Research Progress on Forensic Molecular Phenotyping in the Age of Precision Medicine.

ABSTRACT: With the development of new technologies, such as whole genome sequencing and big data, many countries have constructed large cohorts as strategic resources for precision medicine study. Related studies have significantly promoted research progress in many fields, such as medicine and biology, creating the genetic testing industry. In the forensic field, characterization technologies for features, such as biogeographic ancestry, genetic genealogy and physical appearance have emerged and developed rapidly. Compared with traditional forensic DNA technology, the new technology can search for characteristics and provide clues, quickly becoming an important means to solve difficult cases such as cold cases and accumulated cases. This paper briefly reviews the current status of the construction of large cohorts at home and abroad, and the research progress of forensic molecular phenotyping in the context of precision medicine in the context of precision medicine.

Pigmentation Phenotype Prediction of Chinese Populations from Different Language Families.

ABSTRACT: Objective To predict the pigmentation phenotypes of Chinese populations from different language families, analyze the differences and provide reference data for forensic anthropology and genetics. Methods The HIrisPlex-S multiplex amplification system with 41 loci related to pigmentation phenotypes was constructed in the laboratory, and 2 666 DNA samples of adult males of 17 populations from six language families, including Indo-European, Sino-Tibetan, Altaic, Hmong-Mien, Tai-Kadai and Austro-Asiatic language families distributed in different regions of China were genotyped. The pigmentation phenotype category of each individual was predicted using the online prediction system （https://HIrisPlex.erasmusmc.nl/）, and then the output data were statistically analyzed. Results About 1.92% of the individuals of Asian-European admixed populations from Indo-European and Altaic language families had blue eyes and 34.29% had brown or gold hair. The phenotypes of the color of eyes and hair of other populations had no significant difference, all individuals had brown eyes and black hair. There were differences in skin color of populations of different language families and geographical areas. The Indo-European language family had the lightest skin color, and the Austro-Asiatic language family had the darkest skin color; the southwestern minority populations had a darker skin color than populations in the plain areas. Conclusion The prediction results of pigmentation phenotype of Chinese populations are consistent with the perception of the appearance of each population, proving the reliability of the system. The color of eyes and hair are mainly related to ancestral components, while the skin color shows the differences between language families, and is closely related to geographical distribution of populations.

Age Estimation and Age-related Facial Reconstruction of Xinjiang Uygur Males by Three-dimensional Human Facial Images.

OBJECTIVES: To search age-correlated facial features and construct an age estimation model based on the three-dimensional （3D） facial images of Xinjiang Uygur males, and to structure individual face images of old age and young age. METHODS: Pretreatment was performed to collect 105 3D facial images of Xingjiang Uygur males aged between 17-57 years by Artec Studio software. The facial images were transferred to high-density 3D dot matrix data by FaceAnalysis software, and each image could be represented with 32 251 vertexes. Central correction of the facial images was done and all the data were aligned to a standard coordinate frame by generalized Procrustes analysis （GPA）. The age estimation model was established by partial least square regression （PLSR）. Furthermore, the changes of age-correlated facial features were presented on the heat map of average face, and the reconstruction of facial images at different ages was performed based on this model. RESULTS: With age, the average faces showed a series of changes including the nasolabial sulcus deepening, cheek sinking, cheekbone protruding and eye corner drooping. The Pearson correlation coefficient （r） between estimated age and chronological age was 0.71. The mean absolute deviation （MAD） of age estimation was 6.37 years. The results of age estimation in >30-40 years group showed a best accuracy （MAD=4.27 years）, and the deviations increased with age after 40 years. The composite facial images represented a significant result with age on facial morphological features and aging. CONCLUSIONS: The results of this study reveal the age-correlated facial features and aging markers in Uygur population, which help to construct a reliable age estimation model.

Systematic Review and Meta-Analysis of the Association between Sarcopenia and Dysphagia.

OBJECTIVES: Sarcopenia, an age-related decrease in muscle mass and function, is associated with several potential adverse health outcomes. Its association with another age-related syndrome, dysphagia remains unclear. This systematic review and meta-analysis aims to explore the association between sarcopenia and dysphagia. METHODS: PubMed, Embase, Scopus and CINAHL were searched for cross-sectional, case-control and cohort studies that investigated the association between sarcopenia and dysphagia. First author, publication year, study type, sample size, inclusion and exclusion criteria, participant demographics, definition and measurement for evaluation of sarcopenia and dysphagia, main outcomes were retrieved. The association between sarcopenia and dysphagia were expressed by odds ratio (OR) and 95% confidence interval (CI). RESULTS: 9 studies are eligible in the systematic review, and 5 cross-sectional studies comprising 913 participants which showed dichotomous classification of sarcopenia and dysphagia were included in the meta-analysis. The crude odds ratios (ORs) were extracted from 5 studies, and 4 of them also provide adjusted ORs. The crude ORs between sarcopenia and dysphagia is 6.17 (95% CI, 3.81- 10.00), after adjusting for some confounders, such as age, sex, Barthel Index score, nutritional status, sarcopenia is also have an association with dysphagia (adjusted ORs, 4.06; 95% CI, 2.27-7.29). The subgroup analysis showed that there was no significant difference between different sarcopenia diagnostic criteria, assessment tools of skeletal muscle mass and dysphagia. CONCLUSION: Sarcopenia was positively associated with dysphagia. Prevention and screening of dysphagia is essential among sarcopenic old patients. The causal relationship requires more prospective cohort study for confirmation.

[Assessment of Height Prediction Model Based on SNPs Loci].

OBJECTIVES: To establish a height prediction model of Chinese Han male based on the reported 547 height-associated single nucleotide polymorphisms （SNPs） loci in Europeans, and assess its accuracy for height estimation. METHODS: The DNA typing was analyzed in 59 Han male samples of Shandong province by Affymetrix SNP Array 6.0 chip and HiSeq 4000 sequencing platform. Prediction model was established using 547 height-associated SNPs loci as predictors and weight allele sums （WAS） as computing method. The accuracy of height prediction model was analysed using receiver operating characteristic （ROC） curve and area under curve （AUC）. RESULTS: There was no height-associated SNPs locus was found by genome-wide association studies. In present study, height prediction model was established by WAS and obtained an AUC of 0.67 （95% CI: 0.53-0.90）. CONCLUSIONS: It has reference value for predicting the height of Han male in Shandong province by WAS model based on 547 SNPs loci, while it is still necessary to further promote the accuracy of the prediction model by screening more height-associated SNPs loci with population heterogeneity.

Morphological variability and molecular characterization of Mawsonascaris australis (Johnston & Mawson, 1943) (Nematoda: Ascaridoidea: Acanthocheilidae) from the brown guitarfish Rhinobatos schlegelii Müller & Henle (Rhinopristiformes: Rhinobatidae).

A rare ascaridoid species, Mawsonascaris australis (Johnston & Mawson, 1943), is reported from the Taiwan Strait for the first time, and the brown guitarfish Rhinobatos schlegelii Müller & Henle (Rhinopristiformes: Rhinobatidae) represents a new fish host. The detailed morphology of M. australis was studied using light and scanning electron microscopy (SEM) based on the newly collected specimens. The results revealed some important, but previously unreported or erroneous, morphological features, including the lateral alae starting well posterior to the base of the ventrolateral lips, the presence of one pair of double paracloacal papillae and a single, small precloacal median papilla on the anterior margin of the cloaca. In addition, in order to determine whether the morphometric difference in the relative lengths of the right and left spicules represents intraspecific or interspecific variation, specimens of M. australis were characterized using molecular approaches by sequencing and analysing the ribosomal internal transcribed spacer (ITS) and mitochondrial cox1 genes. The results indicate that this morphometric difference represents intraspecific variability.

Nematode parasites infecting the starry batfish Halieutaea stellata (Vahl) (Lophiiformes: Ogcocephalidae) from the East and South China Sea.

The starry batfish Halieutaea stellata (Vahl) is a small, benthic fish found in Indo-West Pacific Oceans. However, our present knowledge of the helminth parasites of this fish is still fragmentary. In this study, a total of 29 fish collected from the East and South China Sea were examined to determine the prevalence, intensity and species composition of helminth parasites in H. stellata. Using morphological and molecular approaches, four species of nematodes were found parasitic in this fish host, including the adults and fourth-stage larvae of Raphidascaroides nipponensis Yamaguti 1941; adults and third-stage larvae of Raphidascaris lophii (Wu 1949), third- and fourth-stage larvae of Hysterothylacium larval type IV-A of Shamsi, Gasser & Beveridge 2013 and third-stage larvae of Hysterothylacium amoyense (Hsü 1993). Halieutaea stellata represents a new host record for the three last-named nematodes. Raphidascaroides nipponensis with the highest prevalence (82.5%) and intensity (mean = 13.5) of infection was considered as the dominant parasite species in H. stellata. The detailed morphology of the different developmental stages of the four nematode species was studied using light and scanning electron microscopy. All nematode species were also genetically characterized by sequencing and analysing the internal transcribed spacer (ITS) of the ribosomal DNA. This study provides further data on the occurrence of nematode parasites in H. stellata and also contributes to facilitate an accurate and rapid diagnosis of the infection by these little-known nematodes.

Characteristics and functions of human cytomegalovirus UL128 gene/protein.

Human cytomegalovirus (HCMV) ORF UL128 protein is highly conserved among viral field isolates and functions in two different molecular forms, monomeric UL128 protein and in a complex with glycoproteins gH, gL, UL130, and UL131A protein. Monomeric UL128 protein works as soluble chemokine analogue to attract peripheral blood mononuclear cells (PBMCs) and selectively induces expression of interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) in PBMCs. The gH/gL/UL128/UL130/UL131A complex is indispensable for entry into both endothelial and epithelial cells. In conclusion, UL128 plays an important role in HCMV infection.

[High glucose promotes gap junctional communication in cultured neonatal cardiac fibroblasts via AMPK activation].

Cardiac fibroblasts are known to be essential for adaptiveresponses in the patho- genesis of cardiovascular diseases, and increased intercellular communication of myocardial cells and cardiac fibroblasts acts as a crucial factor in maintaining the functional integrity of the heart. AMP-activated kinase (AMPK) is a key stress signaling kinase, which plays an important role in promoting cell survival and improving cell function. However, the underlying link between AMPK and gap junctional communication (GJIC) is still poorly understood. In this study, a connection between AMPK and GJIC in high glucose-mediated neonatal cardiac fibroblasts was assessed using fibroblast migration, measurement of dye transfer and connexin43 (Cx43) expression. 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) and Compound C (CC) were used to regulate AMPK activity. The levels of cell migration and Cx43 protein expression in neonatal cardiac fibroblasts increased during high glucose treatment, accompanied by developed dye transfer. In addition, high glucose induced abundant phosphorylation of AMPK. Suppression of AMPK phosphorylation using CC reduced dye transfer, cell migration and Cx43 protein expression in neonatal cardiac fibroblasts, whereas the activation of AMPK using AICAR mimicked the high glucose-mediated cell migration, Cx43 protein expression and dye transfer enhancement. AMPK appears to participate in regulating GJIC in high-glucose-treated neonatal cardiac fibroblasts, including cell migration, dye transfer, Cx43 expression and distribution.

The effect of maternal infection on cognitive development and hippocampus neuronal apoptosis, proliferation and differentiation in the neonatal rats.

Many epidemiological reports stated a strong association between maternal infection and development of cerebral palsy, which is a major cause of cognitive impairment. The pathophysiological mechanism of intrauterine inflammation is complex. Recently, it was demonstrated that inflammation has a modulating effect on adult neurogenesis. In this study, we discovered the effect of maternal infection to hippocampal neuronal apoptosis, proliferation and differentiation, and cognitive development in the developing brains of neonatal rats. Morris water maze test was used to assess learning and memory. Terminal deoxynucleotidyl transferase dUTP nick end labeling assay was used to determine neuronal apoptosis, immunostaining was conducted to assess neurogenesis, and Western blot for extracellular signal-regulated kinase (ERK), cyclic AMP responsive element-binding protein (CREB) and brain-derived neurotrophic factor (BDNF) expression in the hippocampus. Results demonstrated that maternal infection increased neuronal apoptosis and significantly impaired spatial learning and memory ability. Maternal infection significantly increased cell proliferation, accompanied by an increased expression of ERK (P3-P7), CREB (P3-P7) and BDNF (P3). On P28, there was no significant difference of cell survival and differentiation in two groups. These results suggest that variation in ERK activity and subsequent expression of its downstream targets, including CREB and BDNF might contribute, at least partially, to modulation of inflammation related cell proliferation, survival and differentiation. Maternal infection increased hippocampal neuronal apoptosis and affected cell proliferation and differentiation in neonatal rats, which may be regarded as an etiological factor in cognitive development impairment.

Analysis of GWAS-linked loci in Parkinson disease reaffirms PARK16 as a susceptibility locus.

OBJECTIVE: A genome-wide association study (GWAS) in the Japanese population identified 2 new Parkinson disease (PD) susceptibility loci on 1q32 (PARK16) (OMIM 613164) and BST1. We analyzed single nucleotide polymorphism (SNPs) located at the GWAS-linked loci (PARK16, PARK8, PARK1, and BST1) in a Chinese population and also conducted a meta-analysis in Asians by pooling 2 independent replication studies from Japan. METHODS: We conducted an analysis of 13 SNPs associated with PD GWAS-linked loci in 2 case-control cohorts comprised of 1,349 ethnic Chinese subjects. RESULTS: PARK16, PARK8, and PARK1 loci but not BST1 were found to be associated with PD. PARK16 SNPs were associated with a decreased risk while PARK1 and PARK8 SNPs were associated with an increased risk of PD. A pooled analysis of our Chinese cohorts and 2 Japanese replication cohorts involving 1,366 subjects with PD and 16,669 controls revealed robust association with these 3 loci and also BST1. There was a trend toward a stronger protective effect of SNPs at the PARK16 locus in sporadic PD compared to familial cases and in older compared to younger subjects. CONCLUSIONS: Our study reaffirms the role of GWAS-linked loci in PD in Asian subjects and the strength of association is similar between Chinese and Japanese subjects. Efforts to elucidate the associated gene within PARK16 locus are warranted.