Development, Validation and Characterization of a Novel Portable Closed Fracture Device.

BACKGROUND/AIM: As one of the common clinical diseases, fractures have many causes, mechanisms, healing and influencing factors; especially fracture healing is a long-term and complex process. Animal fracture models can simulate the various states of human fractures, and on this basis, the prevention, mechanism, and treatment of fractures can be studied to further guide clinical practice. MATERIALS AND METHODS: Here, we developed a novel and portable device to create a closed fracture model in mice. We then compared this novel closed fracture model with the traditional open model in multiple dimensions to evaluate the modelling process of establishment and healing. The two models were evaluated by imaging, immunostaining, and behavioral tests, which fully demonstrated the stability, universality and operability of the modified fracture model in mice. RESULTS: Surgical quality assessment revealed that the closed fracture model had a shorter operation time and smaller wound than the open model. X-ray and micro-CT results showed no differences between the two models in the evaluation of radiographic and morphological changes during fracture healing. Histological examination revealed the process of the typical intrachondral osteogenic pathway after fracture. Moreover, animal gait analysis indicated reduced postoperative pain in the closed group compared to the open group. CONCLUSION: This study provides a constructive strategy for a closed fracture model in mice and demonstrates the effectiveness and feasibility of the closed fracture model in studying the typical intrachondral osteogenic pathway of fractures from multiple dimensions.

E2F1-mediated up-regulation of TOP2A promotes viability, migration, and invasion, and inhibits apoptosis of gastric cancer cells.

Previous studies have reported that E2F transcription factor 1 (E2F1) and DNA topoisomerase II alpha (TOP2A) are up-regulated in gastric cancer (GC), the corresponding unexplored regulatory mechanisms and the interactions of which in GC are worth figuring out. In this study, the expressions of E2F1 and TOP2A in GC tissues were assessed by real-time PCR (RT-PCR), and E2F1 binding sites in the TOP2A promoter region were predicted via bioinformatics analyses and confirmed using dual-luciferase reporter and ChIP assays. The viability, apoptosis, migration, and invasion of GC cells after transfection with sh-E2F1 or TOP2A plasmid were measured using methyl thiazolyl tetrazolium (MTT) assay, Hoechst 33258 staining/Annexin V/PI staining, and transwell assay. The expressions of E2F1, TOP2A, Cleaved caspase-3, Proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin, and Vimentin in transfected cells were assessed by reverse transcription quantitative PCR (RT-qPCR) and western blot. Concretely, E2F1, as a transcription factor, has binding sites in the TOP2A promoter region. E2F1 and TOP2A levels were up-regulated and positively correlated with each other in GC tissues, i.e., overexpressed E2F1 increased TOP2A levels and E2F1 silencing decreased TOP2A levels in GC cells. Moreover, E2F1 silencing hindered GC cell viability, migration, and invasion, enhanced apoptosis, diminished the levels of PCNA, N-cadherin and Vimentin, and elevated those of Cleaved caspase-3 and E-cadherin in GC cells. However, overexpressed TOP2A reversed the above effects of E2F1 silencing. To sum up, E2F1-mediated up-regulation of TOP2A promotes the viability, migration, and invasion, and inhibits the apoptosis of GC cells.

Numerical insights into the determinants of stent performance for the management of aneurysm with a visceral vessel attached.

PURPOSE: As the factors affecting the efficacy of the bare-metal stent in the treatment of aneurysm with a visceral vessel attached were not fully understood, we aimed to discuss the effects of different characteristics of the stent on the hemodynamics and flexibility in the treatment of the aneurysm. METHODS: Single-layer (with different strut widths) and multi-layer (with a different number of struts) stent models divided into three porosity groups, with porosities of 72.3, 60.5, and 52.4%, were modeled for a comparison of their hemodynamic isolation and flexibility performance via computational fluid dynamics and finite element methods. RESULTS: The velocity and timeaveraged wall shear stress decreased more noticeably with multi-layer stent interventions. A higher oscillatory shear index and relative residence time occurred at the aneurysmal sac wall after multi-layer stents were employed. Time-averaged wall shear stress on the aneurysmal wall decreased with an increase in the number of struts or a decrease in pore size, but oscillatory shear index and relative residence time increased as the number of struts increased or the pore size decreased. Besides, all stents affect the branch patency slightly. In the bending test, when the porosity exceeded 60.5%, multi-layer stents were more flexible. CONCLUSION: The number of struts or pore size of stent dominated the isolation in the management of the aneurysm and affected the flexibility significantly when the porosity was below 60.5%. These findings may contribute to the special design of the stent in the treatment of such types of aneurysms.

S100A9-containing serum exosomes of burn injury patients promote permeability of pulmonary microvascular endothelial cells.

Microvascular hyperpermeability is a leading mechanism responsible for occurrence of edema in remote organs and tissues in patients with burn injury. Accumulated evidence has shown that exosomes can be transported into target cells, where they are capable of regulating biological functions and physiology. Of exosomal proteins contributing to enhanced inflammation and vascular permeability, S100 calcium binding protein A9 (S100A9) has received increasing attention. Here we hypothesized that S100A9-containing serum exosomes of patients with burn injury contribute to pathogenesis of hyperpermeability of microvascular structure in lung by transferring signaling molecules into it and activating downstream signaling pathways, ultimately leading to disruption of the tight junctions (TJs) and endothelial barrier. A use of enzyme-linked immunosorbent assay revealed that total serum concentrations of S100A9 were significantly augmented in burn injury patients in comparison to normal controls. With use of human pulmonary microvascular endothelial cells (HPMECs) as an in vitro model, we found that patients' serum exosomes were effectively internalized by HPMECs. We further found that serum exosomes of stage II/II burn patients inhibited zonula occludens (ZO-1) and occludin protein levels, which are essential for TJs integrity and endothelial barrier function, but activated p38 MAPK signaling pathway in HPMECs. As expected, such exosomes-mediated effects on HPMECs were reversed by a simultaneous treatment of anti-S100A9 neutralizing antibody. Finally, we found that a recombinant human S100A9 treatment led to inhibition of expression of occludin and ZO-1 but an activation of p38 signaling in HPMECs, and that such effects were reversed when p38 activity was repressed, implying that S100A9 may stimulate p38 activity to inhibit ZO-1 and occludin in HPMECs. Collectively, these data suggest that S100A9-containing serum exosomes may play a critical role in contributing to pulmonary microvascular hyperpermeability, thus supporting that blocking exosomes' access to HPMECs could hold a promise strategy for treatment of lung edema resulting from burn injuries.

Lamivudine-resistant rtL180M and rtM204I/V are persistently dominant during combination rescue therapy with entecavir and adefovir for hepatitis B.

Adefovir (ADV) sequential monotherapy was included in the 2005 Asia-Pacific guidelines for the management of patients with lamivudine (LAM) resistance. However, following the development of ADV resistance, the proportion of resistant variants during combined rescue therapy with ADV and entecavir (ETV) were unknown. The present study characterized the dynamics of resistant variants in patients with chronic hepatitis B (CHB) and LAM-resistant variants during antiviral therapy consisting of ADV monotherapy followed by ADV-ETV combination therapy. A total of 3 patients were selected from a cohort of 55 patients with CHB due to developing ADV resistance. The patients had been previously treated with LAM (100 mg daily) for 21-24 months. At the initiation of sequential monotherapy with ADV, LAM-resistant variants (rtM204V/I and rtL180M) were detected in the three patients. These patients developed ADV resistance during 19-30 months of ADV sequential monotherapy, and then switched their antiviral regimen to ADV-ETV combination therapy. During ADV monotherapy and ADV-ETV combination therapy, the patients were monitored every 3 months for the first year of therapy, and then every 6 months thereafter. A total of 30 serum samples were collected from the patients throughout the monitoring period. In total, 10 mutants that were associated with commonly-used antiviral drugs were detected by pyrosequencing. During ADV sequential monotherapy, LAM-resistant variants were gradually decreased, whereas ADV-resistant rtA181V/T and rtN236T variants gradually increased in the viral population. During 30-41 months of ADV-ETV combination therapy, viral load reduction was 2.59-3.28 log10 copies/ml; ADV-resistant variants rtA181T/V and rtN236T were undetectable following 11-24 months of combination therapy; and rtL180M and rtM204I/V remained dominant in the viral population. In conclusion, the results of the present study suggested that, in patients with LAM and ADV-resistant variants that developed during LAM-ADV sequential monotherapy, ETV-ADV combination therapy may partially inhibit the replication of HBV DNA; however, LAM-resistant rtL180M and rtM204I/V variants remained predominant following 30-41 months combination therapy.

miR-494 inhibits ovarian cancer cell proliferation and promotes apoptosis by targeting FGFR2.

MicroRNAs (miRs) have been reported to be key regulators in numerous types of cancer. The aim of the present study was to investigate the role of miR-494 in ovarian cancer. Expression of miR-494 was analyzed in ovarian cancer tissues and cell lines by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). miR-494 mimic or negative control was transiently transfected into A2780 and SKOV3 cell lines. A cell counting kit-8 assay was performed to assess the effects of miR-494 on cell proliferation, and flow cytometry was used to evaluate the apoptotic rate. The target gene of miR-494 was detected by luciferase assay. Expression of fibroblast growth factor receptor 2 (FGFR2) was identified using RT-qPCR and western blotting. In the present study, decreased expression of miR-494 was observed in ovarian cancer samples and cell lines. Overexpression of miR-494 inhibited ovarian cancer cell proliferation by inducing apoptosis. Additional investigation indicated that FGFR2 was a direct target of miR-494. Taken together, the results of the present study suggested that miR-494 suppressed ovarian cancer cell proliferation by inducing apoptosis via targeting FGFR2.

Use of negative pressure wound therapy as an adjunct to the treatment of extremity soft-tissue sarcoma with ulceration or impending ulceration.

Major wound complications of the extremities, following wide tumor resection and reconstruction for soft-tissue sarcomas (STSs), remain a challenge for limb-sparing surgery. Furthermore, STSs with ulceration or impending ulceration predispose patients to an increased risk of post-operative infection. The present study was conducted to assess the efficacy of negative pressure wound therapy (NPWT) in preventing wound complications associated with surgical treatment of STSs with ulceration or impending ulceration, in patients treated between February 2012 and January 2013. A total of 5 patients, with a mean age of 48 years (range, 24-68 years), were enrolled in the present study. The diagnoses consisted of undifferentiated pleomorphic sarcoma (n=2), leiomyosarcoma (n=1), synovial sarcoma (n=1) and epithelioid sarcoma (n=1). According to American Joint Committee on Cancer criteria, 3 cases were stage III tumors, and the remaining 2 cases were of stages IIA and IIB, respectively. A total of 3 patients exhibited ulceration at diagnosis, and the remaining patients demonstrated impending ulceration. The mean wound area following wide resection of the tumor was 73 cm2 (range, 45-110 cm2). A continuous suction mode, with pressures measuring -200 to -300 mmHg, was used for 7-10 days on the soft-tissue defects as preparation for wound closure. Soft-tissue reconstruction included muscle flaps (n=2) and skin grafts (n=5). No major wound complications occurred. Post-operative functional and cosmetic outcomes were acceptable. A single patient demonstrated local recurrence 12 months after surgery and re-excision of the tumor was performed. All patients remained alive at the conclusion of follow-up, with a mean follow-up time of 26 months (range, 12-36 months). The present study demonstrated that NPWT is effective and safe when used as an adjunct to wound closure following resection of extremity STS with ulceration/impending ulceration.

Chronic myeloid leukaemia following repeated exposure to chest radiography and computed tomography in a patient with pneumothorax: A case report and literature review.

It is well known that radioactive rays may cause damage to the human body. Progress in modern medicine has led to an increased risk of therapeutic and diagnostic radiation exposure of patients. Although clear evidence of a radiation dose-dependent risk of chronic myeloid leukaemia, particularly for patients exposed to radiation at a young age, has been established, it is not known whether radiation exposure during diagnostic imaging also increases the risk of cancer. The present study reports the case of a patient who underwent several diagnostic imaging tests (including repeated chest radiography and computed tomography) for recurrent pneumothorax. At around one year subsequent to these tests, the patient was diagnosed with chronic myeloid leukaemia. The patient exhibited an increase in white blood cell count over time, and a bone marrow smear test showed a myeloid/erythroid ratio of 13.9:1. In addition, the qualitative breakpoint cluster region (BCR)/Abelson (ABL) gene test revealed positive results for BCR/ABL fusion (p210). Based on the data reported in the current case, research aimed at elucidating the potential risks associated with diagnostic radiation is urgently required. It is crucial that medical professionals consider the potential harmful side effects of diagnostic radiation when ordering radiation-based diagnostic imaging examinations.

Giant cell tumor of the tendon sheath originating from the ankle capsule: A case report and literature review.

Giant cell tumor of the tendon sheath (GCTTS), also termed tendosynovial giant cell tumor, is a benign, slow-growing tumor that originates from the tendon sheath or bursa. GCTTS of the foot and ankle is much less frequently reported compared with GCTTS of the hand and knee. However, GCTTS should be considered as a differential diagnosis of soft tissue tumors of the foot and ankle. The optimal treatment strategy for GCTTS in the foot and ankle is controversial due to a scarcity of cases. The present study reports the case of a patient that presented with localized intra-articular GCTTS originating from the capsule of the ankle, which is a rare anatomical location for this tumor. Considering the proximity of the tumor to the adjacent non-tumorous structures, a less radical but complete resection of the tumor was performed, followed by a hydrogen peroxide lavage. There was no evidence of recurrence during a follow-up period of 12 months, and adjuvant radiotherapy was not administered to the patient. A pre-operative diagnosis for GCTTS in the foot and ankle is mainly based on the findings of clinical examination and magnetic resonance imaging, which also facilitates the determination of a surgical strategy. For a localized tumor, an integral resection, as opposed to a radical resection, with a hydrogen peroxide lavage may result in a favorable prognosis. However, the optimal treatment for diffuse GCTTS remains to be identified.

Evolution of entecavir-resistant hepatitis B virus during entecavir and adefovir dipivoxil combination therapy.

The emergence of entecavir (ETV) resistance is rare, particularly in a longitudinal study. The aim of the present study was to characterize the evolution of ETV-resistant variants during antiviral therapy using entecavir monotherapy followed by ETV-adefovir dipivoxil (ADV) combination therapy. The study included a prospective cohort of 53 consecutive chronic hepatitis B (CHB) patients. During the 60-month period of ETV therapy, 2 patients exhibited ETV resistance and their medical records were comprehensively reviewed. A total of 25 consecutive serum samples were regularly collected from the 2 patients. All the samples were used to characterize the evolution of the polymerase gene mutations using pyrosequencing. The linkage of the variants was analyzed from 87 reverse transcriptase sequences of 3 selective samples using clone sequencing. The 2 patients presented with viral breakthrough during ETV monotherapy. In patient A, the rtL180M, rtS202G and rtM204V mutant variants were detected using pyrosequencing prior to virological breakthrough. Although the viral load declined following the administration of ADV, the ETV-resistant variants were persistently dominant in the viral populations. In patient B, the rtL180M, rtM204I and rtM204V mutants were present in ~70, 30 and 10% of the viral populations, respectively, at the time of study entry. In addition, rtT184F was present in ~20% of the viral population during virological breakthrough, at month 24. The rtL180M, rtT184F and rtM204V were predominant during the combination treatment. Clonal analysis further revealed that the rtS202G or rtT184F was in all cases co-localized with rtL180M and rtM204V in any single virus isolate clone. The results of the present study indicate that the addition of ADV therapy with ETV for treating ETV-resistant mutation may not inhibit the replication of ETV-resistant variants that developed previously in lamivudine-treated CHB patients.

Primary intraosseous squamous cell carcinoma in pre-existing keratocystic odontogenic tumor: A case report and literature review.

Primary intraosseous squamous cell carcinoma (PIOSCC) is a rare type of odontogenic carcinoma that arises within the jaws. PIOSCC has no initial connection with oral mucosa and possibly develops from the residues of the odontogenic epithelium or from an odontogenic cyst or tumor. The diagnosis of PIOSCC can be difficult as it must be differentiated from other odontogenic carcinomas, such as malignant ameloblastoma, from SCCs arising from the overlying oral mucosa, from the primary tumors of the maxillary sinus or nasal mucosa, and from the tumors that have metastasized to the jaws from other primary sites. The present study reported a rare case of a 59-year-old male patient with a course of keratocystic odontogenic tumor for 25 years, between 1988 and 2013, which eventually transformed into PIOSCC after at least five recurrences and corresponding treatments. The mandible excision and titanium plate reconstruction was performed. Follow-up examinations have revealed no sign of recurrence thus far. The present study discussed this case from three aspects of clinical history, radiological examination and pathological features.

Identification of a rare case of intra-articular osteochondroma manifesting as three loose bodies in a patient with hereditary multiple osteochondromas: A case report.

Hereditary multiple osteochondromas (HMO) is an autosomal dominant bone disorder characterised by the presence of multiple benign cartilage-capped tumours. Exostosin-1 (EXT1) and EXT2 are the major morbigenous genes associated with HMO, mutations in which are responsible for 90% of all HMO cases. In patients with HMO, osteochondromas arise adjacent to the metaphysis and typically remain in the metaphyseal region of the long bones. Therefore, it is rare for osteochondromas to be identified intra-articularly, although they may manifest as loose bodies. The present study describes a rare case of HMO manifesting as limited flexing range in the right knee joint of a 27-year-old male patient. Computed tomography and magnetic resonance imaging (MRI) revealed three intra-articular osteochondromas located in the intercondylar fossa of the patient's right knee. The intra-articular osteochondromas and protuberant extra-articular osteochondromas around the right knee were resected, resulting in improved right knee function and no postoperative recurrence. Pathological analysis revealed that the intra-articular osteochondromas had a thinner cartilage cap layer than the extra-articular osteochondromas. In addition, genetic analysis of the patient and the patient's mother was conducted. From this, it was determined that a nonsense mutation, c.115G>T (p.E39X) in exon 1 of the EXT1 gene, was the cause of HMO in this case. Thus, it is proposed that osteochondromas with a pedicle within the knee, may tear and become loose intra-articular bodies, resulting in limited joint function and thereby contributing to the progression of HMO.

A comparison of the effects of midazolam, propofol and dexmedetomidine on the antioxidant system: A randomized trial.

Previous studies on the antioxidant activity of sedatives have predominantly been in vitro investigations that are lacking clinical data, resulting in conclusion without cogency. The aim of the present prospective, randomized study was to use single sedative drugs for anesthesia induction to compare their antioxidant properties. The effects on the antioxidant system of midazolam, propofol and dexmedetomidine were assessed using oxidative stress indicators and micronuclei (MN). Forty-nine patients undergoing esophageal cancer radical prostatectomy were selected. Midazolam, propofol and dexmedetomidine were used to induce anesthesia. Venous blood samples were obtained prior to and at 2 and at 24 h after surgery, and oxidative stress indicators were detected using the appropriate kits. The cytokinesis-block micronucleus cytome assay was executed and the frequencies of MN, nucleoplasmic bridges and nuclear buds were examined. It was found that the use of the three sedatives, respectively, led to a marked increase in the levels of free radical indicators at 2 h after surgery, which then decreased at 24 h after surgery. Furthermore, lower levels of oxidative stress were found following the use of propofol and dexmedetomidine compared with those following midazolam administration, and similar results were obtained regarding the level of MN. It is suggested that propofol and dexmedetomidine exhibit a superior antioxidant function to midazolam.

Postoperative infection and survival in osteosarcoma patients: Reconsideration of immunotherapy for osteosarcoma.

There is controversy regarding the impact of infection on long-term prognosis in osteosarcoma patients. Clinical trials and experiments relating to this field could bring reconsideration of immunotherapy for osteosarcoma. The clinical records were reviewed of 125 osteosarcoma patients with a mean follow-up of 5.1±3.9 years (range, 0.5-19.8 years), and a review of the literature was also carried out. Chronic localized infections (but not systemic infection) were determined in 6 patients (4.8%). Similar chemotherapeutic regimens (P=1.00) and histological reactions (P=0.65) were observed in patients with or without infection. Tumor location of proximal tibia (P=0.04) was more common in infected patients. More amputations (P<0.001) were necessitated in infected patients due to uncontrolled infection. The 5-year overall survival rate and event-free survival rate in infected patients were 100%, which were significantly higher than that of the non-infected patients, of whom the rates were 54 and 43% respectively (log-rank test: total survival, P=0.01; tumor-free survival, P=0.01). Distant metastasis was an independent risk factor for survival determined by Cox regression analysis (P<0.001, 95 confidence interval, 1.59-3.98). These findings suggested infection was likely to have positive effects on survival in osteosarcoma patients, however, underlying mechanisms remain to be elucidated. Reconsideration of the association of infection and survival in osteosarcoma patients will help to explore novel therapeutic routes and targets in these patients.

Cbl-b-regulated extracellular signal-regulated kinase signaling is involved in the shikonin-induced apoptosis of lung cancer cells in vitro.

Shikonin (SK), a naturally occurring naphthoquinone, exhibits antitumor activity. However, its precise mechanisms of action are unknown. In the present study, the effects of SK on NCI-H460 human lung cancer cells were investigated. It was found that SK reduced cell viability and induced apoptosis in the NCI-H460 cells. Additionally, SK inhibited extracellular signal-regulated kinase (ERK) activity, which indicates that inhibition of the ERK pathway is probably one of the mechanisms by which SK induced NCI-H460 cell apoptosis. The expression of Cbl-b was significantly increased by treatment with SK for 4 h, and gradually increased to a maximal level at 24 h; the time taken for the upregulation of Cbl-b protein was in accordance to that required for the downregulation of phospho (p)-ERK protein. The Cbl inhibitor Ps341 reversed the SK-induced downregulation of p-ERK and apoptosis of NCI-H460 cells. These results indicate that Cbl-b potentiates the apoptotic action of SK by inhibiting the ERK pathway in lung cancer cells.