OBJECTIVE: To explore the genetic etiology of a Chinese boy affected with Oculo-facio-cardio-dental syndrome (OFCD). METHODS: A child diagnosed with OFCD at West China Hospital of Sichuan University on September 21, 2024 was selected as the study subject. Clinical phenotype of the child was collected through ophthalmologic examination, cardiac ultrasonography, and X-ray imaging. Potential pathogenic variants were detected by trio-whole exome sequencing (Trio-WES). Candidate variant was validated with TA-cloning followed by Sanger sequencing. Mosaic variant was analyzed by ultra-deep sequencing (10,000-fold) and quantitative PCR. This study was approved by the Medical Ethics Committee of the West China Hospital of Sichuan University (Ethics No.: 2019-772 ). RESULTS: The proband had presented with congenital cataracts, mitosis, atrial and ventricular septal defects, dental abnormalities, and right radioulnar synostosis. His mother also exhibited congenital cataracts and dental anomalies, suggesting a diagnosis of OFCD. Trio-WES revealed an novel heterozygous 14-bp deletion (c.4724_4737del) in exon 12 of the BCOR gene in the proband. Deep sequencing identified a mosaic BCOR c.4724_4737del mutation in approximately 3.4% of peripheral leukocytes from his mother. Quantitative PCR analysis also confirmed the presence of this low-level mosaicism. The 14-bp deletion was predicted to cause a frame shift and premature termination (p.Met1575AsnfsTer6). Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic (PVS1+PM2+PP1). CONCLUSION Above findings have expanded the spectrum of BCOR mutations associated with OFCD, which highlighted the role of low-level mosaicism with maternal transmission and provided a basis for genetic counseling and reproductive guidance for the family.
Humans ; Male ; Repressor Proteins/genetics* ; Proto-Oncogene Proteins/genetics* ; Tooth Abnormalities/genetics* ; Eye Abnormalities/genetics* ; Microphthalmos/genetics* ; Child ; Sequence Deletion ; Female ; Cataract/congenital* ; Heart Septal Defects

Background Di(2-ethylhexyl) phthalate (DEHP) is the most prevalent environmental endocrine disruptor among phthalate acid esters (PAEs) worldwide. Previous studies have indicated that exposure to DEHP may disrupt glucose metabolism. Objective To investigate the impact of DEHP on glucose homeostasis in rats, focusing on oxidative stress-induced impairment of autophagy in islet β cells. Methods Forty male SD rats were randomly assigned to four groups, receiving DEHP doses of 0, 187, 375, and 750 mg·kg⁻¹ for 12 weeks. Oral glucose tolerance (OGTT) and insulin tolerance tests (ITT) were conducted 24 h after the final exposure. Pancreatic microstructural alterations were assessed using hematoxylin and eosin (HE) staining and transmission electron microscopy (TEM). Commercial ELISA kits were employed to quantify the levels of insulin, adenosine triphosphate (ATP), and adenosine monophosphate (AMP) in rat serum, as well as the protein expression level of activated caspase-3 in pancreatic tissue. Additionally, commercial microplate kits were utilized to measure the concentration of reduced glutathione (GSH) in serum, the activity of superoxide dismutase (SOD) using water-soluble tetrazolium salt-1, the content of malondialdehyde (MDA) by thiobarbituric acid method, and the level of reactive oxygen species (ROS) in pancreatic tissue by chemical fluorescence method. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure sequestosome1 (SQSTM1/p62), Beclin1, microtubule-associated protein 1 light chain 3 (LC3), and cysteinyl aspartate specific proteinase-8 (Caspase-8) mRNA levels. Western blot analysis was applied to detect the protein relative expression levels of p62, Beclin-1, LC3-I, LC3 II, AMPK, p-AMPK, mTOR, p-mTOR, ULK1, and Caspase-8. Results Compared to the 0 mg·kg⁻¹ DEHP group, the 750 mg·kg⁻¹ DEHP group exhibited a significant increase in fasting blood glucose levels at 2, 4, 6, and 12 weeks (P<0.05). The OGTT showed that, following high-glucose gavage, the 187 mg·kg⁻¹ DEHP group had elevated blood glucose at 30 min (P<0.05), the 375 mg·kg⁻¹ DEHP group showed increased glucose levels at 15, 30, and 180 min (P<0.05), and the 750 mg·kg⁻¹ DEHP group exhibited elevated levels at 15, 30, 60, and 180 min (P<0.05). The 375 and 750 mg·kg⁻¹ DEHP groups demonstrated significantly increased OGTT area under the curve (AUC) values (P<0.05). In contrast, ITT results indicated no significant differences in blood glucose levels or AUC among the DEHP exposure groups at all time points (P>0.05). Compared to the 0 mg·kg⁻¹ DEHP group, the 750 mg·kg⁻¹ DEHP group exhibited significantly higher HOMA-IR levels and markedly lower HOMA-ISI values (P<0.05). HE and TEM showed that in each DEHP exposure group, the number of islet cells decreased, the islet area reduced, and chromatin condensation occurred. The endocrine granules in the cytoplasm of islet β cells decreased, and there were varying degrees of widening of the nuclear membrane gap, flattening and expansion of the Golgi complex, and expansion of the endoplasmic reticulum. Ribosome separation was observed, and autophagosomes were visible. In the 375 and 750 mg·kg⁻¹ DEHP groups, the mitochondria were deformed to varying degrees, and some cristae structures disappeared, presenting vacuolization. Moreover, the chromatin condensation in the nuclei was more severe in the 750 mg·kg⁻¹ DEHP group. The serum SOD activity was significantly elevated in the 750 mg·kg⁻¹ DEHP group (P<0.05). Both the 375 mg·kg⁻¹ and 750 mg·kg⁻¹ DEHP groups exhibited a significant increase in the relative ROS content in pancreatic tissue (P<0.05). In DEHP-treated groups, the MDA content increased (P<0.05), while the GSH content decreased (P<0.05). Additionally, in the 750 mg·kg⁻¹ DEHP group, the AMP/ATP ratio in serum was significantly raised (P<0.05), and the expression of cleaved Caspase-3 protein in pancreatic tissue was also significantly increased (P<0.05). The relative mRNA levels of p62, Beclin-1, LC3, and Caspase-8 in the pancreatic tissue of rats exposed to DEHP were significantly elevated (P<0.05). The relative expression levels of p-AMPK/AMPK, p-ULK1/ULK1, and Beclin-1 proteins in the DEHP-treated groups were significantly increased (P<0.05). In the 375 mg·kg⁻¹ and 750 mg·kg⁻¹ DEHP treatment groups, the relative expression levels of p62, LC3 II/LC1, and Caspase-8 proteins were significantly increased (P<0.05), while the relative expression level of p-mTOR/mTOR was significantly decreased (P<0.05). Conclusion DEHP can disrupt glucose homeostasis by inducing oxidative stress, which subsequently activates autophagy via the ROS/AMPK/ULK1 pathway, impairing autophagic flux and promoting apoptosis of islet β cells, ultimately decreasing their function and number.

Objective:To explore the relationship between the differential genes derived from transcriptome mRNA sequencing and prognosis among heart failure patients with mildly reduced ejection fraction (HFmrEF) and preserved ejection fraction (HFpEF).Methods:This was a case-control study. Ten patients with HFmrEF and 10 patients with HFpEF treated at TEDA International Cardiovascular Disease Hospital from November 2021 to January 2022 were selected and differentially expressed genes were screened by transcriptome mRNA sequencing. Ten healthy people served as control group. In addition, 50 patients with HFmrEF, 62 patients with HFpEF, who were treated at TEDA International Cardiovascular Disease Hospital at the same period, were selected as validation groups, 57 healthy people served as control validation group. Real-time quantitative PCR (RT-qPCR) was used to detect the expression of differential genes in each group. Receiver operating characteristic (ROC) curves and the area under the curve (AUC) were used to assess the differential diagnosis and prognostic value of differential genes in these patients. Patients were followed up regularly to document adverse events within 1 year after discharge including cardiac death and readmission for heart failure. Survival analysis was performed using Kaplan-Meier curves and tested by log rank test. Cox regression analysis was used to explore whether differential mRNA was risk factors for poor prognosis in HFmrEF and HFpEF patients.Results:A total of four genes were differentially expressed (three upregulated and one downregulated gene) between the HFmrEF group and HFpEF group (adjust P<0.05). SLC12A1, C15orf48 and SPP1 were associated with the progress of cardiovascular disease, and selected for validation in the clinical cohort. RT-qPCR results showed that the gene expression of SLC12A1 in the HFmrEF group was significantly higher than that in the HFpEF group ( P<0.001). The AUC for the adjunctive differential diagnostic value of SLC12A1 for HFmrEF and HFpEF was 0.802 ( P<0.001) and the AUC of SLC12A1 with a cut-off value of 6.634 was 0.737 ( P=0.003) in determining poor prognosis in patients with HFpEF. Kaplan-Meier survival analysis showed that patients with SLC12A1≤6.634 had a higher incidence of adverse cardiac events than patients with SLC12A1 >6.634 ( P=0.001). Cox regression analysis showed that the risk of adverse cardiac events in the SLC12A1 ≤6.634 group was 6.787 times higher than in the SLC12A1 >6.634 group ( HR=6.787, P=0.011). Conclusions:Transcriptome mRNA sequencing analysis is valuable for detecting clinical relevant differentially expressed genes in HFmrEF and HFpEF patients, among which SLC12A1 can be used as a novel molecular biomarker to aid the differential diagnosis of HFmrEF and HFpEF. In addition, SLC12A1 may be used as an adjunctive biomarker for the prognosis evaluation in patients with HFpEF.

Objective:To compare the treatment efficacy between the Windowing and Open Book techniques in the treatment of tibial plateau fractures of Schatzker type Ⅱ.Methods:A retrospective study was performed to analyze the clinical data of the 211 patients with tibial plateau fracture of Schatzker type Ⅱ who had been treated by open reduction and internal fixation via the anterolateral knee incision at Department of Traumatic Orthopaedics, The Sixth Hospital of Ningbo from January 2014 to June 2022. There were 142 males and 69 females, with an age of (57.7±14.3) years. Based on the intraoperative reduction techniques, the patients were divided into 2 groups: a Windowing group ( n=107) in which reduction of the split bone fragments was followed by reduction of the depressed articular surface, and an Open Book group ( n=104) in which reduction of the depressed articular surface was followed by reduction of the split bone fragments. The following data were collected and compared between the 2 groups: preoperative general data, operative time, quality of fracture reduction, loss of fracture reduction, fracture healing rate at postoperative 6 months, visual analogue scale (VAS) pain score at postoperative 24 months, post-traumatic arthritis grading at the final follow-up (based on the Resnick-Niwoyam criteria), and Rasmussen classifications of knee function at postoperative 3 and 24 months. Results:There were no significant differences in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). The follow-up period for all patients was (34.0±10.6) months. The operative time was (65.0±8.3) minutes for the Windowing group and (64.2±10.2) minutes for the Open Book group, showing no significant difference ( P>0.05). Postoperative CT scans revealed residual articular depression in 10 cases in the Windowing group and in 25 cases in the Open Book group, showing a significant difference ( P<0.05). However, there was no significant difference in plateau widening or varus/valgus alignment between the 2 groups ( P>0.05). There were no statistically significant differences between the 2 groups in loss of fracture reduction, fracture healing rate at postoperative 6 months, VAS score at postoperative 24 months, post-traumatic arthritis grading at the final follow-up, or Rasmussen classifications of the knee function at postoperative 3 or 24 months ( P>0.05). Conclusions:In the treatment of Schatzker type Ⅱ tibial plateau fractures, the Windowing technique demonstrates superior radiographic outcomes compared to the Open Book technique, but the 2 techniques show similar efficacy in functional evaluation and medium-term prognosis.

Objective:To explore the relationship between the differential genes derived from transcriptome mRNA sequencing and prognosis among heart failure patients with mildly reduced ejection fraction (HFmrEF) and preserved ejection fraction (HFpEF).Methods:This was a case-control study. Ten patients with HFmrEF and 10 patients with HFpEF treated at TEDA International Cardiovascular Disease Hospital from November 2021 to January 2022 were selected and differentially expressed genes were screened by transcriptome mRNA sequencing. Ten healthy people served as control group. In addition, 50 patients with HFmrEF, 62 patients with HFpEF, who were treated at TEDA International Cardiovascular Disease Hospital at the same period, were selected as validation groups, 57 healthy people served as control validation group. Real-time quantitative PCR (RT-qPCR) was used to detect the expression of differential genes in each group. Receiver operating characteristic (ROC) curves and the area under the curve (AUC) were used to assess the differential diagnosis and prognostic value of differential genes in these patients. Patients were followed up regularly to document adverse events within 1 year after discharge including cardiac death and readmission for heart failure. Survival analysis was performed using Kaplan-Meier curves and tested by log rank test. Cox regression analysis was used to explore whether differential mRNA was risk factors for poor prognosis in HFmrEF and HFpEF patients.Results:A total of four genes were differentially expressed (three upregulated and one downregulated gene) between the HFmrEF group and HFpEF group (adjust P<0.05). SLC12A1, C15orf48 and SPP1 were associated with the progress of cardiovascular disease, and selected for validation in the clinical cohort. RT-qPCR results showed that the gene expression of SLC12A1 in the HFmrEF group was significantly higher than that in the HFpEF group ( P<0.001). The AUC for the adjunctive differential diagnostic value of SLC12A1 for HFmrEF and HFpEF was 0.802 ( P<0.001) and the AUC of SLC12A1 with a cut-off value of 6.634 was 0.737 ( P=0.003) in determining poor prognosis in patients with HFpEF. Kaplan-Meier survival analysis showed that patients with SLC12A1≤6.634 had a higher incidence of adverse cardiac events than patients with SLC12A1 >6.634 ( P=0.001). Cox regression analysis showed that the risk of adverse cardiac events in the SLC12A1 ≤6.634 group was 6.787 times higher than in the SLC12A1 >6.634 group ( HR=6.787, P=0.011). Conclusions:Transcriptome mRNA sequencing analysis is valuable for detecting clinical relevant differentially expressed genes in HFmrEF and HFpEF patients, among which SLC12A1 can be used as a novel molecular biomarker to aid the differential diagnosis of HFmrEF and HFpEF. In addition, SLC12A1 may be used as an adjunctive biomarker for the prognosis evaluation in patients with HFpEF.

Objective:To compare the treatment efficacy between the Windowing and Open Book techniques in the treatment of tibial plateau fractures of Schatzker type Ⅱ.Methods:A retrospective study was performed to analyze the clinical data of the 211 patients with tibial plateau fracture of Schatzker type Ⅱ who had been treated by open reduction and internal fixation via the anterolateral knee incision at Department of Traumatic Orthopaedics, The Sixth Hospital of Ningbo from January 2014 to June 2022. There were 142 males and 69 females, with an age of (57.7±14.3) years. Based on the intraoperative reduction techniques, the patients were divided into 2 groups: a Windowing group ( n=107) in which reduction of the split bone fragments was followed by reduction of the depressed articular surface, and an Open Book group ( n=104) in which reduction of the depressed articular surface was followed by reduction of the split bone fragments. The following data were collected and compared between the 2 groups: preoperative general data, operative time, quality of fracture reduction, loss of fracture reduction, fracture healing rate at postoperative 6 months, visual analogue scale (VAS) pain score at postoperative 24 months, post-traumatic arthritis grading at the final follow-up (based on the Resnick-Niwoyam criteria), and Rasmussen classifications of knee function at postoperative 3 and 24 months. Results:There were no significant differences in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). The follow-up period for all patients was (34.0±10.6) months. The operative time was (65.0±8.3) minutes for the Windowing group and (64.2±10.2) minutes for the Open Book group, showing no significant difference ( P>0.05). Postoperative CT scans revealed residual articular depression in 10 cases in the Windowing group and in 25 cases in the Open Book group, showing a significant difference ( P<0.05). However, there was no significant difference in plateau widening or varus/valgus alignment between the 2 groups ( P>0.05). There were no statistically significant differences between the 2 groups in loss of fracture reduction, fracture healing rate at postoperative 6 months, VAS score at postoperative 24 months, post-traumatic arthritis grading at the final follow-up, or Rasmussen classifications of the knee function at postoperative 3 or 24 months ( P>0.05). Conclusions:In the treatment of Schatzker type Ⅱ tibial plateau fractures, the Windowing technique demonstrates superior radiographic outcomes compared to the Open Book technique, but the 2 techniques show similar efficacy in functional evaluation and medium-term prognosis.

Objective:To discuss the effects of moxibustion combined with acupuncture on nasal resistance and quality of life in allergic rhinitis (AR) patients with lung qi deficiency and cold syndrome.Methods:Randomized controlled trial was conducted. Totally 82 patients of AR were divided into two groups according to random number table method, with 41 cases in each group. The control group received routine intervention, and the observation group received moxibustion combined with acupuncture intervention on the basis of routine intervention. The treatment for both groups lasted for 4 weeks. The severity of nasal symptoms was evaluated using the Total Nasal Symptom Score (TNSS) before and after treatment. The Rhinoconjunctival Quality of Life Questionnaire (RQLQ) was used to evaluate the quality of life, patient nasal resistance was measured, and clinical efficacy was evaluated.Results:The total effective rate was 95.12% (39/41) in the observation group and 75.61% (31/41) in the control group, with statistical significance ( χ2=4.78, P=0.029). The scores and total scores of nasal itching, sneezing, runny nose and congestion in the observation group after treatment were lower than those in the control group ( t values were 4.45, 4.73, 4.64, 4.68, 9.09, respectively, P<0.01). The scores of daily activities, sleep, non-nasal/eye symptoms, behavioral problems, nasal symptoms, eye symptoms, and emotion were lower than those in the control group ( t values were 4.83, 4.63, 4.50, 5.32, 5.56, 5.29, 4.84, respectively, P<0.01). The nasal resistance of the observation group after treatment [(0.15±0.03) Pa/(cm?s) vs. (0.21±0.03) Pa/(cm?s), t=9.06] was lower than that of the control group ( P<0.001). Conclusion:Moxibustion combined with acupuncture in patients with AR can further reduce nasal resistance and improve their quality of life and clinical efficacy.

Objective:To compare with video-fluoroscopic swallowing function test(VFSS),so as to evaluate the application value of ultrasonography in the test of dysphagia after stroke.Methods:A total of seventy-two patients with dysphagia after stroke who admitted to The Second Affiliated Hospital of Hainan Medical University from January 2022 to July 2023 were selected as cases group,and a total of 45 healthy aged with normal swallowing function were selected as healthy control group at the same time.All of them underwent video X-ray fluoroscopic examination of swallowing function and quantitative assessment of ultrasound.Quantitative assessment of ultrasound was performed by twice to compare the internal consistency of ultrasound test.Meanwhile,the correlation between ultrasound and VFSS results was tested to verify the validity of ultrasound quantitative assessment.The VFSS was used as gold standard to analyze the sensitivity and specificity of ultrasound assessment.The differences of abnormal grade of Geniohyoid muscle;movement time,movement distance and the change of tongue muscle thickness of semi quantitative description of ultrasound between two groups were compared.Results:The intra group correlation coefficient(ICC),movement time,movement distance and movement speed of grade description of muscle abnormalities of cases group were respectively 0.90,(1.743±0.235)s,(6.323±0.823)mm and(3.826±0.778)mm/s,which intra reliabilities were high correlation(ICC=0.90,0.82,0.87,0.85,P＜0.01),which inter reliabilities of these indicators were high correlation(ICC=0.86,0.85,0.88,0.87,P＜0.01),respectively.The positive results of ultrasound test highly correlated with the results of VFSS examination(r=0.91,P＜0.01).The movement distance,the average movement speed and the changes of the thickness of tongue muscle in patients of cases group were significantly smaller than those of healthy control group,but the movement time was larger than that of healthy control group(t=9.03,30.49,-7.02,22.69,P＜0.05),respectively.The results of the ultrasound on the muscles of all patients in cases group existed abnormality.Conclusion:Ultrasound technique can quantitatively assess dysphagia after stroke.Compared with VFSS technique,ultrasonography can measure and determine the related data of the movement of swallowing muscle,and dynamically record movement parameters of geniohyoid muscle and the changes of the thickness of tongue muscle.At the same time,it can detect the grade of muscle abnormalities of patients with dysphagia,which will contribute to further explore the potential pathological mechanism about muscle,and promote the healthy management for patients with dysphagia.

Objective:To explore the genetic etiology of a child featuring multiple fractures and congenital insensitivity to pain (CIP).Methods:A child who had presented at the West China Hospital of Sichuan University on March 16, 2023 for recurrent fractures and CIP was selected as the study subject. Peripheral blood samples of the child and his parents was collected. Trio-whole exome sequencing was carried out. Candidate variants were verified by Sanger sequencing and bioinformatic analysis. This study has been approved by the Medical Ethics Committee of West China Hospital of Sichuan University (No. 2019-772).Results:Trio-whole exome sequencing revealed that the child has harbored compound heterozygous variants of the SCN9A gene, namely c. 560delC (p.P187Rfs*15) and c. 829C>T (p.R277*), which were respectively inherited from his father and mother. Homozygous c. 829C>T variant had been demonstrated as pathogenic among CIP patients, whilst the c. 560delC (p.P187Rfs*15) variant was unreported previously and predicted to be pathogenic based on the guidelines of the American College of Medical Genetics and Genomics (ACMG). Conclusion:The child was diagnosed with CIP due to the compound heterozygous variants of the SCN9A gene. Above finding has enabled genetic counselling and reproductive guidance for this family.

Objective:Preliminary explore the safety and efficacy of using only vitamin K antagonists without antiplatelet therapy after left ventricular assist devices (LVAD) implantation.Methods:This is a cohort study. Patients who underwent HeartCon LVAD implantation in TEDA International Cardiovascular Hospital from September 2020 to September 2022 were included. Oral warfarin sodium was given on postoperative days 1 to 2, with the target international standardized ratio (INR) of 2.0 to 2.5. Follow-up until September 2022, survival, INR level and occurrence of bleeding and thrombosis were recorded. Survival analysis was performed by the Kaplan-Meier method (censored for heart transplantation).Results:A total of 22 patients, including 16 male patients (72.7%), aged (51.0±13.3) years, were included. The duration of HeartCon LVAD support was (458±166) days and the INR during support was 2.28±0.26. One patient underwent the heart transplant at 307 d after implantation. One patient (4.5%) occured cardiac tamponade, two patients (9.1%) occured hemorrhagic stroke, five patients (22.7%) occured gastrointestinal bleeding, four patients (18.2%) occured gingival hemorrhage, two patients (9.1%) occured epistaxis, one patient (4.5%) occurred ischemic stroke, one patient (4.5%) occured pump thrombosis, and one patient (4.5%) occured aortic valve thrombosis. The survival rates were 100%, 95%, 95%, and 95% at 3 months, 6 months, 1 year, 2 years after implantation respectively.Conclusion:The single antithrombotic strategy using warfarin (target INR 2.0-2.5) without antiplatelet for patients with implantations of HeartCon type LVAD may be safe and effective.