CircPRMT5 promotes progression of osteosarcoma by recruiting CNBP to regulate the translation and stability of CDK6 mRNA.

Research has demonstrated that circular RNAs (circRNAs) exert critical functions in the occurrence and progression of numerous malignant tumors. CircPRMT5 was recently reported to be involved in the pathogenesis of cancers. However, the potential role of circPRMT5 in osteosarcoma needs further investigation. In present study, our results suggested that circPRMT5 was highly upregulated in osteosarcoma cells and mainly localizes in the cytoplasm. CircPRMT5 promoted the proliferation, migration and invasion capacities of osteosarcoma cells, and suppressed cell apoptosis. Knockdown of circPRMT5 exerted the opposite effects. Mechanically, circPRMT5 promoted the binding of CNBP to CDK6 mRNA, which enhanced the stability of CDK6 mRNA and facilitated its translation, thereby promoting the progression of osteosarcoma. Knockdown of CDK6 reversed the promoting effect of circPRMT5 on osteosarcoma cells. These findings suggest that circPRMT5 promotes osteosarcoma cell malignant activity by recruiting CNBP to regulate the translation and stability of CDK6 mRNA. Thus, circPRMT5 may represent a promising therapeutic target for osteosarcoma.

A missense GDF5 variant causes brachydactyly type A1 and multiple-synostoses syndrome 2.

Objective: This study aimed to identify the molecular defects and clinical manifestations in a Chinese family with brachydactyly (BD) type A1 (BDA1) and multiple-synostoses syndrome 2 (SYNS2). Methods: A Chinese family with BDA1 and SYNS2 was enrolled in this study. Whole-exome sequencing was used to analyze the gene variants in the proband. The sequences of the candidate pathogenic variant in GDF5 was validated via Sanger sequencing. I-TASSER and PyMOL were used to analyze the functional domains of the corresponding mutant proteins. Results: The family was found to have an autosomal-dominantly inherited combination of BDA1 and SYNS2 caused by the S475N variant in the GDF5 gene. The variant was located within the functional region, and the mutated residue was found to be highly conserved among species. Via bioinformatic analyses, we predicted this variant to be deleterious, which perturb the protein function. The substitution of the negatively charged amino acid S475 with the neutral N475 was predicted to disrupt the formation of salt bridges with Y487 and impair the structure, stability, and function of the protein, consequently, the abnormalities in cartilage and bone development ensue. Conclusions: A single genetic variant (S475N) which disrupt the formation of salt bridges with Y487, in the interface of the antagonist- and receptor-binding sites of GDF5 concurrently causes two pathological mechanisms. This is the first report of this variant, identified in a Chinese family with BDA1 and SYNS2.

The protective mechanism of action of plantamajoside on a rat model of acute spinal cord injury.

Acute spinal cord injury (ASCI) is a severe traumatic disease of the central nervous system, characterized by a high incidence and high morbidity, for which there are no effective drug therapies in the clinic. A rat model of ASCI was established to study the effects of plantamajoside (PMS) treatment on the expression of apoptotic factors, including caspase-3, caspase-9, poly (ADP-ribose) polymerase (PARP), Bax and Bcl-2. The Allen's weight hit rat ASCI model was used for the present study, and the rats were treated with various concentrations of PMS. The behavior of rats was assessed using the Basso-Beattle-Bresnahan locomotor rating scale (BBB), the histopathologic changes of spinal cord tissue were observed by hematoxylin and eosin staining, the survival of neurons was assessed by TUNEL staining and the expression levels of apoptotic proteins such as caspase-3, caspase-9, PARP, Bcl-2 and Bax was measured using western blot assays and RT-qPCR. It was observed that PMS could reverse the decrease in the BBB score after ASCI, improve the morphological characteristics of the spinal cord, reduce the degree apoptosis and affect the expression of caspase-3, caspase-9, PARP, Bax and Bcl-2 in a concentration dependent manner. In conclusion, PMS protected ASCI rats by inhibiting apoptosis; therefore PMS may be a potential candidate for ASCI therapy.

Effects of Enhanced Recovery After Surgery in Total Knee Arthroplasty for Patients Older Than 65 Years.

OBJECTIVES: To explore the safety and efficacy of the enhanced recovery after surgery (ERAS) program for elderly total knee arthroplasty (TKA) patients. METHODS: A prospective controlled study was conducted for patients older than 65 years, who would undergo unilateral TKA with a minimum follow-up of 2 years. Patients were divided into an ERAS group (n = 106) and a traditional group (n = 141) based on the patients' willingness to participate in the ERAS program. Baseline parameters of American Society of Anesthesiologists classification and comorbidity were recorded. Complication, mortality, knee function assessment using knee society score and knee range of motion, and perioperative clinical outcomes were compared between the two groups. RESULTS: There were no significant differences between the two groups in terms of baseline parameters. Although no significant differences were found in postoperative nausea and vomiting, urinary tract infection, deep venous thrombosis, pulmonary embolism, wound delayed healing, superficial infection, and deep infection, there were significantly fewer total complications in the ERAS group (26/106 vs 52/141; P = 0.039). No significant difference was found in short-term mortality (1/106 vs 3/141; P = 0.836) between the two groups. There were no significant differences in preoperative visual analogue scale (VAS), knee society score (KSS), and range of motion (ROM) between the two groups. Lower VAS scores were found in the ERAS group at time of postoperative day (POD) 1 (P = 0.012) and POD 5 (P = 0.020); no significant differences were observed at time of postoperative month (POM) 1 and final follow-up. Higher KSS scores were found in the ERAS group at time of POD 1 (P = 0.013), and POD 5 (P = 0.011), no significant differences were observed at time of POM 1 and final follow-up. Increased ROM degree was found in the ERAS group at time of POD 1 (P = 0.021); no significant differences were observed at time of POD 5, POM 1 and final follow-up. Decreased intraoperative blood loss (P < 0.001), total blood loss (P < 0.001), transfusion rate (P = 0.004), and length of stay (P < 0.001) were found in the ERAS group; no significant differences were found in operative time and hospitalization costs between the two groups. CONCLUSION: The ERAS program is safer and more efficacious in elderly TKA patients compared to the traditional pathway. It could effectively relieve perioperative pain and improve joint function, and reduce blood transfusion, length of stay, and total complications without increasing short-term mortality.

Change of miRNA expression profiles in patients with knee osteoarthritis before and after celecoxib treatment.

BACKGROUND: This study aimed to investigate the change of circulating miRNA expression profiles in knee osteoarthritis (OA) patients before and after celecoxib treatment. METHODS: Two hundred and eighteen knee OA patients underwent celecoxib treatment for 6 weeks were enrolled. Plasma samples were obtained at baseline (W0) and at W6, and treatment efficacy were assessed by WOMAC index. In the exploration stage, miRNA expression profiles in plasma before and after treatment from 6 patients were detected by microarray. Subsequently, in the validation stage, 10 top differentially expressed miRNAs (DEMs) after and before treatment in microarray were further validated in all 218 patients by qPCR. RESULTS: In the exploration stage, patients after treatment could be distinguished from them before treatment by miRNAs expression profiles by PCA plot and heatmap analysis, and 45 up-regulated and 48 down-regulated miRNAs were identified by volcano plot. In the validation stage, miR-126-5p and miR-320a levels increased at W6 compared to W0, while miR-155-5p and miR-146a-5p levels decreased. WOMAC pain/stiffness/physical function scores were all decreased at W6 compared to W0, and 71% of patients achieved clinical response. The increase of miR-126-5p expression (W6-W0) in clinical responders was much larger compared to nonclinical responders. And miRNA-320a level declined in nonclinical responders while increased in clinical responders. Conversely, miRNA-146a-5p level increased in nonclinical responders while decreased in clinical responders. CONCLUSION: Circulating miRNA expression profiles act as important roles in knee OA patients underwent celecoxib treatment, and miR-126-5p, miR-320a as well as miR-146a-5p might correlate with treatment response to celecoxib.

The Prevalence and Distribution of Vertebral Pedicles in Adolescent Idiopathic Scoliosis in Chinese people: A Computed Tomography-Based Study of 2958 Vertebral Pedicles.

OBJECTIVE: To explore the prevalence and distribution of abnormal vertebral pedicles in adolescent idiopathic scoliosis (AIS) in Chinese people. METHODS: We retrospectively reviewed AIS patients at a single institution between 2011 and 2017. Transverse pedicle widths from T1 to L5 were measured carefully using computed tomography, including cancellous and cortical channels. Pedicle morphology was classified as: type A, a cancellous channel larger than 4 mm; type B, a cancellous channel measuring 2-4 mm; type C, a cancellous channel smaller than 2 mm with an entirely cortical channel of 2 mm or greater; or type D, a cortical channel smaller than 2 mm. Types B, C, and D were defined as abnormal. Prevalence and distribution of abnormal pedicles were assessed. RESULTS: Eighty-seven patients with AIS, with a total of 2958 vertebral pedicles, were carefully measured and classified. The total prevalence of abnormal vertebral pedicles was as high as 65%, with type B comprising 40%, type C comprising 23%, and type D comprising 2%. Pedicles were located between T2 and T10 in 84% of type C and 91% of type D cases. Female sex, proximal thoracic location, major curve greater than 70 degrees, and concave pedicle may be risk factors for type C and D pedicles. CONCLUSIONS: There is a significantly high prevalence of abnormal pedicles in AIS in Chinese people, with a total prevalence of 65%. Female sex, proximal thoracic location, major curve greater than 70 degrees, and concave pedicle may be risk factors for type C and D pedicles.

Isolation, culture and induced differentiation of rabbit mesenchymal stem cells into osteoblasts.

Mesenchymal stem cells (MSCs) may be easily isolated from the bone marrow, and possess multi-lineage differentiation potential and various therapeutic applications. The differentiation of MSCs into osteoblasts is a complex process that is regulated by multiple internal and external factors. In the present study, the differentiation of MSCs isolated from rabbit bone marrow into osteoblasts using different osteoblast inductive media in the presence of dexamethasone, bone morphogenetic protein 2 (BMP-2), 1,25-dihydroxyvitamin D3, transforming growth factor ß (TGFß), platelet lysate and cyclooxygenase 2 (COX2), respectively. Alkaline phosphatase (ALP) activity, mineralization, collagen type (Ct) I and osteocalcin activities, and the mRNA and protein expression levels of vascular endothelial growth factor (VEGF), BMP-2 and Ct II were measured during the differentiation process in MSCs treated with different inducers. Rabbit MSCs were successfully isolated and were observed to be predominantly circular in shape after culture for 24 h. Following subculture for 5 days, the cells demonstrated a spindle shape. ALP, Ct I and osteocalcin activities were higher in cells cultured in dexamethasone, BMP-2 and TGFß compared with the activities in control cells. Following differentiation, the dexamethasone, BMP-2 and TGFß groups demonstrated significantly enhanced mineralization of MSCs detected by Alizarin Red S staining. The mRNA and protein expression levels of VEGF, BMP-2 and Ct II were significantly increased in the same groups compared with the levels in the control group. In conclusion, rabbit MSCs were successfully isolated from bone marrow and differentiated into osteoblasts indicated by raised ALP, Ct I and osteocalcin activities, mineralization and expression of osteogenesis-inducing genes and proteins. The present study revealed that dexamethasone, BMP-2 and TGFß have a positive effect on cell differentiation.