Angiogenesis in a 3D model containing adipose tissue stem cells and endothelial cells is mediated by canonical Wnt signaling

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.

Construction and evaluation of in-house methylation-sensitive SNaPshot system and three classification prediction models for identifying the tissue origin of body fluid

The identification of tissue origin of body fluid can provide clues and evidence for criminal case investigations.To establish an efficient method for identifying body fluid in forensic cases,eight novel body fluid-specific DNA methylation markers were selected in this study,and a multiplex single base extension reaction(SNaPshot)system for these markers was constructed for the identification of five common body fluids(venous blood,saliva,menstrual blood,vaginal fluid,and semen).The results indicated that the in-house system showed good species specificity,sensitivity,and ability to identify mixed biological samples.At the same time,an artificial body fluid prediction model and two machine learning prediction models based on the support vector machine(SVM)and random forest(RF)algorithms were constructed using previous research data,and these models were validated using the detection data obtained in this study(n=95).The accuracy of the prediction model based on experience was 95.79%;the prediction accuracy of the SVM prediction model was 100.00%for four kinds of body fluids except saliva(96.84%);and the prediction accuracy of the RF prediction model was 100.00%for all five kinds of body fluids.In conclusion,the in-house SNaPshot system and RF prediction model could achieve accurate tissue origin identification of body fluids.

“法医系谱学瓜熟蒂落”解读

随着在美国被称为"金州杀手"("golden state killer")的连环奸杀案嫌疑人落网,法医系谱学研究作为一种新型的侦查技术手段受到广泛关注.该技术被美国Science杂志评选为2018年十大科学突破之一.2018年4月, 72岁的美国加州前警察约瑟夫·迪安杰洛(Joseph James DeAngelo)被指控在1974~1986年期间犯下约50起强奸案和12起谋杀案,时隔30多年,警方终于将这个连环杀手缉拿归案(https://www.nytimes.com/2018/04/25/us/golden-state-killer-serial.html).

Genetic diversity analysis of forty-three insertion/deletion loci for forensic individual identification in Han Chinese from Beijing based on a novel panel

Due to the virtues of no stutter peaks,low rates of mutation,and short amplicon sizes,insertion/deletion(InDei)polymorphism is an indispensable tool for analyzing degraded DNA samples from crime scenes for human identifications(Wang et al.,2021).Herein,a self-developed panel of 43 InDei loci constructed previously by our group was utilized to evaluate the genetic diversities and explore the genetic background of the Han Chinese from Beijing(HCB)including 301 random healthy individuals.

An interpretation of the genetic polymorphism and population genetic background of Ankang Han population via a novel InDel panel

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.

Genetic polymorphisms of 44 Y chromosomal genetic markers in the Inner Mongolia Han population and its genetic relationship analysis with other reference populations

Y chromosomal genetic markers in the non-recombining region are commonly used for human evolution research,familial searching,and forensic male differentiation since they strictly follow paternal inheritance.Y chromosomal short tandem repeats(Y-STRs)possess extraordinarily advantages in forensic applications because of their high polymorphisms and special genetic pattern.Here,we assessed the genetic diversities of 41 Y-STRs and three Y chromosomal insertion/deletion(Y-InDels)loci in the Chinese Inner Mongolia Han population;besides,genetic differentiation analyses among the studied Han population and other previously reported populations were conducted based on 27 same Y-STRs.Totally,425 alleles were observed in 324 Inner Mongolia Han individuals for these Y-markers.Gene diversities of these Y-markers distributed from 0.0306 to 0.9634.The haplotype diversity and discriminatory capacity of these Y-markers in the Inner Mongolia Han population were 0.9999 and 0.98457,respectively.Haplotype resolution comparisons of different Y-marker groups in the studied Han population revealed that higher haplotype resolution could be achieved for these 44 Y-markers.Population genetic analyses of the Inner Mongolia Han population and other reference populations demonstrated that the studied Han population had relatively closer genetic affinities with Northern Han Chinese populations than Southern Han and other minority groups.To sum up,these 44 Y-markers can be utilized as a valuable tool for male differentiation in the Inner Mongolia Han population.

Genetic polymorphisms and phylogenetic analyses of the Ü-Tsang Tibetan from Lhasa based on 30 slowly and moderately mutated Y-STR loci

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.

Comprehensive analyses for genetic diversities of 19 autosomal STRs in Chinese Kazak group and its phylogenetic relationships with other continental populations

Short tandem repeats(STRs)play an essential role in forensic genetics due to their high degree of polymorphisms,wide distributions and easy detection method.In this study,allelic frequencies and forensic statistical parameters of the 19 autosomal STR loci in a Kazak ethnic group were calculated,and its genetic relationships with reference populations were assessed in order to understand population structure better and enrich population genetic data for forensic practice in Chinese Kazak ethnic group.There were 226 identified alleles with the corresponding allelic frequencies ranging from 0.0008 to 0.5295 in the 628 unrelated healthy Kazak individuals in Xinjiang Uygur Autonomous Region.All autosomal STRs were conformed to the Hardy-Weinberg equilibrium after Bonferroni’s correction.The cumulative power of discrimination and the combined probability of exclusion of all the 19 autosomal STRs were 0.999999999999999999999997162 and 0.999999994484,respectively.Furthermore,the DA distances and Fixation index values of pairwise populations,principal component analysis,multidimensional scaling analysis,phylogenetic tree analysis and structure analysis were conducted to probe the genetic relationships between the Kazak group and other reference populations.The population genetic results showed that these 19 autosomal STR loci were characterised by high genetic diversities in the Kazak group.Furthermore,the studied Kazak group had close genetic relationships with the Uyghur group and the Uzbek group.The present results may facilitate understanding the genetic background of the Chinese Xinjiang Kazak group.

Genetic and structural characterization of 20 autosomal short tandem repeats in the Chinese Qinghai Han population and its genetic relationships and interpopulation differentiations with other reference populations

China is a multinational country composed of 56 ethnic groups of which the Han Chinese accounts for 91.60%.Qinghai Province is located in the northeastern part of the Qinghai-Tibet Plateau,has an area of 72.12 km2,and is the fourth largest province in China.In the present study,we investigated the genetic polymorphisms of 20 short tandem repeat (STR) loci in a Qinghai Han population,as well as its genetic relationships with other populations.A total of 273 alleles were identified in 2 000 individuals at 20 loci,and the allelic frequency ranged from 0.0002 to 0.5327.The 20 STR loci showed a relatively high polymorphic rate in the studied group.Observed and expected heterozygosities ranged 0.613 0-0.907 5 and 0.614 8-0.920 0,respectively.The combined power of discrimination,and the probability of exclusion in duo and trio cases were 0.999 999 999 999 999 999 999 999 34,0.9999960 and 0.9999999965,respectively.Analyses of interpopulation differentiation revealed that the most significant differences were found between the Qinghai Han and Malaysian,while no significant differences were found between the Qinghai Han and Han people from Shaanxi and Jiangsu.The results of principal component analysis,multidimensional scaling analysis and phylogenetic reconstructions also suggested the close relationships between the Qinghai Han and other two Han populations.The present results,therefore,indicated that these 20 STR loci could be used for paternity testing and individual identification in forensic applications,and may also provide information for the studies of genetic relationships between Qinghai Han and other groups.

Forensic Validation Studies of a Novel 35-InDel Multiplex Polymerase Chain Reaction System

Background:A difficulty associated with forensic applications is the detection of degraded biological materials.Due to the large amplicon sizes of short tandem repeat alleles,valid genotyping results cannot be obtained from degraded biological materials.Recently,insertion/deletion(InDel)polymorphisms have been used in forensic applications for their widespread distributions in the human genome,short amplicon sizes,and low mutation rates.Purpose:Human identification InDel panels have mostly been designed for European populations.Therefore,our laboratory independently developed a multiplex polymerase chain reaction(PCR)system with 35 polymorphic InDel loci to be used for human identification in China.Forensic validation studies were conducted on this novel 35-InDel multiplex PCR system.Methods:The 35 InDel loci were screened in the database,and then used with the traditional PCR amplification and capillary electrophoresis platform combined with five-color fluorescence parallel detection technology.Validation studies were performed on this novel panel,including accuracy,repeatability and reproducibility,species specificity,sensitivity,stability,forensic case sample detection,and mixture studies.In addition,forensic efficiency assessments were conducted in populations from different continents.Results:The data of validated studies indicated that the novel 35-InDel panel was accurate,stable,and efficient for forensic purposes.For human identification,the cumulative power of discrimination values for the these 35 InDel loci in East Asian,South Asian,European,American,and African populations were 0.999999999999995,0.999999999999995,0.999999999999971,0.9999999999999960,and 0.999999999998166,respectively.Conclusions:In this study,a set of 35 InDel loci were conducted in a multiplex amplification system for human identification of degraded DNA sample,and this new assay was efficient and stable.The present results suggested that the 35-InDel panel was a reliable tool for forensic use and could be efficiently used for human identification in the East Asian populations.