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.

BACKGROUND:Titanium alloys lack biological activity when used as orthopedic implants,which can lead to implant loosening and periprosthetic infection.Therefore,it is of great significance to study a titanium alloy surface modification method that combines osteogenic and anti-infection functions.OBJECTIVE:To study the physical and chemical properties of titanium alloy modified with copper and strontium binary doped calcium silicate composite coating,and to evaluate its bone-promoting and antibacterial potential.METHODS:Ball milling and granulation methods were used to prepare composite powder containing copper oxide(CuO),strontium oxide(SrO),and calcium silicate(CS).A copper-strontium binary doped calcium silicate composite coating was prepared on the surface of titanium alloy(Ti6Al4V)through atmospheric plasma spraying technology.The composite coating was characterized.The titanium alloy extract,calcium silicate coating modified titanium alloy extract,copper-doped calcium silicate composite coating modified titanium alloy extract,and copper-strontium binary doped calcium silicate composite coating modified titanium alloy extract were co-cultured with MC3T3-E1 cells to detect the biosafety and osteogenic properties of the materials.Staphylococcus aureus(or Escherichia coli)were co-cultured with titanium alloy,calcium silicate coating modified titanium alloy,copper-doped calcium silicate composite coating modified titanium alloy,and copper-strontium binary doped calcium silicate composite coating modified titanium alloy.The in vitro antibacterial properties of the materials were detected by scanning electron microscopy and plate counting method.RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that a large number of nanostructures existed on the rough surface of the copper-strontium binary doped calcium silicate composite coating.The composite coating was successfully sprayed on the surface of titanium alloy.The composite coating could slowly release Sr2+and Cu2+in vitro.The release concentration of Sr2+was greater than that of Cu2+.(2)CCK-8 assay and cell live/dead staining results showed that the copper-doped calcium silicate composite coating modified titanium alloy had certain cytotoxicity.The calcium silicate coating and the copper-strontium binary doped calcium silicate composite coating modified titanium alloy had good biocompatibility.Alkaline phosphatase and alizarin red staining results showed that compared with titanium alloy and calcium silicate coating modified titanium alloy,copper strontium binary doped calcium silicate composite coating modified titanium alloy showed better osteogenic properties.(3)The results of scanning electron microscopy,bacterial coating,and bacterial counting method showed that compared with titanium alloy and calcium silicate coating modified titanium alloy,copper-doped calcium silicate composite coating and copper strontium binary doped calcium silicate composite coating modified titanium alloy can effectively inhibit the growth of Staphylococcus aureus and Escherichia coli,showing antibacterial potential.(4)The results indicate that copper strontium binary doped calcium silicate composite coating modified titanium sheet has good biocompatibility,osteogenic and antibacterial properties.

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.

Objective:To calculate the reference intervals of immune globulinG (IgG), IgA, IgM, complement 3 (C3) and C4 for adults using Hoffmann and refineR methods, and to compare them with the reference interval reported by WS/T 645 of the People′s Republic of China, exploring the clinical application value of the indirect methods.Methods:Cross-sectional study. The physical examination data were collected at Peking Union Medical College Hospital from 2013 to 2023. Box-Cox was used to normalize the data distribution, and the Tukey method was used to remove outliers. Mann-Whitney test and Kruskal-Wallis test were used to compare the differences between gender and age, respectively. Quantile regression analysis was used to examine the effects of age and gender. Hoffmann and refineR methods were employed to calculate RIs of the five parameters.Results:There were 82 251, 82 483, 49 236, 20 027 and 19 942 test results for IgG, IgA, IgM, C3 and C4, respectively. Gender was significantly correlated with IgG, IgA, IgM, C3, and C4 levels, and age was significantly correlated with IgA, IgM, C3, and C4 levels. Reference intervals were calculated using both Hoffmann and refineR methods, yielding comparable results: IgG (7.58-16.85 g/L; 7.54-16.88 g/L), IgA (0.97-4.26 g/L; 0.97-4.34 g/L), C3 (0.73-1.43 g/L; 0.73-1.44 g/L), and C4 (0.11-0.32 g/L). Gender-specific intervals were established for IgM: women (0.44-2.57 g/L; 0.44-2.62 g/L) and men (0.33-1.91 g/L; 0.31-1.87 g/L). Gender and/or age-specific reference intervals were further calculated according to the bias ratio values. The reference intervals calculated by Hoffmann and refineR are in good agreement. The upper and lower limits of reference intervals for IgA and C3, the lower limits of IgG, the upper limits of IgM and C3 calculated by the indirect methods have good comparability with those reported by standards.Conclusions:The total and gender/age-specific reference intervals of IgG, IgA, IgM, C3 and C4 in our hospital population were established by Hoffmann and refineR methods, and the reference intervals established by the two indirect methods were comparable. The reference intervals calculated using these indirect methods were also comparable with those established using the direct method reported in current health standards.

Abastract:Objective To validate the performance of six different lipoprotein(a)[Lp(a)]immunoassay detection systems,compare the correlation and consistency of the measurement results of different detection systems,and explore their clinical application in the e-valuation of atherosclerotic cardiovascular disease(AsCVD).Methods A total of 150 AsCVD patients attending Peking Union Medi-cal College Hospital were retrospectively selected as the subjects of study group,and 50 individuals of physical examination during the same period were selected as healthy control group.Lp(a)levels were measured in 200 serum samples using six immunoassay detection systems,including two Lp(a)particle concentration assays in nmol/L(Roche and Mindray Ⅱ),and four Lp(a)mass concentration assays in mg/L(Mindray,MedicalSystem,BSBE,and Sekisui).All assays'precisions were evaluated.The results of each assay sys-tem were compared with the mean value of all the Lp(a)assays.Passing-Bablok regression analysis and Bland-Altman bias plots were used to assess the accuracy of assays,and the consistency between different systems was analyzed using the concordance correlation co-efficient(CCC).In addition,the consistencies of different assays in assessing AsCVD in clinical setting were compared using weighted Kappa statistical method,and the positive rates of Lp(a)particle concentration and mass concentration,as well as the overestimation and underestimation of mass concentration were also assessed for both the study group and the healthy control group.Results The pre-cision of the six Lp(a)assays ranged from 0.6％to 2.1％.Passing-Bablok regression analysis showed that the Spearman correlation co-efficients of the regression equations were all greater than 0.970,and the intercepts and slopes of the regression lines were-43.311 to 39.456 and 0.547 to 5.500,respectively.The Bland-Altman bias plots showed that the percent bias of the six assays compared to the mean value of Lp(a)determination was in the range of-25.939％to 40.205％.The results of the Lp(a)mass concentration detection system showed a positive deviation in Mindray and MedicalSystem,and a negative deviation in BSBE and Sekisui.Compared with the mean value of Lp(a),the results of consistency analysis showed that Roche and Mindray Ⅱ had excellent consistency(CCC:0.992 to 0.993).Mindray,MedicalSystem and BSBE had good consistency(CCC:0.950 to 0.986),and Sekisui showed moderate consistency(CCC:0.935).In descending order,the positivity rates of Lp(a)in the study groups were:MedicalSystem＞BSBE＞Mindray＞Roche=Mindray Ⅱ＞Sekisui.The overall concordances of Mindray,MedicalSystem,BSBE and Sekisui compared to Lp(a)particle concentration assay in different groups were 97.33％,93.33％,97.33％and 98.00％with Kappa values of 0.910,0.798,0.912 and 0.927,respec-tively.Conclusion The two assays for Lp(a)particle concentration have fine correlation and consistency,but there were significant differences between the four assays for Lp(a)mass concentration.Compared to the Lp(a)particle concentration assays,the four assay for Lp(a)mass concentration resulted in overestimation or underestimation of Lp(a)levels in the assessment of AsCVD.Accurate de-termination of Lp(a)concentration should be of great importance in accurately assessing the overall risk of AsCVD in patients.

BACKGROUND:Titanium alloys lack biological activity when used as orthopedic implants,which can lead to implant loosening and periprosthetic infection.Therefore,it is of great significance to study a titanium alloy surface modification method that combines osteogenic and anti-infection functions.OBJECTIVE:To study the physical and chemical properties of titanium alloy modified with copper and strontium binary doped calcium silicate composite coating,and to evaluate its bone-promoting and antibacterial potential.METHODS:Ball milling and granulation methods were used to prepare composite powder containing copper oxide(CuO),strontium oxide(SrO),and calcium silicate(CS).A copper-strontium binary doped calcium silicate composite coating was prepared on the surface of titanium alloy(Ti6Al4V)through atmospheric plasma spraying technology.The composite coating was characterized.The titanium alloy extract,calcium silicate coating modified titanium alloy extract,copper-doped calcium silicate composite coating modified titanium alloy extract,and copper-strontium binary doped calcium silicate composite coating modified titanium alloy extract were co-cultured with MC3T3-E1 cells to detect the biosafety and osteogenic properties of the materials.Staphylococcus aureus(or Escherichia coli)were co-cultured with titanium alloy,calcium silicate coating modified titanium alloy,copper-doped calcium silicate composite coating modified titanium alloy,and copper-strontium binary doped calcium silicate composite coating modified titanium alloy.The in vitro antibacterial properties of the materials were detected by scanning electron microscopy and plate counting method.RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that a large number of nanostructures existed on the rough surface of the copper-strontium binary doped calcium silicate composite coating.The composite coating was successfully sprayed on the surface of titanium alloy.The composite coating could slowly release Sr2+and Cu2+in vitro.The release concentration of Sr2+was greater than that of Cu2+.(2)CCK-8 assay and cell live/dead staining results showed that the copper-doped calcium silicate composite coating modified titanium alloy had certain cytotoxicity.The calcium silicate coating and the copper-strontium binary doped calcium silicate composite coating modified titanium alloy had good biocompatibility.Alkaline phosphatase and alizarin red staining results showed that compared with titanium alloy and calcium silicate coating modified titanium alloy,copper strontium binary doped calcium silicate composite coating modified titanium alloy showed better osteogenic properties.(3)The results of scanning electron microscopy,bacterial coating,and bacterial counting method showed that compared with titanium alloy and calcium silicate coating modified titanium alloy,copper-doped calcium silicate composite coating and copper strontium binary doped calcium silicate composite coating modified titanium alloy can effectively inhibit the growth of Staphylococcus aureus and Escherichia coli,showing antibacterial potential.(4)The results indicate that copper strontium binary doped calcium silicate composite coating modified titanium sheet has good biocompatibility,osteogenic and antibacterial properties.