Identification of key genes and pathways associated with gender difference in osteonecrosis of the femoral head based on bioinformatics analysis.

OBJECTIVE: To identify different key genes and pathways between males and females by studying differentially expressed genes (DEGs). METHODS: The gene expression data of GSE123568 were downloaded from GEO database, including osteonecrosis of the femoral head (ONFH) samples from 3 females and 7 males, and DEGs between different gender were identified with R software. Protein-protein interaction (PPI) network was constructed to further analyze the interactions between overlapping DEGs, and finally, GO, KEGG and gene set enrichment analysis (GSEA) were conducted for enrichment analysis. RESULTS: 131 DEGs were identified between ONFH females and ONFH males, including 76 up-regulated genes and 55 down-regulated genes. And 10 hub genes were identified in PPI network, including SLC4A1, GYPA, CXCL8, IFIT1, GBP5, IFI44, IFI44L, IFIT3, KEL and AHSP. Functional enrichment analysis revealed that these genes were mainly enriched in cGMP-PKG signaling pathway, Fatty acid degradation, Non-alcoholic fatty liver disease, Systemic lupus erythematosus, Hematopoietic cell lineage and NO-cGMP-PKG signaling. CONCLUSIONS: NO-cGMP-PKG signaling may play an important role in the occurrence and development of ONFH. SLC4A1, GYPA, CXCL8, GBP5 and AHSP may be key genes associated with gender difference in the progression of ONFH, which may be ideal targets or prognostic markers for the treatment of ONFH.

Identification of key genes and pathways associated with sex difference in osteoarthritis based on bioinformatics analysis.

OBJECTIVES: The present study aimed to identify different key genes and pathways between postmenopausal females and males by studying differentially expressed genes (DEGs). METHODS: GSE32317 and GSE55457 gene expression data were downloaded from the GEO database, and DEGs were discovered using R software to obtain overlapping DEGs. The interaction between overlapping DEGs was further analyzed by establishing the protein-protein interaction (PPI) network. Finally, GO and KEGG were used for enrichment analysis. RESULTS: 924 overlapping DEGs between postmenopausal women and men with osteoarthritis (OA) were identified, including 674 up-regulated genes and 249 down-regulated ones. And 10 hub genes were identified in the PPI network, including BMP4, KDM6A, JMJD1C, NFATC1, PRKX, SRF, ZFX, LAMTOR5, UFD1L and AMBN. The findings of the functional enrichment analysis suggested that these genes were predominantly expressed in MAPK signaling pathway as well as the Thyroid hormone signaling pathway, indicating that those two pathways may be involved in onset and disease progression of OA in postmenopausal patients. CONCLUSION: BMP4, KDM6A, JMJD1C, PRKX, ZFX and LAMTOR5 are expected to play crucial roles in disease development in postmenopausal patients and may be ideal targets or prognostic markers for the treatment of OA.

Growth/differentiation factor-5 induces osteogenic differentiation of human ligamentum flavum cells through activation of ERK1/2 and p38 MAPK.

Ossification of ligamentum flavum (OLF) is a pathological ectopic ossification in the spinal ligament, leading to spinal canal stenosis, but little was known about its pathogenesis. A previous study has found growth/differentiation factor (GDF)-5 expression at ossified sites of the ligaments from OLF patients. This study aimed to investigate the osteogenic effects of GDF-5 on cultured human ligamentum flavum cells (LFCs). LFCs were isolated from human spinal ligamentum flavum, and treated with or without recombinant human (rh) GDF-5. Alkaline phosphatase (ALP) activity was measured. Expression of osteocalcin was assessed by reverse transcriptase-PCR, Western blotting and immunofluorescence. Matrix mineralization was assessed by alizarin red staining. Activation of mitogen-activated protein kinases (MAPK) ERK1/2, p38 and JNK were detected by Western blotting. We found that rhGDF-5 treatment increased ALP activity and osteocalcin expression in a time- and dose-dependent manner, and induced mineralized nodule form. In addition, rhGDF-5 challenge mediated the ERK1/2 and p38 activation but not JNK. Inhibiting this activation pharmacologically, using U0126, a ERK1/2 inhibitor, or SB203580, a p38 inhibitor, resulted in significantly lower ALP activity and osteocalcin protein expression. The present study shows that rhGDF-5 induces osteogenic differentiation of human LFCs through activation of ERK1/2 and p38 MAPK. These findings give some new insight into the pathogenesis of OLF.

The efficacy of a phone assistance nursing program for functional outcomes in patients after shoulder instability surgery: A protocol for randomized controlled trial.

OBJECTIVE: We conduct this research protocol for the assessment of the effect of phone-assisted care programs on functional outcomes in patients receiving shoulder instability surgery. METHODS: This is a randomized controlled, single center trial which will be implemented from October 2020 to December 2021. This trial is conducted according to the SPIRIT Checklist of randomized researches. It was authorized via the Ethics Committee of the First People's Hospital of Xiangyang city affiliated to Hubei Medical College (XY234-026). Ninety participants who undergo shoulder instability surgery are analyzed. Patients are randomly divided into control group (standard management group, with 45 patients) and study group (the phone program group, with 45 patients). In control group, the exercises at home are not monitored. Whereas in study group, patients are asked about their at-home activities, and the extra coaching sessions are provided to patients on self-care, exercise guidance, and the importance of exercise at home, and then answers to their questions. The primary outcome is the range of motion of the shoulder joint, and the pain arcs are determined through the range of motion. The extra assessments include the shoulder functional outcome, pain, and the quality of life. All the analysis needed in this study is implemented with SPSS (IBM, Chicago, USA) for Windows Version 19.0. RESULTS: The clinical outcome variables between groups are shown in Table. CONCLUSION: This investigation can offer a reliable basis for the effectiveness of phone assistance nursing program in patients after shoulder instability surgery. TRIAL REGISTRATION NUMBER: researchregistry6010.

[Progress on the minimally invasive surgical treatment for the acute Achilles tendon rupture].

Acute Achilles tendon rupture is a common sports injury, is currently the best treatment for acute Achilles tendon rupture there are more controversial programs in the clinical, their treatment is divided into conservative treatment and surgical treatment. Conservative treatment for a long time, and then the higher Achilles tendon rupture rate, postoperative recovery slow. There are a number of complications traditional open surgery, and minimally invasive surgery in recent years developed a new technology that minimizes the exposure of the wound, reduce surgical trauma scope, shorten the operation time and reduce wound infection rate increasing importance in clinical practice, worthy of recommendation.

The effect of leptin on the osteoinductive activity of recombinant human bone morphogenetic protein-2 in nude mice.

OBJECTIVE: To investigate the effect of leptin in the model of ectopic bone formation that utilizes subcutaneously implanted recombinant human bone morphogenetic protein-2 (rhBMP-2)-containing type I collagen discs. METHODS: This study was performed in the Clinical Research Center, Southern Medical University, Guangzhou, China from November 2008 to June 2009. A single dose of 20 ul saline (group A), 20 ug leptin (group B), 2 ug rhBMP-2 (group C), 2 ug rhBMP-2 + 20 ug leptin (group D), and type I collagen disks as a carrier were mixed, and subcutaneously implanted into the back of nude mice (n=12). The effect of ectopic bone formation was evaluated by radiography, dual-energy X absorptiometry, biochemical examination of alkaline phosphatase (ALP) activity, histological observation, and semi-quantitative evaluation 4 and 8 weeks after surgery. RESULTS: At 4 and 8 weeks after operation the radiographs, bone mass density, ALP, and histology values showed significant intragroup and intergroup differences, with those at 8 weeks being higher than those at 4 weeks. CONCLUSION: Our results indicated that the sustainedly released material with collagen as a carrier combined treatment with leptin and rhBMP-2 has a very good osteoinductive activity. Leptin is a positive modulator for the osteoinductive efficacy of BMPs, they have synergistic effect. Its mechanisms are probably related to promote the formation of new-vessels and proliferation/ differentiation of many kinds of cells.

[Effect and mechanism of leptin on osteoblastic differentiation of hBMSCs].

OBJECTIVE: To investigate the effect and mechanism of leptin (LEP) on the osteoblastic differentiation of hBMSCs in vitro. METHODS: Whole bone marrow culture method was applied to culture hMBSCs and hBMSCs at passage 3 were divided into groups A, B, C, D, E and F, and when cell attachment was evident, 400, 200, 100 and 50 ng/mL LEP, 100 ng/mL BMP and common nutrient medium were added into each group, respectively. ALP staining and mineralized nodules staining were conducted at 7 and 21 days after culture, respectively. And inverted phase contrast microscope observation was performed. ALP activity and osteocalcin (OCN) level of hBMSCs in each group was detected at 7, 14 and 21 days after culture to select the best induced concentration of LEP on osteoblastic differentiation. For groups of B, E and F at 7 days after culture, RT-PCR was adopted to detect the expression of such osteogenesis-related genes as core-binding factor alpha 1 (Cbf alpha 1), ALP, Col I and OCN mRNA. RESULTS: At 7 days after induced culture, the ALP staining result showed that the endochylema in groups A, B, C, D and E were stained blue and the endochylema in the group F was slightly positive. At 21 days after induced culture, the mineralized nodules staining showed that cells in groups A, B, C, D and E were stained positively and cells in group F were negative. At 7, 14 and 21 days after culture, ALP and OCN activities in group B were less than that of group E (P < 0.05), but significant higher than that of groups A, C, D and F (P < 0.05), the optimal concentration of LEP was 200 ng/mL. At 7 days after culture, group F witnessed no expression of Cbf alpha 1, ALP, Col I and OCN mRNA, while groups B and E witnessed expressions of all those indexes, but the expressions in group B were less than those of group E. CONCLUSION: LEP can stimulate osteoblastic differentiation of hBMSCs in vitro, and the possible mechanism is its role of promoting the expression of osteoblastic related genes.

Leptin expression by heterotopic ossification-isolated tissue in rats with Achilles' tenotomy.

OBJECTIVE: To set up heterotopic ossification (HO) models according to McClure and determine whether leptin messenger ribonucleic acid (mRNA) and protein are expressed in HO-isolated tissue. METHODS: This study was performed in the Department of Spine and Orthopedics, Southern Medical University, Guangzhou, China from November 2007 to June 2008. Thirty-six male rats were randomly divided into sham, partial achilles' tenotomy (PAT), and achilles' tenotomy (AT) groups, with 12 rats in each group. X-ray and histological examination were carried out to ensure the formation of HO at 5 and 10 weeks after operation. Specimens from achilles tendons and surrounding tissue were taken and processed. Meanwhile, the expression of leptin mRNA (5 and 10 weeks) and protein (10 weeks) in achilles tendons and the surrounding tissue were examined respectively using reverse transcription-polymerase chain reaction assay and immunohistochemical methods. RESULTS: There were no leptin mRNA and protein expression in the sham and a weak expression in the PAT of Achilles tendons and surrounding tissue, whereas there was strong expression in the AT group. CONCLUSION: Leptin is involved in the formation of HO, its mechanisms is related to induction of bone formation and maturation through a series of cellular events, including: proliferation/differentiation of many kinds of cells, collagen synthesis, mineralization, and vascularization of the extracellular matrix.