The human genome is diploid, and knowledge of the variants on each chromosome is important for the interpretation of genomic information. Here we report the assembly of a haplotype-resolved diploid genome without using a reference genome. Our pipeline relies on fosmid pooling together with whole-genome shotgun strategies, based solely on next-generation sequencing and hierarchical assembly methods. We applied our sequencing method to the genome of an Asian individual and generated a 5.15-Gb assembled genome with a haplotype N50 of 484 kb. Our analysis identified previously undetected indels and 7.49 Mb of novel coding sequences that could not be aligned to the human reference genome, which include at least six predicted genes. This haplotype-resolved genome represents the most complete de novo human genome assembly to date. Application of our approach to identify individual haplotype differences should aid in translating genotypes to phenotypes for the development of personalized medicine.

No abstract

The replacement of normal endometrial epithelium by fibrotic tissue is the pathological feature of intrauterine adhesion (IUA), which is caused by trauma to the basal layer of the endometrium. COL5A2 is a molecular subtype of collagen V that regulates collagen production in fibrotic tissue. Here, we investigated the roles of Foxf2 and Smad6 in regulating the transcription of COL5A2 and their involvement in the pathogenesis of IUA. Small interference‐mediated Foxf2 (si‐Foxf2) silencing and pcDNA3.1‐mediated Smad6 (pcDNA3.1‐Smad6) up‐regulation were performed in a TGF‐β1‐induced human endometrial stromal cell line (HESC) fibrosis model. Assessment of collagen expression by Western blotting, immunofluorescence and qRT‐PCR showed that COL5A2, COL1A1 and FN were significantly down‐regulated in response to si‐Foxf2 and pcDNA3.1‐Smad6. Transfection of lentivirus vector‐Foxf2 (LV‐Foxf2) and pcDNA3.1‐Smad6 into HESCs and qRT‐PCR showed that Foxf2 promoted COL5A2 expression and Smad6 inhibited Foxf2‐induced COL5A2 expression. Co‐immunoprecipitation, chromatin immunoprecipitation and dual‐luciferase reporter assays to detect the interaction between Foxf2 and Smad6 and their role in COL5A2 transcription showed that Foxf2 interacted with Smad6 and bond the same promoter region of COL5A2. In a rat IUA model, injection of ADV2‐Foxf2‐1810 and ADV4‐Smad6 into the uterine wall showed that Foxf2 down‐regulation and Smad6 up‐regulation decreased fibrosis and the expression of COL5A2 and COL1A1, as detected by haematoxylin/eosin, Masson trichrome staining and immunohistochemistry. Taken together, these results suggested that Foxf2 interacted with Smad6 and co‐regulated COL5A2 transcription in the pathogenesis of IUA, whereas they played opposite roles in fibrosis.

Importance Under‐detection and late diagnosis are major causes of glaucoma‐related visual impairment. Cost‐effective opportunistic glaucoma screening is of great interest in the early identification and prevention of glaucoma. Background To describe the results of a health examination centre‐based opportunistic glaucoma screening and referral model. Design This single centre cross‐sectional study was conducted in a health examination centre affiliated to a tertiary hospital in Shenyang, northeastern China. Participants From 21 March to 30 September 2016, 14 367 individuals aged ≥ 30 years undergoing routine physical examinations were invited for this glaucoma screening. Methods Presenting visual acuity, non‐contact pneumotonometry and non‐mydriatic fundus photography were evaluated. Fundus photographs were classified as non‐glaucoma, possible, probable and definitive glaucoma. Participants with probable and definite glaucomatous discs or intraocular pressure ≥ 24 mmHg were referred for definitive examinations. Main Outcome Measures Detection rate of glaucoma suspects and ocular hypertension (OHT). Cost to identify a single case with suspected and diagnosed glaucoma was also calculated. Results Altogether, 277 glaucoma suspects and 327 ocular hypertension suspects were identified. Among 190 participants with probable/definite glaucomatous discs, 93 (48.9%) accepted further examination. Among these, 78 were diagnosed as glaucoma, seven as suspects and eight were excluded. Only 98 ocular hypertension suspects (30.0%) accepted further examinations: eight had primary angle closure and 23 had confirmed ocular hypertension. The cost to identify a single glaucoma suspect and definite glaucoma case were US$135 and US$857, respectively. Conclusions and Relevance This novel screening model provides opportunities to improve glaucoma detection at low cost. Interventions to improve follow‐up are needed.