CircFOXK2 Promotes the Progression of Osteoarthritis by Regulating the miR-4640-5p/NOTCH2 Axis.

OBJECTIVE: Osteoarthritis (OA) is the most common age-related chronic and disabling joint disease, frequently causing pain and disability in the adult population. Given that there are no proven disease-modifying drugs for OA, it is urgent to gain a deeper understanding of OA pathogenesis. This study intended to uncover the circFOXK2 regulation in OA. METHODS: Firstly, in vitro OA cell model was constructed by treating murine chondrocytes with interleukin (IL)-1ß. Then, a series of functional assays were conducted to evaluate the effect of circFOXK2 on OA progression in murine chondrocytes. Bioinformatics analysis and mechanism investigations were performed to investigate the competitive endogenous RNA (ceRNA) network of circFOXK2 in OA. RESULTS: CircFOXK2 is overexpressed in IL-1ß-treated chondrocyte. We confirmed the cyclic structure and cytoplasmic distribution of circFOXK2. Functionally, circFOXK2 promotes chondrocyte apoptosis and extracellular matrix (ECM) degradation but inhibiting chondrocyte proliferation. Mechanically, circFOXK2 competitively binds to microRNA-4640-5p (miR-4640-5p) to enhance NOTCH2 expression in OA, affecting OA progression. Besides, circFOXK2 could motivate the Notch pathway to accelerate OA progression. CONCLUSION: CircFOXK2/miR-4640-5p/NOTCH2 axis stimulates the Notch pathway to promote the transcription of inflammatory cytokines (IL33, IL17F and IL6), consequently facilitating OA progression in murine chondrocytes.

ß-Ecdysone attenuates cartilage damage in a mouse model of collagenase-induced osteoarthritis via mediating FOXO1/ADAMTS-4/5 signaling axis.

BACKGROUND: ß-Ecdysone has been reported to perform a protective effect to prevent interleukin 1ß (IL-1ß)-induced apoptosis and inflammatory response in chondrocytes. In our study, the chondroprotective effects of ß-Ecdysone were explored in a mouse model of collagenase-induced osteoarthritis (OA). METHODS: Injection of collagenase in the left knee was implemented to establish a mouse model of OA. The histomorphological analysis was detected using safranine O staining. Serum pro-inflammatory cytokines were measured by ELISA assays. Protein expression in the femur and chondrocytes was analyzed using western blot. Chondrocyte apoptosis was evaluated by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) staining. RESULTS: Treatment of OA mice with ß-Ecdysone supplementation significantly inhibited the production of pro-inflammatory cytokines. Histologic examination exhibited that the degradation of proteoglycans and the loss of trabecular bone were observed in collagenase-injected mice. However, OA-like changes were attenuated by ß-Ecdysone administration in collagenase-injected mice. Both in vivo and in vitro models, nuclear forkhead box O1 (FOXO1) protein expression was significantly reduced in the femur of collagenase-treated mice and IL-1ß-stimulated chondrocytes. However, ß-Ecdysone treatment was able to rescue FOXO1 protein expression in the nucleus to inhibit the transcription and translation of a disintegrin-like and metallopeptidase (reprolysin type) with thrombospondin type 1 motif, 4 (ADAMTS-4) and ADAMTS-5. CONCLUSION: The findings suggested that ß-Ecdysone functioned as a FOXO1 activator to protect collagenase-induced cartilage damage. FOXO1 might be a potential molecular target of ß-Ecdysone for the effective prevention and treatment of OA.

[Clinical observation of arthroscopic single channel treatment of carpal tunnel syndrome with self-made instruments].

OBJECTIVE: To investigate the efficacy of self-made arthroscopic single channel in the treatment of carpal tunnel syndrome. METHODS: Sixty patients with primary carpal tunnel syndrome treated from January 2014 to December 2019 were divided into arthroscopic group and traditional open operation group. There were 30 cases in arthroscopic group, including 12 males and 18 females, aged (47.5±4.5) years and the course of disease was (6.6±4.2) months. There were 30 cases in the traditional operation group, including 10 males and 20 females, aged (48.5±3.5) years, and the course of disease was (5.6±4.4) months. Both groups were unilateral. According to the anatomy of wrist joint and the characteristics of transverse carpal ligament and arthroscopy, the instruments including cannula, inner heart and hook knife were designed. The patients in two groups were treated with decompression of transverse carpal ligament using arthroscopy combined with self-made instruments and traditional open sergery. The incision length, operation time, intraoperative bleeding, hospitalization cost, hospitalization time and recovery time of the two groups were observed and compared. Boston Carpal Tunnel Questionnaire (BCTQ) score was used to evaluate the clinical efficacy of arthroscopy combined with self made instruments in the treatment of carpal tunnel syndrome. RESULTS: Compared with the traditional group, the arthroscopic group had significant advantages in incision length, operation time, intraoperative bleeding and hospital stay, but the total cost of hospitalization was increased. The Boston score was significantly higher in the arthroscopic group than that in the traditional group at 1 month after operation, but not at 3 and 6 months after operation. CONCLUSION: Arthroscopy combined with self-made instruments in the treatment of carpal tunnel syndrome is more reliable, minimally invasive and simplified than open surgery, but the patients should be clearly diagnosed and appropriately selected before operation to achieve satisfactory clinical effect.

Association between breastfeeding and osteoporotic hip fracture in women: a dose-response meta-analysis.

OBJECTIVE: Approximately 300 mg of calcium a day is provided into infants to maintain the physical development of infants, and 5 to 10% bone loss occurs in women during breastfeeding. Hip fractures are considered the most serious type of osteoporotic fracture. We performed this meta-analysis to investigate the association between breastfeeding and osteoporotic hip fractures. MATERIAL AND METHODS: PubMed and Embase were searched until May 1, 2019, for studies evaluating the relationship between breastfeeding and osteoporotic hip fracture in women. The quality of the included studies was evaluated by the methodological index for non-randomized studies (MINORS). For the dose-response meta-analysis, we used the "generalized least squares for trend estimation" method proposed by Greenland and Longnecker to take into account the correlation with the log RR estimates across the duration of breastfeeding. RESULTS: Seven studies were moderate or high quality, enrolling a total of 103,898 subjects. The pooled outcomes suggested that breastfeeding can decrease the incidence of osteoporotic hip fracture (RR = 0.64 (95% CI 0.43, 0.95), P = 0.027). Dose-response analysis demonstrated that the incidence of osteoporotic hip fracture decreased with the increase of breastfeeding time. The RR and 95% CI for 3 months, 6 months, 12 months, and 24 months were RR = 0.93, 95% CI 0.88, 0.98; RR = 0.87, 95% CI 0.79, 0.96; RR = 0.79, 95% CI 0.67, 0.92; and RR = 0.76, 95% CI 0.59, 0.98, respectively, whereas no significant relationship was found between them when the duration of breastfeeding time was more than 25 months. CONCLUSIONS: Our meta-analysis demonstrated that the incidence of osteoporotic hip fracture decreased with the extension of breastfeeding time. However, there is no significant relationship between them when the duration of breastfeeding time was more than 25 months.

Comparison of targeted percutaneous vertebroplasty and traditional percutaneous vertebroplasty for the treatment of osteoporotic vertebral compression fractures in the elderly.

OBJECTIVE: To investigate the clinical effect of precise puncture and low-dose bone cement in percutaneous vertebroplasty (PVP). METHODS: Sixty patients with osteoporotic vertebral compression fracture (OVCFs) who were treated with PVP in our hospital from July 2018 to June 2019. These included patients were divided into group A (N = 30) and group B (N = 30). Group A has punctured to the fracture area accurately and injected with a small dose of bone cement, the group B was injected with a conventional dose of bone cement. The operation time, the amount of bone cement injection, the number of X-rays, the VAS scores, the leakage rate of bone cement, and the incidence of adjacent vertebral fractures were compared between the two groups. RESULT: The operation time, fluoroscopic times, and bone cement volume in group A are less than that in group B (P < 0.05). Patients in group A had a lower incidence of cement leakage and adjacent vertebral fracture than that in patients in group B. There was no significant difference in postoperative pain relief between the two groups. CONCLUSIONS: Precise puncture and injection of small doses of bone cement can reduce the number of X-ray fluoroscopy, operation time, amount of bone cement injection, reduce the rate of bone cement leakage and the incidence of adjacent vertebral fractures, which is a safe and effective surgical approach for the treatment for the aged with OVCFs.

[Short term effect of total hip arthroplasty through direct anterior approach for the treatment of ankylosing spondylitis with hip flexion deformity].

OBJECTIVE: To investigate the short term clinical efficacy of direct anterior approach(DAA) total hip arthroplasty for the treatment of ankylosing spondylitis with hip flexion deformity. METHODS: From September 2014 to June 2017, 15 cases of ankylosing spondylitis with flexion deformity of the hip were treated with total hip arthroplasty through DAA approach including 12 males(17 hips) and 3 females(4 hips) with an average age of 34.4 years old ranging from 21 to 57 years old. Harris score system was used before and after operation to evaluate hip function, total hip activity and visual analogue scale (VAS) were used to evaluate the clinical efficacy. RESULTS: All 15 patients were followed up for an average of 26.2 months. In the operation, 1 case of great trochanter avulsion was given wire binding, and 1 case of linear split of the femur were given by wire binding. There were no hematoma, nerve injury and deep vein thrombosis of lower extremity. No prosthesis loosening and sinking were observed in the follow-up of X-ray film after operation. There was no heterotopic ossification after operation. After operation, 18 hips pain were relieved completely, and 3 hips pain were found when walking, which all satisfied with the daily life self-care requirements. Harris hip score, total hip motion and VAS score at 1 week after operation were significantly different from those before operation(P<0.05). There was no significant difference in the scores of HHS, total hip motion and VAS at 1, 6 months after operation(P>0.05). At the final follow-up, the Harris score was 91.2±5.3, the total hip mobility was (217.1±29.7)°, and the postoperative VAS pain score was 1.2±0.5, which was significantly different from the preoperative score(P<0.05). CONCLUSIONS: DAA approach THA has good effect in treating AS hip nonfunctional ankylosis with less trauma, less pain and quick recovery. It has a good short term effect, which can effectively improve the quality of life of patients.

[Comparison of the efficacy of total arthroscopy and traditional surgical treatment for the treatment of popliteal cyst].

OBJECTIVE: To compare the clinical efficacy of total arthroscopic surgery and traditional surgery for popliteal cyst. METHODS: From August 2014 to July 2017, 60 cases of popliteal cyst were treated with total arthroscopy or traditional surgery respectively. In total arthroscopy group, there were 30 patients including 7 males and 23 females with an average age of (55.81±8.53) years old; the duration of the disease was (3.52±1.12) years;according to Rauschning-Lingdgren grading, 7 cases were grade I, 19 cases were grade II, 4 cases were grade III. In traditional surgical group, there were 30 patients including 5 males and 25 females with an average age of (57.93±9.84) years old; the duration of the disease was (3.48±1.34) years; according to Rauschning-Lingdgren grading, 5 cases were grade I, 21 cases were grade II, 4 cases were grade III. Preoperative symptoms involved such as arthralgia and swelling after knee joint and limited mobility. MRI confirmed a popliteal cyst. Arthroscopic surgery and traditional surgical was used respectively. Incision length, operation time, average stay, total hospital cost and Lysholm score of two groups were compared. RESULTS: In the total arthroscopic group, 1 case suffered from nerve injury, and all cases' incision healed well. In the traditional surgical group, there was 1 case of incision infection, 2 cases of poor healing, 1 case of nerve injury and 1 case of recurrence. All 60 cases were followed up for 6 to 30 months with an average of (13.3±6.5) months. After operation, the symptoms of knee joint arthralgia and swelling, discomfort were significantly improved in the two groups. There were statistically significant differences in incision length, operation time, average stay, total hospitalcost and Lysholm score 6 months after surgery(P<0.05). CONCLUSIONS: The total arthroscopic resection of popliteal cyst, via anterior approach to having a knee joint cavity exploration and treatment of meniscus and bursa, while combined with posteromedial approach for cyst excision would promise a minimal surgery and less pain for patients. Patients will have a rapider recovery, lower recurrence rate and less complication. The total arthroscopic resection easy to accepting for the patient and having a better clinical curative effect is obviously superior to the traditional surgery.

Overexpression of miR­214 promotes the progression of human osteosarcoma by regulating the Wnt/ß­catenin signaling pathway.

The aberrant expression of microRNA (miR)­214 contributes to the regulation of normal and cancer cell biology, and is associated with human malignancies, however, it can operate in a contradictory manner. The role of miR­214 in osteosarcoma remains to be fully elucidated. The aim of the present study was to investigate the effects of miR­214 on osteosarcoma progression and tumor cell proliferation, and examine the molecular mechanism underlying osteosarcoma. The level of miR­214 was determined using reverse transcription­quantitative polymerase chain reaction (RT­qPCR) analysis in osteosarcoma and matched paracancerous tissues, and in human osteosarcoma cancer cell lines. The roles of miR­214 in cell proliferation, survival and cell cycle were analyzed using miR­214 lentivirus (LV­miR­214)­infected osteosarcoma cells. In addition, the downstream target proteins in the Wnt/ß­catenin signaling pathway were evaluated using western blot analysis in the LV­miR­214­infected cells. The LV­miR­214­infected MG63 cells were also treated with exogenous ß­catenin for 24, 48 and 72 h, respectively, following which the expression of ß­catenin was measured using western blot analysis and survival was determined using a 3­(4,5­cimethylthiazol­2­yl)­2,5­diphenyl tetrazolium bromide (MTT) assay. The results of the RT­qPCR analysis showed that the expression level of miR­214 was significantly higher in the osteosarcoma tissues, compared with that in the matched paracancerous tissues, and the same was observed in the osteosarcoma cell lines. The MG63, Saos­2 and U2OS cells were infected with the hsa­mir­214 lentivirus for 48 h, and the levels of miR­214 were significantly upregulated in the human osteosarcoma cancer cells. The overexpression of miR­214 in the MG­63 and Saos­2 cells promoted cell growth, and treatment of the cells with specific antisense­microRNA oligonucleotides (AMOs) for miR­214 for indicated durations reversed the effects of miR­214. Additionally, the AMO­treated MG63 cells showed G0/G1 phase arrest, suggesting that miR­214 contributed to regulation of the cell cycle. In addition, the results of western blot analysis showed that, in the miR­214 lentivirus­infected cells, the levels of cyclin­D1, c­myc and lymphoid enhancer­binding factor­1 were significantly increased, compared with those in the control lentivirus­infected cancer cells. Of note, infection with the miR­214 lentivirus did not affect the levels of Wnt1, Wnt2, Wnt4, Axin or glycogen synthase kinase ß in the U2OS cells, whereas the expression levels of ß­catenin in the MG63 cells and Saos­2 cells were significantly increased. The addition of exogenous ß­catenin effectively reversed the efficiency of miR­214­specific AMOs, which was detected using an MTT assay. These data suggested the critical role of miR­214 in human osteosarcoma via regulation of the Wnt/ß­catenin signaling pathway and demonstrated that miR­214 is as an oncogene for human osteosarcoma.

[Expression of hypoxia inducible factor-1alpha in the fetal vertebra development: experiment with mice].

OBJECTIVE: To explore the expression pattern and effects of hypoxia inducible factor-1alpha (HIF-1alpha) in the fetal vertebra development. METHODS: Fetuses at different developmental stages were obtained from C57BL6 mice. The vertebrae of the fetuses were isolated and the development of vertebra was observed by stereoscopic and light microscope. The mRNA expression of HIF-1alpha at different time points was detected by using reverse transcription-polymerase chain reaction (RT-PCR) and the protein expression of HIF-1alpha was detected by immunohistochemistry. RT-PCR was used to detect the mRNA expression the gene of vascular endothelia growth factor (VEGF), the Col2a1, gene for the chondroblast marker collagen type 2, and the gene of the osteoblast marker, osteocalcin (OCN). RESULTS: The cartilaginous spine column began to form at E13.5. The primary ossification center was observed at E15.5, and then the osteogenesis expanded and extended to both sides. HIF-1alpha mRNA began to express at E13.5, and more significantly at E14.5 (P < 0.05), then decreased. The VEGF mRNA was expressed coincidently with HIF-1alpha. while the HIF-1alpha protein expression was observed at E14.5 and lasted a little longer till the time when the fetus was to be born. The Col2a1 mRNA expression was high at E13.3 - E15.5 and decreased at e16.5 - E18.5, however, the OCN mRNA expression was low at E13.5 - E15.5 and became high at E16.5 - 18.5. CONCLUSION: The vertebra development pattern appears to be an endochondral osteogenesis process. There exists a hypoxia microenvironment in the vertebra that may increase the mRNA and protein expression of HIF1alpha and the expression of VEGF, its downstream gene. It shows that HIF1alpha activates the downstream genes and initiates the cascade of endochondral osteogenesis.

[Expression of hypoxia inducible factor-1alpha in long bone development].

OBJECTIVE: To observe dynamically the development of fetal long bone and detect the expression and distribution of HIF-1alpha,to investigate the expression pattern and possible effects of hypoxia inducible factor-1alpha (HIF-1alpha) in fetal long bone development of mouse. METHODS: E12.5, E13.5, E14.5, E15.5, E16.5 and E17.5 pregnant C57BL6 mice were sacrificed. After sacrifice, the embryos were delivered by caesarean section. The development of fetal long bone was dynamically observed by stereoscopic microscope, and the distributional expression of HIF-1alpha protein was detected by using method of immunohistochemistry. The expression of HIF-1alpha mRNA and osteoblast marker gene at various stage were also detected by using methods of reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The cartilaginous long bone began to form and joints outline arised at E13.5, then the primary ossification center was observed at E14.5, showing opaque ossification under stereoscopic microscope,and then the osteogenesis expanded and extended to both sides. Immunohistochemistry demonstrated lots of HIF-1alpha protein positive chondrcytes in the center of primary ossification at E14.5, then they decreased dramatically. HIF-1alpha mRNA expressed at high level from E13.5 to E15.5, and then decreased to low level. CONCLUSION: Fetal long bone development pattern appeared to be endochondral osteogenisis process, existing hypoxia microenviroment may increase HIF-1alpha mRNA expression and thus initiate the cascade of endochondral osteogenisis.