的常规检测结果的参数进行分析研究,了解本地区男性精液的质量情况。方法按照2014年《WHO-5人类精液检查和处理实验室手册》(第五版)中文修订版,应用计算机辅助精子分析仪(CASA),以MACRO计数板做全套相关参数分析,对精子总数、精子密度、精子活动力、精液中有白细胞的感染等多项参数进行统计分析。结果850例精液标本常规检查中,精子密度<15×106/ml占标本总数的12.9%,其中密度为0者占标本数的2.4%;精液量为0.1~9.4 ml;精液量<1.5 ml占标本数的11.8%,精液量>5 ml 150例,占标本总数的17.6%;精子总数(精子密度与精液量的乘积)<39×106/次射精占标本总数的12.9%,精子总数>39×106/次射精占标本总数的87.1%;检测到精液中有白细胞>1×106/ml占15.3%。精子总数变化和精液中有白细胞的感染2组有统计学的差异。排精1 h后不液化或不完全液化有132例,占总数的15.5%,排精后60 min内自行液化718例,占总数的74.5%,2组有统计学差异。结论850例男性精液质量分析中,其中少精子症、感染病例略微增多,其余参数变化不大。精子总数少、精液液化不全或完全不液化、白细胞感染率高。     

酶放大免疫分析法测定环孢霉素和普乐可复的血药浓度

目的 分析环孢霉素A(CSA)和普乐可复(FK506)药物浓度监测(TDM)的室内质控结果.方法 采用酶放大免疫分析法(EMIT)测定CSA和FK506血药浓度的高、低质控品,绘制质控图,并对2007年的随行质控结果进行统计学分析.结果 EMIT法测定CSA和FK506的日内RSD分别为5.89%～9.41%、8.34%～9.30%,日内方法回收率分别为92.8%～98.3%、89.1%～89.5%;日间RSD分别为8.51%～9.74%、8.46%～8.78%,方法回收率分别为91.0%～102.7%、89.6%～99.1%.2007年随行质控测定值的RSD为10.02%～14.41%,符合<中国药典>对生物样品的测定要求.结论 EMIT法测定CSA和FK506血药浓度准确、稳定性良好,适于临床开展TDM,临床应用时应建立合理的质控体系.     

男性不育患者的精子形态与精子密度、活力分析

目的 探讨精子形态与精子密度、活力的关系.方法 492例男性不育患者,采用计算机辅助精液分析方法(CASA)检测精子密度、活力,采用巴氏染色进行精子形态分析.根据密度分为<5×106/ml、5～20×106/ml、>20×106/ml 3组;根据精子活力分为<10%、10%～30%、31%～50%、>50% 4组.比较个组精子形态正常率的差异.结果 随着精子密度、活力的降低,组间比较差异有统计学意义(P<0.05).结论 精子形态与精子密度和活率密切相关.     

吸入麻醉剂浓度及作用时间对人离体精子活率的影响

目的:研究吸入麻醉剂浓度及作用时间对人离体精子活率的影响。方法:选择人精液20份,上游法优化处理后随机分为异氟醚实验组和七氟醚实验组,各10份。分别观测5个时间点(0.5 h、1 h、2 h3、h4、h)及5个浓度水平(0、1.4%、2.8%、4.2%、5.6%)异氟醚和相似浓度七氟醚对精子活率的影响,精子运动功能采用计算机辅助精子分析系统分析。结果:1.4%~5.6%异氟醚作用于精子0.5~4 h后,精子活率显著升高,其变化趋势符合S型曲线,相同浓度异氟醚组,精子活率随放置时间的延长成直线下降。类似浓度七氟醚对精子活率无显著影响。结论:临床浓度异氟醚作用于人离体精子,可以显著升高精子活率。类似浓度七氟醚对精子活率无显著影响。     

Sperm abnormalities and histopathological changes in the testes in Crj:CD(SD)IGS rats

In this study, morphological examination and computer-assisted sperm analysis (CASA) of epididymal spermatozoa in non-treated Crj:CD(SD)IGS rats were performed, and the relationship between the data obtained and the retention of step 19 spermatids in Stage IX to XI seminiferous tubules was examined. Retention of step 19 spermatids in Stage IX to XI seminiferous tubules was observed in all 50 untreated males, and the incidence ranged from 3.3% to 100%. Eighteen animals showed a high incidence of retention (74.7 +/- 14.2%, HIR for short), and the others showed a low incidence (24.9 +/- 11.0%, LIR for short). Although the incidence of retention in Stage X and XI seminiferous tubules was very low in LIR males, it was high in HIR males (1.8 +/- 3.0% vs 58.6 +/- 23.2%). Morphological abnormalities of sperms in the caudal region of the epididymis, mainly amorphous head and no head, were more frequently observed in HIR males than in LIR males (36.2 +/- 28.5% vs 1.8 +/- 1.2%). Sperm analysis also revealed some differences between HIR and LIR males: sperm motility in HIR males was severely lower than that in LIR males, and sperm velocity, beat/cross frequency and amplitude of lateral head displacement in HIR males were lower than the corresponding values in LIR males. In summation, retention of step 19 spermatids frequently occurred in the non-treated Crj:CD(SD)IGS males, and a relationship between the retention of these spermatids and sperm abnormalities, such as morphologically abnormal sperms, low motility and other items revealed by sperm analysis (CASA), was suggested.     

MAR-ASAb计数板计数法与玻片法的对照研究

目的通过比较自拟的计数板计数法和厂商提供的玻片法两种MAR方法,验证计数板计数法的可靠性和实用性。方法借助computer-assisted sperm analysis(CASA)技术定量分析MAR法抗精子抗体,收集男性不育症450例,采用配对试验,分析两种方法抗精子抗体检出的情况。结果玻片法和计数板计数法,阳性率分别为10%和10.44%,灵敏度和特异性分别为93.3%和98.8%,阳性标本结合率分别为头部42.22%和42.55%、中段20%和21.28%、尾部37.78%和36.17%,以头部结合多见。结论MAR-ASAb计数板计数法和玻片法检测抗精子抗体配对比较差异无显著性(P>0.05),计数板计数法能更清晰的显示操作界面,并可量化分析抗精子抗体阳性精子的运动参数、精子密度和结合于精子头部、中段、尾部的阳性精子所占的百分率和密度。     

Effects of progesterone on sperm motility in fathead minnow (Pimephales promelas)

The steroid hormone progesterone (P4) is found at relatively high concentrations (～300 ng/L) in association with concentrated animal feeding operations (CAFOs). In an effort to better understand the potential endocrine disrupting effects of P4 in male fish, computer assisted sperm analysis (CASA) was used to evaluate the effects of this steroid on sperm motility in the fathead minnow (Pimephales promelas). The rationale for focusing on sperm motility is that certain progestins have been shown to bind to surface membrane receptors on fish spermatozoa and increase sperm swimming velocity. It was hypothesized, therefore, that sperm swimming velocity might be a useful indicator of progestin exposure in fish. Adult male fathead minnows (ages 6-12 months) were exposed to environmentally relevant doses of P4, both longer-term (1 week, in vivo exposure) and short-term (minutes, in vitro exposure). Sperm were then video recorded and analyzed by CASA. When fathead minnows were continuously exposed for 1 week to low levels of progesterone in vivo there was a significant dose-dependent reduction in sperm motility. There was no effect of short-term P4 exposure on fathead minnow sperm swimming characteristics. Additional research is required to elucidate the mechanism by which progesterone alters sperm swimming in the fathead minnow. With further validation, the fathead minnow sperm motility assay may be a useful tool to rapidly screen for endocrine disrupting chemicals in the aquatic environment.     

Reproductive performance, blood testosterone, lipid peroxidation and seminal plasma biochemistry of rabbits as affected by feeding Acacia saligna under subtropical conditions.

Thirty-two New Zealand White growing rabbits (eight-week old) were used to determine the effect of feeding acacia-based diets on semen characteristics, plasma testosterone, free radicals, seminal plasma enzymes and lipids. Rabbits were randomly assigned to four equal groups. The first group (control) was fed a basal diet only, and the other three groups were fed other three diets, as follows: 80% of basal diet+20% of acacia leaves (low), 60% of basal diet+40% of acacia leaves (medium) and 40% of basal diet+60% of acacia leaves (high), respectively for 32 week. Semen samples were collected throughout the last 12 week of the experimental period. Rabbits fed on different levels of Acacia showed no significant changes in libido (reaction time), ejaculate volume, sperm concentration, packed sperm volume and initial hydrogen ion concentration compared to control. However, low and/or medium levels of Acacia caused significant (P<0.05) increase in total sperm output (TSO), sperm motility (%), total motile sperm per ejaculate (TMS), normal sperm, total functional sperm fraction (TFSF), semen initial fructose, live sperm and plasma testosterone. On the other hand, high level of Acacia did not show any significant change in TSO, sperm motility (%), TMS, initial fructose, TFSF or testosterone, while live and normal sperm decreased. All levels of Acacia caused a significant decrease in the concentrations of thiobarbituric acid-reactive substances (TBARS) and an increase in the activity of glutathione S-transferease. The activities of lactate dehydrogenase (LDH), aminotransferases and phosphatases were significantly increased in seminal plasma of animals fed low or medium levels of Acacia. Seminal plasma total lipid, triglyceride, low density lipoprotein and free fatty acids were significantly (P<0.05) decreased in low or medium levels of Acacia. On the other hand, total cholesterol, percentage cholesterol (out of total lipids) and high density lipoprotein were significantly (P<0.05) increased in seminal plasma of rabbits fed on low or medium levels of Acacia. High level of Acacia did not cause any changes in the previous parameters. It can be concluded that up to 40% Acacia leaves could be used successfully and safely in the diet of rabbits without adversely affecting their reproductive performance under subtropical conditions.     

苯扎溴胺对精子运动能力的影响

目的：运用计算机辅助精子质量检测系统测定表面活性剂苯扎溴胺对精子运动的影响。方法：新鲜精液液化后，经离心洗涤与不同浓度的苯扎溴胺混匀，30s后滴片，置计算机辅助精子质量检测系统分析。结果：经苯扎溴胺处理的精子的活率和精子活力随着药物剂量的增加而逐渐降低，至0．25mg/ml浓度时，无活动精子，与对照组相比，差异有显著性（P＜0．01）；其直线运动速率VSL、曲线运动速率VCL及平均路径速率VAP，随剂量的增加逐渐降低，且各剂量组与对照组相比，差异均有显著性（P＜0．01）；侧摆幅度ALH与对照组相比，差异无显著性，P＞0．05；而用药组精子的直线笥LIN和前向性STR百分率均低于对照组，但是参数的变化并不规则。结论：苯扎溴胺可显著地影响精子的运动能力，从而使受精能力下降，作为外用避孕药使用是可行的。     

Exposure to bisphenol A (BPA) in Wistar rats reduces sperm quality with disruption of ERK signal pathway

Bisphenol A (BPA) is an estrogenic environmental toxin widely used in the production of plastics and ubiquitous human exposure to this chemical has been proposed to be a potential risk to human health. Exposure to BPA can negatively impact sperm quality. However, the mechanism remains largely unknown. The objectives of this study were to assess the role of BPA on sperm quality and explore the possible mechanisms. The Wistar male rats (aged 28 days) were administered BPA by oral gavage for 28 days at dose of 50, 100 and 200?mg/kg/day; meanwhile, the negative control with corn oil (0?mg/kg/day BPA) and positive control with E2 at the dose of 100?μg/kg/day. The sperm density, sperm activity and sperm survival rate were analyzed byCASA system, and the sperm abnormality rate was analyzed by improved Papanicolaou stained. The protein expression levels of Src/p-Src, ERK1/2, p-ERK1/2 and CREB/p-CREB were detected by Western bolt. The results showed that the body weight gain, testes weight, testis coefficient, sperm density, sperm activity, sperm survival rate and protein expression levels of p-ERK1, p-ERK2 and p-CREB decreased, but the sperm abnormality rate increased with increasing BPA concentrations. There were positive correlations between sperm density, sperm activity and sperm survival rate with protein expression levels of p-ERK1, p-ERK2 and p-CREB, and negative correlations between sperm abnormality rate with the protein expression levels of p-ERK1, p-ERK2 and p-CREB. Results from the structural equation model demonstrated that BPA retained a significant negative effect to p-ERK, whereas p-ERK retained a significant positive effect to sperm quality and acted as the mediate variable. This study provides a novel insight regarding the potential role of p-ERK1 and p-ERK2 protein kinase on reproductive toxicity of BPA. The adverse effects of BPA on adult male sperm quality may be through the induction of the disruption of ERK signal pathway. However, additional research is needed to confirm our findings and to further test the suggested potential mechanisms.     

Variability in measures of reproductive success in laboratory-kept colonies of zebrafish and implications for studies addressing population-level effects of environmental chemicals

Laboratory tests that quantify reproductive success using model fish species are used to investigate for population-level effects of endocrine disrupting chemicals (EDCs) and other chemicals discharged into the environment. Even for the zebrafish (Danio rerio), however, one of the most widely used laboratory models, surprisingly little is known about the normal variability in measures of reproductive success and this information is crucial for robust test design. In this study, the dynamics of breeding and inherent variability in egg output/viability and sperm quality were characterized among individuals/colonies and over time in 34 colonies of laboratory-kept zebrafish over a 20-day study period. For this work, a '6 x 6' (six males and six females) colony size was adopted, as this is both environmentally relevant and optimal when considering egg output and animal welfare combined: an initial experiment showed egg output per female increased with decreasing colony size however, there was also a parallel increase in aggressive behavior. Both egg output and viability in '6 x 6' colonies were highly variable among colonies (with co-efficients of variation (CVs) of 30 and 11%, respectively) and over the 20-day study duration (considering egg output and viability of all the colonies combined, the CVs were 20 and 12%, respectively). The patterns of egg production also differed among the '6 x 6' colonies, and they included a cyclical output, a consistent daily output, an infrequent egg output with intermittent days of very high egg output, and an output with no obvious pattern. Sperm quality, measured as percentage motility and curvilinear velocity (VCL), was variable both among individuals within '6 x 6' colonies and across colonies, with percentage motility being the most variable parameter (mean CVs of 82% inter-individual within colonies and 49% inter-colony). Sperm quality did not, however, vary over a 24h period. A minimum number of six replicate '6 x 6' colonies, assessed daily for a period of 4 days, was required per treatment to detect a 40% change in egg output. The minimum numbers of individual males required per treatment to detect a 40% change in sperm quality using the breeding system adopted were 32 males for percentage motility and 12 males for VCL, equivalent to six and two '6 x 6' colonies, respectively. These data demonstrate the need for high levels of replication when testing for effects of EDCs on reproductive output in the zebrafish model in an environmentally relevant ('6 x 6') breeding matrix.     

弱精子症所致男性不育患者精浆中NO含量测定意义

目的:探讨弱精子症患者精浆中一氧化氮(NO)含量变化及对精子活力的影响。方法:选择弱精子症患者50例为实验组,同时选择正常健康男性30例作对照组。采用计算机辅助精液分析技术测定各组精子活力,采用硝酸还原酶法测定精浆中NO含量。结果:实验组精浆中NO含量为58.16±12.17μmol.L-1,对照组精浆中NO含量为34.07±3.23μmol.L-1,实验组明显高于对照组,两者比较差异具有统计学意义(P<0.05)。实验组精浆中NO水平与精子活动率水平呈负相关性,具有统计学意义(P<0.05)。结论:弱精子症患者精浆中NO含量增高,并与精子活力呈负相关。     

Effect of oral administration of terephthalic acid on testicular functions of rats

To investigate the toxic effect of terephthalic acid (TPA) on testicular functions of rats, male Sprague-Dawley rats were orally administered TPA in diet at the levels 0 (control), 0.2, 1 and 5% for 90 days. Testicular functions were assessed by histopathology, testicular sperm head counts, daily sperm production, sperm motility (measured by computer-assisted sperm analysis, CASA), biochemical indices (marker testicular enzymes), and serum testosterone. Oral feeding with terephthalic acid did not cause body and testes weight loss in TPA-treated groups. Histopathologically, damages of spermatogenic cells and Sertoli cells were observed by electron microscope, testicular sperm head counts, daily sperm production, and activities of sorbitol dehydrogenase (SDH) were decreased significantly in the 5% TPA group. The motility of spermatozoa was reduced significantly in all treated groups, which was correlated with administration doses. Serum testosterone concentrations were not declined in treated groups. In conclusion, TPA can cause impairment of testicular functions. The primary sites of action may be spermatogenic cells and Sertoli cells. The results of the present study provide first information of TPA on testicular functions in male rats.