血浆病毒灭活的临床应用与安全输血

目的：推广亚甲蓝光化学灭活血浆病毒的方法,减少经血液传播感染疾病的发生。方法：用亚甲蓝一次性血浆病毒灭活过滤器和医用病毒灭活箱灭活血浆中病毒。结果：用亚甲蓝光化学法可以灭活血浆中大多脂质包膜病毒,杀灭效果理想。病毒灭活前后血浆球蛋白、清蛋白、血浆Ⅷ因子、纤维蛋白原平均水平对比符合≥50g/L的国家标准;HBV-DNA阳性血浆在经病毒灭活后血浆HBV-DNA转为阴性。结论：亚甲蓝/光化学病毒灭活技术可将血浆中病毒灭活,大大提高了临床输注血浆的安全性。     

人凝血因子Ⅷ血液制品中的病毒灭活与验证

目的 研究人凝血因子Ⅷ制备过程中S/D病毒灭活法和80℃、72 h干热灭活法的病毒灭活工艺的灭活效果。方法 经过S/D法及80℃、72 h干热法双重处理灭活病毒,并通过加入指示病毒(PRV、Sindbis、HIV、EMCV、PPV)验证病毒灭活效果。结果 上述工艺可有效灭活脂包膜和非脂包膜病毒。最终制品中TNBP残余量小于1/10万(10 ppm)、吐温-80(Tween-80)残余量小于1/万(100 ppm),符合安全标准,人凝血因子Ⅷ的生物活性及其他各项指标未发现明显变化。结论 可以作为人凝血因子Ⅷ实验过程中的病毒灭活方法。     

人抗凝血酶Ⅲ生产工艺中病毒灭活/去除效果的验证

目的验证和评价人抗凝血酶Ⅲ(human antithrombin Ⅲ, AT-Ⅲ)生产工艺中有机溶剂/去污剂(solvent/detergent, S/D)法联合纳米膜过滤法灭活/去除病毒的可行性。方法采用S/D法作为AT-Ⅲ病毒灭活工艺, 纳米膜过滤法(20 nm孔径)作为其病毒去除工艺, 分析病毒灭活/去除工艺对AT-Ⅲ活性的影响;以伪狂犬病毒、辛德毕斯病毒、猪细小病毒、脑心肌炎病毒作为指示病毒, 对上述工艺进行病毒灭活/去除工艺效果的验证。结果 S/D法联合纳米膜过滤法可有效灭活/去除脂包膜病毒和非脂包膜病毒, 指示病毒滴度下降均>4 lg;且AT-Ⅲ的生物活性及其他各项指标未发生明显改变, 活性变化<10%。结论研究建立的S/D法联合纳米膜过滤法可有效灭活/去除AT-Ⅲ中的指示病毒, 该方法为凝血因子类制品的病毒灭活验证工艺提供了参考。     

人抗凝血酶Ⅲ生产工艺中病毒灭活／去除效果的验证

目的  验证和评价人抗凝血酶Ⅲ（human antithrombin Ⅲ,AT-Ⅲ）生产工艺中有机溶剂/去污剂（solvent/detergent,S/D）法联合纳米膜过滤法灭活／去除病毒的可行性。方法  采用S/D法作为AT-Ⅲ病毒灭活工艺,纳米膜过滤法（20 nm孔径）作为其病毒去除工艺,分析病毒灭活／去除工艺对AT-Ⅲ活性的影响;以伪狂犬病毒、辛德毕斯病毒、猪细小病毒、脑心肌炎病毒作为指示病毒,对上述工艺进行病毒灭活／去除工艺效果的验证。结果   S/D法联合纳米膜过滤法可有效灭活／去除脂包膜病毒和非脂包膜病毒,指示病毒滴度下降均>4 lg;且AT-Ⅲ的生物活性及其他各项指标未发生明显改变,活性变化<10%。结论  研究建立的S/D法联合纳米膜过滤法可有效灭活／去除AT-Ⅲ中的指示病毒,该方法为凝血因子类制品的病毒灭活验证工艺提供了参考。     

新鲜冰冻血浆中的病毒灭活

血液制品具有传播病毒感染的危险性。本文介绍了溶剂/去污剂法（SD法）和亚甲蓝法（MB法）用于新鲜冰冻血浆（FFP）病毒灭活的研究现状及存在问题,探讨了今后的研究方向。     

病毒灭活血浆亚甲蓝残留量影响因素的分析

目的探讨亚甲蓝光化学法（MB—P）病毒灭活血浆光照后延时过滤以及MB—P病毒灭活血浆留样后延时检测这两个因素对病毒灭活血浆亚甲蓝残留量的影响。方法随机抽取200ml／袋全血20袋,分离血浆后进行病毒灭活,在光照结束后0h、1h、2h、3h过滤留样,进行亚甲蓝残留量检测。随机抽取病毒灭活血浆20袋,留样后样本于室温下放置0h、3h、6h进行亚甲蓝残留量检测。均采用同相萃取法进行检测。结果光照结束后0h、1h、2h、3h过滤的样本亚甲蓝残留量分别为（0．178±0．041）μmol／L、（0．198±0．056）μmol／L、（0．238±0．047）μmol／L、（0．309±0．065）μmol／L,延时过滤2h以上亚甲蓝残留量与0h相比差异有统计学意义（P〈0．01）;留样后样本于室温下放置0h、3h、6h亚甲蓝残留量分别为（0．173±0．058）μmol／L、（0．128±0．049）μmol／L、（0．087±0．043）μmol／L,延时检测6h以上亚甲蓝残留量与0h相比差异有统计学意义（P〈0．01）。结论MB—P病毒灭活光照后延时过滤以及MB—P病毒灭活血浆留样后延时检测均对亚甲蓝残留量有影响,在工作中应引起重视。     

滤除白细胞与MB-P法病毒灭活对血浆生化指标的影响

目的探讨应用滤除白细胞及MB-P病毒灭活技术对血浆生化指标的影响。方法随机采集无偿献血合格者血液标本20份,400ml/份。将每份新鲜血浆在制备过程中平均分为试验组与对照组。所有标本同时进行相关生化指标检测。结果试验组经滤除白细胞后FRUC浓度、FHb及CO2增高明显,与对照组比较有显著性差异(P<0.01)。试验组经MB-P法病毒灭活后FRUC与P3+浓度明显增高,FHb下降较快,与对照组及病毒灭活后比较均有显著性差异(P<0.01)。其它生化指标变化不大。结论滤除白细胞及病毒灭活后的血浆制作,对血液生化指标影响均较小或轻,适用于临床输用血(浆)需求。     

滤除白细胞与MB-P法病毒灭活对血浆胆碱酯酶的影响

目的采用滤除白细胞与MB-P法病毒灭活两种方法处理血浆,探讨2种方法对血浆胆碱酯酶（CHE）的影响与变化规律。方法随机采集无偿献血合格者血液标本20份,每份400ml;将每份新鲜血浆在制备过程中平均分为滤白与病毒灭活组和对照组。所有标本同时进行血浆CHE的测定。结果经滤除白细胞及病毒灭活处理后,实验组CHE活力与对照组比较经方差分析差异无统计学意义（P〉0.05）。结论经滤除白细胞与MB-P法病毒灭活处理后的血浆,其CHE活力影响小,可用于临床有机磷中毒的救治。     

病毒灭活后血浆容量不足分析及亚甲蓝辐照的探讨

目的分析血浆经过病毒灭活后容量不足的原因。方法随机抽取街头无偿献血血液46袋,通过对血液去除白细胞、血浆病毒灭活的过程中容量的检测,分析原因。结果所采血液经过白细胞去除、血浆病毒灭活后的血浆容量很难达到200 m L,其中主要原因是去除白细胞时滤盘内所残留血液量和过滤亚甲蓝时滤盘内所残留血浆的量。结论白细胞过滤、血浆病毒灭活等新技术的应用对血液容量有一定损耗,应采用有效措施保证成分血液的容量。     

滤除白细胞与病毒灭活对血浆酶等生化指标的影响

目的探讨滤除白细胞及MB-P病毒灭活技术对血浆酶学及部分生化指标的影响。方法随机采集无偿献血合格者血液标本20份,400ml/份。将每份新鲜血浆在制备过程中平均分为滤白与灭活实验组与对照组。所有标本同时进行相关酶学及部分生化指标检测。结果经滤除白细胞后,实验组LDH、HBDH、TBA下降明显,与对照组比较有显著性差异(P<0.05)。经MB-P法病毒灭活后,实验组CPK、LDH、HBDH、TBIL、IBIL、DBIL、TBA、CREA、URIC下降较快,与对照组比较均有显著性差异(P<0.05)。结论滤除白细胞及病毒灭活后的血浆制作,对血液酶学及部分生化指标影响均较小,符合临床输用血(浆)的要求。     

Viral Infectivity of Albumin and Plasma Protein Fraction

Original research, reviews, and case reports discussing viral infectivity of blood- and plasma-derived products were reviewed to determine the potential viral infectivity of human serum albumin (HSA) and plasma protein fraction (PPF). Data concerning viral infectivity, viral screening and inactivation procedures, and viral outbreaks associated with blood and plasma products were extracted and evaluated for pertinence to HSA and PPF. The starting material used for fractionation, the manufacturing process, postmanufacturing handling, and immunocompetence of HSA or PPF recipients were assessed to determine risk of symptomatic viral disease after transfusion. Both HSA and PPF are manufactured with pasteurization procedures that have led to an excellent viral safety record based on 50 years of clinical use. One outbreak of hepatitis B was associated with PPF as a result of an unreliable manufacturing process that has been corrected. The pasteurization process is effective in eradicating known viral pathogens when good manufacturing practices are followed. Continued surveillance of such products is warranted for viruses not included in routine screening procedures and for those that are resistant to current inactivation methods.     

Effect of Formulation Variables on the Stability of a Live,Rotavirus (RV3-BB) Vaccine Candidate using in vitro Gastric Digestion Models to Mimic Oral Delivery

In this work, two different in vitro gastric digestion models were used to evaluate the stability of a live attenuated rotavirus vaccine candidate (RV3-BB) under conditions designed to mimic oral delivery in infants. First, a forced-degradation model was established at low pH to assess the buffering capacity of formulation excipients and to screen for RV3-BB stabilizers. Second, a sequential-addition model was implemented to examine RV3-BB stability under conditions more representative of oral administration to infants. RV3-BB rapidly inactivated at < pH 5.0 (37 °C, 1 h) as measured by an infectivity RT-qPCR assay. Pre-neutralization with varying volumes of infant formula (Enfamil®) or antacid (Mylanta®) conferred partial to full protection of RV3-BB. Excipients with sufficient buffering capacity to minimize acidic pH inactivation of RV3-BB were identified (e.g., succinate, acetate, adipate), however, they concomitantly destabilized RV3-BB in accelerated storage stability studies. Both effects were concentration dependent, thus excipient optimization was required to design candidate RV3-BB formulations which minimize acid-induced viral inactivation during oral delivery while not destabilizing the vaccine during long-term 2–8 °C storage. Finally, a statistical Design -of-Experiments (DOE) study examining RV3-BB stability in the in vitro sequential-addition model identified key formulation parameters likely affecting RV3-BB stability during in vivo oral delivery.     

Donepezil Hydrochloride BCS Class Ambiguity: Relevant Aspects to be Considered in Drug Classification

Donepezil hydrochloride (DH) is the most used anti-Alzheimer's disease drug, however, its classification according to the Biopharmaceutics Classification System (BCS) is not clear in the literature. BCS is one of the accepted criteria used to grant biowaiver (waiver of in vivo bioequivalence studies) of new drug products. So, the purpose of this work was to elucidate the BCS classification of DH and to raise the discussion about the possibility of biowaiver for new medicines containing it. The polymorphic form was previously identified as form III of DH. The drug showed high solubility in the entire pH range evaluated (1.2 to 6.8, at 37 °C) with a pH-dependent solubility profile. The effective permeability (P_eff) values obtained with different DH concentrations, using in situ closed-loop perfusion model were statistically similar (p > 0.05), even when compared to high permeability control used (ketoprofen), demonstrating that DH has high permeability which, associated with its high solubility, allows to classify DH as BCS class 1. Relevant data to evaluate for granting a biowaiver for new medicines were also reviewed from the literature. Based on information reunited new immediate-release drug products containing DH should be eligible for BCS-based biowaiver.     

Evaluation of sildenafil pressurized metered dose inhalers as a vasodilator in umbilical blood vessels of chicken egg embryos

Sildenafil citrate is a selective phosphodiesterase-5 inhibitor used for the treatment for erectile dysfunction and pulmonary hypertension. The delivery of sildenafil directly to the lung could have several advantages over conventional treatments for pulmonary hypertension because of the local delivery, a more rapid onset of response, and reduced side effects. The major problem of sildenafil citrate is its limited solubility in water. Sildenafil citrate was complexed with cyclodextrins (CDs) to enhance its water solubility prior to development as an inhaled preparation. Four sildenafil citrate inhaled formulations were prepared with the aid of HP-β-CD (#1), α-CD (#2) and γ-CD (#3) and their effects were compared with the formulations without CDs (#4). The sildenafil citrate pressurized metered dose inhalers (pMDI) used ethanol as a solvent, PEG400 as a stabilizing agent, sorbitan monooleate as a surfactant and HFA-134a as a propellant. All formulations consisted of sildenafil citrate equivalent to a sildenafil content of 20 μg/puff. These products were evaluated according to a standard guideline of inhalation products. Vasodilation testing was performed to investigate the efficacy of sildenafil pMDIs in relieving a vasoconstricted umbilical blood vessel of the chicken egg embryo. The sildenafil contents of the pMDI formulations #1–#3 were within the acceptance criteria (80–120%). The emitted doses (ED) were 102.3 ± 11.5%, the fine particle fractions (FPF) were 60.5 ± 5.6% and the mass median aerodynamic diameters (MMAD) were 2.3 ± 0.3 μm. The vasodilatory activity of those formulations reduced umbilical blood pressure by 67.1–73.7% after treatment by intravenous injection whereas only a 50.1–58.0% reduced blood pressure was obtained after direct spraying of the sildenafil pMDI containing CDs. With sildenafil formulations of a pMDI without CD the blood pressure was reduced by only 39.0% (P-value < 0.05). The available sildenafil in the blood vessels of chicken egg embryos after spraying sildenafil-CDs pMDIs was within the range of 751–825 ng/mL which was much higher than that of a sildenafil only pMDI.     

我国与WHO血液制品去除/灭活病毒技术方法与验证指导原则的比较研究

目的：比较我国现行《血液制品病毒去除/灭活验证指导原则》[国药监注（2002）160号]与世界卫生组织（WHO）《血液制品病毒去除/灭活验证指南》，旨在推动我国血液制品病毒风险控制与管理水平的提高及血液制品的合理应用。方法：从去除/灭活病毒方法的选择、常用去除/灭活病毒方法评价、特定的去除/灭活病毒方法验证、生产工艺去除/灭活病毒能力、去除/灭活病毒方法再验证、临床试验、输血用病毒灭活血浆和正在开发的新型病毒灭活方法等几个方面进行比较，对血浆及其制品生产工艺中病毒灭活/去除的相关方法及其验证进行研究。结果与结论：我国现行《血液制品病毒去除/灭活验证指导原则》与WHO《血液制品病毒去除/灭活验证指南》总体要求基本一致，对我国血液制品去除/灭活病毒技术方法与验证工作仍具有适用性和指导性。     

Consumption of tomato products is associated with lower blood mercury levels in Inuit preschool children

Some evidence suggests that various diet components and nutrients, including vegetables, fruit and food-derived antioxidants, could mitigate contaminant exposure and/or adverse health effects of contaminants. To examine the effect of the consumption of tomato products on blood mercury levels in Inuit preschool children, 155 Inuit children (25.0 ± 9.1 months) were recruited from 2006–2008 in Nunavik childcare centers (northern Québec, Canada). Food frequency questionnaires were completed at home and at the childcare center, and total blood mercury concentration was measured by inductively coupled plasma-mass spectrometry. Multivariate regression analysis was performed after multiple imputation. The median blood concentration of mercury was 9.5 nmol/L. Age, duration of breastfeeding, annual consumption frequency of seal meat, and monthly consumption frequency of tomato products were significant predictors of blood mercury levels, whereas annual consumption frequencies of beluga muktuk, walrus, Arctic char, and caribou meat were not. Each time a participant consumed tomato products during the month before the interview was associated with a 4.6% lower blood mercury level (p = 0.0005). All other significant predictors in the model were positively associated with blood mercury levels. Further studies should explore interactions between consumption of healthy store-bought foods available in Arctic regions and contaminant exposure.     

A latent class analysis of poly-marijuana use among young adults

IntroductionWith more states legalizing marijuana use, the marijuana industry has grown, introducing a variety of marijuana products. Our study explores the use of multiple marijuana products (poly-marijuana use) and the characteristics associated with this behavior.MethodsPast-month marijuana users aged 18–34 years were surveyed online via an existing online panel (n = 2444). Participants answered questions about past-month use of three types of marijuana (plant-based, concentrates, edibles), marijuana use patterns, and driving after use. Latent class analysis was used to identify subgroups of marijuana users.ResultsFour classes of marijuana users were identified: Light plant users, who used only plant-based products infrequently and were unlikely to drive after use (32%); Heavy plant users, who used mainly plant-based products frequently, multiple times per day, and were likely to drive after use (37%); Plant and concentrates users, who used plant-based products heavily and concentrates at least infrequently, used multiple times per day, and were likely to drive after use (20%); Light plant and edibles users, who used both products infrequently and were unlikely to drive after use (10%). Those in legal marijuana states were more likely to belong to the poly-marijuana groups.DiscussionOur findings reflect the increase in popularity of new marijuana products in legal states and suggest that heavy user groups, including concentrates users, are associated with driving after use. As various forms of marijuana use increases, monitoring and surveillance of the use of multiple types of marijuana will be important for determining potential varying impacts on physiological and social consequences.     

Attenuated effect of tungsten carbide nanoparticles on voltage-gated sodium current of hippocampal CA1 pyramidal neurons

Nanomaterials and relevant products are now being widely used in the world, and their safety becomes a great concern for the general public. Tungsten carbide nanoparticles (nano-WC) are widely used in metallurgy, aeronautics and astronautics, however our knowledge regarding the influence of nano-WC on neurons is still lacking. The aim of this study was to investigate the impact of nano-WC on tetrodotoxin (TTX)-sensitive voltage-activated sodium current (I_Na) of hippocampal CA1 pyramidal neurons. Results showed that acute exposure of nano-WC attenuated the peak amplitudes of I_Na in a concentration-dependent manner. The minimal effective concentration was 10^?5 g/ml. The exposure of nano-WC significantly decreased current amplitudes of the current–voltage curves of I_Na from ?50 to +50 mV, shifted the steady-state activation and inactivation curves of I_Na negatively and delayed the recovery of I_Na from inactivation state. After exposure to nano-WC, the peak amplitudes, overshoots and the V-thresholds of action potentials (APs) were markedly reduced. These results suggested that exposure of nano-WC could influence some characteristics of APs evoked from the hippocampal CA1 neurons by modifying the kinetics of voltage-gated sodium channels (VGSCs).