Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts.However,the conventional b...Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts.However,the conventional body fluid identification methods are prone to various limitations,such as time consumption,intensive labor,nonparallel manner,varying degrees of sensitivity and limited specificity.Recently,the analysis of cell-specific messenger RNA expression(mRNA profiling)has been proposed to supplant conventional methods for body fluid identification.Since 2011,the collaborative exercises have been organized by the European DNA Profiling Group(EDNAP)in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification.The major advantages of mRNA profiling,compared to the conventional methods,include higher sensitivity,greater specificity,the ability of detecting several body fluids in one multiplex reaction,and compatibility with current DNA extraction and analysis procedure.In the current review,we provided an overview of the present knowledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possible practical application to forensic casework.展开更多
To further enrich the genetic data of the Chinese Xinjiang Mongolian group,the genetic distribution and forensic parameters of 19 autosomal short tandem repeats(STRs)were investigated.Altogether,249 alleles were obser...To further enrich the genetic data of the Chinese Xinjiang Mongolian group,the genetic distribution and forensic parameters of 19 autosomal short tandem repeats(STRs)were investigated.Altogether,249 alleles were observed in these 19 STRs.The mean values of the polymorphism information content(PIC),match probability(MP),discrimination power(DP),and probability of exclusion(PE)for these 19 STRs were 0.7775,0.0699,0.9301,and 0.6085,respectively.Additionally,the cumulative DP and PE values obtained in the Mongolian group were 0.999 999 999 999 999 999 999 995 67 and 0.999 999 992 163,respectively.展开更多
Adipose-derived stromal cells （ASCs） have gained great attention in regenerative medicine. Progress in our understanding of adult neovascularization further suggests the potential of ASCs in promoting vascular regen...Adipose-derived stromal cells （ASCs） have gained great attention in regenerative medicine. Progress in our understanding of adult neovascularization further suggests the potential of ASCs in promoting vascular regeneration, although the specific cues that stimulate their angiogenic behavior remain controversial In this study, we established a three-dimensional （3D） angiogenesis model by co-culturing ASCs and endothelial cells （ECs） in collagen gel and found that ASC-EC-instructed angiogenesis was regulated by the canonical Wnt pathway. Furthermore, the angiogenesis that occurred in implants collected after injections of our collagen gel- based 3D angiogenesis model into nude mice was confirmed to be functional and also regulated by the canonical Wnt pathway. Wnt regulation of angiogenesis involving changes in vessel length, vessel density, vessel sprout, and connection numbers occurred in our system. Wnt signaling was then shown to regulate ASC- mediated paracrine signaling during angiogenesis through the nuclear translocation of β-catenin after its cytoplasmic accumulation in both ASCs and ECs. This translocation enhanced the expression of nuclear cofactor Lef-1 and cyclin D1 and activated the angiogenic transcription of vascular endothelial growth factor A （VEGFA）, basic fibroblast growth factor （bFGF）, and insulin-like growth factor 1 （IGF-1）. The angiogenesis process in the 3D collagen model appeared to follow canonical Wnt signaling, and this model can help us understand the importance of the canonical Wnt pathway in the use of ASCs in vascular regeneration.展开更多
Bones and teeth often represent the only sources of DNA available for identifying human remains.DNA in bones and teeth is generally better preserved than that in soft tissues because of the presence of hard connective...Bones and teeth often represent the only sources of DNA available for identifying human remains.DNA in bones and teeth is generally better preserved than that in soft tissues because of the presence of hard connective tissue with a high level of calcium.Because of the extensive mineralisation,the choice of an efficient DNA extraction procedure is important to minimise the sampling of a high level of minerals and to remove polymerase chain reaction(PCR)inhibitors.Some protocols are available for DNA extraction from bones and teeth as part of the Qiagen EZ1 DNA Investigator Kit using the EZ1 Advanced XL automated purification platform.To improve the efficiency of DNA extraction from skeletal remains,the present study focuses on a modification to these already available protocols.In this study,different bones and teeth collected between 1 and 50 years after death were subjected to DNA extraction using the standard EZ1 protocol,a supplementary protocol,and a modified protocol.The modified approach included a decalcification step,whereas the Qiagen protocols worked directly on non-decalcified powder.In all three procedures,150 mg samples were used for DNA extraction.We evaluated the quantity of DNA recovered from samples,the presence of any PCR inhibitors co-extracted,the level of DNA degradation,the quality of short tandem repeat(STR)profiles,and the reproducibility of the modified procedure.When compared with the other protocols,the modified protocol resulted in the best recovery of DNA that was free of PCR inhibitors.Additionally,the STR profiles were reliable and of high quality.In our opinion,the decalcification step increases DNA recovery by softening tissues,which allows lysis solutions to act more effectively.Furthermore,the use of two lysis solutions and the variation added to the EZ1 purification step allow for DNA recovery with quality and quantity superior to those of the previously available Qiagen-based protocols.These findings may be helpful solutions to the problems commonly encountered when dealing with difficult samples,such as bones and teeth.展开更多
Genetic profiling is a standard procedure for human identification,i.e.in criminal cases and mass disasters,and has been proven to be an important part in the process in the repatriation of victims to their relatives....Genetic profiling is a standard procedure for human identification,i.e.in criminal cases and mass disasters,and has been proven to be an important part in the process in the repatriation of victims to their relatives.In the event of a catastrophe whether it be a natural disaster,terror attack or accident,fatalities of many nationalities may be a consequence and international collaboration becomes necessary.Current DNA techniques used on a routine basis at forensic laboratories world-wide are very useful,and results reported from different labs are compared,making it possible to be matched in order to declare the identification of a victim.Statistical calculations of possibilities of a random match are achievable since population data from many parts of the world are available.However,decomposition and degradation of the remains are not uncommon in the aftermath of a catastrophe and hence it may be difficult to retrieve detailed DNA profiles from such samples.Massive parallel sequencing(MPS)is a technique capable of producing a vast amount of DNA sequence data in a high-through put manner,and panels of single nucleotide polymorphism(SNP)markers allow the amplification of small DNA fragments,often seen in compromised samples.Here,we report the results from a set of 10 samples from missing person identification cases,analyzed with an MPS based method comprising 131 SNP markers and compared with direct reference material or buccal swab samples collected from relatives of the deceased.We assess the weight of evidence of a match by statistical calculation.Furthermore,we compare results reported on different platforms using different SNP panels,and conclude that more work has to be done if results from missing person identification cases analyzed on MPS with SNP panels at different laboratories are to be fully reliable and thus comparable.展开更多
Epigenetics governs a chromatin state regulatory system through five key mechanisms:DNA modification,histone modification,RNA modification,chromatin remodeling,and non-coding RNA regulation.These mechanisms and their ...Epigenetics governs a chromatin state regulatory system through five key mechanisms:DNA modification,histone modification,RNA modification,chromatin remodeling,and non-coding RNA regulation.These mechanisms and their associated enzymes convey genetic information independently of DNA base sequences,playing essential roles in organismal development and homeostasis.Conversely,disruptions in epigenetic landscapes critically influence the pathogenesis of various human diseases.This understanding has laid a robust theoretical groundwork for developing drugs that target epigenetics-modifying enzymes in pathological conditions.Over the past two decades,a growing array of small molecule drugs targeting epigenetic enzymes such as DNA methyltransferase,histone deacetylase,isocitrate dehydrogenase,and enhancer of zeste homolog 2,have been thoroughly investigated and implemented as therapeutic options,particularly in oncology.Additionally,numerous epigenetics-targeted drugs are undergoing clinical trials,offering promising prospects for clinical benefits.This review delineates the roles of epigenetics in physiological and pathological contexts and underscores pioneering studies on the discovery and clinical implementation of epigenetics-targeted drugs.These include inhibitors,agonists,degraders,and multitarget agents,aiming to identify practical challenges and promising avenues for future research.Ultimately,this review aims to deepen the understanding of epigeneticsoriented therapeutic strategies and their further application in clinical settings.展开更多
To evaluate the promising advantages of massively parallel sequencing(MPS)in our casework,we analysed a total of 33 Y-chromosomal short tandem repeats(Y-STRs)with traditional capillary electrophoresis(CE)and 25 Y-STRs...To evaluate the promising advantages of massively parallel sequencing(MPS)in our casework,we analysed a total of 33 Y-chromosomal short tandem repeats(Y-STRs)with traditional capillary electrophoresis(CE)and 25 Y-STRs using the newer MPS technology.We studied the outcome of both technologies in 64 father-son pairs using stock and custom-designed kits.Current MPS technology confirmed the 13 mutational events observed with CE and improved our understanding of the complex nature of STR mutations.By detecting isometric sequence variants between unrelated males,we show that sequencing Y-STRs using MPS can boost discrimination power.展开更多
We describe the formation of the major axon pathways in the embryonic central and peripheral nervous systems of the amphipod crustacean Orchestia cavimana Heller,1865 by means of antibody staining against acetylated a...We describe the formation of the major axon pathways in the embryonic central and peripheral nervous systems of the amphipod crustacean Orchestia cavimana Heller,1865 by means of antibody staining against acetylated alphatubulin.The data add to a long list of previous studies of various other aspects of development in Orchestia and provide a basis for future studies of neurogenesis on a deeper cellular and molecular level.Orchestia exhibits a tripartite dorsal brain,which is a characteristic feature of euarthropods.Its anlagen are the first detectable structures in the developing nervous system and can be traced back to distinct neuronal cell clusters in the early embryo.The development of the ventral nervous system proceeds with an anteroposterior gradient of development.In each trunk segment,the longitudinal connectives and the anterior commissure form first,followed by the intersegmental nerve,the posterior commissure and segmental nerves,respectively.A single commissure of a vestigial seventh pleonal segment is found.In the peripheral nervous system we observe a spatial and temporal pattern of leg innervation,which is strikingly similar in both limb types,the uniramous pereopods and the biramous pleopods.A proximal leg nerve splitting distally into two separated nerves probably reflects a general feature of crustaceans.展开更多
In this research,genotyping data of 43 InDel loci in 311 Han individuals in Ankang City,Shaanxi Province,China were detected using a self-developed five-dye multiplex amplification panel.The allelic frequencies and fo...In this research,genotyping data of 43 InDel loci in 311 Han individuals in Ankang City,Shaanxi Province,China were detected using a self-developed five-dye multiplex amplification panel.The allelic frequencies and forensic parameters of all InDel loci were calculated.The combined power of discrimination and probability of exclusion values were 0.99999999999999999882739 and 0.999887424,respectively,which demonstrated that this 43-InDel panel was powerful for individual identifications in Ankang Han population.Moreover,genetic distances,pairwise F_(ST)values,principal component analyses,phylogenetic trees and STRUCTURE analyses were performed to investigate the genetic affinities between Ankang Han and reference groups.Population genetic investigations indicated that Ankang Han population had a close genetic relationship with Southern Han population compared with other reference groups.展开更多
As a result of the expansion of old Tibet on the Qinghai-Tibet Plateau,Tibetans diverged into three main branches,Ü-Tsang,Amdo,and Kham Tibetan.Ü-Tsang Tibetans are geographically distributed across the wide...As a result of the expansion of old Tibet on the Qinghai-Tibet Plateau,Tibetans diverged into three main branches,Ü-Tsang,Amdo,and Kham Tibetan.Ü-Tsang Tibetans are geographically distributed across the wide central and western portions of the Qinghai-Tibet Plateau while Lhasa is the central gathering place for Tibetan culture.The AGCU Y30,a 6-dye fluorescence kit including 30 slowly and moderately mutated Y-STR loci,has been validated for its stability and sensitivity in different biomaterials and diverse Chinese populations(Han and other minorities),and widely used in the practical work of forensic science.However,the 30 Y-STR profiling of Tibetan,especially forÜ-Tsang Tibetan,were insufficient.We utilized the AGCU Y30 to genotype 577Ü-Tsang Tibetan unrelated males from Lhasa in the Tibet Autonomous Region of China to fill up the full and accurate Y-STR profiles.A total of 552 haplotypes were observed,536(97.10%)of which were unique.One hundred and ninety-four alleles were observed at 26 single copy loci and the allelic frequencies ranged from 0.0017 to 0.8180.For the two multi-copy loci DYS385a/b and DYS527a/b,64 and 36 allelic combinations were observed,respectively.The gene diversity(GD)values ranged from 0.3079 at DYS391 to 0.9142 at DYS385a/b and the overall haplotype diversity(HD)was 0.9998,and its discrimination capacity(DC)was 0.9567.The population genetic analyses demonstrated that LhasaÜ-Tsang Tibetan had close relationships with other Tibetan populations from Tibet and Qinghai,especially withÜ-Tsang Tibetan.From the perspective of Y haplogroups,the admixture of the southward Qiang people with dominant haplogroup O-M122 and the northward migrations of the initial settlers of East Asia with haplogroup D-M175 hinted the Sino-Tibetan homologous,thus,we could not ignore the gene flows with other Sino-Tibetan populations,especially for Han Chinese,to characterize the forensic genetic landscape of Tibetan.展开更多
The custom-designed single nucleotide polymorphism(SNP)panel amplified 231 autosomal SNPs in one PCR reaction and subsequently sequenced with massively parallel sequencing(MPS)technology and Ion Torrent personal genom...The custom-designed single nucleotide polymorphism(SNP)panel amplified 231 autosomal SNPs in one PCR reaction and subsequently sequenced with massively parallel sequencing(MPS)technology and Ion Torrent personal genome machine(PGM).SNPs were chosen from SNPforID,IISNP,HapMap,dbSNP,and related published literatures.Full concordance was obtained between available MPS calling and Sanger sequencing with 9947A and 9948 controls.Ten SNPs(rs4606077,rs334355,rs430046,rs2920816,rs4530059,rs1478829,rs1498553,rs7141285,rs12714757 and rs2189011)with low coverage or heterozygote imbalance should be optimized or excluded from the panel.Sequence data had sufficiently high coverage and gave reliable SNP calling for the remaining 221 loci with the custom MPS-SNP panel.A default DNA input amount of 10 ng per reaction was recommended by Ampliseq technology but sensitivity testing revealed positive results from as little as 1 ng input DNA.Mixture testing with this panel is possible through analysis of the F MAR(frequency of major allele reads)values at most loci with enough high coverage depth and low level of sequencing noise.These results indicate the potential advantage of the custom MPS-SNP assays and Ion Torrent PGM platform for forensic study.展开更多
Background:In forensic investigations,accurate estimation of the postmortem interval(PMI)is an important task,but also an ongoing challenge.Especially in cases where the cadaver has been specially treated,for example,...Background:In forensic investigations,accurate estimation of the postmortem interval(PMI)is an important task,but also an ongoing challenge.Especially in cases where the cadaver has been specially treated,for example,by boiling,the determination of PMI becomes extremely difficult.Previous studies have shown that the succession of the microbial community after decomposition of the cadaver can be used to infer PMI.However,the feasibility of determining the PMI of boiled cadavers has not yet been demonstrated.Aims and Objectives:The main objective of this study was to test whether we can infer PMI of boiled cadavers based on the succession of microbial communities.Materials and Methods:SD rats were killed by cervical dislocation.Subsequently,the rat cadavers were divided into the case(boiled cadavers)and control(unboiled cadavers)groups.Rectal samples were collected from the rats for 45 days and at nine time points.High-throughput sequencing of the 16S rRNA gene was performed to characterize the microbial community in the rectum.Results:The results showed that the composition and relative abundance of bacterial communities at the phylum level were significantly different between the case and control groups.The alpha diversity of the microbial community showed a decreasing trend with the decomposition process.Principal coordinate analysis showed that the case and control groups had obvious patterns along the succession of microbial communities.The rectal microbial communities showed a significant linear trend in the time course of decomposition.A random forest model was used to infer PMI.The goodness-of-fit(R2)of the model was 68.00%and 84.00%,and the mean absolute errors were 2.05 and 1.48 days within 45 days of decomposition for the case and control groups,respectively.Conclusions:Our results suggest that microbial community succession could be a potential method to infer PMI of boiled cadavers.展开更多
The field of forensic DNA typing,often referred to as“DNA fingerprinting,”has evolved and expanded considerably since its beginnings in the mid-1980s.Originally,forensic DNA typing was primarily used for individual ...The field of forensic DNA typing,often referred to as“DNA fingerprinting,”has evolved and expanded considerably since its beginnings in the mid-1980s.Originally,forensic DNA typing was primarily used for individual identification and criminal investigations,but it has evolved into a versatile discipline with a wide range of applications.This article addresses the growing scope of forensic genetics,which includes advances in DNA sequencing technologies,mixture analysis,body fluid identification,phenotypic profiling,forensic genealogy,microbiological analysis,exploration of novel markers,and ethical and legal considerations.These developments have enabled the analysis of difficult samples and provided comprehensive insights into the origins of biological evidence.In an ever-evolving landscape,forensic genetics continues to shape the future of forensic science by providing new tools and techniques that help deliver justice in an increasingly complex world.展开更多
Dear Editor,Short tandem repeats(STRs),polymorphic DNA regions with a variable number of repeated units(2–6 base pairs),are attractive to forensic applications such as human identification and parentage testing[1].No...Dear Editor,Short tandem repeats(STRs),polymorphic DNA regions with a variable number of repeated units(2–6 base pairs),are attractive to forensic applications such as human identification and parentage testing[1].Nowadays,most of the commercial STR kits are designed based on STRs from the combined DNA index system(CODIS),European Standard Set(ESS),expanded CODIS,and extended ESS[2].In this study,we evaluated 21 STRs from GoldeneyeTM DNA ID 22NC kit(PeopleSpot Inc.,Beijing,China),which including 20 polymorphic non-CODIS STR loci(i.e.D1S1656,D2S441,D3S1744,D3S3045,D4S2366,D5S2500,D6S477,D7S1517,D7S3048,D8S1132,D10S1248,D10S1435,D11S2368,D13S325,D14S608,D15S659,D17S1290,D18S535,D19S253,D22GATA198B05)and a CODIS STR locus(D3S1358),in five ethnic groups(i.e.Eastern Han,Ningxia Hui,Xinjiang Uygur,Xizang Tibetan,and Inner Mongolia Mongolian)of China.The forensic genetic investigation of above loci may provide more genetic information in complex kinship testing and population studies[3,4].展开更多
In forensic practice,the age of suspects or victims is crucial information that aids in the resolution of cases.In recent years,age estimation based on DNA methylation has gained significant attention in forensic scie...In forensic practice,the age of suspects or victims is crucial information that aids in the resolution of cases.In recent years,age estimation based on DNA methylation has gained significant attention in forensic science.DNA methylation,an epigenetic marker,undergoes specific changes with age,making it a valuable tool for inferring the age of samples left at crime scenes.Therefore,the identification of age-related DNA methylation markers and the development of novel age estimation models are of great importance in forensic medicine.Numerous studies in the past decade have successfully established age estimation models based on DNA methylation,demonstrating excellent sensitivity and accuracy.To provide a comprehensive review,the authors of this paper conducted a systematic review of relevant articles published from 2012 to the present.We used keywords such as“forensic,”“DNA methylation,”and“age estimation”to retrieve pertinent articles from the Web of Science database.The review covers various aspects,including the sources of sample tissues used for age estimation,DNA methylation conversion methods,and different techniques for DNA methylation detection.In addition,the paper reviews the modeling methods for age estimation based on DNA methylation and factors that can influence DNA methylation.Overall,this review serves as a valuable reference for forensic genetics,offering insights into the latest advancements in age estimation using DNA methylation.As the field of forensic science continues to evolve,the integration of DNA methylation-based age estimation into practice is expected to enhance the accuracy and reliability of age determination in criminal investigations.展开更多
Formalin-fixed and paraffin-embedded(FFPE)tissues provide a wealth of pathological information crucial for clinical and forensic examinations.Formalin induces robust complexes between DNA and proteins,impacting DNA ex...Formalin-fixed and paraffin-embedded(FFPE)tissues provide a wealth of pathological information crucial for clinical and forensic examinations.Formalin induces robust complexes between DNA and proteins,impacting DNA extraction and complicating short tandem repeat(STR)typing for personal identification and paternity testing.Here,we present a case of paternity testing involving one FFPE tissue and one blood specimen.We compared four DNA extraction methods and analyzed the obtained products from the most successful approach.To ensure robust statistical support,we used a combination of three STR kits for the analysis.This case demonstrates the viability of using multiple kits in tandem for STR profiling of FFPE tissues.展开更多
Cervical cancer remains a significant global public health issue due to its high incidence and mortality.Current clinical guidelines recommend screening for high-risk human papillomavirus(hrHPV)-DNA alongside a Thinpr...Cervical cancer remains a significant global public health issue due to its high incidence and mortality.Current clinical guidelines recommend screening for high-risk human papillomavirus(hrHPV)-DNA alongside a Thinprep cytologic test(TCT)before further medical evaluation[1].展开更多
基金supported by the grants from the National Key Technology Research&Development Program of the Ministry of Science and Technology of People’s Republic of China(2012BAK16B01)the National Natural Science Foundation of Peo-ple’s Republic of China(No.81222041,81172908)
文摘Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts.However,the conventional body fluid identification methods are prone to various limitations,such as time consumption,intensive labor,nonparallel manner,varying degrees of sensitivity and limited specificity.Recently,the analysis of cell-specific messenger RNA expression(mRNA profiling)has been proposed to supplant conventional methods for body fluid identification.Since 2011,the collaborative exercises have been organized by the European DNA Profiling Group(EDNAP)in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification.The major advantages of mRNA profiling,compared to the conventional methods,include higher sensitivity,greater specificity,the ability of detecting several body fluids in one multiplex reaction,and compatibility with current DNA extraction and analysis procedure.In the current review,we provided an overview of the present knowledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possible practical application to forensic casework.
基金Project supported by the National Natural Science Foundation of China(No.81460286)the Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme(GDUPS)(2017)China
文摘To further enrich the genetic data of the Chinese Xinjiang Mongolian group,the genetic distribution and forensic parameters of 19 autosomal short tandem repeats(STRs)were investigated.Altogether,249 alleles were observed in these 19 STRs.The mean values of the polymorphism information content(PIC),match probability(MP),discrimination power(DP),and probability of exclusion(PE)for these 19 STRs were 0.7775,0.0699,0.9301,and 0.6085,respectively.Additionally,the cumulative DP and PE values obtained in the Mongolian group were 0.999 999 999 999 999 999 999 995 67 and 0.999 999 992 163,respectively.
基金funded by the National Natural Science Foundation of China(81771125,81471803,81671031)the Sichuan Province Youth Science and Technology Innovation Team(2014TD0001)
文摘Adipose-derived stromal cells （ASCs） have gained great attention in regenerative medicine. Progress in our understanding of adult neovascularization further suggests the potential of ASCs in promoting vascular regeneration, although the specific cues that stimulate their angiogenic behavior remain controversial In this study, we established a three-dimensional （3D） angiogenesis model by co-culturing ASCs and endothelial cells （ECs） in collagen gel and found that ASC-EC-instructed angiogenesis was regulated by the canonical Wnt pathway. Furthermore, the angiogenesis that occurred in implants collected after injections of our collagen gel- based 3D angiogenesis model into nude mice was confirmed to be functional and also regulated by the canonical Wnt pathway. Wnt regulation of angiogenesis involving changes in vessel length, vessel density, vessel sprout, and connection numbers occurred in our system. Wnt signaling was then shown to regulate ASC- mediated paracrine signaling during angiogenesis through the nuclear translocation of β-catenin after its cytoplasmic accumulation in both ASCs and ECs. This translocation enhanced the expression of nuclear cofactor Lef-1 and cyclin D1 and activated the angiogenic transcription of vascular endothelial growth factor A （VEGFA）, basic fibroblast growth factor （bFGF）, and insulin-like growth factor 1 （IGF-1）. The angiogenesis process in the 3D collagen model appeared to follow canonical Wnt signaling, and this model can help us understand the importance of the canonical Wnt pathway in the use of ASCs in vascular regeneration.
文摘Bones and teeth often represent the only sources of DNA available for identifying human remains.DNA in bones and teeth is generally better preserved than that in soft tissues because of the presence of hard connective tissue with a high level of calcium.Because of the extensive mineralisation,the choice of an efficient DNA extraction procedure is important to minimise the sampling of a high level of minerals and to remove polymerase chain reaction(PCR)inhibitors.Some protocols are available for DNA extraction from bones and teeth as part of the Qiagen EZ1 DNA Investigator Kit using the EZ1 Advanced XL automated purification platform.To improve the efficiency of DNA extraction from skeletal remains,the present study focuses on a modification to these already available protocols.In this study,different bones and teeth collected between 1 and 50 years after death were subjected to DNA extraction using the standard EZ1 protocol,a supplementary protocol,and a modified protocol.The modified approach included a decalcification step,whereas the Qiagen protocols worked directly on non-decalcified powder.In all three procedures,150 mg samples were used for DNA extraction.We evaluated the quantity of DNA recovered from samples,the presence of any PCR inhibitors co-extracted,the level of DNA degradation,the quality of short tandem repeat(STR)profiles,and the reproducibility of the modified procedure.When compared with the other protocols,the modified protocol resulted in the best recovery of DNA that was free of PCR inhibitors.Additionally,the STR profiles were reliable and of high quality.In our opinion,the decalcification step increases DNA recovery by softening tissues,which allows lysis solutions to act more effectively.Furthermore,the use of two lysis solutions and the variation added to the EZ1 purification step allow for DNA recovery with quality and quantity superior to those of the previously available Qiagen-based protocols.These findings may be helpful solutions to the problems commonly encountered when dealing with difficult samples,such as bones and teeth.
文摘Genetic profiling is a standard procedure for human identification,i.e.in criminal cases and mass disasters,and has been proven to be an important part in the process in the repatriation of victims to their relatives.In the event of a catastrophe whether it be a natural disaster,terror attack or accident,fatalities of many nationalities may be a consequence and international collaboration becomes necessary.Current DNA techniques used on a routine basis at forensic laboratories world-wide are very useful,and results reported from different labs are compared,making it possible to be matched in order to declare the identification of a victim.Statistical calculations of possibilities of a random match are achievable since population data from many parts of the world are available.However,decomposition and degradation of the remains are not uncommon in the aftermath of a catastrophe and hence it may be difficult to retrieve detailed DNA profiles from such samples.Massive parallel sequencing(MPS)is a technique capable of producing a vast amount of DNA sequence data in a high-through put manner,and panels of single nucleotide polymorphism(SNP)markers allow the amplification of small DNA fragments,often seen in compromised samples.Here,we report the results from a set of 10 samples from missing person identification cases,analyzed with an MPS based method comprising 131 SNP markers and compared with direct reference material or buccal swab samples collected from relatives of the deceased.We assess the weight of evidence of a match by statistical calculation.Furthermore,we compare results reported on different platforms using different SNP panels,and conclude that more work has to be done if results from missing person identification cases analyzed on MPS with SNP panels at different laboratories are to be fully reliable and thus comparable.
基金supported by the National Natural Science Foundation grant of China(No.82371647,82071607,82071651,82403489)National Key Research and Development Program(2022YFC3600304,2022YFC2704700)+9 种基金Science and Technology Department of Sichuan Province(23ZDYF2489)Cadre Health Care Committee of Sichuan Province,China(2023-1701)Chengdu Science and Technology Bureau(2021-YF05-02106-SN,2017-GH02-00030-HZ)Outstanding Scientific Fund of Shengjing Hospital(No.202003)Sichuan University 0-1 Innovation Research project(2023SCUH0019)The Frontiers Medical Center,Tianfu Jincheng Laboratory Foundation(T FJC2023010001)China Postdoctoral Science Foundation(No.2023M733902,2023M743909)PHD Research Initiated Fund Project in Liaoning Province(No.2023-BSBA-331,2023-BSBA-352)Science and Technology Plan of Liaoning Province(2022JH2/20200066)345 Talent Project of Shengjing Hospital of China Medical University(No.M1344).
文摘Epigenetics governs a chromatin state regulatory system through five key mechanisms:DNA modification,histone modification,RNA modification,chromatin remodeling,and non-coding RNA regulation.These mechanisms and their associated enzymes convey genetic information independently of DNA base sequences,playing essential roles in organismal development and homeostasis.Conversely,disruptions in epigenetic landscapes critically influence the pathogenesis of various human diseases.This understanding has laid a robust theoretical groundwork for developing drugs that target epigenetics-modifying enzymes in pathological conditions.Over the past two decades,a growing array of small molecule drugs targeting epigenetic enzymes such as DNA methyltransferase,histone deacetylase,isocitrate dehydrogenase,and enhancer of zeste homolog 2,have been thoroughly investigated and implemented as therapeutic options,particularly in oncology.Additionally,numerous epigenetics-targeted drugs are undergoing clinical trials,offering promising prospects for clinical benefits.This review delineates the roles of epigenetics in physiological and pathological contexts and underscores pioneering studies on the discovery and clinical implementation of epigenetics-targeted drugs.These include inhibitors,agonists,degraders,and multitarget agents,aiming to identify practical challenges and promising avenues for future research.Ultimately,this review aims to deepen the understanding of epigeneticsoriented therapeutic strategies and their further application in clinical settings.
基金supported by the Internal Security Funding Police Program of the European Commission-Directorate General MigrationHome Affairs under the European Commission[grant number HOME/2014/ISFP/AG/LAWX/4000007135].
文摘To evaluate the promising advantages of massively parallel sequencing(MPS)in our casework,we analysed a total of 33 Y-chromosomal short tandem repeats(Y-STRs)with traditional capillary electrophoresis(CE)and 25 Y-STRs using the newer MPS technology.We studied the outcome of both technologies in 64 father-son pairs using stock and custom-designed kits.Current MPS technology confirmed the 13 mutational events observed with CE and improved our understanding of the complex nature of STR mutations.By detecting isometric sequence variants between unrelated males,we show that sequencing Y-STRs using MPS can boost discrimination power.
基金We thank the plant physiology section at Humboldt University,Berlin for help with the CLSM.We are very grateful to Caterina Biffis,Georg Brenneis and the 2 anonymous reviewers for the helpful advice.We also thank Stephen Rossiter for improving the English.
文摘We describe the formation of the major axon pathways in the embryonic central and peripheral nervous systems of the amphipod crustacean Orchestia cavimana Heller,1865 by means of antibody staining against acetylated alphatubulin.The data add to a long list of previous studies of various other aspects of development in Orchestia and provide a basis for future studies of neurogenesis on a deeper cellular and molecular level.Orchestia exhibits a tripartite dorsal brain,which is a characteristic feature of euarthropods.Its anlagen are the first detectable structures in the developing nervous system and can be traced back to distinct neuronal cell clusters in the early embryo.The development of the ventral nervous system proceeds with an anteroposterior gradient of development.In each trunk segment,the longitudinal connectives and the anterior commissure form first,followed by the intersegmental nerve,the posterior commissure and segmental nerves,respectively.A single commissure of a vestigial seventh pleonal segment is found.In the peripheral nervous system we observe a spatial and temporal pattern of leg innervation,which is strikingly similar in both limb types,the uniramous pereopods and the biramous pleopods.A proximal leg nerve splitting distally into two separated nerves probably reflects a general feature of crustaceans.
基金This study was supported by National Natural Science Foundation of China(NSFC,81930055 and 81772031)Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme(GDUPS,2017).
文摘In this research,genotyping data of 43 InDel loci in 311 Han individuals in Ankang City,Shaanxi Province,China were detected using a self-developed five-dye multiplex amplification panel.The allelic frequencies and forensic parameters of all InDel loci were calculated.The combined power of discrimination and probability of exclusion values were 0.99999999999999999882739 and 0.999887424,respectively,which demonstrated that this 43-InDel panel was powerful for individual identifications in Ankang Han population.Moreover,genetic distances,pairwise F_(ST)values,principal component analyses,phylogenetic trees and STRUCTURE analyses were performed to investigate the genetic affinities between Ankang Han and reference groups.Population genetic investigations indicated that Ankang Han population had a close genetic relationship with Southern Han population compared with other reference groups.
基金supported by the Shanghai Key Laboratory of Forensic Medicine(Academy of Forensic Science)Open Project Foundation[grant number KF1812]the National Natural Science Foundation of China[grant number 81971786].
文摘As a result of the expansion of old Tibet on the Qinghai-Tibet Plateau,Tibetans diverged into three main branches,Ü-Tsang,Amdo,and Kham Tibetan.Ü-Tsang Tibetans are geographically distributed across the wide central and western portions of the Qinghai-Tibet Plateau while Lhasa is the central gathering place for Tibetan culture.The AGCU Y30,a 6-dye fluorescence kit including 30 slowly and moderately mutated Y-STR loci,has been validated for its stability and sensitivity in different biomaterials and diverse Chinese populations(Han and other minorities),and widely used in the practical work of forensic science.However,the 30 Y-STR profiling of Tibetan,especially forÜ-Tsang Tibetan,were insufficient.We utilized the AGCU Y30 to genotype 577Ü-Tsang Tibetan unrelated males from Lhasa in the Tibet Autonomous Region of China to fill up the full and accurate Y-STR profiles.A total of 552 haplotypes were observed,536(97.10%)of which were unique.One hundred and ninety-four alleles were observed at 26 single copy loci and the allelic frequencies ranged from 0.0017 to 0.8180.For the two multi-copy loci DYS385a/b and DYS527a/b,64 and 36 allelic combinations were observed,respectively.The gene diversity(GD)values ranged from 0.3079 at DYS391 to 0.9142 at DYS385a/b and the overall haplotype diversity(HD)was 0.9998,and its discrimination capacity(DC)was 0.9567.The population genetic analyses demonstrated that LhasaÜ-Tsang Tibetan had close relationships with other Tibetan populations from Tibet and Qinghai,especially withÜ-Tsang Tibetan.From the perspective of Y haplogroups,the admixture of the southward Qiang people with dominant haplogroup O-M122 and the northward migrations of the initial settlers of East Asia with haplogroup D-M175 hinted the Sino-Tibetan homologous,thus,we could not ignore the gene flows with other Sino-Tibetan populations,especially for Han Chinese,to characterize the forensic genetic landscape of Tibetan.
基金supported by grants from the National Natu-ral Science Foundation of China[grant number 81330073],[grant number 81302620]the Ministry of Science and Technology of China[grant number 2016YFC0800703]the Science and Technology Commission of Shanghai Municipality[grant number 14DZ2270800].
文摘The custom-designed single nucleotide polymorphism(SNP)panel amplified 231 autosomal SNPs in one PCR reaction and subsequently sequenced with massively parallel sequencing(MPS)technology and Ion Torrent personal genome machine(PGM).SNPs were chosen from SNPforID,IISNP,HapMap,dbSNP,and related published literatures.Full concordance was obtained between available MPS calling and Sanger sequencing with 9947A and 9948 controls.Ten SNPs(rs4606077,rs334355,rs430046,rs2920816,rs4530059,rs1478829,rs1498553,rs7141285,rs12714757 and rs2189011)with low coverage or heterozygote imbalance should be optimized or excluded from the panel.Sequence data had sufficiently high coverage and gave reliable SNP calling for the remaining 221 loci with the custom MPS-SNP panel.A default DNA input amount of 10 ng per reaction was recommended by Ampliseq technology but sensitivity testing revealed positive results from as little as 1 ng input DNA.Mixture testing with this panel is possible through analysis of the F MAR(frequency of major allele reads)values at most loci with enough high coverage depth and low level of sequencing noise.These results indicate the potential advantage of the custom MPS-SNP assays and Ion Torrent PGM platform for forensic study.
基金supported by the National Natural Science Foundation of China(82030058,82101977,82130056).
文摘Background:In forensic investigations,accurate estimation of the postmortem interval(PMI)is an important task,but also an ongoing challenge.Especially in cases where the cadaver has been specially treated,for example,by boiling,the determination of PMI becomes extremely difficult.Previous studies have shown that the succession of the microbial community after decomposition of the cadaver can be used to infer PMI.However,the feasibility of determining the PMI of boiled cadavers has not yet been demonstrated.Aims and Objectives:The main objective of this study was to test whether we can infer PMI of boiled cadavers based on the succession of microbial communities.Materials and Methods:SD rats were killed by cervical dislocation.Subsequently,the rat cadavers were divided into the case(boiled cadavers)and control(unboiled cadavers)groups.Rectal samples were collected from the rats for 45 days and at nine time points.High-throughput sequencing of the 16S rRNA gene was performed to characterize the microbial community in the rectum.Results:The results showed that the composition and relative abundance of bacterial communities at the phylum level were significantly different between the case and control groups.The alpha diversity of the microbial community showed a decreasing trend with the decomposition process.Principal coordinate analysis showed that the case and control groups had obvious patterns along the succession of microbial communities.The rectal microbial communities showed a significant linear trend in the time course of decomposition.A random forest model was used to infer PMI.The goodness-of-fit(R2)of the model was 68.00%and 84.00%,and the mean absolute errors were 2.05 and 1.48 days within 45 days of decomposition for the case and control groups,respectively.Conclusions:Our results suggest that microbial community succession could be a potential method to infer PMI of boiled cadavers.
基金supported by grants from the National Natural Science Foundation of China(No.82030058).
文摘The field of forensic DNA typing,often referred to as“DNA fingerprinting,”has evolved and expanded considerably since its beginnings in the mid-1980s.Originally,forensic DNA typing was primarily used for individual identification and criminal investigations,but it has evolved into a versatile discipline with a wide range of applications.This article addresses the growing scope of forensic genetics,which includes advances in DNA sequencing technologies,mixture analysis,body fluid identification,phenotypic profiling,forensic genealogy,microbiological analysis,exploration of novel markers,and ethical and legal considerations.These developments have enabled the analysis of difficult samples and provided comprehensive insights into the origins of biological evidence.In an ever-evolving landscape,forensic genetics continues to shape the future of forensic science by providing new tools and techniques that help deliver justice in an increasingly complex world.
基金supported by the National Key R&D Program of China[grant number 2016YFC0800703]the National Natural Science Fund for Distinguished Young Scholars[grant number 81625013]+2 种基金the Standard Program of Shanghai Municipality[grant number 16DZ0501600]the Public Interest Research Grant Program of National Research Institutes[grant number GY2017D-2]General Program of National Natural Science Foundation of China.
文摘Dear Editor,Short tandem repeats(STRs),polymorphic DNA regions with a variable number of repeated units(2–6 base pairs),are attractive to forensic applications such as human identification and parentage testing[1].Nowadays,most of the commercial STR kits are designed based on STRs from the combined DNA index system(CODIS),European Standard Set(ESS),expanded CODIS,and extended ESS[2].In this study,we evaluated 21 STRs from GoldeneyeTM DNA ID 22NC kit(PeopleSpot Inc.,Beijing,China),which including 20 polymorphic non-CODIS STR loci(i.e.D1S1656,D2S441,D3S1744,D3S3045,D4S2366,D5S2500,D6S477,D7S1517,D7S3048,D8S1132,D10S1248,D10S1435,D11S2368,D13S325,D14S608,D15S659,D17S1290,D18S535,D19S253,D22GATA198B05)and a CODIS STR locus(D3S1358),in five ethnic groups(i.e.Eastern Han,Ningxia Hui,Xinjiang Uygur,Xizang Tibetan,and Inner Mongolia Mongolian)of China.The forensic genetic investigation of above loci may provide more genetic information in complex kinship testing and population studies[3,4].
基金supported by the Fundamental Research Funds for the Central Universities,Sichuan University(No.2023SCU12036).
文摘In forensic practice,the age of suspects or victims is crucial information that aids in the resolution of cases.In recent years,age estimation based on DNA methylation has gained significant attention in forensic science.DNA methylation,an epigenetic marker,undergoes specific changes with age,making it a valuable tool for inferring the age of samples left at crime scenes.Therefore,the identification of age-related DNA methylation markers and the development of novel age estimation models are of great importance in forensic medicine.Numerous studies in the past decade have successfully established age estimation models based on DNA methylation,demonstrating excellent sensitivity and accuracy.To provide a comprehensive review,the authors of this paper conducted a systematic review of relevant articles published from 2012 to the present.We used keywords such as“forensic,”“DNA methylation,”and“age estimation”to retrieve pertinent articles from the Web of Science database.The review covers various aspects,including the sources of sample tissues used for age estimation,DNA methylation conversion methods,and different techniques for DNA methylation detection.In addition,the paper reviews the modeling methods for age estimation based on DNA methylation and factors that can influence DNA methylation.Overall,this review serves as a valuable reference for forensic genetics,offering insights into the latest advancements in age estimation using DNA methylation.As the field of forensic science continues to evolve,the integration of DNA methylation-based age estimation into practice is expected to enhance the accuracy and reliability of age determination in criminal investigations.
基金supported by the Chengdu Science and Technology Program(No.2022-YF05-02026-SN).
文摘Formalin-fixed and paraffin-embedded(FFPE)tissues provide a wealth of pathological information crucial for clinical and forensic examinations.Formalin induces robust complexes between DNA and proteins,impacting DNA extraction and complicating short tandem repeat(STR)typing for personal identification and paternity testing.Here,we present a case of paternity testing involving one FFPE tissue and one blood specimen.We compared four DNA extraction methods and analyzed the obtained products from the most successful approach.To ensure robust statistical support,we used a combination of three STR kits for the analysis.This case demonstrates the viability of using multiple kits in tandem for STR profiling of FFPE tissues.
基金supported by the National Natural Science Foundation of China(82071651)Chengdu Science and Technology Bureau(2017-GH02-00030-HZ)+4 种基金National Key Research and Development Program of China(2022YFC3600304,2022YFC2704700)Science and Technology Department of Sichuan Province(23ZDYF2489)Sichuan Province Cadre Health Care Scientific Research Project(CGY2023-1701)the Chengdu Technological Innovation R&D Project(2021-YF05-00595-SN)partially sponsored by Lisen Imprinting Diagnostics,Inc.
文摘Cervical cancer remains a significant global public health issue due to its high incidence and mortality.Current clinical guidelines recommend screening for high-risk human papillomavirus(hrHPV)-DNA alongside a Thinprep cytologic test(TCT)before further medical evaluation[1].
