1.Research Progress on Individual Identification Using Forensic Imaging Data under the Influence of Evidence Rule.
Jian Jun WANG ; Jun Chang PEI ; Yun Liang QIU
Journal of Forensic Medicine 2016;32(5):367-372
With the progress and development of the DNA test and imaging technique, and the evolution of evidence rule which bring the discussions about whether the individual identification using imaging data is outdated, and other disputes such as whether radiologic evidence could be suitable for contemporary evidence and be used to solve the posture difference of imaging test. This article summaries the domestic and foreign researches of individual identification using imaging data in the past 20 years and reviews the problems above.
DNA/analysis*
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Forensic Genetics/standards*
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Humans
2.Identification of Cannabis Sativa L. Based on rbcL Sequence.
Ruo Cheng XIA ; Xiao Chun ZHANG ; Xiao Xiao WANG ; Qi YANG ; Chong CHEN ; Huan YU ; Yi Ling QU ; Zi Wei WANG ; Yan SHI ; Ping XIANG ; Su Hua ZHANG ; Cheng Tao LI
Journal of Forensic Medicine 2021;37(2):187-191
Objective To assess the feasibility of the rbcL sequence of chloroplast DNA as a genetic marker to identify Cannabis sativa L. Methods The rbcL sequences in 62 Cannabis sativa L. samples, 10 Humulus lupulus samples and 10 Humulus scandens DNA samples were detected, and 96 rbcL sequences of the Cannabaceae family were downloaded from Genbank. Sequence alignment was performed by MEGA X software, the intraspecific and interspecific Kimura-2-Parameter (K2P) genetic distances were calculated, and the system clustering tree was constructed. Results The rbcL sequence length acquired by sequencing of Cannabis sativa L. and Humulus scandens were 617 bp and 649 bp, respectively, and two haplotypes of Cannabis sativa L. were observed in the samples. The BLAST similarity search results showed that the highest similarity between the sequences acquired by sequencing and Cannabis sativa L. rbcL sequences available from Genbank was 100%. The genetic distance analysis showed that the maximum intraspecific genetic distance (0.004 9) of Cannabis sativa L. was less than the minimum interspecific genetic distance (0.012 9). The results of median-joining network and system clustering tree analysis showed that Cannabis sativa L. and other members of the Cannabaceae family were located in different branches. Conclusion The rbcL sequence could be used as a DNA barcode for identifying Cannabis sativa L., and combined with comparative analysis of the rbcL sequence and system cluster analysis could be a reliable and effective detection method for Cannabis sativa L. identification in forensic investigation.
Cannabis/genetics*
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Genetic Markers
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Sequence Analysis, DNA
3.Identification of Dendrobii Caulis basing on ITS sequence.
Zi YE ; Ye LU ; Zheng-Tao WANG ; Hong XU ; Zhi-Bi HU
China Journal of Chinese Materia Medica 2014;39(20):3928-3935
Isolation of high-quality genomic DNA from dried and processed crude drug is the key for the DNA identification of Dendrobii Caulis. The DNA extract of Dendrobii Caulis was firstly compared using different method to isolate genomic DNA from dried and processed crude drug, including commercial DNA extracted kit and CTAB method. Using modified CTAB method (extracted from large samples), the genomic DNA was successfully isolated from Dendrobii Caulis, including Huangcao and Fengdou. The ITS regions were amplified using the purified DNA as template, and then cloned and sequenced. These ITS sequences were compared with data from Genbank database and our lab, 14 Dendrobium species and five similar species were identified from "Huangcao" and "Huangcao" slice, while six species and three similar species from "Fengdou" according to their sequence similarity. The study demonstrated that the dried Dendrobii Caulis could be identified using DNA molecular method, which could overcome deficiencies and limitations of traditional identification method and has a certain application prospects.
DNA, Intergenic
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genetics
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DNA, Plant
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genetics
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Dendrobium
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classification
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genetics
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Sequence Analysis, DNA
4.Clinical feature and ATP8B1 mutation analysis of a patient with progressive familial intrahepatic cholestasis type I.
Ying CHENG ; Li GUO ; Yuan-Zong SONG
Chinese Journal of Contemporary Pediatrics 2016;18(8):751-756
Progressive familial intrahepatic cholestasis type I (PFIC1) is an autosomal recessive disorder caused by biallelic mutations of ATP8B1 gene, with progressive cholestasis as the main clinical manifestation. This paper reports the clinical and genetic features of a PFIC1 patient definitely diagnosed by ATP8B1 genetic analysis. The patient, a boy aged 14 months, was referred to the hospital with the complaint of jaundiced skin and sclera over 10 months. The patient had been managed in different hospitals, but the therapeutic effects were unsatisfactory due to undetermined etiology. On physical examination, hepatosplenomegaly was discovered in addition to jaundice of the skin and sclera. The liver was palpable 4 cm below the right subcostal margin and 2 cm below the xiphoid while the spleen 2 cm below the left subcostal margin. The liver function test revealed elevated levels of serum total bile acids, bilirubin, and transaminases; however, the γ-glutamyl transferase level was normal. The diagnosis was genetic cholestasis of undetermined origin. At the age of 1 year and 8 months, a Roux-en-Y cholecystocolonic bypass operation was performed, and thereafter the jaundice disappeared. At 5 years and 1 month, via whole genome sequencing analysis and Sanger sequencing confirmation, the boy was found to be a homozygote of mutation c.2081T>A(p.I694N) of ATP8B1 gene, and thus PFIC1 was definitely diagnosed. The boy was followed up until he was 6 years, and jaundice did not recur, but the long-term outcome remains to be observed.
Adenosine Triphosphatases
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genetics
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Cholestasis, Intrahepatic
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genetics
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DNA Mutational Analysis
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Humans
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Infant
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Male
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Mutation
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Sequence Analysis, DNA
5.Applying DNA barcoding technique to identify menthae haplocalycis herba.
Xiaohui PANG ; Haibin XU ; Jianping HAN ; Jingyuan SONG
China Journal of Chinese Materia Medica 2012;37(8):1114-1117
OBJECTIVETo identify Menthae Haplocalycis Herba and its closely related species using DNA barcoding technique.
METHODTotal genomic DNA was isolated from Mentha canadensis and its closely related species. Nuclear DNA ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner V3.0. The Kimura 2-Parameter (K2P) distances were calculated using software MEGA 5.0. Identification analyses were performed using BLAST1, Nearest Distance and neighbor-joining (NJ) methods.
RESULTThe intra-specific genetic distances of M. canadensis were ranged from 0 to 0.006, which were lower than inter-specific genetic distances between M. canadensis and its closely related species (0.071-0.231). All the three methods showed that ITS2 could discriminate M. canadensis from its closely related species correctly.
CONCLUSIONThe ITS2 region is an efficient barcode for identification of Menthae Haplocalycis Herba, which provides a scientific basis for fast and accurate identification of the herb.
DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; genetics ; Plants, Medicinal ; classification ; genetics ; Sequence Analysis, DNA ; methods
6.DNA barcoding identification of original plants of a rare medicinal material Resina Draconis and related Dracaena species.
Yue ZHANG ; Mei-Fang SONG ; Hai-Tao LI ; Hui-Fang SUN ; Zhong-Lian ZHANG
China Journal of Chinese Materia Medica 2021;46(9):2173-2181
Resina Draconis, a rare and precious traditional medicine in China, is known as the "holy medicine for promoting blood circulation". According to the national drug standard, it's derived from the resin extracted from the wood of Dracaena cochinchinensis, a Liliaceae plant. In addition, a variety of Dracaena species all over the world can form red resins, and there is currently no molecular identification method that can efficiently identify the origin of Dracaena medicinal materials. In this study, seven species of Dracaena distributed in China were selected as the research objects. Four commonly used DNA barcodes(ITS2, matK, rbcL and psbA-trnH), and four highly variable regions(trnP-psaJ, psbK-psbI, trnT-trnL, clpP) in chloroplast genome were used to evaluate the identification efficiency of Dracaena species. The results showed that clpP sequence fragment could accurately identify seven species of Dracaena plants. However, due to the long sequence of clpP fragment, there were potential problems in the practical application process. We found that the combined fragment "psbK-psbI+ trnP-psaJ" can also be used for accurate molecular identification of the Resina Draconis origin plants and relative species of Dracaena, which were both relatively short sequences in the combined fragment, showing high success rates of amplification and sequencing. Therefore, the "psbK-psbI+ trnP-psaJ" combined fragment can be used as the DNA barcode fragments for molecular identification of Resina Dracon's origin plants and relative species of Dracaena. Research on the identification of Dracaena species, the results of this study can be used to accurately identify the original material of Resina Draconis, and providing effective means for identification, rational development and application of Resina Draconis base source.
China
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DNA Barcoding, Taxonomic
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DNA, Plant/genetics*
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Dracaena/genetics*
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Plants
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Resins, Plant
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Sequence Analysis, DNA
7.Study on sequence characterized amplified region (SCAR) markers of Polygonum capitatum.
Tao ZHOU ; Yu XIE ; Li-Yan ZHANG ; Sheng-Hua WEI ; Yan-Lei JIN
China Journal of Chinese Materia Medica 2013;38(16):2577-2580
OBJECTIVETo establish sequence characterized amplified region markers of Polygonum capitatum.
METHODThe random primer was screened through RAPD to obtain the specific RAPD marker band, and the band was separated, extracted, cloned and sequenced. The specific primers were designed for conventional PCR reaction on the basis of the specific band, and the SCAR marker was acquired.
RESULTScreening from 50 RAPD primer, only C29 primer had 2 specific bands could distinguish P. capitatum from P. nepalense, then 4 pairs of specific primers were designed based on the 2 sequences of RAPD marker bands, and only 1 pair primer (Z1-2) was successfully converted into SCAR marker after repeated tests.
CONCLUSIONThe Z1-2 primer, could be used as an effective SCAR mark to identify Z300 DNA for P. capitatum. The SCAR mark was established and can be used as a molecular marker to distinguish P. capitatum from P. nepalense
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; Genetic Markers ; genetics ; Polygonum ; classification ; genetics ; Random Amplified Polymorphic DNA Technique ; Sequence Analysis, DNA
9.Analysis of synonymous codon usage and evolution of begomoviruses.
Xiao-zhong XU ; Qing-po LIU ; Long-jiang FAN ; Xiao-feng CUI ; Xue-ping ZHOU
Journal of Zhejiang University. Science. B 2008;9(9):667-674
Begomoviruses are single-stranded DNA viruses and cause severe diseases in major crop plants worldwide. Based on current genome sequence analyses, we found that synonymous codon usage variations in the protein-coding genes of begomoviruses are mainly influenced by mutation bias. Base composition analysis suggested that the codon usage bias of AV1 and BV1 genes is significant and their expressions are high. Fourteen codons were determined as translational optimal ones according to the comparison of codon usage patterns between highly and lowly expressed genes. Interestingly the codon usages between begomoviruses from the Old and the New Worlds are apparently different, which supports the idea that the bipartite begomoviruses of the New World might originate from bipartite ones of the Old World, whereas the latter evolve from the Old World monopartite begomoviruses.
Begomovirus
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genetics
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Biological Evolution
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Chromosome Mapping
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Codon
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genetics
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DNA Mutational Analysis
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DNA, Viral
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genetics
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Evolution, Molecular
10.Analysis of 28S rRNA and COⅠ Gene Sequence of Nine Necrophagous Calliphorid Flies from Luoyang.
Lin Lin ZHAO ; Xian Dun ZHAI ; Zhe ZHENG ; Zhou LÜ ; Yong Lin LI ; Yao Nan MO
Journal of Forensic Medicine 2019;35(2):181-186
Objective To assess the feasibility of using 28S ribosomal RNA (28S rRNA) and mitochondrial cytochrome c oxidase subunit Ⅰ (COⅠ) gene sequences of nine necrophagous Calliphorid flies for the identification of common necrophagous Calliphorid flies, and to provide technical support for postmortem interval (PMI) estimation. Methods Twenty-three Calliphorid flies were collected and identified morphologically, and DNA were extracted from legs. The gene fragments of 28S rRNA and COⅠ were amplified and sequenced, then the sequence alignment was performed with BLAST. The composition of obtained sequences was analyzed and evolutionary divergence rate between species and intraspecies were established. The phylogeny tree was constructed with neighbor-joining method. Results The 23 necrophagous Calliphorid flies were identified to 9 species of 5 genera. The 715 bp from 28S rRNA and 637 bp from COⅠ gene were obtained and the online BLAST result showed more than 99% of similarity. The phylogeny tree showed that the necrophagous flies could cluster well into 9 groups, which was consistent with morphological identification results. The intraspecific difference in 28S rRNA was 0 and the interspecific difference was 0.001-0.033. The intraspecific difference in COⅠ was 0-0.008 and the interspecific difference was 0.006-0.101. Conclusion Combined use of 28S rRNA and COⅠ gene sequence fragments can effectively identify the nine Calliphorid flies in this study. However, for closely related blowfly species, more genetic markers should be explored and used in combination in future.
Animals
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DNA, Mitochondrial/genetics*
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Diptera/genetics*
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Phylogeny
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RNA, Ribosomal, 28S/genetics*
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Sequence Analysis, DNA
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Species Specificity