Short-term outcome of total knee replacement in a patient with hemophilia: A case report and review of literature.

BACKGROUND: Hemophilia A is a rare inherited bleeding disorder caused by mutations in the factor VIII gene. This clotting factor plays an intrinsic role in the blood coagulation pathway. Patients with hemophilia may develop orthopedic manifestations such as hemarthrosis, but multiple malunion of fractures over the knee is rare and difficult to treat. CASE SUMMARY: We report a patient with hemophilia A who developed severe knee osteoarthritis along with fracture malunion and nonunion. Total knee replacement was performed using a custom-made modular hinged knee prosthesis (cemented) equipped with extended distal and proximal stems. At 3 years' follow-up, the patient exhibited excellent clinical function and remained satisfied with the surgical outcome. Surgical intervention was accompanied by rigorous coagulation factor replacement. CONCLUSION: This case highlights various unique scenarios specific to individuals with hemophilia and fracture deformity.

Epidermal stem cells participate in the repair of scalds via Nanog and Myc regulation.

Epidermal stem cells (EpSCs) with high expression of regulatory factor Nanog can promote wound healing. The aim of the present study was to investigate the effectiveness and mechanism of epidermal stem cells (EpSCs) in healing scalds and the underlying molecular mechanism. Mouse EpSCs were isolated from skin tissues and cultured in vitro. First, the proliferative ability of EpSCs was determined via the upregulation and downregulation of Nanog expression levels in EpSCs using the MTS­assay. Second, a wound healing assay of the EpSCs with different Nanog expression levels was performed to investigate cell migratory capacities. Third, the protein expression levels of various proteins in EpSCs with Nanog overexpression or knockdown, were determined. Finally, the transfected EpSCs were applied to the rat scald model to observe their effect on scald healing. Subsequently, wound scores, re­epithelialization and capillary density were determined histologically. The results demonstrated that Nanog overexpression enhanced the proliferative ability of EpSCs via cellular (c)­Myc. Moreover, the LV­Nanog group of EpSCs with increased Nanog expression levels exhibited improved healing abilities in the wound healing test than control group. Using western blotting, it was demonstrated that EpSCs that were transfected with a Nanog­overexpression vector expressed high Nanog protein expression levels, whereas small interfering RNA­Nanog­transfected EpSCs exhibited low Nanog protein expression levels. Furthermore, c­Myc expression was synchronized with Nanog expression. It was also revealed that as the expression levels of c­Myc increased, p53 expression levels also increased. In the rat scald model, Nanog­overexpressing EpSCs enhanced wound closure and re­epithelialization. The EpSCs with Nanog knockdown exhibited the opposite effects. The present study therefore indicated that Nanog may have a positive effect on scald healing in rats, which supports its use in EpSC­based treatments against scalds. Furthermore, it was suggested that c­Myc potentially serves a key role in this process and that this process avoids cancerization by relying on the supervision of p53.

Antitumor Activity of a Mitochondrial-Targeted HSP90 Inhibitor in Gliomas.

PURPOSE: To investigate the antitumor activity of a mitochondrial-localized HSP90 inhibitor, Gamitrinib, in multiple glioma models, and to elucidate the antitumor mechanisms of Gamitrinib in gliomas. EXPERIMENTAL DESIGN: A broad panel of primary and temozolomide (TMZ)-resistant human glioma cell lines were screened by cell viability assays, flow cytometry, and crystal violet assays to investigate the therapeutic efficacy of Gamitrinib. Seahorse assays were used to measure the mitochondrial respiration of glioma cells. Integrated analyses of RNA sequencing (RNAseq) and reverse phase protein array (RPPA) data were performed to reveal the potential antitumor mechanisms of Gamitrinib. Neurospheres, patient-derived organoids (PDO), cell line-derived xenografts (CDX), and patient-derived xenografts (PDX) models were generated to further evaluate the therapeutic efficacy of Gamitrinib. RESULTS: Gamitrinib inhibited cell proliferation and induced cell apoptosis and death in 17 primary glioma cell lines, 6 TMZ-resistant glioma cell lines, 4 neurospheres, and 3 PDOs. Importantly, Gamitrinib significantly delayed the tumor growth and improved survival of mice in both CDX and PDX models in which tumors were either subcutaneously or intracranially implanted. Integrated computational analyses of RNAseq and RPPA data revealed that Gamitrinib exhibited its antitumor activity via (i) suppressing mitochondrial biogenesis, OXPHOS, and cell-cycle progression and (ii) activating the energy-sensing AMP-activated kinase, DNA damage, and stress response. CONCLUSIONS: These preclinical findings established the therapeutic role of Gamitrinib in gliomas and revealed the inhibition of mitochondrial biogenesis and tumor bioenergetics as the primary antitumor mechanisms in gliomas.

Impacts of Fracture Types on Success Rate of Closed Reduction and Percutaneous Pinning in Pediatric Lateral Condyle Humerus Fractures Displaced >4 mm.

BACKGROUND: Closed reduction and percutaneous pinning (CRPP) is a promising treatment for pediatric lateral condyle humerus fractures (LCHFs) displaced >4 mm. However, few studies discussed roles of fracture types on success of CRPP in LCHFs. This study aimed to analyze the impacts of types of LCHFs displaced >4 mm on the success rate of CRPP. METHODS: We retrospectively reviewed 66 consecutive pediatric LCHFs attempted CRPP at our center. Song, Milch, Jakob, and Weiss classification were used to classify LCHFs. The fracture gap ≤2 mm and step of articular surface ≤2 mm were deemed as a successful CRPP. Otherwise, open reduction and internal fixation (ORIF) would be performed. Different fracture types and preoperative displacement were analyzed for their roles on success rate of CRPP in treating LCHFs displaced >4 mm. RESULTS: Fifty patients met the inclusion criteria were finally included in this study. Results showed that Milch type II LCHFs had a higher success rate of CRPP than type I LCHFs (P=0.03, <0.05). Correlation was found between Milch types and success rate of CRPP displaced >4 mm. No difference was found between Song stage 4 and 5 LCHFs displaced >4 mm in success rate of CRPP (P=0.90, >0.05), also no difference was found in pre-operative displacement between CRPP group and ORIF group. CONCLUSIONS: Milch classification is more important than Song classification in the success rate of CRPP when treating LCHFs displaced >4 mm. Milch type II LCHFs are recommended to be treated with CRPP, while Milch type I LCHFs are recommended to be treated with ORIF. The current study confirm Song's initial report that closed reduction can be successful and should be attempted for fractures with such displacement rather than going directly to ORIF. LEVEL OF EVIDENCE: Level III.

Celecoxib reverses the glioblastoma chemo-resistance to temozolomide through mitochondrial metabolism.

Temozolomide (TMZ) is used for the treatment of high-grade gliomas. Acquired chemoresistance is a serious limitation to the therapy with more than 90% of recurrent gliomas showing little response to a second line of chemotherapy. Therefore, it is necessary to explore an alternative strategy to enhance the sensitivity of glioblastoma (GBM) to TMZ in neuro-oncology. Celecoxib is well known and widely used in anti-inflammatory and analgesic. Cyclooxygenase-2 (COX-2) expression has been linked to the prognosis, angiogenesis, and radiation sensitivity of many malignancies such as primitive neuroectodermal tumor and advanced melanoma. The objective of this study was to explore the chemotherapy-sensitizing effect of celecoxib on TMZ in GBM cells and its potential mechanisms. From the study, we found that the combination therapy (TMZ 250uM+celecoxib 30uM) showed excellent inhibitory effect to the GBM, the LN229 and LN18, which were the TMZ resistant GBM cell lines. Our data suggest that the combination therapy may inhibits cell proliferation, increases apoptosis, and increases the autophagy on LN229 and LN18. The potential molecular mechanisms were related to mitochondrial metabolism and respiratory chain inhibition.

Nanog down-regulates the Wnt signaling pathway via ß-catenin phosphorylation during epidermal stem cell proliferation and differentiation.

BACKGROUND: Skin tissue homeostasis is maintained by a balance between the proliferation and differentiation of epidermal stem cells (EpSCs). EpSC proliferation and differentiation are complex processes regulated by many factors and signaling pathways. This study aimed to explore the connection between the Nanog and the Wnt/ß-catenin pathway in the proliferation and differentiation of EpSCs. RESULTS: Our results demonstrated that during the study period, EpSC underwent differentiation when incubated in the presence neuropeptide substance P (SP), there was an opposing expression trend of Nanog and ß-catenin after SP treatment, which could be antagonized by the Wnt antagonist, Dkk-1. The transduced EpSCs had a greater proliferative ability than the SP treatment group and they did not undergo differentiation upon SP treatment. More important, ß-catenin expression was down-regulated but phosphorylated ß-catenin expression and phosphorylated GSK-3ß expression was up-regulated upon Nanog overexpression. CONCLUSIONS: These results strongly suggest that Nanog plays an important role in maintaining the proliferation and differentiation homeostasis of EpSCs by promoting ß-catenin phosphorylation via GSK-3ß to inhibit the activity of the Wnt/ß-catenin signaling pathway. This is important for precise regulation of proliferation and differentiation of EpSC in the application of tissue engineering.

HOXA10 expression is decreased by testosterone in luteinized granulosa cells in vitro.

Polycystic ovary syndrome (PCOS) is perhaps the most prevalent endocrine disorder in women of reproductive age, characterized by elevated levels of circulating androgens or clinical manifestations of androgen excess. The specific cytokine profile of PCOS patients is probably related to the lower implantation rate, since follicular fluid appears to function as an embryotrophic agent. For a better understanding of the local regulation of human follicles, the present study investigated the protein expression levels and cellular localization of HOXA10 in granulosa cells (GCs) from women with normal ovarian function undergoing IVF due to their husbands suffering from azoospermia. We demonstrated by immunohistochemical studies that the expression of HOXA10 was mainly localized in the cytoplasm of GCs. Our data indicate that these alterations were associated with changes in the expression of ovarian transcription factors of HOXA10. GC dose-responsive decreases in HOXA10 protein were observed in response to physiological or supraphysiological concentrations (10-4 to 10-7 M) of testosterone. These data reveal that testosterone may be involved in HOXA10 gene regulation in GCs. Decreased HOXA10 expression in GCs treated with testosterone suggest that this androgen is responsible for the decreased expression of HOXA10 in PCOS patients.

Nanog and ß-catenin: a new convergence point in EpSC proliferation and differentiation.

Skin tissue homeostasis is maintained by the balanced proliferation and differentiation of certain types of proliferating cells such as epidermal stem cells (EpSCs). The proliferation and differentiation of EpSCs are complex processes which are not well understood. This study aimed to find the internal relationship between the Nanog pathway and the Wnt/ß-catenin pathway in the proliferation and differentiation process of EpSCs. In brief, EpSCs were isolated from rat epidermis and cultured. The MTT assay, western blotting, polymerase chain reaction (PCR) and immunocytochemistry were performed during the proliferation and differentiation process of EpSCs. Our results showed that 10⁻7 M neuropeptide substance P could effectively stimulate proliferation of EpSCs and that a possible link exists between the Nanog pathway and the Wnt/ß-catenin pathway.

Plumbagin induces apoptosis via the p53 pathway and generation of reactive oxygen species in human osteosarcoma cells.

Osteosarcoma, which is the most common primary bone tumor, occurs most frequently in adolescents. A number of studies have indicated that plumbagin (PL) (5-hydroxy-2-methyl-1, 4-naphthoquinone), a compound found in the plants of the Plumbaginaceae and Droseraceae families, possesses anticancer activity. However, its anticancer effects and mechanisms against osteosarcoma have not been explored. To determine the anticancer effect of PL on osteosarcoma cell lines MG-63 and U2OS, cell viability, apoptosis, cell cycle distribution, caspase-3 and caspase-9 activity and intracellular reactive oxygen species (ROS) generation were measured, and Western blot analyses were performed. PL significantly inhibited the growth of osteosarcoma cells, particularly U2OS cells. PL up-regulated the expression of p53 in U2OS cells and p21 in the two osteosarcoma cell lines causing cell cycle arrest by decreasing the expression of murine double minute 2 (MDM2)/cyclin B1 and cyclin D1. Furthermore, PL altered the ratio of Bax/Bcl-2, and may have triggered the mitochondrial apoptotic pathway, resulting in caspase-3 and caspase-9 activation. We also found that PL induced the generation of ROS in osteosarcoma cell lines. To conclude, PL exerted anticancer activity on osteosarcoma cells by inducing pro-apoptotic signaling and modulating the intracellular ROS that causes induction of apoptosis. These effects may relate to the p53 status.