A rare ACAN non-canonical splicing-site intron variant results in familial short stature.

OBJECTIVE: ACAN gene variants, prevalent monogenic defects linked to short stature, are characterized by impaired cartilage generation in growth plates. We aimed to unravel the genetic basis of short stature in a specific pedigree by investigating the role of a novel non-canonical splicing-site variant, c.630-13G > A, within the ACAN gene. METHOD: Sanger sequencing was used for pedigree verification, and the effects of this variant on mRNA splicing were analyzed through minigene assay. RESULTS: The study revealed that this variant led to the creation of a previously unreported splice site in the fourth intron, resulting in the incorporation of an 11 bp sequence from the intron into the final transcript. This alteration led to a frameshift and formation of a premature termination codon, impacting the structure of the aggrecan protein. CONCLUSIONS: We document the pathogenicity of an ACAN non-canonical splicing-site variant, emphasizing the significance of considering intronic variants during genetic testing.

Effectiveness of health management team program to enhance prevention of mother-to-child transmission of hepatitis B virus in Ningxia, China.

BACKGROUND: Hepatitis B mother-to-child transmission interruption (PMTCT) poses a formidable challenge in underdeveloped regions of China. This study aims to evaluate the effectiveness of PMTCT and the health management team (HMT) model in Ningxia, China, as well as the risk factors for adverse outcomes. METHODS: The PMTCT + HMT model was established, and 360 pregnant women diagnosed with HBV infection in 2020-2022 were selected and divided into the control and the study groups based on different intervention modes. HBV serum markers and HBV DNA levels were assessed, the indicators of compliance behaviors and adverse outcomes were compared, and the factors influencing adverse outcomes were analyzed. RESULTS: The majority of subjects were residents of the local city, married, with secondary school or higher education, and employees of public sectors. The proportion of ethnic minorities was 40.8% and 34.2% in the control group and study group. HBeAg positivity was 23.3% and 26.3%, and the proportion with HBV DNA levels ≥ 2 × 105 IU/mL was 9.2% and 7.1%. Compared with the control group (PMTCT alone), the PMTCT + HMT model led to improved maternal knowledge (17.5% vs. 57.1%), voluntary counseling (34.2% vs. 63.3%), and testing (37.5% vs. 70.4%). The incidence of adverse pregnancy outcomes ((including miscarriage, preterm birth) decreased significantly (17.5% vs. 6.2%), as did adverse neonatal outcomes (low birth weight and congenital HBV) (26.9% vs. 10.5%). Adverse outcomes were associated with low educational attainment, non-locals, unmarried status, and ethnic minority identity. Additionally, HBeAg positivity and HBV DNA levels ≥ 2 × 105 IU/mL were risk factors for adverse outcomes. CONCLUSIONS: The PMTCT + HMT model demonstrates significant effectiveness in preventing mother-to-child transmission of hepatitis B in Ningxia. The unique demographic structure of Ningxia region is closely linked to poor outcomes, emphasizing the importance of monitoring HBeAg status and HBV DNA viral load level.

TIFA contributes to periodontitis in diabetic mice via activating the NF­κB signaling pathway.

Diabetic periodontitis (DP) refers to destruction of periodontal tissue and absorption of bone tissue in diabetic patients. Tumor necrosis factor receptor­associated factor (TRAF)­interacting protein with forkhead­associated domain (TIFA) as a crucial regulator of inflammation activates the NF­κB signaling pathway to regulate cell biological behavior. However, the function and mechanism of TIFA on DP suffer from a lack of research. In the present study, TIFA was upregulated in the periodontal tissue of a DP mouse model. In addition, the expression of TIFA in RAW264.7 cells was induced by high glucose (HG) culture and increased by lipopolysaccharide (LPS) from Porphyromonas gingivalis treatment in a time­dependent manner. Knockdown of TIFA significantly reduced the levels of inflammatory cytokines, including TNF­α, IL­6, IL­1ß and monocyte chemoattractant protein­1, in HG and LPS­induced RAW264.7 cells. The nuclear translocation of NF­κB p65 was induced by HG and LPS and was clearly suppressed by absence of TIFA. The expression of downstream factors Nod­like receptor family pyrin domain­containing 3 and apoptosis­associated speck­like protein was inhibited by silencing TIFA. Moreover, TIFA was increased by receptor activator of NF­κB (RANK) ligand (RANKL) in a concentration dependent manner. The expression of cathepsin K, MMP9 and nuclear factor of activated T cells cytoplasmic 1 was downregulated by depletion of TIFA. RANKL­induced osteoclast differentiation was inhibited by silencing of TIFA. Meanwhile, the decrease of TIFA blocked activation of the NF­κB pathway in RANKL­treated RAW264.7 cells. In conclusion, TIFA as a promoter regulates the inflammation and osteoclast differentiation via activating the NF­κB signaling pathway.

1,25-Dihydroxyvitamin D3: A Positive Factor for the Osteogenic Differentiation of hPDLSCs and for the Tissue Regenerative Activity of Cell Sheets.

This study aims to investigate the effects of 1,25-dihydroxyvitamin D3 (1,25(OH)2VitD3) on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and the activity of hPDLSC sheets and the differences in the tissue regeneration activity of hPDLSC sheets on tooth root fragment treated by different methods. Healthy caries-free premolars were collected. The hPDLSCs were obtained by enzymatic digestion. Surface markers of stem cells were analyzed by flow cytometry and the multidirectional differentiation ability of hPDLSCs was detected. During the osteogenic differentiation of hPDLSCs, 1,25(OH)2VitD3 was added and the effect of 1,25(OH)2VitD3 on osteogenic differentiation of hPDLSCs was assessed using Western blotting, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay, cell staining, and immunofluorescence. After hPDLSC sheets were prepared, histology and immunofluorescence analysis of the effect of 1,25(OH)2VitD3 on sheet activity were performed. In addition, root fragments were prepared and treated with scaling, 24% EDTA (ethylenediamide tetraacetic acid), and Er,Cr:YSGG lasers, respectively, and the tissue regeneration activity of hPDLSC sheets on different root fragments were observed. 1,25(OH)2VitD3 promoted the high gene and protein expressions of osteogenic markers ALP (alkaline phosphatase), Runx2, and OPN (osteopontin antibody) in hPDLSCs, along with enhanced ALP activity and staining, alizarin red staining, and immunofluorescence staining, indicating that the osteogenic differentiation ability of hPDLSCs was improved. Extracellular matrix secretion was increased in hPDLSC sheets, along with the positive expressions of the protein markers fibronectin and collagen I, suggesting that 1,25(OH)2VitD3 could enhance these effects. In addition, the root fragments treated by Er,Cr:YSGG laser were more suitable for the attachment and regeneration of hPDLSC sheets, demonstrating that 1,25(OH)2VitD3 could improve the tissue regeneration performance of these sheets. 1,25(OH)2VitD3 can promote osteogenic differentiation of hPDLSCs and thus plays an active role in hPDLSC sheet formation and tissue regeneration. In addition, the Er,Cr:YSGG laser can be used as the recommended treatment method for the root surface regenerated by hPDLSCs.

Roles of human periodontal ligament stem cells in osteogenesis and inflammation in periodontitis models: Effect of 1α,25-dihydroxyvitamin D3.

Periodontitis is a chronic inflammatory disease caused by Porphyromonas gingivalis and other bacteria, and human periodontal ligament stem cells (hPDLSCs) are a promising candidate for the treatment of periodontal supporting tissue defects. This study aimed to investigate the effect of 1α,25-dihydroxyvitamin D3 [1,25(OH)2VitD3] on osteogenic differentiation of hPDLSCs in an in vitro periodontitis model and whether it can improve inflammatory status. hPDLSCs were in vitro isolated and identified. After treatment with 1,25(OH)2VitD3 and ultrapure pure Porphyromonas gingivalis lipopolysaccharide (LPS-G), the viability of hPDLSCs was detected using Cell Counting Kit-8, the expressions of osteogenic markers and inflammatory genes using Western blotting and quantitative reverse transcription PCR (qRT-PCR), the levels of inflammatory factors in cells using enzyme linked immunosorbent assay (ELISA), and the fluorescence signal intensity of osteoblastic markers and inflammatory genes in cells using immunofluorescence assay. It was found that 1,25(OH)2VitD3 reversed the inhibition of hPDLSCs proliferation by LPS-G; LPS-G exhibited inhibitory effect on ALP, Runx2, and OPN expressions, and such inhibitory effect was significantly weakened when co-acting with 1,25(OH)2VitD3. Meanwhile, LPS-G upregulated the expressions of inflammatory genes IL-1ß and Casp1, whereas 1,25(OH)2VitD3 antagonized such an effect and improved the inflammatory status. In conclusion, 1,25(OH)2VitD3 can reverse the inhibitory effect of LPS-G on hPDLSCs proliferation and osteogenic differentiation and suppress LPS-G-induced upregulation of inflammatory gene expressions.

Nonalcoholic fatty pancreas disease: An emerging clinical challenge.

Nonalcoholic fatty pancreas disease (NAFPD) is an emerging disease that has gained an increasing amount of attention in recent years. It describes fat accumulation in the pancreas with insignificant alcohol consumption, but the pathogenesis is largely unknown. A wide range of terms have been used to describe the phenomenon of pancreatic fat accumulation, but NAFPD remains an under-recognized and non-independent disorder. Obesity, age, sex, race, and unhealthy lifestyle are established independent risk factors for NAFPD, which is strongly associated with metabolic syndrome, type 2 diabetes, pancreatitis, pancreatic fistula, pancreatic cancer, and nonalcoholic fatty liver disease. At present, imaging techniques are common diagnostic aids, but uniform criteria and consensus are lacking. Therapeutically, healthy diet, weight loss, and exercise are the mainstays to reduce pancreatic fat accumulation. It can be seen that there is a limited understanding of NAFPD at this stage and further exploration is needed. Previous studies have revealed that NAFPD may directly affect diagnosis and clinical decision-making. Therefore, exploring the pathophysiological mechanism and clinical associations of NAFPD is a major challenge for researchers and clinicians.

Comparative Analysis of the Temporomandibular Joints in Patients with Chronic Periodontitis Using Cone-Beam Computed Tomography (CBCT).

INTRODUCTION: Chronic periodontitis is a common disorder in adults causing periodontal destruction and loss of teeth. These clinical presentations may lead to temporomandibular joint disorders (TMDs). This study aimed to examine the anatomic structures of the temporomandibular joints (TMJs) using cone-beam computed tomography (CBCT) in patients with chronic periodontitis. METHODS: Fifty patients with chronic periodontitis were enrolled in the study. Based on the severity of chronic periodontitis, these patients were divided into the mild, moderate, and severe groups. CBCT images of TMJs were acquired and reconstructed. Several indices on the reconstructed CBCT images were collected and analyzed, such as the oblique joint space parallel to the long axis of the condyle, the long axis diameter of the condyle, the vertical angle of the condyle, the inclination of the articular eminence vertical to the long axis of condyle at the oblique and sagittal positions, the depth of the fossa, and the horizontal angle of the cross-sections. The measurements between right and left sides of each patient were compared. Statistical analysis (paired samples t test) was performed. RESULTS: The differences of the joint space vertical to the bilateral condyles were statistically significant (P < 0.05). Additionally, in the severe periodontitis group, the distances between the inner and outer poles of the condyles were statistically different (P < 0.05). CONCLUSION: In patients with chronic periodontitis, TMJ space vertical to the condyles and the distances between the outer and inner poles of the condyle may change over time. These two indices can potentially be used as indicators for diagnosis and further comparative analyses.

[Expression and significance of retinoid-related orphan receptor gamma t and 4-1BB/4-1BBL in oral lichen planus].

OBJECTIVE: To investigate the expression of the peripheral blood retinoid-related orphan receptor gamma t (RORgamma t) and 4-1BB/4-1BBL mRNA in oral lichen planus (OLP) patients. METHODS: The expression levels of peripheral blood' RORyt mRNA and 4-1BB/4-1BBL mRNA of 30 samples of OLP patients and 30 cases of healthy human were detected using real-time fluorescent quantitative-polymerase chain reaction (FQ-PCR) technology. RESULTS: In the OLP group, the expression level of RORgamma t, 4-1BB, 4-1BBL mRNA were higher than those in the control group (P < 0.05). After the conversion of the 2-delta delta CT, the expression levels of three kinds of mRNA in the OLP group were 3.087, 3.320 and 4.005 times of the control group respectively. In the OLP group, the expression levels of RORgamma t mRNA and 4-1BB mRNA had no obvious correlation (P > 0.05), the expression levels of 4-1BB and 4-1BBL mRNA had a positive correlation (r = 0.455 0, P < 0.05). CONCLUSION: The abnormal expression of RORyt, 4-1BB and 4-1BBL mRNA may play certain roles in the pathogenesis of OLP.