新型冠状病毒灭活操作对替考拉宁血药浓度的影响及一致性评价

摘要：目的:考察不同的新型冠状病毒灭活方式对替考拉宁血药浓度的影响,评价未经灭活操作与经灭活操作后替考拉宁血药浓度结果的一致性。方法:以替考拉宁质控对照品、质控血浆样品和临床实测样本作为考察对象,分别经56℃水浴加热30 min或经紫外照射60 min后,考察质控对照品和质控血浆样品的稳定性;并采用组内相关系数、Passing-Bablok回归法和Bland-Altman分析法,对灭活前后临床实测标本浓度结果进行统计分析。结果:紫外灭活条件下,替考拉宁的质控对照品不稳定;湿热灭活法条件下,替考拉宁质控对照品和质控血浆样品均稳定。52例临床样本经湿热灭活与未灭活浓度结果的组内相关系数为0.994（P<0.001）;Passing-Bablok回归方程为Y=0.9994X-0.214 2（R2=0.996,n=52）;96.15%（50/52）的检测数据在95%一致性界限内。3种统计结果均表明未经灭活操作与经湿热灭活法操作后,两者血药浓度结果的相关性与一致性良好。结论:替考拉宁的临床血药浓度检测标本可采用湿热病毒灭活法进行预处理,以减少实验室工作人员的病毒感染风险。     

葡萄籽原花青素对酒精性肝损伤小鼠的保护作用

目的 考察葡萄籽原花青素（grape seed procyanidin,GSP）对酒精性肝损伤小鼠的保护作用及其抗炎机制。方法 ICR小鼠60只,♂,随机分成正常对照组、模型组（56%乙醇）、水飞蓟素组（56%乙醇+90 mg·kg^-1·d^-1水飞蓟素）及GSP高、中、低剂量组（56%乙醇+400,200,100 mg·kg^-1·d^-1GSP）。8周后用分光光度法检测血清中ALT、AST水平,肝MDA含量和SOD、GSH-Px活力;酶联免疫法检测肝NF-κB、TNF-α、IL-1β的含量;HE染色检测肝脏病理改变。结果 与模型组比较,GSP可以降低酒精性肝损伤小鼠血清中ALT、AST含量及肝组织中MDA、NF-κB、TNF-α、IL-1β含量（P<0.05）,提高SOD、GSH-PX活力（P<0.05）,减轻肝脏的病理损伤程度（P<0.05）。结论 GSP对酒精性肝损伤小鼠具有保护作用,其机制可能是通过抗氧化和降炎症反应来发挥作用。     

56℃加热30min灭活血液标本对选择性5-羟色胺再摄取抑制剂治疗药物监测的影响

目的:评估新型冠状病毒肺炎疫情期间56℃加热30 min灭活血液标本对5-羟色胺再摄取抑制剂治疗药物监测的影响。方法:血清样品用乙腈蛋白沉淀法前处理,用液相色谱串联质谱法测定艾司西酞普兰、氟伏沙明、氟西汀、去甲氟西汀、帕罗西汀和舍曲林血清药物浓度,考察血液标本灭活前后基质效应、血清质控样品灭活后药物稳定性和临床血液标本灭活后对血清药物浓度的影响。结果:6个化合物灭活前后内标归一化基质效应无明显差别,艾司西酞普兰、氟伏沙明、氟西汀、去甲氟西汀和帕罗西汀在该灭活条件下可保持稳定,血液标本灭活后浓度无明显改变,舍曲林灭活后出现降解,而血液标本由于加热后溶血导致细胞内舍曲林扩散至血清中,其灭活后血清浓度反而增加。结论:艾司西酞普兰、氟伏沙明、氟西汀、去甲氟西汀和帕罗西汀的血液标本可用56℃加热30 min灭活处理,而舍曲林在该灭活条件下检测结果不准确。     

血清灭活对小鼠淋巴瘤试验结果的影响

目的 探索小鼠淋巴瘤胸腺激酶（tk）位点基因突变试验中血清灭活对结果的影响。方法 取对数生长期的小鼠淋巴瘤细胞（L5178Y3.7.2c-tk^+/-）设立阴性对照组和阳性对照组,阴性对照组加入100 μL DMSO,阳性对照组加入100 μL 10 μg/mL 4-硝基喹啉（4-NQO）诱导突变,测定细胞悬浮增长率（SG）、相对悬浮增长率（RSG）、平板效率（PE）、相对存活率（RS）和相对总生长率（RTG）等指标。根据血清灭活与否和选择突变剂三氟胸苷（TFT）的含量,在筛选突变集落过程中,将阴性对照组和阳性对照组各分为：灭活组、未灭活+1.5倍TFT组、未灭活组,测定突变频率（MF）,对试验结果进行评价。结果 阴性对照组SG在8~32,细胞倍增速度符合试验要求;PE在65%~120%,符合试验成立条件。阳性对照的灭活组、未灭活+1.5倍TFT组、未灭活组中MF均比阴性对照组至少增加300×10^-6且小克隆突变频率至少占40%,符合试验要求。阴性对照灭活组的MF数值在标准范围内,未灭活组的MF数值明显高于灭活组,超过标准数值范围,且96孔板中的大小克隆不易区分;阴性对照未灭活+1.5倍TFT组使得MF数值在标准范围内,但远高于灭活组。结论 在小鼠淋巴瘤试验中,测定MF的平板一定要用灭活的血清,未灭活血清可能通过抵消TFT作用对结果产生很大影响。     

轻中度创伤性脑损伤患者记忆受损与血清Tau蛋白的关系

目的 探讨轻中度创伤性脑损伤（TBI）患者记忆受损与血清Tau蛋白的关系。方法 选取安徽医科大学第一附属医院2016年6月至2017年6月收治的TBI患者60例为研究对象,依据患者入院时格拉斯哥昏迷评分（GCS）分为轻度TBI组（30例）与中度TBI组（30例）。另选取20名志愿者为健康对照组。在入院时、伤后第72小时、2周、6周和3个月时,采用ELISA法检测受试对象的血清Tau蛋白浓度,同时运用神经心理学测试量表对受试对象记忆认知特点进行评估。结果 入院时,TBI组患者血清Tau蛋白浓度为（574.3±270.1） pg/mL、健康对照组为（79.9±36.3） pg/mL,两组差异有统计学意义（t=13.807,P<0.05）。伤后第72小时,中度TBI组患者血清Tau蛋白浓度为（1 051.2±333.9） pg/mL、轻度TBI组为（805.2±400.2） pg/mL,两组差异有统计学意义（t=2.585,P=0.012）。伤后3个月,TBI组患者血清Tau蛋白浓度为（100.8±35.6） pg/mL,与健康对照组相比,差异无统计学意义（t=2.001,P=0.051）。TBI组患者记忆受损明显,且伤后第72小时患者记忆认知水平处于最低,至伤后6周,TBI组患者记忆受损逐渐恢复至健康对照组水平,两组差异无统计学意义（P>0.05）。伤后第72小时和伤后2周,TBI组患者血清Tau蛋白浓度与神经心理学测试评分呈负相关（P<0.05）。结论 早期检测TBI患者的血清Tau蛋白浓度不仅可以评估患者的受伤程度,而且能够有效地反映患者记忆受损的程度。     

抗肿瘤药物职业暴露对PIVAS工作人员的健康危害评估★

目的 采用环境检测与生物检测方法评估静脉药物配置中心（PIVAS）工作人员抗肿瘤药物职业暴露情况。方法 于2020年4月随机选取本院PIVAS工作人员50例作为研究对象，根据是否接触过抗肿瘤药物分为暴露组（25例）和非暴露组（25例）。以5-氟尿嘧啶（5-Fu）及环磷酰胺（CTX）为检测标志物，采用超高效液相串联质谱法（UHPLC-MS）连续2年（2020年、2021年）测定PIVAS环境中不同物体擦拭样本及两组研究对象血液、尿液中的5-Fu及CTX浓度；流式细胞术测定外周血细胞凋亡率；酶联免疫吸附法测定8-羟化脱氧鸟苷（8-OHdG）水平。结果 工作人员一般情况比较，差异无统计学意义。PIVAS环境中物体表面存在不同程度的5-Fu和CTX污染，配置区振荡器、传递仓内把手及电话表面的5-Fu平均浓度分别为2 286.26、976.82、202.42 ng·mL^-1，CTX平均浓度分别为2 991.26、88.12、155.40 ng·mL^-1；5-Fu及CTX储存箱存在CTX残留。2021年流式结果显示，暴露组单核细胞早期及总凋亡率均显著高于非暴露组（P=0.024， P=0.024）；暴露组中性粒细胞早、晚期凋亡率均高于非暴露组（P=0.004， P<0.001）。两年均在研究对象尿液中检出了CTX残留，暴露组CTX尿液检出率高于非暴露组（χ² =11.52， P<0.001）。暴露组CTX血药浓度高于非暴露组［Z=-7.500， P<0.001 （2020）； Z=-4.500， P<0.001 （2021）］。8-OHdG是公认的评估DNA损伤和氧化应激的敏感生物标志物，暴露组8-OHdG水平明显高于非暴露组（Z=-4.605， P<0.001），提示暴露组DNA损伤程度明显高于非暴露组。结论 PIVAS存在一定程度抗肿瘤药物污染，应重视抗肿瘤药物职业暴露对人体健康的影响并加强防护。     

新型冠状病毒灭活方式对高效液相色谱法监测万古霉素血药浓度的影响

目的考察新型冠状病毒不同灭活方式对高效液相色谱(HPLC)法监测万古霉素血药浓度的影响。方法经56℃水浴加热0 min和30 min,以及紫外照射0,30,60 min后,采用HPLC法测定质控液和患者血样中万古霉素的质量浓度。结果56℃水浴加热30 min不会影响监测结果;紫外照射30 min和60 min后,质控液和患者血样中万古霉素均降解,对监测结果影响较大。结论采用HPLC法监测新型冠状病毒肺炎(COVID-19)患者万古霉素血药浓度时,可用湿热法56℃水浴30 min灭活病毒,不可用紫外照射灭活病毒,且万古霉素质控液与患者血样均要注意避免紫外照射。     

针刺麻醉对老年直肠癌根治术患者术后认知功能和血清S100β、MMP-9、BDNF蛋白水平的影响

目的 观察针刺麻醉对老年直肠癌根治术患者术后认知功能及血清S100β、基质金属蛋白酶-9（MMP-9）、脑源性神经生长因子（BDNF）水平的影响。方法 将90例行直肠癌根治术的老年患者随机分为对照组和观察组,每组45例。两组均采取全凭静脉麻醉,观察组于麻醉诱导前30 min开始予以针刺麻醉并持续至术毕。比较两组术中全身麻醉药物用量及术后不同时间点简易精神状态量表（MMSE）评分,统计两组术后7 d认知功能障碍（POCD）发生率,并于手术前后行血清S100β、MMP-9、BDNF检测。结果 两组术中维库溴铵、丙泊酚用量比较,差异无统计学意义（P>0.05）,而观察组瑞芬太尼用量明显低于对照组（P<0.05）。术后1 d、3 d、7 d,观察组MMSE评分均明显高于对照组（P<0.05）。术后7 d内,观察组患者POCD发生率为11.11%,明显低于对照组的31.11%,差异有统计学意义（P<0.05）。两组术后血清S100β、MMP-9水平相比术前均明显上升（P<0.05）,血清BDNF水平相比术前明显降低（P<0.05）,但观察组上述指标的变化幅度小于对照组（P<0.05）。结论 针刺麻醉能够改善老年直肠癌根治术患者的认知功能,降低POCD发生率,其机制可能与降低血清S100β、MMP-9水平及升高血清BDNF水平有关。     

快速解剖标测方式构建肺静脉对环肺静脉电隔离术中主要质量指标的影响

目的 探讨快速解剖标测方式构建肺静脉在环肺静脉电隔离术中应用价值。方法 收集2016年2月至2016年10月在皖南医学院弋矶山医院心内科接受导管消融治疗的60例房颤患者,依据肺静脉造影方式分为快速解剖标测组30例和非选择性肺静脉造影组30例。评价两组患者总手术时间、X线曝光时间、预定消融径线外消融点数、肺静脉隔离率等方面差异。结果 快速解剖标测组与非选择性肺静脉造影组相比：曝光时间减少[（16.18±4.26） min vs（45.64±6.48） min,P<0.05],总手术时间降低[（122.34±13.82） min vs（180.19±12.56） min,P<0.05）],预定消融径线外消融点数减少[（3±1）个vs（16±4）个,P<0.05],肺静脉隔离率差异无统计学意义（96.67%vs 93.33%,P>0.05）。结论 快速解剖标测方式构建肺静脉能明显提高肺静脉隔离术的手术质量。     

齐拉西酮对精神分裂症患者血清胆碱酯酶浓度的影响

目的 探讨齐拉西酮对精神分裂症患者血清胆碱酯酶浓度（CHE）浓度的影响。方法 将126例精神分裂症患者随机分为观察组和对照组,观察组给予齐拉西酮治疗,对照组给予氯氮平治疗,进行阳性和阴性症状量表（PANSS）评定,并检测治疗后7、14、28 d的CHE浓度。结果 ①治疗后观察组和对照组PANSS得分均有不同程度的降低,但对照组PANSS得分显著低于观察组[阳性症状：（15.3±4.7）vs（22.4±4.8）,P<0.01;阴性症状：（14.6±4.5）vs（21.8±5.2）,P<0.01;一般精神症状：（13.3±3.4）vs（19.2±3.9）,P<0.01）]。②齐拉西酮或氯氮平治疗均能明显降低CHE的浓度;两种药物分别治疗28 d后,对照组CHE浓度较观察组明显降低[（8 339±2 106）vs（7 366±2 024）,P<0.01]。③治疗中两组均有不良反应,观察组不良反应率为34.9%,对照组不良反应率为71.4%,组间比较有显著性差异（χ²=16 862,P<0.001）。结论 齐拉西酮能对CHE活性有抑制作用,其抑制强度较氯氮平弱,从而可能使齐拉西酮对精神分裂症患者疗效低于氯氮平,但其安全性明显高于氯氮平。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.