ABSTRACT
【Objective】 To study the metabolism and morphology changes of platelets in vitro using traditional Chinese medicine named Qingkailing injection as the additive solution, and to explore the viability of Qingkailing in the extension of platelet storage. 【Methods】 Apheresis platelets, adding 1% final concentration of Qingkailing injection, were taken as experiment groups, and sampled on 1, 3, 5, 8, 10 and 14 days(6 time points)of storage. Apheresis platelets without adding Qingkailing injection were taken as the control, and sampled on 1, 3 and 5 days(3 time points)of storage. The platelet count, morphology scores, biochemical parameters, pH and response rate of hypotonic shock during agitated storage(22 ℃) were tested. 【Results】 1) No significant change in platelet count was noticed in both experimental group(within 14 days) and the control(within 5 days)(P>0.05). The MPV and PDW of both groups increased at any sampling time within 5 days(P<0.05). 2) The morphology score of experimental groups and the control all decreased within the storage period(P<0.05), but its decrease of the control was greater than that of the experimental groups, especially on day 8(P<0.05). 3)Glucose, lactate dehydrogenase, lactate, Na+, and K+ values increased or decreased in varying degrees(P<0.05), while Cl- value stayed almost the same during 14 days(P>0.05). Glucose, lactate dehydrogenase, lactate and Na+ value changed significantly in the control within 5 days(P<0.05), while K+ and Cl- value did not(P>0.05). Within 5-day storage, the glucose consumption, lactate dehydrogenase and lactate generation in the control were significantly greater than those in experimental groups(P<0.05), but the added value of Na+, K+ and Cl- showed no significant difference(P>0.05). 4) pH value, relative to the baseline of day 1, decreased in both groups within 5 days, and its decreasing trend was significant in the control (P<0.05), but not in the experimental group(P>0.05). No significant difference was noticed in the response rate of hypotonic shock in experimental groups within 8 days, while significant decrease was noticed in the control within 5days(P<0.05). The response rate of hypotonic shock in experimental groups were significantly higher than that in the control on day 3 and day 5(P<0.05). 【Conclusion】 The comparison of apheresis platelets, stored in vitro, in terms of platelet count, morphology scores, biochemical parameters, pH and response rate of hypotonic shock showed that platelets, adding 1% final concentration of Qingkailing injection, could prolong the platelet storage to at least 10 days in vitro.
ABSTRACT
Uma das biotecnologias com propósito relevante à saúde humana é o plasma rico em plaquetas (PRP) o qual, não há registro protocolar padronizado para Rattus novergicus (Wistar). De fato, a falta de padronização do método leva pesquisadores à dúvida sob qual protocolo seguir para os melhores resultados referente as concentrações e mudanças morfológicas plaquetárias. Tal indeterminação promove morosidade nas pesquisas científicas. Frente a conjectura, usou-se o mínimo de animais possíveis em experimentação duplo-cego, retirando-se 3,2 mL de sangue de cada animal por pulsão cardíaca o qual posteriormente, foi testado em três protocolos distintos. Cada protocolo foi designado como P1; P2; P3 e o grupo controle designado PSP. Desta forma, i. P1 foi submetido a uma única centrifugação de 160G por 6min resultando em 100 µL de PRP; ii. P2 foi resignado a dupla centrifugação sendo a primeira etapa 160G por 20min e a segunda etapa 400G por 15 min, resultando em 100 µL de PRP e iii. P3 destinado a dupla centrifugação sendo a primeira etapa 300G por 10 min e a segunda etapa 640G por 10 min, resultando em 100 µL de PRP. Obtivemos maiores concentrações plaquetárias dos protocolos P2 e P3 com concentrações superiores a 519%. Quanto a visualização morfológica plaquetária por esfregaço, os três protocolos apresentaram boa visualização, no entanto P3 sobressaiu-se aos demais quanto ao tempo global de preparo e menor inclusão eritrocitária. Assim, padronizamos um modelo animal in vivo para futuras investigações científicas interessadas na utilização do PRP de Rattus novergicus (Wistar)(AU)
One of the biotechnologies with a relevant human health purpose is platelet rich plasma (PRP), which has no standardized protocol record for Rattus novergicus (Wistar). In fact, the lack of standardization of the method leads researchers to doubt which protocol to follow for the best results regarding platelet concentrations and morphological changes. Such indeterminacy promotes slowness in scientific research. In view of the conjecture, how few animals were used in a double-blind experiment and 3.2 mL of blood was drawn from each animal by heartbeat and subsequently tested in three separate protocols. Each protocol was designated as P1; P2; P3 is the control group designated PSP. i. P1 was subjected to a single 160G centrifugation for 6min resulting in 100 µL of PRP, ii. P2 was resigned to double centrifugation with the first stage 160G for 20min and the second stage for 400G for 15 min, resulting in 100 µL of PRP and iii. P3 intended for double centrifugation with the first stage of 300G for 10 min and the second stage of 640G for 10 min, resulting in 100 µL of PRP. We obtained higher platelet concentrations of protocols P2 and P3 with concentrations higher than 519%. Regarding the morphological visualization of platelet smears, the three protocols presented good visualization, but P3 stood out for the others in terms of total preparation time and less erythrocyte invasion. Thus, we standardized an in vivo animal model for future scientific investigations interested in the use of PRP Rattus novergicus (Wistar)(AU)
Subject(s)
Animals , Rats , Rats, Wistar , Guidelines as Topic , Platelet-Rich Plasma , Reference Standards , Blood Platelets , BiotechnologyABSTRACT
Las pruebas de laboratorio son un complemento importante para que el médico homeópata recopile la totalidad de los datos con que se manifiesta la enfermedad de sus pacientes. Uno de los exámenes más habituales a los que se puede recurrir es la biometría hemática, el cual ofrece una orientación conveniente sobre los mecanismos que se activan en el organismo ante un estímulo nocivo determinado,mediante información detallada sobre los elementos formes de la sangre: serie blanca, serie roja y plaquetas. Se presentan a continuación las consideraciones más importantes sobre las variables que se estudian mediante un hemograma, pues aunque ninguno delos valores que nos puede proporcionar se encuentra en los repertorios clásicos,es evidente que nos ofrece información sobre signos tan importantes o comunescomo anemia, hemorragias, trombosis, tromboflebitis o abscesos, todos ellos ensus diferentes modalidades.
Laboratory tests are an important complement to the homeopath. They help to collect all the data manifested by the patient´s disease. One of the most common tests to which the homeopath can turn to is the Cell Blood Count, which offers a convenient guidance of the mechanisms that are activated in the body to a determined noxious stimulus, through detailed information about the blood cells: white,red and platelets count.The following are the most important considerations of the variables that are studied by a CBC, as though none of the values that we can find is in the classical repertoire, it is evident that offers very important information on common signs such as anemia, bleeding, thrombosis, thrombophlebitis or abscess, all in their different ways.