RESUMEN
OBJECTIVES: To investigate the technical performance of IDentifier DNA typing kit (YanHuang34) and evaluate its forensic application value. METHODS: Following the Criterion of Forensic Science Human Fluorescence STR Multiplex Amplification Reagent (GB/T 37226-2018), IDentifier DNA typing kit (YanHuang34) was verified in 11 aspects of species specificity, veracity, sensibility, adaptability, inhibitor tolerance, consistency, balance, reaction condition verification, mixed samples, stability and inter batch consistency. The system efficiency of IDentifier DNA typing kit (YanHuang34) was compared with the PowerPlex® Fusion 6C System, VersaPlex® 27PY System and VeriFilerTM Plus PCR Amplification Kit. The IDentifier DNA typing kit (YanHuang34) was used to detect the swabs of biological samples in daily cases and the STR performances were observed. RESULTS: IDentifier DNA typing kit (YanHuang34) had good species specificity, veracity, adaptability, inhibitor tolerance and balance. The sensibility was up to 0.062 5 ng. It was able to detect different types of samples, degraded samples and inhibitor mixed samples. Complete DNA typing could be obtained for samples with the mixture ratio less than 4â¶1. The system efficiency of IDentifier DNA typing kit (YanHuang34) was very high, with TDP up to 1-1.08×10-37, CPEtrio and CPEduo up to 1-5.47×10-14 and 1-6.43×10-9, respectively. For the touched biological samples in actual cases, the effective detection rate was 21.05%. The system efficiency of kinship, single parent and full sibling identifications was effectively improved. CONCLUSIONS: The IDentifier DNA typing kit (YanHuang34) is adaptive to the GB/T 37226-2018 requirements. It can be used for individual identification and paternity identification, and is suitable for application in the field of forensic science.
Asunto(s)
Dermatoglifia del ADN , Repeticiones de Microsatélite , Humanos , Reacción en Cadena de la Polimerasa , Paternidad , Especificidad de la EspecieRESUMEN
Glycyrrhiza polysaccharide liposome (GPSL) was prepared by reverse-phase evaporation method and optimized with response surface methodology. As a result, the optimum preparation conditions were as follows: the ratio of soybean phosphatide to Glycyrrhiza polysaccharide (GPS) of 24:1, temperature of drug incubation of 46°C, the ultrasound time of 16min. Confirmation experiments and transmission electron micrograph (SEM) exhibited the entrapment efficiency of liposome as 78.33±0.25%, with spherical shape and uniform sizes. Furthermore, proliferation assay, allogeneic mixed lymphocyte reaction and supernatant cytokines of chBM-DCs were performed by MTT and ELISA methods to investigate the immune-modulating effects of GPSL and GPS. The results showed that GPSL could significantly promote the proliferation of immature chBM-DCs, enhance the ability of mature chBM-DCs to induce T-cell proliferation and modulate the mature chBM-DCs to secret cytokines such as IL-2, IFN-γ and IL-10. These evidences indicated that the immune-modulating activity of GPS was significantly improved after encapsulated with liposome.
Asunto(s)
Glycyrrhiza/química , Factores Inmunológicos/administración & dosificación , Factores Inmunológicos/farmacología , Polisacáridos/administración & dosificación , Polisacáridos/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Pollos , Citocinas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Liposomas , Masculino , Modelos Teóricos , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Temperatura , Ondas UltrasónicasRESUMEN
One acidic polysaccharide named EPS-1 was isolated from the aqueous extract of the leaves of Epimedium acuminatum Franch. It may be composed of 1,4-linked α-d-GalpA, 1,3,4-linked α-d-GalpA, 1,6-linked ß-d-Galp and terminal α-l-Rhap residues in a molar ratio of 11.0:1.0:1.0:1.0 by chemical and spectroscopic analysis. EPS-1 possessed immune modulation effects on peripheral T lymphocyte and immature chBM-DCs such as promoting the proliferation and cytokine secretion of these cells and increasing the phagocytosis ability of immature chBM-DCs.
Asunto(s)
Epimedium/metabolismo , Polisacáridos/química , Animales , Células de la Médula Ósea/citología , Proliferación Celular/efectos de los fármacos , Pollos , Cromatografía Líquida de Alta Presión , Citocinas/metabolismo , Células Dendríticas/citología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Cromatografía de Gases y Espectrometría de Masas , Masculino , Microscopía Electrónica de Rastreo , Fagocitosis/efectos de los fármacos , Hojas de la Planta/metabolismo , Polisacáridos/análisis , Polisacáridos/farmacologíaRESUMEN
Garlic polysaccharide (GPS) was modified in selenylation respectively by nitric acid-sodium selenite (NA-SS), glacial acetic acid-selenous acid (GA-SA), glacial acetic acid-sodium selenite (GA-SS) and selenium oxychloride (SOC) methods each under nine modification conditions of L9(3(4)) orthogonal design and each to obtain nine selenizing GPSs (sGPSs). Their structures were identified, yields and selenium contents were determined, selenium yields were calculated, and the immune-enhancing activities of four sGPSs with higher selenium yields were compared taking unmodified GPS as control. The results showed that among four methods the selenylation efficiency of NA-SS method were the highest, the activity of sGPS5 was the strongest and significantly stronger than that of unmodified GPS. This indicates that selenylation modification can significantly enhance the immune-enhancing activity of GPS, NA-SS method is the best method and the optimal conditions are 0.8:1 weight ratio of sodium selenite to GPS, reaction temperature of 70 °C and reaction time of 10h.
Asunto(s)
Ajo/química , Factores Inmunológicos/química , Polisacáridos/química , Selenio/química , Animales , Células Cultivadas , Pollos , Citocinas/metabolismo , Factores Inmunológicos/farmacología , Linfocitos/efectos de los fármacos , Linfocitos/inmunología , Polisacáridos/farmacologíaRESUMEN
The effects of two selenizing polysaccharides (sCAP2 and sGPS6) on immune function of murine peritoneal macrophages taking two non-selenizing polysaccharides (CAP and GPS) and modifier Na2SeO3 as control. In vitro test, the changes of selenizing polysaccharides, non-selenizing polysaccharides and Na2SeO3 on murine macrophages function were evaluated by phagocytosis and nitric oxide (NO) secretion tests. In vivo test, the mice were injected respectively with 0.2, 0.4 and 0.6 mg of sCAP2, sGPS6, CAP and GPS, or Na2SeO3 80 µg or normal saline 0.4 mL. The peritoneal macrophages were collected and cultured to determine the contents of TNF-α, IL-6 and IL-10 in supernatants by enzyme-linked immunosorbent assay. The results showed that sCAP2 and sGPS6 could significantly promote the phagocytosis and secretion of NO and three cytokines of macrophages in comparison with CAP and GPS. sCAP2 possessed the strongest activity. This indicates that selenylation modification can further improve the immune-enhancing activity of polysaccharide, and sCAP2 could be as a new immunopotentiator.
Asunto(s)
Antígenos de Plantas/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Polisacáridos/farmacología , Óxidos de Selenio/farmacología , Angelica/inmunología , Animales , Antígenos de Plantas/química , Células Cultivadas , Citocinas/metabolismo , Femenino , Ajo/inmunología , Inmunidad Celular/efectos de los fármacos , Inmunización , Mediadores de Inflamación/metabolismo , Macrófagos Peritoneales/inmunología , Ratones , Ratones Endogámicos ICR , Óxido Nítrico/metabolismo , Fagocitosis/efectos de los fármacos , Polisacáridos/químicaRESUMEN
The Atractylodes macrocephala polysaccharide (AMP) was extracted purified and modified in selenylation by Nitric acid-sodium selenite method to get nine selenizing AMPs (sAMPs), sAMP(1)-sAMP(9). In vitro test their effects on chicken peripheral lymphocyte proliferation were determined by MTT assay. The results showed that nine sAMPs and AMP at five concentrations could significantly promote lymphocyte proliferation, the actions in six sAMPs were significantly stronger than that in AMP, and in sAMP(9) was the strongest. In vivo test, 14-day-old chickens vaccinated with ND vaccine were injected respectively with sAMP(9) and AMP, the peripheral lymphocytes proliferation, serum antibody titer, IFN-γ, IL-2 and IL-6 contents were determined. The results displayed that the sAMP could significantly promote lymphocyte proliferation and elevate the antibody titers and content of IFN-γ, IL-2 and IL-6 in comparison with unmodified AMP. These results indicate that selenylation modification can significantly enhance the immune-enhancing activity of AMP.
Asunto(s)
Atractylodes/química , Factores Inmunológicos/farmacología , Polisacáridos/inmunología , Selenio/metabolismo , Animales , Anticuerpos/sangre , Proliferación Celular/efectos de los fármacos , Pollos , Citocinas/sangre , Linfocitos/citología , Linfocitos/efectos de los fármacos , Masculino , Selenio/análisis , Selenio/químicaRESUMEN
OBJECTIVE: To explore a new method in order to extract DNA from bones and teeth automatically. METHODS: Samples of 33 bones and 15 teeth were acquired by freeze-mill method and manual method, respectively. DNA materials were extracted and quantified from the triturated samples by AutoMate Express forensic DNA extraction system. RESULTS: DNA extraction from bones and teeth were completed in 3 hours using the AutoMate Express forensic DNA extraction system. There was no statistical difference between the two methods in the DNA concentration of bones. Both bones and teeth got the good STR typing by freeze-mill method, and the DNA concentration of teeth was higher than those by manual method. CONCLUSION: AutoMate Express forensic DNA extraction system is a new method to extract DNA from bones and teeth, which can be applied in forensic practice.
Asunto(s)
Huesos/química , Dermatoglifia del ADN/métodos , ADN/aislamiento & purificación , Medicina Legal/métodos , Diente/química , Automatización , Dermatoglifia del ADN/instrumentación , Humanos , Repeticiones de Microsatélite , Reacción en Cadena de la Polimerasa , Manejo de Especímenes/métodosRESUMEN
With the development of molecular biology, the evidences of genetics has been used widely in forensic sciences. DNA technology has played an important role in individual identification and paternity testing, RNA technology is showing more and more wide application in prospect. This article reviews the application and progress of RNA in forensic science including estimation of postmortem interval, bloodstain age, wound age, as well as determination of cause of death and the source of body fluids.