[Identification of pathogenic variants in three Chinese patients with McCune-Albright syndrome].

OBJECTIVE: To explore the genetic basis for three Chinese patients with McCune-Albright syndrome (MAS). METHODS: Three children who had respectively presented at Shandong Provincial Hospital in April 2019 and Peking Union Medical College Hospital in August 2020 and May 2021 were selected as the research subjects. Peripheral blood samples of the probands and their family members were taken for the extraction of genomic DNA. Potential variants were screened by whole exome sequencing (WES), and candidate variants were validated by Sanger sequencing of the patients and their family members. RESULTS: The proband from family 1 was found to harbor a heterozygous c.601C>T (p.R201C) missense variant in exon 8 of the GNAS gene, whilst the probands from families 2 and 3 were both found to harbor a heterozygous c.602G>A (p.R201H) missense variant in exon 8 of the GNAS gene. Both variants were known to be pathogenic, and all probands were found to be mosaics for the corresponding variants but with various degrees. CONSLUSION: WES can effectively diagnose MAS and other somatic genetic disorders. In this study, the combined WES and Sanger sequencing have verified the degree of mosaicisms of pathogenic variants in the three MAS patients, albeit no apparent correlation was found between the degree of mosaicisms and the phenotype of patients. Above finding has provided a basis for genetic counseling and prenatal diagnosis for the affected families.

[Genetic analysis of two families with Short-rib thoracic dysplasia type 3].

OBJECTIVE: To explore the pathogenic variants and clinical classification of two fetuses with Short-rib thoracic dysplasia with or without polydactyly (SRTD). METHODS: With informed consent obtained, the phenotypic characteristics of the fetuses were comprehensively examined, and genomic DNA was extracted from fetal skin tissue and peripheral blood samples of the parents with conventional phenol-chloroform method. Whole exome sequencing (WES) was carried out on both fetuses, and the candidate variants were validated by Sanger sequencing. The pathogenicity of the candidate variants was analyzed using bioinformatic software VarCards, and the impact of the variants on the protein structure was predicted with Swiss-Pdb-viewer. RESULTS: Both fetuses were found to harbor compound heterozygous variants of the DYNC2H1 gene, including c.515C>A (p.Pro172Gln) and c.5983G>A (p.Ala1995Thr) in fetus 1, and c.5920G>T (pGly1974) and c.9908T>C (p.He3303Thr) in fetus 2. The parents of both fetuses were heterozygous carriers. CONCLUSION: The compound heterozygous variants of the DYNC2H1 gene probably underlay the SRTD3 in the two fetuses.

Identifying the role of array electrodes in improving the compost quality of food waste during electric field-assisted aerobic composting.

Low composting temperature and long maturation periods are two major problems during food waste composting. In this study, a novel array-based electric field-assisted aerobic composting (Pin-EAC) process was tested on food waste compost. Pin-EAC increase the composting temperature to 69.3 °C, and improved the germination index by 15%. The Pin-EAC took at least 40% less time to reach the standard compost maturity. The fluorescent spectroscopy results showed that Pin-EAC could increase humic acid and fulvic acid by 33% and 37%, respectively. Pin-EAC could increase the diversity of thermophilic bacteria during composting. The co-occurrence network shown that Pin-EAC are more closely related to oxygen and temperature. This work has initially shown that the use of an electric field could improve food waste composting quality, suggesting that the Pin-EAC process is an effective strategy for high-water and high-oil organic solid waste aerobic composting.

CRISPR/Cas9 correction of a dominant cis-double-variant in COL1A1 isolated from a patient with osteogenesis imperfecta increases the osteogenic capacity of induced pluripotent stem cells.

Osteogenesis imperfecta (OI) is a hereditary skeletal disorder that is mainly caused by variants in COL1A1/2. So far, no specific treatment has been developed to correct its underlying etiology. We aimed to gain a better understanding of the pathological mechanisms of OI and develop gene therapies to correct OI-causing variants. A de novel cis-double-variant c.[175C>T; 187T>A] in COL1A1 was identified from a 5-year-old OI patient by whole-exome sequencing (WES). Three peptide nucleic acids (PNAs) were designed and then transfected patient-derived fibroblasts. PNA2 affected the translational strand and induced an optimal interfering effect at 0.25µM concentration, proved by Sanger sequencing, qPCR, Western blot, and immunostaining. Additionally, induced pluripotent stem cells (iPSCs) were cultured from patient-derived fibroblasts. Clones of iPSCs with c.187T>A variant and those with both variants largely restored their osteogenic capacities after CRISPR/Cas9 gene editing, which corrected the variants. Importantly, correcting c.187T>A variant alone in CRISPR-edited iPSCs was sufficient to alleviate OI phenotypes, as indicated by increased levels of COL1A1, COL1A2, ALP mRNAs, and COL1A1 protein. Our findings suggest that c.187T>A is the dominant variant of cis-double-variant in COL1A1 that led to OI, and PNA interference and CRISPR/Cas9 gene editing may be new therapeutic tools for OI treatment. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Deciphering the structural characteristics and molecular transformation of dissolved organic matter during the electrolytic oxygen aerobic composting process.

Electrolytic oxygen aerobic composting (EOAC) effectively treats organic solid waste by using in-situ electrolytic oxygen for aeration. However, the fundamental mechanism of compost maturity is still unclear. Therefore, we comprehensively characterized dissolved organic matter (DOM) transformation closely related to compost maturity during EOAC. Excitation-emission matrix-parallel factor (EEM-PARAFAC) and Fourier transform infrared (FTIR) analysis confirmed that EOAC quickly decreased organic matter and increased humus substances, accelerating the compost humification process compared with conventional aerobic composting. Electrospray ionization (ESI) Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) analysis reveals that the double bound equivalent and aromaticity index during EOAC are higher than in conventional aerobic composting (CAC), suggesting more aromatic compounds in EOAC. DOM's detailed transformation investigation suggested that low O/C and high H/C compounds were preferentially decomposed during EOAC. Our investigation firstly extends the in-depth molecular mechanisms of humification during EOAC, and reveals its practical engineering applications.

In situ generated oxygen distribution causes maturity differentiation during electrolytic oxygen aerobic composting.

Electrolytic oxygen aerobic composting (EOAC) is an effective treatment with greater technical superiority and cost advantages for organic solid waste using in situ electrolytic oxygen as a feasible strategy to replace conventional aeration. However, the unclear effects of distribution and variation of in situ electrolytic oxygen on compost maturation in different depth zones of EOAC need further exploration. This study demonstrated that the humification of organic matter was faster at the bottom than in the middle and at the top. The main reason was that the higher oxygen content and lower moisture content in the bottom promoted microbial degradation and heat production, resulting in higher temperatures. The microbial analysis showed that the abundance of typical thermophilic bacteria (such as Cerasibacillus, Lactobacillus, and Pseudogracilibacillus) that could promote compost maturation was higher at the bottom than in the middle and at the top. The finding provided in-depth molecular insights into differentiated humification from bottom to top in EOAC and revealed its further practical engineering applications.

Moisture migration driven by the electric field causes the directional differentiation of compost maturity.

Electric field-assisted aerobic composting (EAC) has been recently believed as a novel and effective process for the resource utilization of organic solid waste. However, the effect of electric field in composting process needs to be further clarified. Herein, moisture migration and compost maturity along electric-field-direction (from anode to cathode) in EAC was first to be explored. It was found that moisture content and compost maturity changed regularly from anode to cathode. At the end of composting, the moisture content of S3 (cathodic zone) was 30% and 62% higher than that of S2 (middle zone) and S1 (anodic zone), respectively. The germination index (a key parameter for compost maturity) in S3 (138.92%) was significantly higher than that of S2 (104.98%) and S1 (84.45%). However, temperatures in S3 were lower than that of S1 and S2, indicating the moisture content played a more important role than temperature for compost maturity in EAC. Furthermore, the microbial activities in S3 were also higher than that of S1 and S2, supporting the trend of compost maturity. This pioneering study demonstrates the electric field can drive moisture gradient migration to control the directional differentiation of compost maturity, showing a great application potential in aerobic composting.

Derivation of a human induced pluripotent stem cell (hiPSC) line (PUMCSMi001-A) from peripheral blood mononuclear cells of one healthy female donor.

Human induced pluripotent stem cells (hiPSCs) are valuable models for studying the molecular pathogenesis of inherited diseases and key tools for cell therapy. We derived peripheral blood mononuclear cells (PBMCs) isolated from a 30-year-old healthy female donor using integration-free Sendai virus to generate a hiPSC line, PUMCSMi001-A, which can serve as a control for the studies of pathogenesis of cardiovascular disease and drug screening. Quality assessment confirmed that the cell line PUMCSMi001-A expressed pluripotency genes, had normal karyotype and differentiation potential into three layers in vivo.

Genotypic and Phenotypic Analysis in Chinese Cohort With Autosomal Recessive Osteogenesis Imperfecta.

Osteogenesis imperfecta (OI) is a rare heritable skeletal disorder which is mainly caused by defected type I collagen. Autosomal recessive OI (AR-OI) is caused by mutations of genes that are responsible for type I collagen modification and folding, and is often associated with more severe phenotypes. Due to the limited number of recessive OI patients, it has been difficult to study the mutation spectrum as well as the correlation of genotype and phenotype. This study recruited a Chinese cohort of 74 AR-OI families, aiming to establish the mutation spectrum and to examine the genotypic and phenotypic correlation. We identified 82 variants including 25 novel variants and 57 HGMD reported variants in these AR-OI patients, using whole exome sequencing/panel sequencing combined with Sanger sequencing. Pathogenic mutations were found at WNT1 (n = 30, 40.54%), SERPINF1 (n = 22, 29.73%), FKBP10 (n = 10, 13.51%), CRTAP (n = 3, 4.05%), P3H1 (n = 3, 4.05%), SERPINH1 (n = 2, 2.70%), SEC24D (n = 3, 4.05%), and PLOD2 (n = 1, 1.35%) respectively. Thus, WNT1 represents the most frequent pathogenic gene of AR-OI in Chinese population. The most common clinical manifestations of AR-OI patients include walking problem (72.86%), scoliosis (65.28%) and frequent fractures (fractures ≥2/year) (54.05%). Interestingly, ptosis represents a unique phenotype of patients carrying WNT1 variants, and it was rare in patients harboring other pathogenic genes. Our study expanded the mutation spectrum of AR-OI and enriched the knowledge of genotypic and phenotypic correlation in Chinese cohort with AR-OI.

Climate change leads to a doubling of turbidity in a rapidly expanding Tibetan lake.

Recent climate change is causing most lakes on the Tibetan Plateau to grow at an unprecedented rate. Changes in the physical properties and water storage of the lakes are now relatively well documented. Yet the impacts on their water quality remain poorly understood. Turbidity is a well-established optical water-quality indicator related to suspended particulate matter concentration which can affect vertical light attenuation and ecosystem functioning. Here, we use remotely sensed data to assess the seasonal and long-term variations in turbidity in Siling Lake, one of the fastest growing lakes on the Tibetan Plateau, and to identify potential driving mechanisms of this change. The lake experiences two distinct peaks of turbidity during the year: one in August (warm season) caused by the seasonal influx of sediments from the Zagya Zangbo River, and one in December (cold season) caused by the wind-driven resuspension of sediments along the lakes' shorelines. The analysis further revealed a persistent increasing trend that doubled the average lake turbidity between 2000 and 2017. Evidence suggests this rise in turbidity results from a climate-driven increase in sediment supply from the Zagya Zangbo River, and from sediment resuspension associated with the erosion of shorelines recently submerged during the rapid expansion of the lake (paleoshorelines). Our results highlight the vulnerability of the Tibetan Lakes' water quality to climate change.

Selective attentional biases towards a self-related facial feature among orthognathic patients.

The present study aimed to determine what kind of face and which area of the face (eyes or mouth) rapidly capture attention among patients with dentomaxillofacial deformities (DDs). In the present experiments, faces were categorized as highly or lowly attractive (HA and LA, respectively) and as a face with dentomaxillofacial deformities (DD face). A dot-probe task and a change-detection task were utilized to examine the attentional bias to the faces and facial features, respectively. In Study 1, we found that DD patients showed an attentional bias to DD male faces compared with HA male faces, while controls showed an opposite attentional pattern. In Study 2, we found that patients responded faster to the changes in the mouths of DD male faces than those of HA and LA faces. Moreover, patients paid less attention to the mouths of HA faces than to the region of the eyes, and they paid less attention than did the controls. These results indicated that DD patients selectively directed their attention to DD male faces and the mouths of these faces. These findings were not evident in female faces. Furthermore, patients showed avoidance of the mouths of HA faces regardless of the gender of the faces.

[Influence of sociocultural factors on the selection of orthognathic surgery in patients with dental and maxillofacial deformities].

PURPOSE: To investigate social and cultural influence on preference of orthognathic surgery in patients with maxillofacial deformities through establishment of models, in order to provide motivational supports, improve treatment coordination and patients' satisfaction. METHODS: The orthognathic group included 137 patients with maxillofacial malformations, while the control group included 245 students from one university in Guizhou province. Acceptance of cosmetic surgery scale(ACSS), Orthognathic quality of life questionnaire(OQLQ) and sociocultural attitudes towards appearance questionnaire-3(SATAQ-3) were used in two groups. SPSS 24.0 software package was used to analyze the data with hierarchical regression function. RESULTS: The results showed that there was a strong correlation between the studied variables (R2=0.26). Sociocultural factors could be used to predict patients' preference to orthognathic surgery. Patients in the orthognathic group could be predicted based on upward physical appearance comparison (t=2.28, P<0.024), while subjects in the control group could be predicted based on upward physical appearance comparison (t=2.47, P<0.014) and the appearance media pressure (t=2.30, P<0.022). Orthognathic related social pressure significantly explained the considerations of plastic surgery in patients with dento-maxillofacial deformities (P<0.000); However, this factor had no significant explanation for the controls (P<0.779). CONCLUSIONS: Social and cultural factors are proven to contribute to the selection of orthognathic surgery for patients with dental and/or maxillofacial malformations, and may affect patients' expectation on the final results.