Ghrelin up-regulates cartilage-specific genes via the ERK/STAT3 pathway in chondrocytes of patients with adolescent idiopathic scoliosis.

Adolescent idiopathic scoliosis (AIS) is a severe spinal deformity that often occurs during puberty. The occurrence of AIS is suggested to be related to abnormal development of cartilage. Our previous study found increased serum ghrelin levels in AIS patients that may linked to the development of AIS. However, whether ghrelin affects cartilage in AIS patients is unclear. We used quantitative real-time PCR (qRT-PCR) and immunohistochemistry to detect the expression of cartilage-specific genes and the ghrelin receptor, growth hormone secretagogue receptor (GHSR). The mRNA and protein levels of collagen II (COLII), SOX9, AGGRECAN (ACAN) and GHSR were higher in AIS patients than in controls. In addition, the protein levels of GHSR downstream signaling pathway members p-STAT3 (Ser727), and p-ERK1/2 were increased. Furthermore, we treated chondrocytes from AIS patients with 100 nM ghrelin, the cell proliferation assay and Western blotting showed that ghrelin promotes chondrocyte proliferation and enhances COLII, SOX9, ACAN, p-ERK1/2 and p-STAT3 expression, respectively. Interestingly, all these observed alterations were abolished by ghrelin + [D-Lys3]-GHRP-6 (a ghrelin receptor inhibitor) treatment. And after U0126 (an inhibitor of ERK1/2 phosphorylation) treatment, ERK1/2 and STAT3 (Ser727) phosphorylation was simultaneously suppressed indicating that ERK1/2 is an upstream pathway protein of STAT3 (Ser727). In conclusion, ghrelin plays an important role in upregulating cartilage-specific genes on AIS primary chondrocytes by activating ERK/STAT3 signaling pathway.

Adiponectin regulates bone mass in AIS osteopenia via RANKL/OPG and IL6 pathway.

BACKGROUND: Osteopenia have been well documented in adolescent idiopathic scoliosis (AIS). Adiponectin has been shown to be inversely proportional to body mass index and to affect bone metabolism. However, the circulating levels of adiponectin and the relationship between adiponectin and low bone mass in AIS remain unclear. METHODS: A total of 563 AIS and 281 age-matched controls were recruited for this study. Anthropometry and bone mass were measured in all participants. Plasma adiponectin levels were determined by enzyme-linked immunosorbent assay (ELISA) in the AIS and control groups. An improved multiplex ligation detection reaction was performed to study on single nucleotide polymorphism. Facet joints were collected and used to measure the microstructure, the expression of RANKL, OPG, osteoblast-related genes, inflammatory factors, adiponectin and its receptors by qPCR, western blotting and immunohistochemistry. Furthermore, primary cells were extracted from facet joints to observe the reaction after adiponectin stimulation. RESULTS: Compared with the controls, lower body mass index and a marked increase in circulating adiponectin were observed in AIS osteopenia (17.09 ± 1.09 kg/m2 and 21.63 ± 10.30 mg/L). A significant difference in the presence of rs7639352was detected in the AIS osteopenia, AIS normal bone mass and control groups. The T allele showed a significant higher proportion in AIS osteopenia than AIS normal bone mass and control groups (41.75% vs 31.3% vs 25.7%, p < 0.05). micro-CT demonstrated that the AIS convex side had a significant lower bone volume than concave side. RNA and protein analyses showed that in cancellous bone, higher RANKL/OPG and adipoR1 levels and lower runx2 levels were observed, and in cartilage, higher adipoR1 and IL6 levels were observed in AIS. Furthermore, convex side had higher RANKL/OPG, IL6 and adipoR1 than concave side. Compared with normal primary cells, convex side primary cells showed the most acute action, and concave side primary cells showed the second-most acute action when exposed under same adiponectin concentration gradient. CONCLUSION: Our results indicated that high circulating adiponectin levels may result from gene variations in AIS osteopenia. Adiponectin has a negative effect on bone metabolism, and this negative effect might be mediated by the ADR1-RANKL/OPG and ADR1-IL6 pathways.

Analysis of risk factors associated with pre-myopia among primary school students in the Mianyang Science City.

Objectives To find out the prevalence rate of pre-myopia among primary school students in the Mianyang Science City Area, analyze its related risk factors, and thus provide a reference for local authorities to formulate policies on the prevention and control of myopia for primary school students. Methods From September to October 2021, Cluster sampling was adopted by our research group to obtain the vision levels of primary school students employing a diopter test in the Science City Area. In addition, questionnaires were distributed to help us find the risk factors associated with pre-myopia. Through the statistical analysis, we identify the main risk factors for pre-myopia and propose appropriate interventions. Results The prevalence rate of pre-myopia among primary school students in the Science City Area was 45.27% (1020/2253), of which 43.82% were boys and 46.92% were girls, with no statistically significant difference in the prevalence rate of myopia between boys and girls (2 =2.171, P=0.141). The results of the linear trend test showed that the prevalence rate of pre-myopia tends to decrease with increasing age (Z=296.521, P=0.000). Logistic regression analysis demonstrated that the main risk factors for pre-myopia were having at least one parent with myopia, spending less than 2 hours a day outdoors, using the eyes continuously for more than 1 hour, looking at electronic screens for more than 2 hours, and having an improper reading and writing posture. Conclusion The Science City Area has a high prevalence rate of pre-myopia among primary school students. It is proposed that students, schools, families, and local authorities work together to increase the time spent outdoors, reduce digital screens and develop scientific use of eye habits.

Deformed Complex Vertebral Osteotomy Technique for Management of Severe Congenital Spinal Angular Kyphotic Deformity.

OBJECTIVES: To (i) introduce the deformed complex vertebral osteotomy (DCVO) technique for the treatment of severe congenital angular spinal kyphosis; (ii) evaluate the sagittal correction efficacy of the DCVO technique; and (iii) discuss the advantages and limitations of the DCVO technique. METHODS: Multiple malformed vertebrae were considered a malformed complex, and large-range and angle wedge osteotomy was performed within the complex using the DCVO technique. Patients with local kyphosis greater than 80° who were treated with DCVO and did not have tumors, infections, or a history of surgery were included. A retrospective case study was performed in these patients with severe angular kyphosis who underwent the DCVO technique from 2008 to 2016. Demographic data, the operating time, and the volume of intraoperative blood loss were collected. Spinopelvic parameters (pelvic incidence [PI], pelvic tilt [PT], and sacral slope [SS]), local and global sagittal parameters (deformity angle, thoracic kyphosis [TK], and lumbar lordosis [LL]), visual analog scale (VAS) score, and Oswestry disability index (ODI) score were recorded pre- and postoperatively. Paired t-tests (α = 0.05) were used for all data (to compare the mean preoperative value with the mean postoperative and most recent follow-up values). P < 0.05 was considered statistically significant. RESULTS: Twenty-nine patients with a mean age of 34 years (range, 15-55) were included in the final analysis. Seventeen patients were male, and 12 were female. The mean follow-up was 44 months (range, 26-62). The mean operating time was 299 min (range, 260-320 min). The mean blood loss was 2110 mL (range, 1500-2900 mL). Three patients had T7 -T8 deformities (3/29, 10.3%), six had T8 -T9 deformities (6/29, 20.7%), six had T9 -T10 deformities (6/29, 20.7%), 10 had T10 -T11 deformities (10/29, 34.5%), three had T11 -T12 deformities (3/29, 10.3%), and one had T9 -T11 deformities (1/29, 3.4%). The mean local deformity angle significantly improved from 94.9° ± 10.8° to 24.0° ± 2.3° through the DCVO technique, with no significant loss at the follow-up. Moreover, the global sagittal parameters and spinopelvic parameters exhibited ideal magnitudes of improvement; TK decreased from 86.1° ± 12.1° to 28.7° ± 2.5°, LL improved from 94.5° ± 4.1° to 46.1° ± 3.0°, and PI minus LL improved from -60.9° ± 6.5° to -13.7° ± 2.6°. Both the VAS and ODI scores significantly improved at the last follow-up. CSF fistula and neural injury did not occur during the perioperative period. At the last follow-up, fixation failure was not observed. CONCLUSION: The DCVO technique provides an alternative and effective method for the treatment of congenital severe angular spinal kyphotic deformities and may decrease the occurrence of perioperative complications.

Opposite Function of ERα and ERß in Controlling 17ß-Estradiol-mediated Osteogenesis in Osteoblasts.

Estrogen receptor plays critical roles in osteogenesis but the underlying mechanism remains unclear. In order to determine the effect of ERα and ERß on several critical factors in regulating osteogenesis in human osteoblasts. Cell based assy, RT-PCR and immunoblot analyses were used in the research. Both RT-PCR and immunoblot showed that gene expression of OPG, MBP2, TGF-ß, RUNX2, IGF-1 was significantly reduced while expression of RANKL was drastically increased after shRNA-based depletion of ERα in MG-63 osteoblasts. Surprisingly, 17ß-estradiol (E2) treatment led to remarkably reduced RANKL compared with that in E2 untreated cells. In contrast, ERß plays an opposite role in regulating gene expression of OPG, MBP2, TGF-ß, RUNX2, IGF-1 and RANKL. However, double depletion of ERα and ERß could not rescue the gene expression of these factors in vitro. Our results provide a novel mechanism of estrogen receptor in controlling osteogenesis in human cells as well as a potential clinic therapeutic target in human osteoporosis.