Paternal genetic structure of Kyrgyz ethnic group in China revealed by high-resolution Y-chromosome STRs and SNPs

Kyrgyz ethnic group is one of the nomads in China, with the majority in Xinjiang and a small part of them living in Heilongjiang province. Historically, they have went through five migrations westward due to the wars. The name “Kyrgyz” means 40 tribes, originating from the primary groups of Kyrgyz. However, it is a largely understudied population, especially from the Y chromosome. In this study, we used a previously validated high-resolution Y-chromosome single nucleotide polymorphisms (Y-SNPs) and short tandem repeats (Y-STRs) system to study Kyrgyz ethnic group. A total of 314 male samples of Kyrgyz ethnic group were genotyped by 173 Y-SNPs and 27 Y-STRs. After data analysis, the results unveiled that Kyrgyz ethnic group was a population with high percentage of both haplogroup C2a1a3a1d∼-F10091 (91/134) and R1a1a1b2a2-Z2124 (109/134), which has never been reported. This implied that Kyrgyz ethnic group might have gone through bottleneck effects twice, with these two main lineages left. Mismatch analysis indicated that the biggest mismatch number in haplogroup C2a1a3a1d∼-F10091 was 10, while that of haplogroup R1a1a1b2a2-Z2124 was 20. This huge difference reflected the different substructure in two lineages, suggesting that haplogroup C2a1a3a1d∼-F10091 might have the least admixture compared to the other two lineages. After admixture modelling with other datasets, the conclusion could be drawn that Kyrgyz ethnic group had great genetic affinity with Punjabi from Lahore, Pakistan, which supported that Kyrgyz ethnic group in China was close to central Asian.     

Exploitation of a novel slowly mutating Y-STRs set and evaluation of slowly mutating Y-STRs plus Y-SNPs typing strategy in forensic genetics and evolutionary research

The Y-chromosome short tandem repeats (Y-STRs) loci with different mutation rates existing in the Y chromosome non-recombination region (NRY) allow to be applied in human forensics, genealogical researches, historical investigations and evolutionary studies. Currently, there is a high demand for pedigree search to narrow the scope of crime investigations. However, the commonly used Y-STRs kits generally contain Y-STRs with high mutation rates that could cause individuals from the same pedigree to display different haplotypes. Herein, we put forward a new strategy of Slowly Mutating (SM) Y-STRs plus Y-SNPs typing, which could not only improve the resolution and accuracy of pedigree search, but also be applicable to evolutionary research. First, we developed a nine SM Y-STRs assay by evaluating their mutation rates in 210 pedigrees. Then the gene diversity and efficiency of the SM Y-STRs and 172 Y-SNPs sets were investigated by 2304 unrelated males from 24 populations. Furthermore, network and time estimation analyses were performed to evaluate the new strategy's capability to reconstruct phylogenetic tree and reliability to infer the time to the most recent common ancestor (TMRCA). The nine SM Y-STRs assay even had a higher resolution and a comparable capacity of revealing population genetic differentiation compared to 172 Y-SNPs system. This new strategy could optimize the phylogenetic tree generated by commonly used Y-STR panels and obtain a quite consistent time estimations with the published dating.     

Development and validation of a custom panel including 183 Y-SNPs for Chinese Y-chromosomal haplogroups dissection using a MALDI-TOF MS system

Y-chromosome SNP haplogroups exhibit geographic structuring in many populations around the world. Therefore, Y-chromosome haplogroups have been widely used to infer paternal biogeographical ancestry and high-resolution paternal lineage classification. In the present study, we designed a customized panel containing 183 Y-SNPs based on previous studies and evaluated the genotyping performance and repeatability, concordance, sensitivity, and ability of analysing case-type samples using a MALDI-TOF MS platform. The average call rate for duplicate typing of any one SNP in the panel was 97.0%. In the concordance and accuracy study, the results of haplogroup designation obtained from MALDI-TOF MS platform were fully consistent with those obtained from the next-generation sequencing (NGS) platform. The optimal amount of template DNA in the PCR seemed to be 10 ng. However, if less DNA (≥156.25 pg) was available, it was still possible to obtain meaningful haplogroup information. For the application part, this panel could be applied for the detection of blood, semen, and buccal swabs samples. Particularly, blood stain on FTA card samples could be dissected by direct PCR amplification on the MALDI-TOF MS platform. Besides, 371 unrelated male individuals from four Chinese ethnic groups (Han, Hui, Mongolian, and Kazak) were detected using this panel. Total 78 terminal haplogroups were found and the haplogroup diversity was 0.933576. The results demonstrate that this panel could be an accurate, fast, and cost-effective method for database construction where the amount of sample material is less of a concern and when the cost of the assay is taken into consideration.     

Developmental validation of the Yfiler Platinum PCR Amplification Kit for forensic genetic caseworks and databases

Y chromosome kits are successfully applied in cases where human biological material exists. With the development of genotyping ability, more Y chromosomal markers are needed for finer identification of male individuals and lineages. In this study, a developmental validation of a newly emerged Y chromosome kit that combines two different kinds of markers: 38 Y-STRs and 3 Y-indels are conducted. The results show that this kit has high sensitivity when there is a small amount of DNA (125 pg), more than one male (minor:major = 1:7), or a mixture of males and females (male:female = 125pg:1875pg), inhibited substances (800 μM hematin and more than 1600 ng/μL humic acid). The kit exhibits high precision level with a standard deviation of allele size no more than 0.14 nt. Locus DYS481 shows the largest stutter rate, with three stutters per true allele. Population samples are well identified (MP of 0.001106), and mutations can be observed in father–son pairs (46 mutations in 70 pairs, 10 in locus DYS627). Out of all the population samples, 13.2% belong to haplogroup M117-O2a2b1a1, with their ethnic group being Han Chinese. The results show that this kit can improve the performance of identifying male individuals, obtaining more unique haplotypes (increasing from 894 to 918 of 1000 male samples) and higher discrimination capacity (increasing from 0.942 to 0.955) in this study compared to previous widely used Yfiler Plus kit. Besides, it gives information about their paternal lineages in forensic genetic casework and genealogical database construction.     

Population genetics and forensic efficiency of twenty‐one novel microsatellite loci of Chinese Yi ethnic group

In this study, we investigated polymorphic distributions of allelic frequencies and forensic genetic parameters of 21 novel autosomal microsatellite loci from 110 unrelated healthy individuals of Chinese Yi ethnic group. Expected heterozygosity, power of discrimination, and polymorphic information content ranged from 0.617 to 0.812, 0.777 to 0.936 and 0.560 to 0.790. The microsatellite loci showed high forensic efficiency. The total discrimination power and cumulate probability of exclusion were 0.99999999999999999986902 and 0.999998818, respectively. Locus‐by‐locus allelic frequencies were compared using analysis of molecular variance (AMOVA) method, and the statistically significant differences were observed between Yi group and Russian, Tujia, Kazak, Bai, Ningxia Han, Salar, Tibetan, and Uigur groups at 5, 6, 7, 7, 7, 8, 12, and 13 loci, respectively. The results of genetic distance comparisons, genetic structure analyses, and principal component analysis all indicated that the Yi group showed relatively short genetic relationships with Russian, Salar, and Bai group. The experimental results showed that the 21 loci in the multiplex system provided highly polymorphic information and forensic efficiency for forensic individual identification and paternity testing, also basic population data for population genetics and anthropological research.     

Inference of population structure and admixture proportion from Y chromosomal data of Chinese population

In the past two decades, Y chromosome data has been generated for human population genetic studies. These Y chromosome datasets were produced with various testing methods and markers, thus difficult to combine them for a comprehensive analysis. In this study, we combine four human Y chromosomal datasets of Han, Tibetan, Hui, and Li ethnic groups. The dataset contains 27 microsatellites and 137 single nucleotide polymorphisms these populations share in common. We assembled a single dataset containing 2439 individuals from 25 nationwide populations in China. A systematic analysis of genetic distance and clustering was performed. To determine the gene flow of the studied population with worldwide populations, we modeled the ancestry informative markers. The reference panel was regarded as a mixture of South Asian (SAS), East Asian (EAS), European (EUR), African (AFR), and American (AMR) populations from 1000 Genomes data of Y chromosome using nonlinear data-fitting. We then calculated the admixture proportion of these four studied populations with 26 worldwide populations. The results showed that the Han and Hui have great genetic affinity, and Hui is the most admixed ethnic group, with 61.53% EAS, 34.65% SAS, 1.91% AFR, 1.56% AMR, and 0.04% EUR ancestry component (the AMR is highly admixed and thus should be ignored). All the other three ethnic groups contained more than 97% EAS ancestry component. The Li is the least admixed population in this study. The combined dataset in this study is the largest of this kind reported to date and proposes reference population data for use in future paternal genetic studies and forensic genealogical identification.     

SNPlexing the human Y-chromosome: a single-assay system for major haplogroup screening

SNPs are one of the main sources of DNA variation among humans. Their unique properties make them useful polymorphic markers for a wide range of fields, such as medicine, forensics, and population genetics. Although several high-throughput techniques have been (and are being) developed for the vast typing of SNPs in the medical context, population genetic studies involve the typing of few and select SNPs for targeted research. This results in SNPs having to be typed in multiple reactions, consuming large amounts of time and of DNA. In order to improve the current situation in the area of human Y-chromosome diversity studies, we decided to employ a system based on a multiplex oligo ligation assay/PCR (OLA/PCR) followed by CE to create a Y multiplex capable of distinguishing, in a single reaction, all the major haplogroups and as many subhaplogroups on the Y-chromosome phylogeny as possible. Our efforts resulted in the creation of a robust and accurate 35plex (35 SNPs in a single reaction) that when tested on 165 human DNA samples from different geographic areas, proved capable of assigning samples to their corresponding haplogroup.     

Developmental validation of the Microreader Group Y Direct ID System: A novel six-dye typing system with 54 Y-chromosomal loci for forensic application

Y-chromosome, as a gender-determined biological marker, is inherited only between fathers and sons. The Y-chromosome short tandem repeats (Y-STRs) play an essential role in paternity lineage tracing as well as sexual assault cases. The Microreader Group Y Direct ID System as a six-dye multiplex amplification kit, including 53 Y-STR and one Y-Indel locus, would improve performance and aid in obtaining more information through a greater number of loci with high polymorphism. In the present study, to verify the accuracy and efficiency of the kit, developmental validation was conducted by investigating sensitivity, species specificity, PCR inhibition, male–male and male–female mixtures, and reproducibility. The kit was tested using 311 male samples from Han and Qiang populations in Sichuan Province. The results showed that this kit had fairly high power for forensic discrimination (Han: haplotype diversity [HD] = 1, Qiang: HD = 0.999944). Additionally, 44 confirmed father–son pairs were also genotyped, among which 69 distinct haplotypes could be obtained. These father–son pairs cannot be distinguished by commonly used Y-STR panels, indicating that adding these extra Y-STRs to a single panel can achieve better discrimination performance. Collectively, the Microreader Group Y Direct ID System is robust and informative for forensic applications.     

Introduction of an single nucleodite polymorphism-based "Major Y-chromosome haplogroup typing kit" suitable for predicting the geographical origin of male lineages

The European Consortium "High-throughput analysis of single nucleotide polymorphisms for the forensic identification of persons--SNPforID", has performed a selection of candidate Y-chromosome single nucleotide polymorphisms (SNPs) for making inferences on the geographic origin of an unknown sample. From more than 200 SNPs compiled in the phylogenetic tree published by the Y-Chromosome Consortium, and looking at the population studies previously published, a package of 29 SNPs has been selected for the identification of major population haplogroups. A "Major Y-chromosome haplogroup typing kit" has been developed, which allows the multiplex amplification of all 29 SNPs in a single reaction. Allele genotyping was performed with a single base extension reaction (minisequencing) detected by CE. The validation of the multiplex was performed in a total of 1126 unrelated males distributed among 12 worldwide populations. The approach takes advantage of the specific geographic distribution of the Y-chromosome haplogroups and demonstrates the utility of binary polymorphisms to infer the origin of a male lineage.     

Development a multiplex panel of AISNPs,multi-allelic InDels,microhaplotypes, and Y-SNP/InDel loci for multiple forensic purposes via the NGS

Recently, next generation sequencing has shown the promising application value in forensic research. In this study, we constructed a multiplex amplification system of different molecular genetic markers based on the previous selected ancestry informative single nucleotide polymorphisms (SNPs), multi-allelic insertion/deletion (InDel) polymorphisms, microhaplotypes, and Y-chromosomal SNP/InDel loci, and evaluated forensic efficiencies of the system in Chinese Shaanxi Han, Chinese Hui, and Chinese Mongolian groups via the next generation sequencing platform. Ancestry information analyses of Shaanxi Han, Hui, and Mongolian groups revealed that most Mongolian individuals could be differentiated from Shaanxi Hans and Huis based on the selected ancestry informative SNPs. Multi-allelic InDels and microhaplotypes showed the multiple allele variations and possessed relatively high genetic polymorphisms in these three groups, indicating these loci could provide higher forensic efficiencies for individual identification and paternity testing. Based on Y-chromosomal SNPs, different haplogroup distributions were observed among Shaanxi Han, Hui, and Mongolian groups. In conclusion, the self-developed system could be used to simultaneously carry out the individual identification, paternity analysis, mixture deconvolution, forensic ancestry information analysis, and Y-chromosomal haplogroup inference, which could provide more valuable investigative clues in forensic practices.     

Equilibria between alpha- and beta-agostic stabilized rotamers of secondary alkyl niobium complexes.

The isopropyl chloro complex Tp(Me2)NbCl(i-Pr)(PhC&tbd1;CMe) (2) [Tp(Me2) = hydrotris(3,5-dimethylpyrazolyl)borate] exhibits a beta-agostic structure in the crystal. The conformation of the alkyl group is such that the agostic methyl group lies in the Calpha-Nb-Cl plane and the nonagostic one, in a wedge formed by two pyrazole rings. As observed by solution NMR spectroscopy, restricted rotation about the Nb-C bond allows the observation of an equilibrium between this species, 2beta, and a minor alpha-agostic rotamer 2alpha. A putative third rotamer which would have the secondary hydrogen in the wedge is not observed. Similar behavior is observed for related Tp'NbCl(i-Pr)(R(2)C=CMe) [Tp' = Tp(Me2), R(2) = Me (3); Tp' = Tp(Me2,4Cl), R(2) = Ph (4)]. The two diastereomers of the sec-butyl complex Tp(Me2)NbCl(sec-Bu)(MeC=CMe) (5) have been separated. In the crystal, 5CR-AS has a beta-agostic methyl group with the ethyl group located in the wedge formed by two pyrazole rings. The same single beta-agostic species is observed in solution. The other diastereomer, 5AR-CS has a beta-agostic methylene group in the solid state, and the methyl group sits in the wedge. In solution, an equilibrium between this beta-agostic methylene complex 5AR-CSbeta and a minor alpha-agostic species 5AR-CSalpha, where the ethyl substituent of the sec-Bu group is located in the wedge between two pyrazole rings, is observed. NMR techniques have provided thermodynamic parameters for these equilibria (K = 2beta/2alpha = 4.0 +/- 0.1 at 193 K, DeltaG(o)(193) = -2.2 +/- 0.1, DeltaH(o) = -7.4 +/- 0.1 kJ mol(-)(1), and DeltaS(o) = -27 +/- 1 J K(-)(1) mol(-)(1)), as well as kinetic parameters for the rotation about the Nb-C bond (at 193 K, DeltaG(2)= 47.5 +/- 2.5, DeltaH= 58.8 +/- 2.5 kJ mol(-)(1), and DeltaS = 59.0 +/- 10 J K(-)(1) mol(-)(1)). Upon selective deuteration of the beta-methyl protons in Tp(Me2)NbCl[CH(CD(3))(2)](PhC=CMe) (2-d(6)), an expected isotope effect that displaces the equilibrium toward the alpha-agostic rotamer is observed (K = 2-d(6)beta/2-d(6)alpha = 3.1 +/- 0.1 at 193 K, DeltaG(o)(193) = -1.8 +/- 0.1, DeltaH(o) = -8.3 +/- 0.4 kJ mol(-)(1) and DeltaS(o)= -34 +/- 2 J K(-)(1) mol(-)(1)). The anomalous values for DeltaH(o) and DeltaS(o) are discussed. Hybrid quantum mechanics/molecular mechanics calculations (IMOMM (B3LYP:MM3)) on the realistic model Tp(Me2)NbCl(i-Pr)(HC=CMe) have reproduced the energy differences between the alpha- and beta-agostic species with remarkable accuracy. Similar calculations show that Tp(Me2)NbCl(CH(2)Me)(HC=CMe) is alpha-agostic only and that Tp(5)(-)(Me)NbCl(CH(2)Me)(HC=CMe), which has no methyl groups at the 3-positions of the pyrazole rings, is beta-agostic only. Analysis and discussion of the computational and experimental data indicate that the unique behavior observed for the secondary alkyl complexes stems from competition between electronic effects favoring a beta-agostic structure and steric effects directing a bulky substituent in the wedge between two pyrazole rings of Tp(Me2). All of the secondary alkyl complexes thermally rearrange to the corresponding linear alkyl complexes via a first-order reaction.     

Single nucleotide polymorphisms of mitochondrial DNA HVS‐I and HVS‐II in Chinese Bai ethnic group

For forensic and population genetic purposes, a total of 125 unrelated volunteers’ blood samples were collected from Chinese Bai ethnic minority group to analyze sequence variation of two hypervariable segments (HVS‐I and HVS‐II) in the mitochondrial DNA control region. Comparing the HVS‐I and HVS‐II sequences of the 125 Chinese Bais to the Anderson reference sequence, we found 86 polymorphic loci in HVS‐I and 40 in HVS‐II in mitochondrial DNA sequences of the Chinese Bai ethnic minority group, which defined 93 and 53 different haplotypes, respectively. Haplotype diversity and the mean pairwise differences were 0.992 ± 0.003 and 6.553 in HVS‐I, and 0.877 ± 0.027 and 2.407 in HVS‐II, respectively. We defined four macrohaplogroups R, M, N and D with the proportions ranging from 9.6% to 40.0%. With the analysis of the hypervariable domain from nucleotide 16 180–16 193 in HVS‐I, our study revealed new haplotypes of sequence variations. In addition, the Fst metric, phylogenetic tree, and principal component analysis demonstrated a close genetic relationship between the Bai group and Chinese Han populations from South China, Changsha, and Guangdong. The results support that the Bai group is a multiorigin ethnic minority that has merged with the Chinese Han population.     

The Y-STR landscape of coastal southeastern Han: Forensic characteristics,haplotype analyses,mutation rates,and population genetics

The Y-STR landscape of Coastal Southeastern Han (CSEH) living in Chinese southeast areas (including Guangdong, Fujian, and Zhejiang provinces) is still unclear. We investigated 62 Y-STR markers in a reasonably large number of 1021 unrelated males and 1027 DNA-confirmed father-son pairs to broaden the genetic backgrounds of CSEH. In total, 85 null alleles, 121 off-ladder alleles, and 95 copy number variants were observed, and 1012 distinct haplotypes were determined with the overall HD and DC values of 0.999974 and 0.9912. We observed 369 mutations in 76 099 meiotic transfers, and the average estimated Y-STR mutation rate was 4.85 × 10^–3 (95% CI, 4.4 × 10^–3–5.4 × 10^–3). The Spearman correlation analyses indicated that GD values (R² = 0.6548) and average allele sizes (R² = 0.5989) have positive correlations with Y-STR mutation rates. Our RM Y-STR set including 8 candidate RM Y-STRs, of which DYS534, DYS630, and DYS713 are new candidates in CSEH, distinguished 18.52% of father–son pairs. This study also clarified the population structures of CSEH which isolated in population-mixed South China relatively. The strategy, SM Y-STRs for familial searching and RM Y-STRs for individual identification regionally, could be applicable based on enough knowledge of the Y-STR mutability of different populations.     

Chiral Phosphine Ligands Derived from Sugars8. Crystal and Molecular Structure of Diphenyl (methyl 4', 6'-O-benzylidene-2'-deoxy-α-D-altropyranoside-2-)phosphine Oxide

1INTRODUCTIONChiralphosphinesareusefulligandsinenantioselectivecatalysiswithtransitionmetalcomplexes"',someofthemderivedfromsugarsarealsoofPOtentialapplicationsinbiphasiccatalysis"'andmedicinet3i.Bythenucleophilicreplacementreaction,itisdifficulttolinkthephosphorusatomtothesecondarycarbonofpyrano--,orfurano--sugarderivativesbecauseofthecompetingeliminationreaction[4i.Wehavefoundaneffectivemethodtoincorporateaphosphorusintoapyranosering"',viaregioselectiveandstereospecificring--openingreac…     

Hyperfine interactions in the A3Φ4 and X3Φ4 states of iridium monofluoride, IrF

Laser induced-fluorescence spectra of the 1-0 band of the A(3)Φ(4)-X(3)Φ(4) transition of iridium monofluoride, IrF, have been obtained at near natural linewidth resolution using supersonic molecular beam techniques. The spectra show a complex, clearly resolved hyperfine structure which has significant contributions from the magnetic and quadrupole hyperfine terms in (193)Ir and (191)Ir, both with I = 3/2, and the fluorine magnetic hyperfine term (I = 1∕2). The spectra of both (193)IrF and (191)IrF isotopologues have been assigned and analyzed. The hyperfine structure was interpreted with the aid of atomic hyperfine parameters, which were used to determine the configurational composition of the ground state and to estimate the individual molecular hyperfine parameters.     

Cross-correlated relaxation enhanced 1H[bond]13C NMR spectroscopy of methyl groups in very high molecular weight proteins and protein complexes

A comparison of HSQC and HMQC pulse schemes for recording (1)H[bond](13)C correlation maps of protonated methyl groups in highly deuterated proteins is presented. It is shown that HMQC correlation maps can be as much as a factor of 3 more sensitive than their HSQC counterparts and that the sensitivity gains result from a TROSY effect that involves cancellation of intra-methyl dipolar relaxation interactions. (1)H[bond](13)C correlation spectra are recorded on U-[(15)N,(2)H], Ile delta 1-[(13)C,(1)H] samples of (i) malate synthase G, a 723 residue protein, at 37 and 5 degrees C, and of (ii) the protease ClpP, comprising 14 identical subunits, each with 193 residues (305 kDa), at 5 degrees C. The high quality of HMQC spectra obtained in short measuring times strongly suggests that methyl groups will be useful probes of structure and dynamics in supramolecular complexes.     

含铜类水滑石催化材料热分解过程的研究

共沉淀法合成了Cu0.13Mg0 6Al0.27(OH)2(CO3)0.135·xH2O类水滑石物质 (CuHTlc) ,采用XRD、DTA TG、BET、TEM和27AlMASNMR技术对其热分解过程进行了表征。结果表明 ,在较低焙烧温度时 (低于300℃ ),氢氧根和层间水部分脱除 ,但水滑石仍保持其层状结构 ;500℃时 ,其层状结构被完全破坏 ,出现氧化镁晶相结构 ,随着焙烧温度的进一步升高 ,尖晶石晶相生成。500℃时的焙烧产物具有最大比表面 (193m2·g-1)。当温度高于500℃ ,焙烧产物组成可表示为Cu0.13Mg0.6Al0.27O0.135,CuHTlc的热分解过程可表示为 :Cu0.13Mg0.6Al0.27(OH)2(CO3)0.135·xH2O→Cu0.13Mg0.6Al0.27O0.135 (1 x)H2O 0.135CO2。     

Determination of urinary calcium-oxalate formation risk with BONN-Risk-Index and EQUIL applied to a family

Apart from environmental and acquired risk factors, a person's genetic predisposition may have a distinct influence on the probability of the onset of urolithiasis. To investigate the family related development of calcium oxalate, CaOx, crystallization risk, we studied urines from three generations of the same family. The paternal line has been suffering from CaOx-urolithiasis for at least two generations; no case of urolithiasis has been reported from the maternal line and the youngest generation.We applied the BONN-Risk-Index and the computer program EQUIL to determine the crystallization risk of each family member (n = 7). We clearly verified by probability calculations of the existence of the two risk groups within the family and showed that one of the siblings of the youngest generation may have inherited the stone-formation risk from its paternal relatives as this person clearly reflects a high risk pattern.     

Mitochondrial SNPs associated with Japanese centenarians, Alzheimer's patients, and Parkinson's patients

In this paper we examined the relations between three classes of people (96 Japanese centenarians, 96 Japanese Alzheimer's disease (AD) patients and 96 Japanese Parkinson's disease (PD) patients) and their mitochondrial single nucleotide polymorphism (mtSNP) frequencies at individual mitochondrial DNA (mtDNA) positions of the entire mt-genome by using the radial basis function (RBF) networks. As a result, we got new findings of mtSNPs for representing characteristics of individual classes. These mtSNPs show distinct differences for three classes of people. That is, individual classes of people are characterized by unique mtSNPs. Interestingly, Japanese centenarians are closely associated with haplogroup D4, Japanese AD patients with haplogroup G2a, and Japanese PD patients with haplogroup M7a. These characteristics of mtSNPs are different from those of previously reported works. As the amino acid replacement mtSNPs were at four mtDNA positions, it is indicated that mtSNPs of synonymous nucleotide substitutions as well as those of nonsynonymous nucleotide substitutions may play important roles in mitochondrial functions.     

Independent development and validation of a novel six-color fluorescence multiplex panel including 61 diallelic DIPs and 2 miniSTRs for forensic degradation sample

Current forensic DNA profiles are obtained based on analyses of PCR product sizes or DNA sequence polymorphisms. Sometimes routine forensic analysis using short tandem repeat (STR) generates unsuccessful DNA testing result if the biological sample encountered is excessively degraded and low-template DNA. Herein, a new six-color fluorescence labeling system, including 59 autosomal diallelic deletion or insertion polymorphisms (DIPs), 2 miniSTRs, 2 Y-chromosome DIPs, and 1 Amelogenin gene with the amplicon sizes of less than 200 bp, was self-developed. According to the validation guidelines for DNA analysis methods formulated by the Scientific Working Group on DNA Analysis Methods, the validation studies have also been carried out for the multiplex system. This novel panel possessed the features of strong stability, high sensitivity, and good specificity, which was especially suitable for the forensic degraded and mixed sample detections. The cumulative power of exclusion and cumulative matching probability of the system were 0.9999978 and 9.833E-28, respectively, in Han Chinese in Hunan, China. Moreover, this system will be an effective new tool that can be independently applied to forensic personal identification and paternity testing in the populations from the East Asia region, even from the South Asia, America, and Europe regions. The system can also contribute to population phylogenetic affinity and genetic structure analyses among different populations.