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OBJECTIVE: Platelet-rich plasma (PRP) has been reported to alleviate degenerative pathological damage to joint cartilage. This study aimed to investigate the effect of PRP on Wnt/ß-catenin signaling in rabbit chondrocytes. METHODS: Using 3-month-old New Zealand white rabbits, PRP was prepared from venous blood, and chondrocytes were cultured from knee joint cartilage and identified by staining for type II collagen and proteoglycan. The effects of PRP on chondrocyte viability were measured. The chondrocytes were divided into 5 groups: control, IL-1ß, PRP (100-fold dilution), Dkk-1 (100ng/mL) and Dkk-1+PRP. The IL-1ß, PRP, Dkk-1 and Dkk-1+PRP groups were treated with interleukin (IL)-1ß (50µL, 10µg/mL) for24h. Chondrocyte morphology was observed by electron microscopy. Levels of carboxy terminal peptide (CTX-II) and cartilage oligomeric matrix protein (COMP) in culture media were measured by ELISA. Wnt-1, ß-catenin and GSK-3ß mRNA and protein expression were determined by RT-PCR and western blot respectively. RESULTS: PRP enhanced chondrocyte proliferation. Chondrocytes in the IL-1ß group showed ultrastructural abnormalities that were less pronounced in the PRP, Dkk-1 and Dkk-1+PRP groups. CTX-II and COMP concentrations were higher in the IL-1ß group than in the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). The IL-1ß group had higher mRNA and protein Wnt1 and ß-catenin levels and lower GSK-3ß levels than the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). CONCLUSION: PRP may protect chondrocytes activated by IL-1ß via inhibiting Wnt/ß-catenin signaling.
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Condrocitos/fisiología , Articulación de la Rodilla/citología , Plasma Rico en Plaquetas/metabolismo , Animales , Proteína de la Matriz Oligomérica del Cartílago/metabolismo , Supervivencia Celular , Células Cultivadas , Colágeno Tipo II/metabolismo , Glucógeno Sintasa Quinasa 3 beta/genética , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Interleucina-1beta/inmunología , Osteoartritis , Conejos , Vía de Señalización Wnt , Proteína Wnt1/genética , Proteína Wnt1/metabolismo , beta Catenina/genética , beta Catenina/metabolismoRESUMEN
To understand the function of the encoded proteins, we need to be able to know the subcellular location of a protein. The most common method used for determining subcellular location is fluorescence microscopy which allows subcellular localizations to be imaged in high throughput. Image feature calculation has proven invaluable in the automated analysis of cellular images. This article proposes a novel method named LDPs for feature extraction based on invariant of translation and rotation from given images, the nature which is to count the local difference features of images, and the difference features are given by calculating the D-value between the gray value of the central pixel c and the gray values of eight pixels in the neighborhood. The novel method is tested on two image sets, the first set is which fluorescently tagged protein was endogenously expressed in 10 sebcellular locations, and the second set is which protein was transfected in 11 locations. A SVM was trained and tested for each image set and classification accuracies of 96.7 and 92.3 % were obtained on the endogenous and transfected sets respectively.
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Biología Computacional/métodos , Reconocimiento de Normas Patrones Automatizadas/métodos , Proteínas/química , Proteínas/metabolismo , Fracciones Subcelulares/metabolismo , Biología Computacional/instrumentación , Microscopía Fluorescente , Transporte de Proteínas , Proteínas/genética , Fracciones Subcelulares/química , Máquina de Vectores de SoporteRESUMEN
OBJECTIVE: To explore the choice and effect of internal fixation in treating upper cervical vertebrae instability through anterior approach. METHODS: From March 2000 to September 2010,83 patients with upper cervical vertebrae instability were treated with internal fixation through anterior approach. There were 59 males and 24 females with a mean age of 42 years old (ranged, 20 to 68). Among these patients, 36 patients were treated with odontoid screw fixation, 16 patients with C1,2 transarticular screw fixation, 23 patients with C2,3 steel plate fixation, 5 patients with odontoid screw and transarticular screw fixation,2 patients with odontoid screw and C2.3 steel plate fixation, 1 patient with C1,2 transarticular screw and C2,3 steel plate fixation. RESULTS: One patient with completely cervical vertebrae cord injury died of pulmonary infection after C1,2 transarticular screw fixation. Other patients were followed up from 8 to 36 months with an average of 15 months. Upper cervical vertebrae stability were restored without vertebral artery and spinal cord injury. Thirty-six patients were treated with odontoid screw fixation and 5 patients were treated with screw combined with transarticular screw fixation obtained bone union in the dentations without bone graft. Among the 16 patients treated with C1,2 transarticular screw fixation, 13 patients obtained bone union after bone graft; 1 patient died of pulmonary infection after surgery; 1 patient with comminuted odontoid fracture of type II C and atlantoaxial anterior dislocation did not obtain bone union after bone graft,but the fibrous healing was strong enough to maintain the atlantoaixal joint stability; 1 patient with obsolete atlantoaxial anterior dislocation were re-treated with Brooks stainless steel wire fixation and bone graft through posterior approach, and finally obtained bone union. CONCLUSION: It could obtain satisfactory effects depending on the difference of cervical vertebrae instability to choose the correctly surgical method.
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Vértebras Cervicales/cirugía , Fijación Interna de Fracturas/métodos , Inestabilidad de la Articulación/cirugía , Adulto , Anciano , Placas Óseas , Tornillos Óseos , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Osteosarcoma (OS) is the most common primary malignant bone tumor, particularly in adolescents and young adults. Early diagnosis remains a significant problem in the clinical treatment of OS as we remain far from a comprehensive understanding of the molecular genetic mechanisms and the biology involved. In addition, microRNAs (miRNAs or miRs), a large family of small non-coding RNAs, may provide a greater understanding of OS as they play a complex role in gene expression regulation in vitro and in vivo. In the current study, the differential expression profiles of miRNAs between OS and osteoblast cell lines were investigated by miRNA microarrays and real-time quantitative PCR (RT-qPCR). A total of 268 miRNAs were identified that were significantly dysregulated in OS compared with the osteoblast cell line, including miR-9, miR-99, miR-195, miR-148a and miR-181a, which had been validated as overexpressed, and miR-143, miR-145, miR-335 and miR-539, which were confirmed to be downregulated. This differential expression may aid future OS diagnosis and prognosis prediction and illustration of the potential mechanisms in the oncogenesis, development and metastasis of OS. Bioinformatic research on these differentially expressed miRNAs suggests that they are able to regulate the biological behaviors of OS in a complex and effective manner. Further study on the function of these miRNAs is likely to provide new insights into OS biology and treatment.
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OBJECTIVE: To evaluate the feasibility and efficacy of staged treatment of patients with long bone shaft fractures and severe thoracic trauma. METHODS: A retrospective study was performed on 41 patients with long bone shaft fractures and severe thoracic trauma who underwent staged treatment under the guidance of damage control orthopedics from January 2005 to October 2009 in our department. Among the patients,29 patients were male and 12 patients were female,with an average age of (37.2 +/- 6.1 ) years (ranged, 17 to 65 years). On admission, the value of injury severity score (ISS) for these patients was above 16 and the average value was 27.4. The therapy process included three stages: first stage,controlling thoracic symptoms and provisionally fixing fracture; second stage, resuscitating and optimizing physiological condition in intensive care unit;third stage, operating for fractures when pathogenic condition was equable. RESULTS: There was no dead case. Pathogenic condition of thorax in all patients regained equable in 5 d to 23 d and long bone shaft fractures were fixed in 10 d to 25 d after injury. There was no seriously operative or postoperative complication. All the patients were followed up, the duration ranged from 6 to 36 months, with an average of 19.4 months. Their respiratory functions were good. The long bone shaft fractures were healed in 10 to 16 weeks among 38 patients. The bone nonunion happened in 1 patient with closed fracture and 2 patients with opened fracture. CONCLUSION: Damage control orthopedics is a safe strategy for treatment of long bone shaft fractures with severe thoracic trauma. It is suggested that the surgeons should take a staged approach to perform damage control and definitive surgery.
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Fracturas Óseas/cirugía , Traumatismos Torácicos/cirugía , Adolescente , Adulto , Anciano , Femenino , Fijación de Fractura , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
OBJECTIVE: To design ABC damage variable and positioning system for acetabular fracture and explore the feasibility and clinical practical value of the system through the multi-center analysis of 1122 acetabular fractures. METHODS: According to acetabular three-column conception, and pelvic ring lesions damage direction caused by acetabular fracture domino effect and injury degree of proximal femur joint, it defined class A as any column acetabular fracture; class B as any two-column acetabular fracture; class C as front, dome and posterior mixture acetabular fracture. Lower case English letters a, m, p represented front, dome, posterior fracture, respectively. Acetabular damage variables: 1 was simple displaced fractures; 2 was comminuted fractures; 3 was compression fractures. Pelvic ring lesions damage variables: alpha was sacroiliac joints or sacroiliac fracture horizontal separation deflection; beta was sacroiliac joints or sacroiliac fracture vertical separation deflection; gamma was pubic symphysis separation/superior and inferior ramus of pubis fracture deflection; alpha beta gamma delta was compound floating damage. Proximal humerus joint damage variables: I was femoral head fracture; II was femoral neck fracture; II was intertrochanteric fractures of femur; IV was I to III compound fracture. The ABC damage variable positioning system for acetabular fracture was made up by the above-mentioned variables. The statistics from March 1997 to February 2010 showed 1122 cases acetabular fractures with 18 cases of double side acetabular fracture and 1140 cases of acetabular fractures. The pelvics anterior-posterior view, ilium and obturator oblique view, and 2/3D-CT materials were analyzed and researched. RESULTS: Each damage variables distribution situation in 1140 cases of acetabular fracture involved A in 237 cases (20.8%), B in 605 cases (53.1%), C in 298 cases (26.1%);front column fracture in 808 cases(70.9%), dome fracture in 507 cases (44.5%), posterior fracture in 1026 cases (90%). Acetabular variables: variabe 1 in 203 cases of simple displaced fracture (17.8%); variabe 2 in 516 cases of comminuted fracture(45.3%); variabe 3 in 421 cases of compression fracture (36.9%); 249 cases of pelvic ring lesions damage (21.8%), 75 cases femoral head fracture (6.6%); 18 cases of double side acetabular fracture and relative pelvic ring and proximal humerus joint variables (1.58%). Key part and curative effect elements of 1140 cases acetabular fracture: 507 cases of dome or posterior acetabular fracture (44.5%); 421 cases of compression fracture (36.9%); 249 cases of pelvic ring variables (21.8%); 75 cases of proximal humerus joint variables (6.6%); 486 cases of simple Aa/pl/2,Bapl/2 acetabular fracture (42.6% ). CONCLUSION: Compression fracture, especially defected compression fracture, takes important part in acetabular damage variables, and also presents that acetabular fracture with pelvic ring and proximal femoral damage variables are not rare at all. The relationship of the acetabular fracture damage variables, and its percentage shows the key points and elements in clinical treatment: weight-bearing to dome accounts for 44.5%; compression to defects account for 36.9%, pelvic ring to float accounts for 21.8%; dome fracture to double side fracture account for 6.6%. The system has significant guiding effects on clinic in terms of evaluation of injury severity, anatomic localization, difficulty index, alternative strategy, operative approach, effect of treatment,and prognosis. And the most important thing is that the system creates the comparison of damage variables in same type of fracture and the communication of homo-language and explores a new method.