Loss-of-function variants in GLMN are associated with generalized skin hyperpigmentation with or without glomuvenous malformation.

BACKGROUNDS: Inherited hyperpigmented skin disorders comprise a group of entities with considerable clinical and genetic heterogenicity. The genetic basis of a majority of these disorders remains to be elucidated. OBJECTIVES: This study aimed to identify the underlying gene for an unclarified disorder of autosomal-dominant generalized skin hyperpigmentation with or without glomuvenous malformation. METHODS: Whole-exome sequencing was performed in five unrelated families with autosomal-dominant generalized skin hyperpigmentation. Variants were confirmed using Sanger sequencing and a minigene assay was employed to evaluate the splicing alteration. Immunofluorescence and transmission electron microscopy (TEM) were used to determine the quantity of melanocytes and melanosomes in hyperpigmented skin lesions. GLMN knockdown by siRNA assays was performed in human MNT-1 cells to examine melanin concentration and the underlying molecular mechanism. RESULTS: We identified five variants in GLMN in five unrelated families, including c.995_996insAACA(p.Ser333Thrfs*11), c.632 + 4delA, c.1470_1473dup(p.Thr492fs*12), c.1319G > A(p.Trp440*), and c.1613_1614insTA(Thr540*). The minigene assay confirmed that the c.632 + 4delA mutant resulted in an abolishment of the canonical donor splice site. Although the number of melanocytes remained unchanged in skin lesions as demonstrated by immunofluorescent staining of tyrosinase and premelanosome protein (PMEL), TEM revealed an increased quantity of melanosomes in the skin lesion of a patient. The GLMN-knockdown MNT-1 cells demonstrated a higher melanin concentration, a higher proportion of stage III and IV melanosomes, upregulation of MITF and tyrosinase, and downregulation of phosphorylated p70S6 K, compared to mock-transfected cells. CONCLUSIONS: We found loss-of-function variants in GLMN are associated with generalized skin hyperpigmentation with or without glomuvenous malformation. Our study implicates a potential role of glomulin in human skin melanogenesis, in addition to vascular morphogenesis.

Biallelic mutations in LSS in autosomal-recessive mutilating palmoplantar keratoderma.

Mutilating palmoplantar keratoderma (PPK) is a heterogeneous genetic disease that poses enormous challenges to clinical diagnosis and genetic counselling. Lanosterol synthase (LSS) gene encodes LSS involved in the biosynthesis pathway of cholesterol. Biallelic mutations in LSS were found to be related to diseases such as cataracts, hypotrichosis and palmoplantar keratoderma-congenital alopecia syndrome. The aim of this study was to investigate the contribution of the LSS mutation to mutilating PPK in a Chinese patient. The clinical and molecular characteristics of the patient were evaluated. A 38-year-old male patient with mutilating PPK was recruited in this study. We identified biallelic variants in the LSS gene (c.683C > T, p.Thr228Ile and c.779G > A, p.Arg260His). Immunoblotting revealed that the Arg260His mutant showed a significantly reduced expression level while Thr228Ile showed an expression level similar to that of the wild type. Thin layer chromatography revealed that mutant Thr228Ile retained partial enzymatic activity and mutant Arg260His did not show any catalytic activity. Our findings show the correlation between LSS mutations and mutilating PPK.

Variants in KLK11, affecting signal peptide cleavage of kallikrein-related peptidase 11, cause an autosomal-dominant cornification disorder.

BACKGROUND: Mendelian disorders of cornification (MeDOC) are a group of heterogeneous genodermatoses with different genetic bases. The pathogenesis of a substantial group of MeDOC remains to be elucidated. OBJECTIVES: To identify a new causative gene and the pathogenesis of a previously undescribed autosomal-dominant cornification disorder. METHODS: Whole-exome sequencing was performed in three families with the novel cornification disorder to identify the disease-causing variants. As the variants were located around the signal peptide (SP) cleavage site of a kallikrein-related peptidase, SP cleavage, subcellular localization and extracellular secretion of the variants were evaluated in eukaryotic overexpression systems by Western blotting or immunocytochemistry. Then the trypsin-like and chymotrypsin-like proteolytic activity of the peptidase and degradation of its catalytic substrate were assayed using the patients' stratum corneum (SC) samples. The morphology of the lamellar bodies and corneodesmosomes (CDs) in the patients' SC was ultrastructurally examined. A mouse model harbouring the equivalent variant was constructed and evaluated histologically. RESULTS: We identified two heterozygous variants affecting Gly50 in kallikrein-related peptidase (KLK)11 in a familial case and two sporadic cases with the new disorder, which is characterized by early-onset ichthyosiform erythroderma or erythrokeratoderma. KLK11 belongs to the family of kallikrein-related peptidases participating in skin desquamation by decomposing CDs, a process essential for shedding of the SC. In vitro experiments demonstrated that the variants perturbed the SP cleavage of KLK11, leading to subcellular mislocalization and impaired extracellular secretion of the KLK11 Gly50Glu variant. Both trypsin-like and chymotrypsin-like proteolytic activities were significantly decreased in the patients' SC samples. Reduced proteolysis of desmoglein 1 and delayed degeneration of CDs were detected in patients' SC, indicating delayed skin desquamation. Consistently, the patients showed a thickened, dense SC, indicating abnormal skin desquamation. Mice harbouring the homozygous c.131G>A (p.Gly44Glu) Klk11 variant, which is equivalent to KLK11 c.149G>A (p.Gly50Glu) in humans, exhibited hyperkeratosis and abnormal desquamation, partially recapitulating the phenotype. CONCLUSIONS: We provide evidence that variants at Gly50 affecting the SP cleavage of KLK11 cause a new autosomal-dominant cornification disorder with abnormal desquamation. Our findings highlight the essential role of KLKs in maintaining homeostasis of skin keratinization and desquamation.

Insights into As accumulation in soil-groundwater-wheat-hair system of suburban farmland: Distribution, transfer and potential health risk.

Health risks caused by arsenic (As) contamination in soils and its migration in environmental media have attracted much attention. In this study, suburban farmland of KF city in the ecotone of the Yellow River and Huaihe River Basin was taken as the research area. A series of samples including topsoils (246), profile soils (280), matched wheat grains (22 groups), groundwater (26) and human hair (355) were collected. As distribution and transfer in soil-groundwater-wheat-hair (SGWH) system in typical sites were explored, and comprehensive health risk of As in SGWH system was assessed based on US EPA model and local exposure parameters. The results showed that spatial distribution of total As presented a significant high value area, and higher As contents (in the range of 0.45-29.86 mg kg-1) and bioavailability was mainly in topsoils, which indicated that anthropogenic sources have led to As enrichment in studied area. Also, it was found that the As contents in 95 % of wheat grain samples were higher than that in the control soils, and 9 % groundwater samples were above national Class I standards. Especially, average As content in hair in typical sites was obviously influenced by that in soil, wheat and groundwater. Moreover, As migration curve along soil → wheat (groundwater) → hair appeared an irregular 'V' shape, and transfer coefficients of Tf water/soil (10-5), Tf wheat/soil (10-3), Tf hair/soil (10-2), Tf hair/wheat (101) and Tf hair/water (104) presented an obvious increasing trend of magnitude, implying that human body has a higher As enrichment risk. Furthermore, comprehensive health risks for children and adults in typical sites were significant, while wheat is the main risk medium. In general, arsenic accumulation in human hair is good consistent with EPA health risk model, and their combination can better evaluate environmental exposure risk of As.

Alterations of spontaneous brain activity in type 2 diabetes mellitus without mild cognitive impairment: a resting-state functional magnetic resonance study.

Background: Type 2 diabetes mellitus (T2DM) has been demonstrated an increased risk factor of cognitive impairment or even dementia. Kinds of resting-state functional magnetic resonance imaging indices have been proposed and used to investigate the brain mechanism underlying diabetic cognitive impairment. This study aimed to explore the early changes in spontaneous neural activity among T2DM patients without cognitive impairment by means of multiple rs-fMRI indices. Methods: T2DM patients without cognitive impairment and age-, sex-, and education matched control subjects were included in this study. Three rs-fMRI indices, namely amplitude of low-frequency fluctuation (ALFF), regional homogeneity (ReHo) and voxel-mirrored homotopic connectivity (VMHC) were computed after image pre-processing. The Montreal Cognitive Assessment (MoCA) was performed to distinguish normal cognition. Brain volume was also evaluated. Correlation analyses were conducted to explore any relationship among rs-fMRI indices and clinical characteristics. Results: The T2DM patients were detected significantly decreased neural activity in right angular and left prefrontal gyrus including middle and superior frontal gyrus. Increased activities were also observed in left caudate and the supplementary motor area. No correlation between rs-fMRI indices and clinical characteristics was survived after multiple comparison correction. But we observed a significant, but decreased correlation between ALFF and ReHo values in the reported brain areas. Conclusion: The combination of ALFF, ReHo and VMHC analyses demonstrated abnormal spontaneous neural activity in brain regions which were reported in T2DM patients without cognitive impairment. These results may enhance our understanding of the diabetic brain changes at the early stage.

A New Sight of Influencing Effects of Major Factors on Cd Transfer from Soil to Wheat (Triticum aestivum L.): Based on Threshold Regression Model.

Due to the high toxicity and potential health risk of cadmium (Cd), the influencing effects of major factors (like pH, OM, and clay, etc.) on Cd bioaccumulation and transfer from soil to crop grains are highly concerned. Multiple linear regression models were usually applied in previous literature, but these linear models could not reflect the threshold effects of major factors on Cd transfer under different soil environmental conditions. Soil pH and other factors on Cd transfer in a soil-plant system might pose different or even contrary effects under different soil Cd exposure levels. For this purpose, we try to apply a threshold regression model to analyze the effects of key soil parameters on Cd bioaccumulation and transfer from soil to wheat. The results showed that under different soil pH or Cd levels, several factors, including soil pH, organic matter, exchangeable Cd, clay, P, Zn, and Ca showed obvious threshold effects, and caused different or even contrary impacts on Cd bioaccumulation in wheat grains. Notably, the increase of soil pH inhibited Cd accumulation when pH > 7.98, but had a promotional effect when pH ≤ 7.98. Thus, threshold regression analysis could provide a new insight that can lead to a more integrated understanding of the relevant factors on Cd accumulation and transfer from soil to wheat. In addition, it might give us a new thought on setting regulatory limits on Cd contents in wheat grains, or the inhibitory factors of Cd transfer.

Resistivity Response of Thermally Treated Granite During the Compression Test.

To determine the stress and damage state of rock mass is important for many geotechnical engineering. To study the feasibility of resistivity measurement in characterizing the damage and stress state, the resistivity measurement, uniaxial compression test, and incremental loading-unloading compression test were carried out on granite samples with different porosities (induced by different treatment temperatures). Results show that the resistivity is very sensitive to thermal damage and mechanical damage during compression. The evolution of resistivity can not only quantify thermal damage but also clearly indicate the critical stress (crack closure stress and crack damage stress) and damage stage during compression. In addition, the resistivity evolution was quite different in the pore closure stage, elastic deformation stage, and unstable cracking stage during the loading-unloading process, which is useful in field stress and damage state identification for field monitoring. The conductive mechanism variation during compression was discussed using the Archie equation considering crack volume strain evolution during the mechanical damage process. Overall, the resistivity measurement holds great potential in geotechnical engineering for field monitoring.

Biallelic Variants in Lanosterol Synthase (LSS) Cause Palmoplantar Keratoderma-Congenital Alopecia Syndrome Type 2.

Palmoplantar keratoderma-congenital alopecia syndrome type 2 is an autosomal recessive disorder with an unknown genetic basis. In this study, we identified biallelic variants in the LSS gene in two unrelated palmoplantar keratoderma-congenital alopecia syndrome type 2 cases (c.3G>A, p.Met1? and c.1025T>G, p.Ile342Ser in patient 1; c.1522G>T, p.Gly508Trp and c.428+42T>A in patient 2) presenting with additional clinical features, including early-onset cataracts, pseudoainhum, and agenesis of the corpus callosum. LSS encodes lanosterol synthase (LSS), which functions in the cholesterol biosynthesis pathway by converting (S)-2,3-oxidosqualene to lanosterol. The c.3G>A variant resulted in an alternative translation initiation at residue Met81, producing an N-terminal truncated protein (LSS-ΔN80), as shown by immunoblotting. The c.428+42T>A variant introduced a potential splicing site, leading to a premature stop codon. Ex vivo studies revealed downregulation of LSS in both patients. Remarkably decreased lanosterol levels were found in vitro in three LSS variants, LSS-ΔN80, p.Ile342Ser, and p.Gly508Trp, suggesting a loss of enzymatic activity. Transmission electron microscopy and immunofluorescence showed abnormal cornified envelope formation in the stratum corneum of the patients. Taken together, our findings indicate LSS as a causative gene for palmoplantar keratoderma-congenital alopecia syndrome type 2, which emphasizes the importance of the cholesterol synthesis pathway in human skin cornification.

Non-contrast CT-based radiomic signature of pericoronary adipose tissue for screening non-calcified plaque.

Objective.To develop two combined clinical-radiomics models of pericoronary adipose tissue (PCAT) for the presence and characterization of non-calcified plaques on non-contrast CT scan.Approach.Altogether, 431 patients undergoing Coronary Computed Tomography Angiography from March 2019 to June 2021 who had complete data were enrolled, including 173 patients with non-calcified plaques of the right coronary artery(RCA) and 258 with no abnormality. PCAT was segmented around the proximal RCA on non-contrast CT scan (calcium score acquisition). Two best models were established by screening features and classifiers respectively using FeAture Explorer software. Model 1 distinguished normal coronary arteries from those with non-calcified plaques, and model 2 distinguished vulnerable plaques in non-calcified plaques. Performance was assessed by the area under the receiver operating characteristic curve (AUC-ROC).Main results.4 and 9 features were selected for the establishment of the radiomics model respectively through Model 1 and 2. In the test group, the AUC values, sensitivity, specificity and accuracy were 0.833%, 78.3%, 80.8%, 76.6% and 0.7467%, 75.0%, 77.8%, 73.5%, respectively. The combined model including radiomics features and independent clinical factors yielded an AUC, sensitivity, specificity and accuracy of 0.896%, 81.4%, 86.5%, 77.9% for model 1 and 0.752%, 75.0%, 77.8%, 73.5% for model 2.Significance.The combined clinical-radiomics models based on non-contrast CT images of PCAT had good diagnostic efficacy for non-calcified and vulnerable plaques.

Combination of UNMIX, PMF model and Pb-Zn-Cu isotopic compositions for quantitative source apportionment of heavy metals in suburban agricultural soils.

Quantitative identification of heavy metals (HM) sources in soils is key to prevention and control of heavy metal pollution. In this study, UNMIX, PMF (Positive matrix factorization) model and Pb-Zn-Cu isotopic compositions were combined to quantitatively identify heavy metal sources in a suburban agricultural area of Kaifeng, China. Using multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS) and ICP-MS, we measured Pb, Zn and Cu stable isotopic compositions, HM concentrations and HM chemical fractions in studied soils, as well as potential sources around the highly polluted site, including total suspended particle, compound fertilizer, irrigated river water and sediments. The results showed that total contents and chemical fractions of heavy metals, as well as Pb-Zn-Cu isotopic compositions presented great variation in different sites, which implied that heavy metal accumulation was obviously affected by local anthropogenic pollution source. UNMIX and PMF presented good agreement on source apportionment that industrial and agricultural activities (61.74% and 60.75% for UNMIX and PMF, respectively) were the major contributors to heavy metal accumulation in the study area. Especially, sewage irrigation and atmosphere deposition accounted for a large proportion (28.14% and 41.03% for UNMIX and PMF, respectively). Moreover, isotopic compositions of Pb, Zn and Cu in highly polluted soils and environment media gave further confirmation that sewage irrigation and atmosphere deposition were primary anthropogenic source. Therefore, combination of UNMIX, PMF model and Pb-Zn-Cu isotopic compositions showed good coordination in quantitative and specific source identification of heavy metals in agricultural soils.

Somatic frameshift mutation in PIK3CA causes CLOVES syndrome by provoking PI3K/AKT/mTOR pathway.

BACKGROUND: CLOVES syndrome (OMIM# 612918) is a rare overgrowth disorder resulted from mosaic gain-of-function mutations in the PIK3CA gene. All the reported CLOVES-associated PIK3CA mutations are missense mutations affecting certain residues. We aim to investigate underlying mutation and its pathogenicity in a patient with CLOVES syndrome and to evaluate the inhibitory effects of the PI3K/AKT/mTOR pathway inhibitors. RESULTS: We performed whole-exome sequencing (WES) and Sanger sequencing to detect underlying somatic mutations in the skin lesion of the patient. Quantitative real-time PCR (qRT-PCR) was employed to evaluate the mRNA abundance of PIK3CA in the patient's skin lesion. AKT phosphorylation level assessed by immunoblotting of lysates from transiently transfected cells was performed to evaluate the PIK3CA mutations and inhibitory effects of PI3K/AKT/mTOR pathway inhibitors. A somatic frameshift mutation c.3206_3207insG (p.X1069Trpfs*4) in PIK3CA was identified in the genomic DNA extracted from the vascular malformation sample of the patient. This mutation affects the canonical stop codon of PIK3CA (NM_006218.4) and is predicted to produce a prolonged protein with four additional residues. qRT-PCR demonstrated that the mRNA expression levels of the patient's affected skin tissue were comparable compared to the normal control. In vitro studies revealed that p.X1069Trpfs*4 mutant exhibited increased AKT phosphorylation significantly to that of the wildtype, which could be inhibited by PI3K/AKT/mTOR pathway inhibitors. CONCLUSIONS: We have identified the first frameshift mutation in PIK3CA that causes CLOVES syndrome, which was confirmed to overactive PI3K/AKT/mTOR pathway by transient transfection assays. We also provided more evidence of ARQ092 to be a potential therapeutic option for PROS in vitro.

Effect of Methylene Blue on White Matter Injury after Ischemic Stroke.

Methylene blue, the FDA-grandfathered drug was proved to be neuroprotective in ischemic stroke in rat. However, the mechanism of the protective effect was unknown. In this study, we used different animal models to investigate the effect of MB administration given within and beyond the therapeutic time window on behavioral deficits and infarct volume and related mechanism about the white matter protection. Middle cerebral artery occlusion and reperfusion (MCAO) and photothrombotic middle cerebral artery occlusion (PT-MCAO) models were used. Behavioral deficits and infarct volume were measured by foot fault test, Garcia neurological score, and TTC staining. Black gold staining and western blot were used to evaluate the brain white matter injury. We found that intraperitoneal administration of MB immediately or 24 h after the MCAO or PT-MCAO surgery reduced infarct volume, improved the neurological deficits, and reduced the white matter injury via myelin basic protein (BMP) protection. These findings suggested that MB relieved the white matter injury besides neuronal protection and has potential therapeutic effects on ischemic stroke.

Difference Between Medical and Nonmedical Students on Knowledge, Practice, and Attitude Towards the Human Papillomavirus Vaccine in China: a Cross-Sectional Study.

HPV vaccine can prevent HPV infection effectively. The college student's vaccination status is unclear in mainland China. We assessed the knowledge, practice, and attitude towards HPV vaccine and compared the differences between medical and nonmedical students. It was a cross-sectional study using self-administered anonymous questionnaires. Nine-hundred sixty full-time college students were recruited randomly at Peking University in China. The medical students had higher level of knowledge of HPV and its vaccine than the nonmedical students (p < 0.001). The vaccinated female students were 9.0%. The high-grade clinical students had a higher uptake rate than the nonmedical students (19.5 vs 8.6%, p < 0.05). Awareness of HPV (p < 0.01), awareness of the vaccine (p < 0.001), and vaccinated family members or friends (p < 0.001) were related to the nonmedical students' vaccination. Vaccinated family members or friends were significant predictor for students' vaccination status (p < 0.001). Medical students knew more about HPV and its vaccine than nonmedical students. Female students' vaccinated rate was low, and the high-grade clinical students had a higher uptake rate than the nonmedical students.

Differential Diagnostic Models Between Vasovagal Syncope and Psychogenic Pseudosyncope in Children.

Objective: We aimed to establish useful models for the clinical differential diagnosis between vasovagal syncope (VVS) and psychogenic pseudosyncope (PPS). Methods: This bicentric study included 176 patients (150 VVS and 26 PPS cases) for model development. Based on the results of univariate and multivariate analyses, a logistic regression model and a scoring model were established and their abilities to differentiate VVS from PPS were tested. Another 78 patients (53 VVS and 25 PPS) were used for external validation. Results: In the logistic regression model, the outcome indicated that the QT-dispersion (QTd) (P < 0.001), syncope duration (P < 0.001), and upright posture (P < 0.001) acted as independent factors for the differentiation of VVS from PPS, which generated an area under the curve (AUC) of 0.892. A cutoff value of 0.234 yielded a sensitivity and specificity of 89.3 and 80.8%, respectively, for the differentiation between VVS and PPS in the logistic regression model. In the scoring model which consists of three variables, a cutoff score of three points yielded a sensitivity and specificity of 91.3 and 76.9%, respectively, with an AUC of 0.909. The external validation test indicated that the negative and positive predictive values of the scoring model were 78.8 and 91.7%, respectively, and the accuracy was 80.8%. Conclusion: The scoring model consisting of three variables is an easy-to-perform, inexpensive, and non-invasive measure for initial differential diagnosis between VVS and PPS.