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Precision medicine relies on high-accuracy individual-level genotype data. However, the whole-genome sequencing (WGS) is currently not suitable for studies with very large sample sizes due to budget constraints. It is particularly important to construct highly accurate haplotype reference panel for genotype imputation. In this study, we selected 9,950 individuals from the China Kadoorie Biobank (CKB) cohort and 50 Chinese samples from the 1000 Genome Project (1KGP) for medium-depth WGS to construct a CKB reference panel. The results of imputing microarray datasets showed that the CKB panel outperformed the extended high coverage 1KGP, TOPMed, ChinaMAP, and NuyWa panels in terms of both the number of well-imputed variants and imputation accuracy. In addition, we have completed the imputation of over 100,000 CKB microarray data with the CKB panel, and the after-imputed genotype data is the hitherto largest whole genome data of the Chinese population. Finally, we developed an online server for offering free genotype imputation service based on the CKB reference panel (https://db.cngb.org/imputation/). We believe that the constructed CKB reference panel is of great value for imputing microarray or low-depth genotype data of Chinese population. The imputation-completed 100,000 microarray data are fundamental resources of population genetic studies for complex traits and diseases in the Chinese population.

Hg(II) ions in wastewater are highly toxic to the environment and human health, yet many materials to remove the ions exhibit lower adsorption efficiency, and few studies report the reuse of Hg(II)-loaded waste materials. Here, a cheap and efficient adsorbent was prepared for the removal of Hg(II) based on corn bracts (CB) loading copper sulfide (CuS), and the Hg(II)-adsorbed material was reused as a photocatalyst. By changing the adsorption variables such as pH, adsorbent dosage, Hg(II) concentration, contact time and coexisting ions, the optimum adsorption conditions were obtained. The study indicated the adsorption capacity and removal rate of CB/CuS reached 249.58 mg/g and 99.83% at pH 6 with 20 mg CB/CuS, 50 mL Hg(II) concentration (100 mg/L) and 60 min, and coexisting ions did not affect the uptake of Hg(II). The adsorption behavior of CB/CuS toward Hg(II) followed pseudo-second-order and Langmuir models, with the theoretical maximum adsorption capacity of 316.46 mg/g. Finally, we explored an alternative strategy to dispose of spent adsorbents by converting the CB/CuS/HgS into a photocatalyst for the degradation of rhodamine B, with a removal rate of 98%. Overall, this work not only develops a promising material for the treatment of Hg(II)-containing wastewater, but opens up a new approach for the use of the waste adsorbent. HIGHLIGHTS Corn bracts as a carrier of CuS was designed for rapid uptake of Hg(II).; The removal rate of the adsorbent could reach 99.83% in only 60 min.; The theoretical maximum could reach 316.46 mg/g.; Coexisting ions had no significant effect on selective Hg(II) ion removal.; The spent adsorbent was used for photodegradation reaction.

// Yun Wang 1, * , Abir Garraoui 2, 3, * , Liuzhi Zeng 4, * , Mingying Lai 1 , Fen He 1 , Huaizhou Wang 5 , Chongyi Jiang 2 , Yulan Chen 2 , Lanlan Dai 2 , Ning Fan 1 , Huanming Yang 2, 6 , Jianguo Zhang 2, 7 and Xuyang Liu 1 1 Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Shenzhen University, Shenzhen, China 2 BGI-Shenzhen, Shenzhen, China 3 Laboratory of Human Molecular Genetics, Faculty of Medicine, University of Sfax, Sfax, Tunisia 4 The First People's Hospital of Chengdu, Chengdu, China 5 Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Beijing Ophthalmology & Visual Sciences Key Laboratory, Capital Medical University, Beijing, China 6 James D. Watson Institute of Genome Sciences, Hangzhou, China 7 Shenzhen Key Laboratory of Neurogenomics, BGI-Shenzhen, Shenzhen, China * These authors have contributed equally to this work Correspondence to: Xuyang Liu, email: xliu1213@126.com Jiangguo Zhang, email: Jianguozhang@genomics.cn Keywords: Bardet-Biedl syndrome, retinitis pigmentosa, FBN3 , whole exome sequencing Received: March 27, 2017 Accepted: August 29, 2017 Published: September 30, 2017 ABSTRACT Purpose: This study was designed to evaluate the molecular genetics of a Chinese family with Bardet-Biedl syndrome (BBS). Methods: All the family members underwent medical history evaluation, ophthalmologic and physical examinations. Whole exome sequencing was performed on two affected individuals and their parents. All variants were verified in all family members by PCR amplification and Sanger sequencing. Results: Patients in this family were diagnosed as Bardet-Biedl syndrome, with an inheritance pattern of autosomal recessive. Compound heterozygous mutations of the FBN3 gene (c.3616G>A and c.6037C>T) were identified by whole exome sequencing. Results from Sanger sequencing showed co-segregation of these compound heterozygous mutations in the FBN3 gene with BBS disease in the family. Conclusion: Novel compound heterozygous mutations c.3616G>A and c.6037C>T of FBN3 were identified in all affected individuals but not in the unaffected family members. This is the first time to the best of our knowledge, that the FBN3 gene is involved in the pathogenesis of BBS. This study will expand our understanding about the gene spectrum related to this genetically heterogeneous disorder.

Inherited retinal diseases (IRDs) are a common cause of visual impairment. IRD covers a set of genetically highly heterogeneous disorders with more than 150 genes associated with one or more clinical forms of IRD. Molecular genetic diagnosis has become increasingly important especially due to expanding number of gene therapy strategies under development. Next generation sequencing (NGS) of gene panels has proven a valuable diagnostic tool in IRD. We present the molecular findings of 677 individuals, residing in Denmark, with IRD and report 806 variants of which 187 are novel. We found that deletions and duplications spanning one or more exons can explain 3% of the cases, and thus copy number variation (CNV) analysis is important in molecular genetic diagnostics of IRD. Seven percent of the individuals have variants classified as pathogenic or likely-pathogenic in more than one gene. Possible Danish founder variants in EYS and RP1 are reported. A significant number of variants were classified as variants with unknown significance; reporting of these will hopefully contribute to the elucidation of the actual clinical consequence making the classification less troublesome in the future. In conclusion, this study underlines the relevance of performing targeted sequencing of IRD including CNV analysis as well as the importance of interaction with clinical diagnoses.

The objective of this study was to identify novel causal genes involved in the pathogenesis of Kashin-Beck disease (KBD). A representative grade III KBD sib pair with serious skeletal growth and development failure was subjected to exome sequencing using the Illumina Hiseq2000 platform. The detected gene mutations were then filtered against the data of 1000 Genome Project, dbSNP database, and BGI inhouse database, and replicated by a genome-wide association study (GWAS) of KBD. Ninety grade II or III KBD patients with extreme KBD phenotypes and 1627 healthy controls were enrolled in the GWAS. Affymetrix Genome-Wide Human SNP Array 6.0 was applied for genotyping. PLINK software was used for association analysis. We identified a novel 106T>C at the 3'UTR of the FGF12 gene, which has not been reported by now. Sequence alignment observed high conversation at the mutated 3'UTR+106T>C locus across various vertebrates. In the GWAS of KBD, we detected nine SNPs of the FGF12 gene showing association evidence (P value

As a powerful tool to identify the molecular pathogenesis of Mendelian disorders, exome sequencing was used to identify the genetic basis of two siblings with hearing loss and hypotrichosis and clarify the diagnosis. No pathogenic mutations in GJB2, GJB3 and GJB6 genes were found in the siblings. By analysis of exome of the proband, we identified a novel missense (p.R306C) mutation and a nonsense (p.R186*) mutation in the BCS1L gene. Mutations were confirmed by Sanger sequencing. The siblings were compound heterozygotes, and the inheritance mode of autosomal recessive was postulated. BCS1L is the causative gene of Björnstad syndrome, which is characterized by sensorineural hearing loss and pili torti. The longitudinal gutters along the hair shaft were found by scanning electron microscopy in our patient. Therefore the diagnosis of Björnstad syndrome was eventually made for the patients. Our study extends the phenotypic spectrum of Björnstad syndrome and highlights the clinical applicability of exome sequencing as a diagnostic tool for atypical Mendelian disorders.

Calpastatin is an endogenous specific inhibitor of calpain, a calcium-dependent cysteine protease. Here we show that loss-of-function mutations in calpastatin (CAST) are the genetic causes of an autosomal-recessive condition characterized by generalized peeling skin, leukonychia, acral punctate keratoses, cheilitis, and knuckle pads, which we propose to be given the acronym PLACK syndrome. In affected individuals with PLACK syndrome from three families of different ethnicities, we identified homozygous mutations (c.607dup, c.424A>T, and c.1750delG) in CAST, all of which were predicted to encode truncated proteins (p.Ile203Asnfs∗8, p.Lys142∗, and p.Val584Trpfs∗37). Immunohistochemistry shows that staining of calpastatin is reduced in skin from affected individuals. Transmission electron microscopy revealed widening of intercellular spaces with chromatin condensation and margination in the upper stratum spinosum in lesional skin, suggesting impaired intercellular adhesion as well as keratinocyte apoptosis. A significant increase of apoptotic keratinocytes was also observed in TUNEL assays. In vitro studies utilizing siRNA-mediated CAST knockdown revealed a role for calpastatin in keratinocyte adhesion. In summary, we describe PLACK syndrome, as a clinical entity of defective epidermal adhesion, caused by loss-of-function mutations in CAST.

Keratoderma-hypotrichosis-leukonychia totalis syndrome (KHLS) is an extremely rare, autosomal-dominant disorder characterized by severe skin hyperkeratosis, congenital alopecia and leukonychia totalis. The genetic defect underlying KHLS remained undetermined. By performing whole-exome sequencing in a family with KHLS, we identified a heterozygous mutation (c.23G>T [p.Gly8Val]) in GJA1, which cosegregated with the phenotype in the family. In an additional affected individual, we also found the identical de novo mutation which was absent in his unaffected family members. GJA1 encodes a gap junction protein connexin 43 (Cx43) which is ubiquitously expressed in various organs, including the epidermis and hair follicles. In vitro studies on HEK293 cells expressing Cx43(Gly8Val) found that the protein formed gap junction plaques between adjacent transfected cells, as observed in the wild-type. Dye-transfer experiments by microinjection of Lucifer yellow displayed functional gap junction of the Cx43(Gly8Val) mutant. Using patch clamp and Ca(2+) imaging methods, we observed that the Cx43(Gly8Val) hemichannel had significantly more openings than Cx43(WT), facilitating Ca(2+) influx at resting potential. Such gain-of-function effect might result in cytoplasmic Ca(2+) overload, accelerated apoptosis of keratinocytes and subsequent skin hyperkeratosis. Taken together, our results demonstrated that, with probably enhanced hemichannel activities, a mutation in GJA1 is linked to KHLS without extracutaneous involvement.

Pure hair and nail ectodermal dysplasia (PHNED) is a congenital condition characterized by hypotrichosis and nail dystrophy. Autosomal-recessive PHNED has previously been mapped to chromosomal region 12q12-q14.1, which contains the type II hair keratin and HOXC clusters. Hoxc13-null mice are known to develop hair and nail defects very similar to those seen in human PHNED. We performed whole-exome sequencing in a consanguineous Chinese family affected by PHNED and identified a homozygous nonsense mutation (c.390CA [p.Tyr130(∗)]) in HOXC13 in all affected individuals. In an additional affected female from a consanguineous Afghan family, we found a 27.6 kb homozygous microdeletion involving the first exon of HOXC13. We examined HOXC13 expression in scalp specimen obtained from the index individual of the Chinese family and detected dramatically reduced mRNA levels in skin tissue and nearly absent protein staining in hair follicles, suggesting a mechanism of nonsense-mediated mRNA decay. We also observed markedly decreased expression of four HOXC13 target genes in the specimen. Taken together, our results demonstrate that loss-of-function mutations in HOXC13 cause autosomal-recessive PHNED and further highlight the importance of HOXC13 in hair and nail development.