SSR marker-based DNA fingerprinting of Sub1 introgressed lines in the background of traditional rice varieties of Assam India-Supplementary data

Rice varieties are usually characterized by agro-morphological descriptors used for seed certification and seed characterization by following distinctiveness, uniformity, and stability (DUS) test. But in fact, these primary distinguishing morphological descriptors among rice varieties are very limited and hence face problems to distinguish germplasm accessions. Germplasm certification in NBPGR requires a DNA fingerprinting profile to explain germplasm uniqueness compared to existing varieties. Varietal identification has gained a key role worldwide, particularly in plant variety protection. Sixty-two morphological descriptors studies have shown the Sub1 introgressed advanced lines E-6, C-210, C-196, 1189-1 and 1160-1 are distinct from the other varieties for more than 15morphological traits, based on these variations the lines were selected for DNA fingerprinting. About68 SSRs markers were used for DNA fingerprinting in seven genotypes, two of which were parents (Ranjit, Bahadur) and three Sub1 introgressed advanced lines (E6, C210, C196) in Ranjit background, and two Sub1 introgressed advanced lines (1189-1, 1160-1) in Bahadur background. DNA fingerprinting was done on these genotypes of rice using SSR markers. Among the 68 SSR markers, total 65 markers were amplified and three were found not amplified. Out of 65 markersfour of them viz. RM 152, RM 172, RM 251, and RM 346 showed better polymorphism with amplicon size ranges from 155-163 bp, 150-159 bp, 137-147 bp, and 166-175 bp, respectively, and remaining 61 showed monomorphic amplification. Therefore, SSR (Simple-sequence repeats) based DNA fingerprinting helped to differentiate Ranjit, Bahadur, E-6, C-210, C-196, 1189-1, and 1160-1. Hence, the research reveals that newly developed high-yielding Sub1 introgressed advanced lines in the background of traditional Assam rice varieties (Ranjit and Bahadur) are unique in their identity.

SSR marker-based DNA fingerprinting of Sub1 introgressed lines in the background of traditional rice varieties of Assam India-Supplementary data

Rice varieties are usually characterized by agro-morphological descriptors used for seed certification and seed characterization by following distinctiveness, uniformity, and stability (DUS) test. But in fact, these primary distinguishing morphological descriptors among rice varieties are very limited and hence face problems to distinguish germplasm accessions. Germplasm certification in NBPGR requires a DNA fingerprinting profile to explain germplasm uniqueness compared to existing varieties. Varietal identification has gained a key role worldwide, particularly in plant variety protection. Sixty-two morphological descriptors studies have shown the Sub1 introgressed advanced lines E-6, C-210, C-196, 1189-1 and 1160-1 are distinct from the other varieties for more than 15morphological traits, based on these variations the lines were selected for DNA fingerprinting. About68 SSRs markers were used for DNA fingerprinting in seven genotypes, two of which were parents (Ranjit, Bahadur) and three Sub1 introgressed advanced lines (E6, C210, C196) in Ranjit background, and two Sub1 introgressed advanced lines (1189-1, 1160-1) in Bahadur background. DNA fingerprinting was done on these genotypes of rice using SSR markers. Among the 68 SSR markers, total 65 markers were amplified and three were found not amplified. Out of 65 markersfour of them viz. RM 152, RM 172, RM 251, and RM 346 showed better polymorphism with amplicon size ranges from 155-163 bp, 150-159 bp, 137-147 bp, and 166-175 bp, respectively, and remaining 61 showed monomorphic amplification. Therefore, SSR (Simple-sequence repeats) based DNA fingerprinting helped to differentiate Ranjit, Bahadur, E-6, C-210, C-196, 1189-1, and 1160-1. Hence, the research reveals that newly developed high-yielding Sub1 introgressed advanced lines in the background of traditional Assam rice varieties (Ranjit and Bahadur) are unique in their identity.

SSR marker-based DNA fingerprinting of Sub1 introgressed lines in the background of traditional rice varieties of Assam India

Rice varieties are usually characterized by agro-morphological descriptors used for seed certification and seed characterization by following distinctiveness, uniformity, and stability (DUS) test. But in fact, these primary distinguishing morphological descriptors among rice varieties are very limited and hence face problems to distinguish germplasm accessions. Germplasm certification in NBPGR requires a DNA fingerprinting profile to explain germplasm uniqueness compared to existing varieties. Varietal identification has gained a key role worldwide, particularly in plant variety protection. Sixty-two morphological descriptors studies have shown the Sub1 introgressed advanced lines E-6, C-210, C-196, 1189-1 and 1160-1 are distinct from the other varieties for more than 15morphological traits, based on these variations the lines were selected for DNA fingerprinting. About68 SSRs markers were used for DNA fingerprinting in seven genotypes, two of which were parents (Ranjit, Bahadur) and three Sub1 introgressed advanced lines (E6, C210, C196) in Ranjit background, and two Sub1 introgressed advanced lines (1189-1, 1160-1) in Bahadur background. DNA fingerprinting was done on these genotypes of rice using SSR markers. Among the 68 SSR markers, total 65 markers were amplified and three were found not amplified. Out of 65 markersfour of them viz. RM 152, RM 172, RM 251, and RM 346 showed better polymorphism with amplicon size ranges from 155-163 bp, 150-159 bp, 137-147 bp, and 166-175 bp, respectively, and remaining 61 showed monomorphic amplification. Therefore, SSR (Simple-sequence repeats) based DNA fingerprinting helped to differentiate Ranjit, Bahadur, E-6, C-210, C-196, 1189-1, and 1160-1. Hence, the research reveals that newly developed high-yielding Sub1 introgressed advanced lines in the background of traditional Assam rice varieties (Ranjit and Bahadur) are unique in their identity.

Assessment of genotyping markers in the molecular characterization of a population of clinical isolates of Fusarium in Colombia / Evaluación de marcadores de genotipificación en la caracterización molecular de una población de aislamientos clínicos de Fusarium en Colombia

Introduction: Fusarium is a very heterogeneous group of fungi, difficult to classify, with a wide range of living styles, acting as saprophytes, parasites of plants, or pathogens for humans and animals. Prevalence of clinical fusariosis and lack of effective treatments have increased the interest in the precise diagnosis, which implies a molecular characterization of Fusarium populations. Objective: We compared different genotyping markers in their assessment of the genetic variability and molecular identification of clinical isolates of Fusarium. Materials and methods: We evaluated the performance of the fingerprinting produced by two random primers: M13, which amplifies a minisatellite sequence, and (GACA)4, which corresponds to a simple repetitive DNA sequence. Using the Hunter Gaston Discriminatory Index (HGDI), an analysis of molecular variance (AMOVA), and a Mantel test, the resolution of these markers was compared to the reference sequencing-based and PCR genotyping methods. Results: The highest HGDI value was associated with the M13 marker followed by (GACA)4. AMOVA and the Mantel tests supported a strong correlation between the M13 classification and the reference method given by the partial sequencing of the transcription elongation factor 1-alpha (TEF1-α) and rDNA 28S. Conclusion: The strong correlation between the M13 classification and the sequencing-based reference together with its higher resolution demonstrates its adequacy for the characterization of Fusarium populations.

Introducción. Fusarium es un grupo heterogéneo de hongos, difícil de clasificar y con una amplia gama de estilos de vida, que actúa como saprófito, parásito de plantas o patógeno de humanos y animales. La prevalencia de la fusariosis clínica y la falta de tratamientos han incrementado el interés en su diagnóstico preciso, lo que conlleva la caracterización molecular de las poblaciones. Objetivo. Comparar marcadores de genotipificación en la evaluación de la variabilidad genética e identificación de aislamientos clínicos de Fusarium. Materiales y métodos. Se evaluó la huella genética producida por dos cebadores aleatorios: M13, que amplifica una secuencia minisatélite, y (GACA)4, que corresponde a una secuencia repetitiva de ADN. Utilizando el índice discriminatorio de Hunter Gaston (HGDI), el análisis de varianza molecular (AMOVA) y una prueba de Mantel, se comparó la resolución de estos marcadores con métodos de genotipificación basados en secuenciación y PCR. Resultados. El mayor HGDI se asoció con el marcador M13, seguido de (GACA)4. Las pruebas AMOVA y Mantel mostraron correlación entre las clasificaciones obtenidas con M13 y la referencia basada en la secuenciación parcial del factor de elongación de transcripción 1-alfa (TEF1-α) y el ADNr 28S. Conclusión. La fuerte correlación entre la clasificación obtenida con M13 y el método de referencia, así como su alta resolución, demuestran su idoneidad para la caracterización de poblaciones de Fusarium.

First Genetically Differentiation between Upupa epops major and Upupa epops epops (Family: Upupidae)

Abstract Hoopoe has been traditionally treated as a single species within the order Coraciiformes. Presently, however, various authors have suggested separating the hoopoe into two or more species and even its order, Bucerotiformes. So, this work aimed to use the RAPD PCR and DNA sequences of the COI gene barcodes to confirm and to assess whether the Egyptian hoopoe is a different species named Upupa epops major from the European hoopoe called Upupa epops epops, and to determine the relationships among them. Five primers were used in this technique. Two hoopoes were taken in this work as studying birds, migratory and resident one. The results showed the highest genetic distance between them using different random primers while genetic identity was in general low, overall primers. DNA fingerprinting detected greater genetic distance between Upupa epops major and Upupa epops epops and low genetic identity, this may indicate that both hoopoes fall into two separate species. Furthermore, using mitochondrial cytochrome oxidase subunit I (COI) sequences in this study suggests the separation of Upupa epops major into a new species.

Genetic traceability of Staphylococcus aureus strains isolated from primiparous dairy cows mastitis, humans and environment in the Northeast region of Brazil / Rastreabilidade genética de cepas de Staphylococcus aureus isoladas de mastite de vacas primíparas, ordenhadores e ambiente de ordenha na região Nordeste do Brasil

ABSTRACT: This research aimed to investigate the genotypic relatedness of 18 Staphylococcus aureus strains isolated from intramammary infections in primiparous cows and extramammary sites on five dairy herds by rep-PCR using RW3A primers, and by PFGE using the endonuclease SmaI. The isolates were also evaluated in vitro for the susceptibility against beta-lactam antimicrobials drugs (penicillin and oxacillin), considering that beta-lactams are frequently used for treating staphylococcal intrammamary infections. The rep-PCR typing was highly discriminatory (D value= 0.9804) and a total of 15 patterns were detected. The PFGE method was also highly discriminatory (D value= 0.9667) and a total of 13 patterns were observed. A total of 15 out of 18 (83%) isolates were resistant to penicillin and one out of 18 (6%) to oxacillin. In conclusion, these findings confirmed the occurrence of a high genetic diversity of S. aureus strains at the herds and the presence of clonally-related strains only at the same herd, emphasizing a variety of genotypic profiles among the isolates.

RESUMO: Objetivou-se com este estudo investigar a correlação genética de 18 cepas de Staphylococcus aureus isoladas de infecções intramamárias em vacas primíparas e de locais extramamários em cinco propriedades leiteiras através das técnicas de PCR por sequências palindrômicas extragênicas repetitivas (rep-PCR), usando iniciadores RW3A, e de eletroforese em gel de campo pulsado (PFGE), usando a endonuclease SmaI. Os isolados também foram avaliados in vitro quanto à suscetibilidade aos antimicrobianos beta-lactâmicos (penicilina e oxacilina). A tipagem por rep-PCR foi altamente discriminatória (valor D = 0,9804) e um total de 15 padrões foram detectados. Os isolados de S. aureus foram agrupados em três grupos diferentes (A a C), com 80% de similaridade. A técnica de PFGE também foi altamente discriminatória (valor D = 0,9667) e um total de 13 padrões foi observado. A análise do dendrograma com um coeficiente de similaridade de 80% gerou dois grupos diferentes (A e B). Além disso, cepas clonais isoladas do leite foram identificadas na mesma propriedade pelos dois métodos de tipificação e, apesar da presença de cepas dominantes, nossos resultados sugerem uma alta diversidade genética dentre as cepas de S. aureus analisadas. Um total de 15, dos 18 (83%) isolados, eram resistentes à penicilina e um dos 18 (6%) à oxacilina. Assim, esses achados confirmam a ocorrência de uma alta diversidade genética de cepas de S. aureus nas propriedades e a presença de cepas clonalmente relacionadas apenas na mesma propriedade, enfatizando uma variedade de perfis genotípicos entre os isolados.

Twins from different fathers: A heteropaternal superfecundation case report in Colombia / Gemelos de diferentes padres: un caso de superfecundación heteropaternal en Colombia

Abstract: Heteropaternal superfecundation is an extremely rare phenomenon that occurs when a second ova released during the same menstrual cycle is additionally fertilized by the sperm cells of a different man in separate sexual intercourse. In August, 2018, the Grupo de Genética de Poblaciones e Identificación at Universidad Nacional de Colombia received a request to establish the paternity of a pair of male twins with genetic markers. The following analyses were performed: amelogenin gene, autosomal short tandem repeat (STR), and Y-STR analyses by means of human identification commercial kits, paternity index, and the probability of paternity calculation and interpretation. A paternity index of 2.5134E+7 and a probability of paternity of 99.9999% for twin 2 were obtained while 14 out of 17 Y-chromosome markers and 14 out of 21 autosomal short tandem repeats were excluded for twin 1. The results indicated that the twins have different biological fathers. Although heteropaternal superfecundation is rarely observed among humans given its low frequency, in paternity disputes for dizygotic twins it is mandatory to demand the presence of the two twins in the testing to avoid wrong conclusions.

Resumen: La superfecundación heteropaternal es un fenómeno extremadamente raro que se produce cuando un segundo óvulo, liberado durante el mismo ciclo menstrual, es fertilizado por un espermatozoide de un hombre diferente en relaciones sexuales separadas. En agosto de 2018, el Grupo de Genética de Poblaciones e Identificación de la Universidad Nacional de Colombia recibió una solicitud para establecer la paternidad mediante marcadores genéticos de un par de mellizos varones, en quienes se hizo el análisis del gen de amelogenina, el análisis de repeticiones cortas en tándem (Short Tandem Repeats, STR) autosómicas y del cromosoma Y (Y-STR) mediante kits comerciales de identificación humana y cálculos e interpretación del índice de paternidad y probabilidad de paternidad. Se obtuvo un índice de paternidad de 2,5134E+7 y una probabilidad de paternidad de 99,9999 % para el gemelo 2, en tanto que en el gemelo 1 se excluyeron 14 de los 17 marcadores del cromosoma Y y 14 de los 21 sistemas STR autosómicos evaluados. Los resultados indicaron que los gemelos tienen diferentes padres biológicos. A pesar de que la superfecundación heteropaternal rara vez se observa en humanos debido a su baja frecuencia, en las disputas de paternidad para los gemelos dicigóticos, es obligatorio exigir en la prueba la presencia de los dos gemelos para evitar conclusiones incorrectas.

Use of molecular markers SSR and SCAR for identification of olive accessions / Uso de marcadores moleculares SSR e SCAR para identificação de acessos de oliveira

Morphological and agronomical describers are traditionally used in plant characterization. However, the usage of these describers have some limitations such as susceptibility to abiotic and biotic stress and environmental factors. Furthermore, the describers are not stable over time and many can only be evaluated during the adult phase of the plants, which requires time and physical space. Molecular markers offer numerous advantages compared to the conventional alternatives based on phenotype: they are stable and detectable in all vegetable tissues, and are independent of the environment and development phase. One of the main advantages of the use of molecular markers is the time reduction in the identification of genetic diversity among the studied subjects, as the genotypes may even be described for the seed or seedling phase. Many countries have already adopted molecular markers to identify olive cultivars more accurately. The aim of this study was to evaluate the genetic identity of eight olive accessions supposedly belonging to cultivar Arbequina by using microsatellite (SSR) and Sequence Characterized Amplified Region (SCAR) markers. One accession corresponding to the cultivar was also incorporated into the analysis as a reference genotype. The molecular marker data were analyzed on the software GENALEX6.The markers generated an accumulated PI and PE of 1.26 x 10-6 and 0.949, respectively.The results supported the hypothesis that all accessions belong to the cultivar Arbequina, and the markers can therefore be applied to other varieties of olive species.

Descritores morfológicos e agronômicos são tradicionalmente utilizados na caracterização de plantas. Apesar de recomendado, o emprego destes descritores apresenta algumas limitações como a influência a estresses abióticos e bióticos e aos efeitos do ambiente. Além disso, não são estáveis ao longo do tempo e muitos só podem ser avaliados durante a fase adulta das plantas, o que requer tempo e espaço físico para as avaliações. Os marcadores moleculares oferecem numerosas vantagens relativamente às alternativas convencionais baseadas no fenótipo, pois são estáveis e detectáveis em todos os tecidos vegetais, independente do ambiente e fase de desenvolvimento e uma das principais vantagens da utilização destes é proporcionar a redução do tempo na identificação da diversidade genética entre os indivíduos trabalhados, podendo ser avaliadas genótipos ainda na fase de semente ou de plântula. O objetivo deste estudo foi avaliar a identidade genética de oito acessos de oliveira supostamente pertencentes a cultivar Arbequina usando microssatélites (SSR) e Sequence Characterized Amplified Region (SCAR) marcadores. Um acesso correspondente a cultivar também foi incorporado na análise como o genótipo de referência. Os dados de marcadores moleculares foram analisados com o software GENALEX 6. Como resultado, os marcadores SSR geraram um PI acumulada e PE de 1,26 x 10- 6 e 0,949, respectivamente. Os resultados suportam a hipótese de que todos os acessos pertencem a cultivar Arbequina, e, por conseguinte, esses marcadores podem ser aplicados em situações semelhantes em outras variedades de espécies de oliveira.

DNA fingerprinting and genetic diversity in a threatened alpine medicinal plant, Gentiana crassicaulis (Gentianaceae) using AFLP / 药学学报

Jieji Nabao is a common Tibetan herb. According to our ethnobotanical studies, one of its original plants is identified as Gentiana crassicaulis Duthie ex Burk. (Gentianaceae). Endemic to the Qinghai-Tibet Plateau, this medicinal alpine plant is a threatened species. In this study, 163 individuals from 20 populations of G. crassicaulis were collected throughout its geographical range and amplified fragment length polymorphism (AFLP) was used to investigate genetic variation in this species. A cluster analysis was performed on the AFLP data with Halenia elliptica and Gentiana straminea as the outgroups. From 64 pairs of AFLP primer combinations, 12 pairs were selected for amplification and a total of 315 bands were amplified, of which 254 bands were polymorphic, accounting for 80.63%. High genetic differentiation was detected between populations (87%), and low within populations (13%). The UPGMA (unweighted pair-group method with arithmetic means) tree was topologically consistent with the traditional taxonomic treatments at the species level, and the populations of G. crassicaulis were divided into two branches: one from Yunnan and Guizhou, the other from Tibet, Qinghai, Sichuan and Gansu. PCA analysis and the Mantel test showed that there was a positive correlation between genetic distance and geographical distance. In addition, combined with SSR and SNP markers within cpDNA, the genetic differentiation within the Sichuan population S1 was validated.

Pharmacognostical study of Ochrosia elliptica Labill. (Apocynaceae)

Ochrosia elliptica Labill. is a small shrub belonging to family Apocynaceae and well-known as a promising anticancer agent. Botanical study of the plant was done for the young and old stems, stem bark, and leaves. Laticiferoustubes with yellowish brown content, isodiametric stone cells (sclereids), sclerenchyma (rod-like), and calcium oxalateclusters are the main diagnostic elements observed in this plant. Furthermore, DNA fingerprinting was done usingrapid amplified (RAPD) and inter simple sequence repeat (ISSR) techniques with the identification of a total of 30RAPD markers and 17 ISSR markers. Carbohydrates, sterols, catechol tannins, flavonoids, and alkaloids were presentin all the organs under investigation. This study could be valuable for quality control of the plant.

Teeth in forensics: A review

With the ever-increasing crime rate in our society, the field of forensic sciences has become highly evolved. Forensic dentists play a pivotal role in various areas of crime scene investigations and thereby help solve innumerable mysteries. Teeth appear to be vital pieces of evidence in several such investigations. Teeth are preserved in the closed cavities of the mouth and are generally resistant to the threatening environmental conditions that may be associated with the death of an individual, making them very useful in postmortem analysis. Teeth thus obtained may be useful in age estimation of the deceased victim or in determining his blood group. Identification of individuals in mass disasters can also be performed based on the unique morphological characteristics of the human dentition and through dental DNA fingerprinting. Again teeth play an all important role in catching a culprit through the positive correlation of the bite marks left behind at the crime scene and the suspect's own teeth marks. Thus, teeth prove to be an important adjunct in forensics. Its scope is ever-increasing with time, and a great amount of research is being carried out to implement the same. A PubMed, MEDLINE, and Scopus search was conducted of the past 70 years using several search terms like “Forensic odontology,” “history of forensic odontology,” “dental DNA fingerprinting,” “forensic age estimation,” “age estimation from teeth” and “bitemarks.” Other articles and textbook references which were considered to be important were also included in this study. The articles gathered were divided into the following groups: history of forensic odontology, teeth and DNA (dental DNA fingerprinting), teeth and blood grouping, teeth and age estimation, and teeth in bite marks.

Succession Law of Intestinal Flora after Death in SD Rats / 法医学杂志

OBJECTIVES@#To initially explore the sequential changes in the intestinal flora of corpse for the estimation of postmortem interval （PMI）.@*METHODS@#Rats were sacrificed by cervical dislocation, and samples were taken from their intestines using cotton swab to extract the DNA of intestinal flora. The 16S rRNA V3 universal primers were selected for PCR, and the PCR products were used for denatured gradient gel electrophoresis. The diversity and similarity analysis of intestinal flora were analyzed between groups, and the bands were cut from denaturing gradient gel electrophoresis. After purification, PCR and sequencing, the percentage of major bacteria in each group was obtained.@*RESULTS@#The flora diversity showed a reduced tendency from 1st to 30th day after death （ P<0.05）, while the intra-group similarity showed a downward trend （ P<0.05）. The number of bands and intra-group similarity coefficient （Cs） on the first day was higher than that of other groups （ P<0.05）. The intra-group Cs of the 25th and 30th day had a significant difference compared with the 5th day （ P<0.05）. At the genus level, the intestinal flora was mainly composed of Enterococcus sp. on the 1th and 5th day after death, Bacillus thuringienssis was the dominant species on the 10th, 15th and 20th day, and Enterococcus faecalis became the dominant species on the 25th and 30th day.@*CONCLUSIONS@#The composition and structure of intestinal flora change significantly in rats with the time after death, which indicates that the succession of intestinal flora is related to the postmortem interval.

Comparison of Bacteria ERIC-PCR Fingerprints of Index Fingers and Contactants / 法医学杂志

Objective To explore the bacteria relevance between index fingers and contactant' surfaces (mobile phone touch screen and desktop of personal office table). Methods Bacteria were collected from the index fingers, mobile phone touch screen and desktop of personal office table of 10 volunteers. Enterobacterial repetitive intergenic consensus(ERIC)-PCR fingerprint was established by PCR amplifi-cation technique of metagenome. Results There were 7 volunteers' ERIC-PCR fingerprints of index fin-gers matched that took from the mobile phone touch screens, and different from each other. There were 3 volunteers' ERIC-PCR fingerprints of index fingers matched that took from desk top of personal office table, and other 7 volunteers' ERIC-PCR fingerprints did not match perfectly with that took from desk top of personal office table,but had at least one similar band for both. Conclusion The bacteria on index finger shows individual specificity, which on mobile phone touching screen and personal desktop may be a new biological sample of forensic identification.

Preliminary study on serum pharmacochemistry of Shijing pill / 国际中医中药杂志

Objective To conduct serum pharmacochemistry study onShijing pill. Methods HPLC fingerprints of serum in rats after takingShijingpill were established,the serun samples after takingShijing pill, decoction without one of the component drugs and single crude drug were compared,the transitional constituents absorbed into the blood and the original crude after takingShijing pill was determined.Results Fourteen transitional constituents were detected in rat blood after takingShijing pill,four of which were permanent component of blood,seven of which were prototype constituents,three of which were metabolites. Conclusions Some of chemical composition ofShijing pill was clarified preliminarily,that contributes to further studying pharmacodynamic meterial foundation ofShijing pill.

Preliminary study on serum pharmacochemistry of Shijing pill / 国际中医中药杂志

Objective To conduct serum pharmacochemistry study onShijing pill. Methods HPLC fingerprints of serum in rats after takingShijingpill were established,the serun samples after takingShijing pill, decoction without one of the component drugs and single crude drug were compared,the transitional constituents absorbed into the blood and the original crude after takingShijing pill was determined.Results Fourteen transitional constituents were detected in rat blood after takingShijing pill,four of which were permanent component of blood,seven of which were prototype constituents,three of which were metabolites. Conclusions Some of chemical composition ofShijing pill was clarified preliminarily,that contributes to further studying pharmacodynamic meterial foundation ofShijing pill.

Assessment of genetic diversity on Gentiana straminea based on ISSR markers / 中草药

Objective: To assess the genetic diversity of Gentiana straminea (Gentianaceae). Methods: Intersimple sequence repeat (ISSR) markers were used. Twenty-eight populations (83 individuals) of G. straminea were sampled from Sichuan, Qinghai, Gansu provinces, and Autonomous Region of Tibet. ISSR data were analyzed with the program POPGEN, and a UPGMA dendrogram was constructed. Results: PCR products corresponding to 183 alleles at 95 loci, amplified by the seven primers were scored. Eighty-eight loci were polymorphic. Expected heterozygosity (He) and Shannon's information index (Im) were 0.288 2 and 0.437 1, respectively. Nei's coefficient of genetic differentiation (Gst) and gene flow (Nm) were 0.678 3 and 0.237 1 at the population level, respectively. The genetic distance varied from 0.074 3 to 0.490 0. G. straminea populations were mainly divided into two braches by UPGMA tree. Conclusion: Genetic diversity level of G. straminea is high, and genetic diversity level among populations of G. straminea is higher than that in one population; Its autogenous variation mainly exist among populations; Hereditary capacity has certain correlation with geographical distribution. This work contributes basic information for variety identification, species in situ conservation, and enviroment investigation on the influence of heredity differentiated and Chinese Materia Medica genuine study. The congruence between genetic distance and geographic distances is high.

Recognition of Y Fragment Deletion by Genotyping Graphs after Amplified by PowerPlex® 21 Detection Kit / 法医学杂志

OBJECTIVES@#To recognize the possibility of Y fragment deletion of Amelogenin gene intuitively and simply according to the genotyping graphs.@*METHODS@#By calculating the ratio of total peak height of genotyping graphs, the statistics of equilibrium distribution between Amelogenin and D3S1358 loci, Amelogenin X-gene and Amelogenin Y-gene, and different alleles of D3S1358 loci from 1 968 individuals was analyzed after amplified by PowerPlex® 21 detection kit.@*RESULTS@#Sum of peak height of Amelogenin X allele was not less than 60% that of D3S1358 loci alleles in 90.8% female samples, and sum of peak height of Amelogenin X allele was not higher than 70% that of D3S1358 loci alleles in 94.9% male samples.@*CONCLUSIONS@#The result of genotyping after amplified by PowerPlex® 21 detection kit shows that the possibility of Y fragment deletion should be considered when only Amelogenin X-gene of Amelogenin is detected and the peak height of Amelogenin X-gene is not higher than 70% of the total peak height of D3S1358 loci.

Recognition of Y Fragment Deletion by Genotyping Graphs after Amplified by PowerPlex?21 Detection Kit / 法医学杂志

Objective To recognize the possibility of Y fragment deletion of Amelogenin gene intuitively and simply according to the genotyping graphs.Methods By calculating the ratio of total peak height of genotyping graphs, the statistics of equilibrium distribution between Amelogenin and D3S1358loci,Amelo-genin X-gene and Amelogenin Y-gene, and different alleles of D3S1358loci from 1968 individuals was analyzed after amplified by PowerPlex? 21 detection kit.Results Sum of peak height of Amelogenin X allele was not less than 60% that of D3S1358loci alleles in 90.8% female samples, and sum of peak height of Amelogenin X allele was not higher than 70% that of D3S1358loci alleles in 94.9% male samples.Conclusion The result of genotyping after amplified by PowerPlex? 21 detection kit shows that the possibility of Y fragment deletion should be considered when only Amelogenin X-gene of Amelo-geninis detected and the peak height of AmelogeninX-gene is not higher than 70% of the total peak height of D3S1358loci.

Research on HPLC fingerprint ofJinjing solid beverage / 国际中医中药杂志

Objective To establish an HPLC fingerprint ofJinjing solid beverage,and provide experimental evidence of evaluating its quality.Methods Ten batches ofJinjing solid beverage were analyzed by HPLC under the gradient elution condition. A Kromasil C18 column(4.6 mm×250 mm, 5μm) was used, and the flow phase was acetonitrile-H2O(acidified to 0.1% with phosphoric acid) with gradient elution, and the detection wavelength was 327 nm, and the flow rate was 1.0 ml/min, and the column temperature was 35℃. The data were evaluated by the similarity evaluation software for TCM fingerprint.Results The HPLC fingerprint ofJinjing solid beverage were established. Twelve common peaks including chlorogenic acid were identified with similarity of more than 0.9.Conclusion HPLC method is a reliable, available and quick method, that provides a means for controlling and evaluating the quality ofJinjing solid beverage.

Factors Affecting DNA Yields from Serum and Plasma Samples Used for Personal Identification Testing / 대한법의학회지

Serum or plasma is free of cellular components. As DNA is in the nucleus or mitochondria of a cell, it can be presumed that serum/plasma is DNA free. However, there are cases wherein serum/plasma is the only resource available for identification analysis, yet no sufficient data are available regarding whether reliable DNA testing can be applied to such cases, and what the influencing factors are when testing is a valid course of action. The aim of this study is to illustrate the factors that can be used in the genetic testing of serum/plasma when identifying an individual. The results showed that the concentration of serum DNA significantly increased over time in 4℃ storage, and the DNA yields from samples stored in heparin tubes were overall higher than from samples stored in ethylenediaminetetraacetic acid tubes. We observed that the concentration of DNA in serum successfully matched 100% to the short tandem repeat data of blood DNA.