Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Cardiac fibrosis is characterized by an elevated amount of extracellular matrix (ECM) within the heart. However, the persistence of cardiac fibrosis ultimately diminishes contractility and precipitates cardiac dysfunction. Circular RNAs (circRNAs) are emerging as important regulators of cardiac fibrosis. Here, we elucidate the functional role of a specific circular RNA CELF1 in cardiac fibrosis and delineate a novel feedback loop mechanism. Functionally, circ-CELF1 was involved in enhancing fibrosis-related markers' expression and promoting the proliferation of cardiac fibroblasts (CFs), thereby exacerbating cardiac fibrosis. Mechanistically, circ-CELF1 reduced the ubiquitination-degradation rate of BRPF3, leading to an elevation of BRPF3 protein levels. Additionally, BRPF3 acted as a modular scaffold for the recruitment of histone acetyltransferase KAT7 to facilitate the induction of H3K14 acetylation within the promoters of the Celf1 gene. Thus, the transcription of Celf1 was dramatically activated, thereby inhibiting the subsequent response of their downstream target gene Smad7 expression to promote cardiac fibrosis. Moreover, Celf1 further promoted Celf1 pre-mRNA transcription and back-splicing, thereby establishing a feedback loop for circ-CELF1 production. Consequently, a novel feedback loop involving CELF1/circ-CELF1/BRPF3/KAT7 was established, suggesting that circ-CELF1 may serve as a potential novel therapeutic target for cardiac fibrosis.

Objective Accurate speed perception is crucial for tasks such as man-controlled rendezvous and docking,and teleoperation of space manipulator.Therefore,it is necessary to conduct in-orbit experiments to explore the influence of long-term spaceflight on human speed perception characteristics.Methods The Time-to-Collision(TTC)paradigm was selected to develop experimental software,using a tablet computer for stimulus presentation.Human speed perception characteristics were evaluated based on the subjects'keystroke response data on the keyboard.Through ground-based experiments,the usability and reliability of the paradigm were explored,and the gravity internal model effect was quantitatively analyzed.Through in-orbit experiments on space station tasks,the influence of long-term spaceflight on human speed perception characteristics was further investigated.Results Under the 1G environment on the ground,the TTC paradigm has a high test-retest reliability(r>0.8),and indicators such as average deviation rate and absolute value of average deviation rate show no practice effect.In addition,ground experiments found that compared to vertical upward movement,vertical downward movement is estimated to be faster(i.e.,keystroke time is advanced),showing the existence of the gravity internal model effect.In the microgravity environment of spaceflight,there are no significant differences in average deviation rate and absolute value of average deviation rate among three stages(pre-flight,in-flight,post-flight)and seven tests,indicating that no obvious changes in astronauts'speed perception ability were found at the existing test time points and paradigms.However,the gravity internal model effect(difference between vertical downward and vertical upward)showed a trend of fading in the early stage of astronauts entering orbit.Conclusion Based on the computer screen TTC estimation paradigm,no significant changes in human speed perception ability were found during long-term spaceflight,but microgravity may weaken the human brain's gravity internal model.

Objective:To develop a deep learning-based segmentation model for anterior segment ultrasound biomicroscopy (UBM) images to automatically segment the anterior segment tissues of patients with primary angle-closure glaucoma (PACG).Methods:A single-center retrospective case series was conducted.A small-scale dataset comprised 468 UBM images of the anterior chamber angle closure from 156 patients with PACG who underwent the UBM examination at Tianjin Medical University Eye Hospital between July 12, 2022, and February 20, 2023.The UBM images were randomly split into a training dataset of 228 images and a testing dataset of 152 images using a random seed method in a ratio of 6∶4.The models were trained using the PSPNet model with MobileNet V2 and ResNet50 as backbones, the DeepLab v3+ model with MobileNet V2 and Xception as backbones, and the SegFormer model with MiT-B0 and MiT-B2 as backbones.The testing dataset was used for result prediction and to achieve segmentation of four regions: the cornea and sclera, iris, ciliary body, and anterior lens surface.To evaluate the performance of the models in segmenting the anterior segment structures, multiple metrics were assessed, including the mean intersection over union (mIoU), Dice coefficient, precision, recall, false negative rate, and specificity.A comparative analysis of the test results across the different models was subsequently performed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Tianjin Medical University Eye Hospital (No.2023KY-05).Results:The two models with the best segmentation performance were PSPNet and DeepLab v3+ .The PSPNet model with ResNet50 as the backbone achieved the mIoU of 85.11%, Dice coefficient of 91.38%, precision of 91.83%, recall of 90.94%, false negative rate of 9.06%, and specificity of 98.89%.The DeepLab v3+ model with MobileNet V2 as the backbone achieved an mIoU of 85.84%, Dice coefficient of 92.01%, precision of 92.67%, recall of 91.36%, false negative rate of 8.64%, and specificity of 98.90%.Among the five key metrics, mIoU, Dice coefficient, recall, false negative rate, and specificity, DeepLab v3+ exhibited the best segmentation performance.In addition, the DeepLab v3+ model with Xception as the backbone had the highest precision among all models, reaching 92.77%.Conclusions:The deep learning-based DeepLab v3+ model achieves precise segmentation of anterior segment tissue structures in PACG anterior segment UBM image segmentation, providing auxiliary support for clinical diagnosis.

Objective:To develop a deep learning-based segmentation model for anterior segment ultrasound biomicroscopy (UBM) images to automatically segment the anterior segment tissues of patients with primary angle-closure glaucoma (PACG).Methods:A single-center retrospective case series was conducted.A small-scale dataset comprised 468 UBM images of the anterior chamber angle closure from 156 patients with PACG who underwent the UBM examination at Tianjin Medical University Eye Hospital between July 12, 2022, and February 20, 2023.The UBM images were randomly split into a training dataset of 228 images and a testing dataset of 152 images using a random seed method in a ratio of 6∶4.The models were trained using the PSPNet model with MobileNet V2 and ResNet50 as backbones, the DeepLab v3+ model with MobileNet V2 and Xception as backbones, and the SegFormer model with MiT-B0 and MiT-B2 as backbones.The testing dataset was used for result prediction and to achieve segmentation of four regions: the cornea and sclera, iris, ciliary body, and anterior lens surface.To evaluate the performance of the models in segmenting the anterior segment structures, multiple metrics were assessed, including the mean intersection over union (mIoU), Dice coefficient, precision, recall, false negative rate, and specificity.A comparative analysis of the test results across the different models was subsequently performed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Tianjin Medical University Eye Hospital (No.2023KY-05).Results:The two models with the best segmentation performance were PSPNet and DeepLab v3+ .The PSPNet model with ResNet50 as the backbone achieved the mIoU of 85.11%, Dice coefficient of 91.38%, precision of 91.83%, recall of 90.94%, false negative rate of 9.06%, and specificity of 98.89%.The DeepLab v3+ model with MobileNet V2 as the backbone achieved an mIoU of 85.84%, Dice coefficient of 92.01%, precision of 92.67%, recall of 91.36%, false negative rate of 8.64%, and specificity of 98.90%.Among the five key metrics, mIoU, Dice coefficient, recall, false negative rate, and specificity, DeepLab v3+ exhibited the best segmentation performance.In addition, the DeepLab v3+ model with Xception as the backbone had the highest precision among all models, reaching 92.77%.Conclusions:The deep learning-based DeepLab v3+ model achieves precise segmentation of anterior segment tissue structures in PACG anterior segment UBM image segmentation, providing auxiliary support for clinical diagnosis.

Background: Twenty-four-hour urinary free cortisol (UFC) measurement is the initial diagnostic test for Cushing’s syndrome (CS). We compared UFC determination by both direct and extraction immunoassays using Abbott Architect, Siemens Atellica Solution, and Beckman DxI800 with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, we evaluated the value of 24-hr UFC measured by six methods for diagnosing CS. Methods: Residual 24-hr urine samples of 94 CS and 246 non-CS patients were collected.A laboratory-developed LC-MS/MS method was used as reference. UFC was measured by direct assays (D) using Abbott, Siemens, and Beckman platforms and by extraction assays (E) using Siemens and Beckman platforms. Method was compared using Passing–Bablok regression and Bland–Altman plot analyses. Cut-off values for the six assays and corresponding sensitivities and specificities were calculated by ROC analysis. Results: Abbott-D, Beckman-E, Siemens-E, and Siemens-D showed strong correlations with LC-MS/MS (Spearman coefficient r = 0.965, 0.922, 0.922, and 0.897, respectively), while Beckman-D showed weaker correlation (r = 0.755). All immunoassays showed proportionally positive bias. The areas under the curve were 0.975 for Abbott-D, 0.972 for LCMS/MS, 0.966 for Siemens-E, 0.948 for Siemens-D, 0.955 for Beckman-E, and 0.877 for Beckman-D. The cut-off values varied significantly (154.8–1,321.5 nmol/24 hrs). Assay sensitivity and specificity ranged from 76.1% to 93.2% and from 93.0% to 97.1%, respectively. Conclusions Commercially available immunoassays for measuring UFC show different levels of analytical consistency compared to LC-MS/MS. Abbott-D, Siemens-E, and Beckman-E have high diagnostic accuracy for CS.