Evaluating cell metabolism is crucial during pluripotent stem cell (PSC) differentiation and somatic cell reprogramming as it affects cell fate. As cultured stem cells are heterogeneous, a comparative analysis of relative metabolism using existing metabolic analysis methods is difficult, resulting in inaccuracies. In this study, we measured human PSC basal metabolic levels using a Seahorse analyzer. We used fibroblasts, human induced PSCs, and human embryonic stem cells to monitor changes in basal metabolic levels according to cell number and determine the number of cells suitable for analysis. We evaluated normalization methods using glucose and selected the most suitable for the metabolic analysis of heterogeneous PSCs during the reprogramming stage. The response of fibroblasts to glucose increased with starvation time, with oxygen consumption rate and extracellular acidification rate responding most effectively to glucose 4 hours after starvation and declining after 5 hours of starvation. Fibroblasts and PSCs achieved appropriate responses to glucose without damaging their metabolism 2∼4 and 2∼3 hours after starvation, respectively. We developed a novel method for comparing basal metabolic rates of fibroblasts and PSCs, focusing on quantitative analysis of glycolysis and oxidative phosphorylation using glucose without enzyme inhibitors. This protocol enables efficient comparison of energy metabolism among cell types, including undifferentiated PSCs, differentiated cells, and cells undergoing cellular reprogramming, and addresses critical issues, such as differences in basal metabolic levels and sensitivity to normalization, providing valuable insights into cellular energetics.

Background: In Korea, there have been no reports comparing the prevalence of major congenital anomalies with other countries and no reports on surgical treatment and longterm mortality. We investigated the prevalence of 67 major congenital anomalies in Korea and compared the prevalence with that of the European network of population-based registries for the epidemiological surveillance of congenital anomalies (EUROCAT). We also investigated the mortality and age at death, the proportion of preterm births, and the surgical rate for the 67 major congenital anomalies. Methods: Korean National Health Insurance claim data were obtained for neonates born in 2013–2014 and admitted within one-year-old. Sixty-seven major congenital anomalies were defined by medical diagnoses classified by International Classification of Diseases-10 codes according to the EUROCAT definition version 2014. Mortality and surgery were defined if any death or surgery claim code was confirmed until 2020. Poisson distribution was used to calculate the 95% confidence interval of the congenital anomaly prevalence. Results: The total prevalence of the 67 major anomalies was 433.5/10,000 livebirths. When compared with the prevalence of each major anomaly in EUROCAT, the prevalence of spina bifida, atrial septal defect (ASD), congenital megacolon, hip dislocation and/or dysplasia and skeletal dysplasia were more than five times higher in Korea. In contrast, the prevalence of aortic atresia/interrupted aortic arch and gastroschisis was less than one-fifth in Korea. The proportion of preterm births was 15.7%; however, more than 40% of infants with anencephaly, annular pancreas and gastroschisis were preterm infants. Additionally, 29.2% of the major anomalies were admitted to the neonatal intensive care units at birth, and 25.6% received surgical operation. The mortality rate was 1.7%, and 78.2% of the deaths occurred within the first year of life. However, in neonates with tricuspid valve atresia and stenosis, duodenal atresia or stenosis, and diaphragmatic hernia, more than half died within their first month of life. ASD and ventricular septal defect were the most common anomalies, and trisomy 18 and hypoplastic left heart syndrome were the most fatal anomalies. All infants with aortic atresia/interrupted aortic arch and conjoined twins received surgery. Conclusion The proportion of surgeries, preterm births and mortality was high in infants with major congenital anomalies. The establishment of a national registry of congenital anomalies and systematic support by national medical policies are needed for infants with major congenital anomalies in Korea.

Background: Infants with congenital anomalies of the digestive system and abdominal wall defects requiring surgery are at risk of growth and developmental delays. The aim of this study was to analyze long-term growth and developmental outcomes for infants with congenital anomalies of the digestive system and abdominal wall defects who underwent surgery in Korea. Methods: We extracted data from the Korean National Health Insurance Service database for the years 2013–2019. Major congenital anomalies were defined according to the International Classification of Diseases-10 and surgery insurance claim codes. The χ 2 test and the CochranArmitage trend test were performed for data analysis. Results: A total of 4,574 infants with major congenital anomalies in the digestive system and abodminal wall defects, who had undergone surgey, were reviewed. Anorectal obstruction/ stenosis was the most prevalent anomaly (4.9 per 10,000 live births). The prevalence of congenital anomalies of the digestive system was 15.5 per 10,000 live births, and that of abdominal wall defects was 1.5 per 10,000 live births. Seven percent of infants with congenital anomalies in the digestive system died, of which those with diaphragmatic hernia had the highest mortality rate (18.8%). Among 12,336 examinations at 6, 12, 24, 36, 48, 60, and 72 months of age, 16.7% showed a weight below the 10th percentile, 15.8% had a height below the 10th percentile, and 13.2% had a head circumference below the 10th percentile.Abnormal developmental screening results were observed in 23.0% of infants. Infants with esophageal atresia with/without tracheoesophageal fistula most often had poor growth and development. Delayed development and cerebral palsy were observed in 490 (10.7%) and 130 (2.8%) infants respectively. Comparing the results of infants born in 2013 between their 24- and 72-month health examinations, the proportions of infants with poor height and head circumference growth increased by 6.5% and 5.3%, respectively, whereas those with poor weight growth and abnormal developmental results did not markedly change between the two examinations. Conclusion Infants with congenital anomalies of the digestive system and abdominal wall defects exhibit poor growth and developmental outcomes until 72 months of age. Close monitoring and careful consideration of their growth and development after discharge are required.

Background: Dipeptidyl peptidase-4 (DPP-4) inhibitor has been reported to have kidney-protective benefits. To elucidate how antidiabetic agents prevent diabetic kidney disease progression, it is important to investigate their effect on the kidney environment in type 2 diabetes mellitus (DM) patients. Herein, we investigated the expression pattern of urinary exosome-derived microRNA (miRNA) in patients taking a combination of DPP-4 inhibitor and metformin (DPP-4 inhibitor group) and compared them with patients taking a combination of sulfonylurea and metformin (sulfonylurea group). Methods: This was a prospective study involving 57 patients with type 2 DM (DPP-4 inhibitor group, n = 34; sulfonylurea group, n = 23) and healthy volunteers (n = 7). We measured urinary exosomal miRNA using the NanoString nCounter miRNA array (NanoString Technologies) across the three groups (n = 4 per each group) and validated findings using real-time polymerase chain reaction. Results: Twenty-one differentially expressed candidate miRNAs were identified, and six (let-7c-5p, miR-23a-3p, miR-26a-3p, miR-30d, miR-205, and miR-200a) were selected for validation. Validation showed no significant difference in miRNA expression between the DPP-4 inhibitor and sulfonylurea groups. Only miR-23a-3p was significantly overexpressed in the diabetes group compared with the control group (DPP-4 inhibitor vs. control, p = 0.01; sulfonylurea vs. control, p = 0.007). This trend was consistent even after adjusting for age, sex, and body mass index. Conclusion There was no significant difference in urine exosome miRNA expression between diabetic participants taking DPP-4 inhibitor and those taking sulfonylurea. The miR-23a levels were higher in diabetic participants than in nondiabetic controls.

Phosphoinositide 3-kinase (PI3K) is considered as a promising therapeutic target for rheumatoid arthritis (RA) because of its involvement in inflammatory processes. However, limited studies have reported the involvement of PI3KC2γ in RA, and the underlying mechanism remains largely unknown. Therefore, we investigated the role of PI3KC2γ as a novel therapeutic target for RA and the effect of its selective inhibitor, PBT-6. In this study, we observed that PI3KC2γ was markedly increased in the synovial fluid and tissue as well as the PBMCs of patients with RA. PBT-6, a novel PI3KC2γ inhibitor, decreased the cell growth of TNF-mediated synovial fibroblasts and LPS-mediated macrophages. Furthermore, PBT-6 inhibited the PI3KC2γ expression and PI3K/AKT signaling pathway in both synovial fibroblasts and macrophages. In addition, PBT-6 suppressed macrophage migration via CCL2 and osteoclastogenesis. In CIA mice, it significantly inhibited the progression and development of RA by decreasing arthritis scores and paw swelling. Three-dimensional micro-computed tomography confirmed that PBT-6 enhanced the joint structures in CIA mice. Taken together, our findings suggest that PI3KC2γ is a therapeutic target for RA, and PBT-6 could be developed as a novel PI3KC2γ inhibitor to target inflammatory diseases including RA.

Improved approaches for promoting umbilical cord blood (CB) hematopoietic stem cell (HSC) homing are clinically important to enhance engraftment of CB-HSCs. Clinical transplantation of CB-HSCs is used to treat a wide range of disorders. However, an improved understanding of HSC chemotaxis is needed for facilitation of the engraftment process. We found that ectopic overexpression of miR-9 and antisense-miR-9 respectively down- and up-regulated C-X-C chemokine receptor type 4 (CXCR4) expression in CB-CD34⁺ cells as well as in 293T and TF-1 cell lines. Since CXCR4 is a specific receptor for the stromal cell derived factor-1 (SDF-1) chemotactic factor, we investigated whether sense miR-9 and antisense miR-9 influenced CXCR4-mediated chemotactic mobility of primary CB CD34⁺ cells and TF-1 cells. Ectopic overexpression of sense miR-9 and antisense miR-9 respectively down- and up-regulated SDF-1-mediated chemotactic cell mobility. To our knowledge, this study is the first to report that miR-9 may play a role in regulating CXCR4 expression and SDF-1-mediated chemotactic activity of CB CD34⁺ cells.
Cell Line ; Cell Movement ; Chemotaxis ; Fetal Blood ; Hematopoietic Stem Cells ; MicroRNAs ; Stromal Cells

PURPOSE: To evaluate the interstitial and appositional growth of greater trochanter post-screw apophysiodesis in Legg-Calve-Perthes (LCP) disease. MATERIALS AND METHODS: A total of 17 patients, who were diagnosed with LCP and underwent greater trochanter screw apophysiodesis and metal removal between December 2003 and December 2012, and were followed-up for at least 4 years, were selected. Anterioposterior radiologic images were taken in each process of apophysiodesis, metal removal, and last follow-up. From such images, articulotrochanter tip distance (ATD), trochanter tip-screw distance (TSD), trochanter tip-trochanter lower margin distance (TLD), and screw-trochanter lower margin distance (SLD) were measured. Appositional growth and greater trochanter growth rates were compared using paired t-test, independent t-test, and correlation analysis. RESULTS: The average ATD of the affected and unaffected sides was 14.2 and 16.8 mm, respectively at apophysiodesis and 9.2 and 14.8 mm at the last follow-up, with a significantly greater decrease observed on the affected side the unaffected side (p=0.030). TLD of the affected side during the follow-up increased 11.0 mm, from an average of 30.8 to 41.8 mm, while the un-affected side increased 14.3 mm, from an average of 26.7 to 41.0 mm. The growth of greater trochanter after the operation in the affected side was 76.7% of that in unaffected side. The ratio of TLD of the affected side to the unaffected side was significantly reduced, from 1.15 to 1.02 (p=0.014) at the final follow-up. TSD was significantly increased from 4.5 to 14.4 mm at metal removal (p < 0.001) and increased to 17.0 mm at the last follow-up. Moreover, the ratio of TSD to SLD was significantly increased from 0.20 to 0.74 at metal removal (p < 0.001) and increased to 0.84 at the final follow-up. CONCLUSION: The results of this study showed that screw apophysiodesis can suppress the overall growth, but not the appositional growth of the greater trochanter. Therefore, screw apophysiodesis may not be a good procedure to inhibit the growth of greater trochanter.
Femur ; Follow-Up Studies ; Humans ; Legg-Calve-Perthes Disease

BACKGROUND: To determine the relationship between superior disc-endplate complex injury and correction loss after surgery in a group of young adult patients with a stable thoracolumbar burst fracture. METHODS: The study group was comprised of young adult patients who had undergone short-segment posterior fixation and bone grafting under the diagnosis of a stable thoracolumbar burst fracture from March 2008 to February 2014. Follow-up was available for more than 1 year. Before surgery, magnetic resonance imaging was performed to determine injury to the anterior longitudinal ligament, posterior longitudinal ligament, and superior and inferior intervertebral discs and endplates. Correction loss was evaluated by the Cobb angle, intervertebral disc height, upper intervertebral disc angle, vertebral wedge angle, and vertebral body height. RESULTS: No significant relation was noted between correction loss and an injury to the anterior longitudinal ligament, posterior longitudinal ligament, inferior intervertebral disc/endplate, and fracture site, whereas an injury to the superior endplate alone and superior disc-endplate complex showed a significant association. Specifically, a superior intervertebral disc-endplate complex injury showed statistically significant relation to postoperative changes in Cobb angle (p = 0.026) and vertebral wedge angle (p = 0.047). CONCLUSIONS: A superior intervertebral disc-endplate complex injury may have an influence on the prognosis after short-segment fixation in young adult patients with a stable thoracolumbar burst fracture.
Body Height ; Bone Transplantation ; Diagnosis ; Follow-Up Studies ; Humans ; Intervertebral Disc ; Longitudinal Ligaments ; Magnetic Resonance Imaging ; Prognosis ; Young Adult*

PURPOSE: Lumbar fusion surgery was performed on transfusion-free patients and hemodynamic changes were analyzed. MATERIALS AND METHODS: A total of 36 transfusion-free patients who had undergone lumbar fusion surgery using recombinant human erythropoietin (rHuEPO) before surgery from April 2007 to March 2014 were included in the study. Hemoglobin and hematocrit levels were measured before surgery, immediately after surgery, 12 hours after surgery, on day 1, day 2, day 3, and day 7. Changes in levels were investigated and the factors affecting the changes in hemoglobin levels were analyzed. RESULTS: Changes in hemoglobin and hematocrit were -18.11% before surgery and -22.92% on day 7, respectively, and they tended to recover from day 2 after surgery. Depending on the patient's age, gender, body mass index, blood loss, and surgery method (presence of lumbar interbody fusion), the changes in hemoglobin level did not show statistically significant differences; however, significant differences were observed in the surgical time and extent of the operation. CONCLUSION: Transfusion alternatives during lumbar spinal fusion are deemed safe methods, leading to good, postoperative hemodynamic outcomes. However, the surgical time and extent of the operation must be determined before surgery.
Blood Substitutes ; Blood Transfusion* ; Body Mass Index ; Erythropoietin ; Hematocrit ; Hemodynamics* ; Humans ; Methods ; Operative Time ; Spinal Fusion ; Spine*

STUDY DESIGN: Retrospective study. OBJECTIVES: To determine the influence of trauma on the neurologic course in patients who have undergone surgery for cervical myelopathy. SUMMARY OF LITERATURE REVIEW: The postsurgical outcomes were worse from trauma in patients who had a cervical ossification of the posterior longitudinal ligament (OPLL) or cervical canal stenosis, in comparison with patients who did not. MATERIALS AND METHODS: The study was conducted on 70 patients who had undergone surgery due to cervical myelopathy from January 2004 to December 2013 and had at least 1 year of follow-up. Depending on trauma history, the patients were divided into two groups, and their radiological (simple radiographic, computed tomographic, and magnetic resonance imaging) and clinical (Japanese Orthopaedic Association [JOA] score, motor power of upper extremities) results were compared retrospectively. RESULTS: Among 70 patients in total, 18 patients were in the trauma group and 52 were in the non-trauma group, and all cases in the trauma group had a history of minor trauma (11 cases of drivers traffic accidents, 4 cases of slipping and falling, 2 cases of minor pedestrian accidents, and 1 case of falling). Radiologically narrower diameter of the spinal canal showed statistically significant difference between two groups (p=0.042). The JOA scores before and after surgery and the recovery rate did not have a clinically meaningful difference with trauma. However, the degree of motor improvement was significantly higher for the trauma group within 1 week after surgery (p=0.040). CONCLUSIONS: Minor trauma itself may adversely affect the patients' clinical courses.
Accidents, Traffic ; Constriction, Pathologic ; Follow-Up Studies ; Humans ; Longitudinal Ligaments ; Retrospective Studies ; Spinal Canal ; Spinal Cord Diseases*