Chondroitin sulfate (CS) proteoglycans (PGs) are a family of complex molecules in the extracellular matrix and cell surface that regulate axon growth and guidance during development of the central nervous system. In this study, the expression of CSPGs was investigated in the mouse spinal cord at late embryonic and neonatal stages using CS-56 antibody. CS immunoreactivity was observed abundantly in ventral regions of spinal cord of embryonic day (E) 15 embryos. At E16 to E18, CS expression spread dorsally, but never reached the superficial layers of the dorsal horn. This pattern was maintained until postnatal day 4, the latest stage examined. Antibodies against calcitonin gene related peptide (CGRP) and parvalbumin (PV) were employed to label primary afferents from nociceptors and proprioceptors, respectively. CGRP-immunoreactive fibers terminated in the superficial regions of the dorsal horn where CSPGs were weakly expressed, whereas PV-immunoreactive fibers were found in CSPG-rich regions in the ventral horn. Therefore, we conclude that CS expression is spatiotemporally regulated in the spinal cord, which correlates to the termination of sensory afferents. This pattern suggests a role of CSPGs on patterning afferents in the spinal cord, probably through a differential response of axons to these growth inhibitory molecules.

We quantified differences in trabecular volumetric bone mineral density (BMD) of the femoral head between patients with proximal femoral fractures and healthy subjects in the control group by using quantitative computed tomography (QCT) with the purpose of providing guidance for the choice of head screw in the intramedullary nail fixation. Participants suffering from intertrochanteric fractures (n=536 patients) were recruited. In addition, 497 fracture-free, age-matched cases were considered as the control group. The volumetric BMD of different regions of interest (ROI) in the proximal femur scanned by QCT were analyzed between the fracture and control groups. BMD of proximal femur in the fracture group was markedly lower than that in the control group. There were significant differences at distinct regions for male and female between the two groups. Furthermore, the trend of BMD changes among the femoral head, femoral neck and intertrochanter were not parallels in terms of their average value. In conclusion, osteoporosis has been demonstrated to be a main risk factor of the proximal femoral fracture. BMD value of proximal femur was often inconsistent with that of femoral head. Given this, preoperative QCT assessment plays an important role in choosing proper head screw in the intramedullary nail fixation.

Objective To investigate the effects of fracture displacement and operation method on perioperative blood loss of femoral neck fractures. Methods From December 2013 to October 2014, 130 cases (58 males and 72 females, aged from 18 to 91 years, with an average age of 71.1±12.9 years) with femoral neck fractures were retrospectively analyzed for the periopera?tive blood loss. The degree of displacement was described according to Garden's grades. According to the degree of fracture dis?placement, the patients were divided into two groups:GardenⅠ-Ⅱgroup and Garden Ⅲ-Ⅳgroup, and patients' preoperative hidden blood loss was compared between two groups. According to the degree of fracture displacement and the method of opera?tion, the patients were divided into four groups:Group 1 indicates the group in which patients received cannulated screws fixation for Garden gradeⅠ-Ⅱ;Group 2 in which patients received hemiarthroplasty for Garden gradeⅠ-Ⅱ;Group 3 in which patients received cannulated screws fixation for Garden grade Ⅲ-Ⅳ; Group 4 in which patients received hemiarthroplasty for Garden grade Ⅲ-Ⅳ; and variation in the following four parameters was analyzed: the dominant blood loss, postoperative hidden blood loss, total hidden blood loss, total blood loss in the four different groups. Results For the Garden gradeⅢ-Ⅳfemoral neck frac?ture group, the preoperative hidden blood loss was significantly higher than that of the Garden gradeⅠ-Ⅱfemoral neck fracture group (t=2.267, P=0.001). The dominant blood loss volume, postoperative hidden blood loss volume, total hidden blood loss vol?ume and total blood loss volume of hemiarthroplasty groups (402.1 ± 36.8 ml, 641.3 ± 53.2 ml, 880.7 ± 61.7 ml, 1 246.1 ± 76.7 ml) were higher than those of the cannulated screws fixation group (45.8±34.9 ml, 301.9±50.6 ml, 436.6±58.6 ml, 478.5±72.9 ml). Conclusion The perioperative hidden blood loss is mainly related with the degree of fracture displacement, the greater the de?gree of fracture displacement, the more the preoperative hidden blood loss. While the dominant blood loss volume, postoperative hidden blood loss volume, total hidden blood loss volume and total blood loss volume are mainly related to the method of operation, the blood loss in which patients received hemiarthroplasty should be increased significantly.

The goal of this study was to assess the incidence of soft tissue injury in the tibial plateau fracture by magnetic resonance image (MRI) and reveal the relationship between the articular widening/depression and the risk of meniscus and ligament disorder. A total of 54 patients with tibial plateau fracture were indicated for operative intervention. Soft tissue injuries were assessed by MRI. Meniscus, anterior/posterior cruciate ligaments and medial/lateral collateral ligaments injuries on MRI were evaluated. The articular widening/depression was measured in picture archiving and communication systems. Schatzker classification of fracture types was not significantly associated with soft tissue injuries. The rates of soft tissue injury in types IV and II (respectively, 85.7 and 74.1 %) were higher than those in other types. The meniscus injury was the most common soft tissue damage, and the incidence of meniscus injury was 55.6 %. When LPDCT and LPWCT were, respectively, about 7.6 mm and 10.1 mm and LPDX-ray and LPWX-ray, respectively, 5.6 and 7.4 mm, more attention should be paid on the collateral and cruciate ligament injuries in types I, II and III. Furthermore, when LPWCT and LPWX-ray were, respectively, about 10.3 and 8.6 mm, the collateral and cruciate ligaments were susceptible to injury in types IV and V. In conclusion, tibial plateau fracture can occur high morbidity of soft tissue injury, including meniscus and ligament disorder. X-ray and CT scan had different predicting standards for soft tissue injury, and the articular widening/depression in the tibial plateau was associated with meniscus and ligament injuries.

Objective To explore the frequencies and morphological characteristics of posteromedial and posterolateral fragments in acute tibial plateau fractures.Methods A retrospective analysis was conducted of the radiographic and computed tomographic (CT) data of the 298 tibial plateau fractures from November 2012 to June 2014 at our department.They involved 212 males and 86 females with an average age of 47.0 years (range,from 18 to 82 years).The left side was affected in 157 cases,and the right side in 141.By the Schatzker classification,there were 12 cases of type Ⅰ,139 ones of type Ⅱ,12 ones of type Ⅲ,40 ones of type Ⅳ,62 ones of type Ⅴ,and 33 ones of type Ⅵ.Frequencies of the posteromedial and posterolateral fragments were calculated on the CT images using the Picture Archiving and Communication System.Of the fragments,actual major fracture angle (AMFA),surface area percentage (SAP),sagittal angle (SA),fracture height (FH),maximum displacement (MD) were measured on CT images to characterize the imageological morphology of posteromedial and posterolateral fragments in acute tibial plateau fractures.Results Of the 298 tibial plateau fractures,the posteromedial fragments were found in 56 (18.8％),the posterolateral fragments in 60 (20.1％),and both the posteromedial and the posterolateral fragments in 24 (8.1％).Of the posteromedial and posterolateral fragments,respectively,AMFA was 59.6° ± 58.5° and 92.2°±53.9°,SAP was 21.4％ ±9.0％ and 15.4％ ±5.9％,SA was 64.9° ± 14.3° and 72.4° ±21.3°,FH was 33.3±8.2 mm and 27.0 ±9.8 mm,and MD was 4.3±6.3 mm and 3.7±6.5 mm,showing significant differences between the posteromedial and posterolateral fragments (P ＜ 0.05).Conclusions Incidence of posteromedial and posterolateral fragments may not be low in tibial plateau fractures.Due to the fibular support,the area and displacement of posterolateral fragments are significantly lower than those of posteromedial fragments.Appropriate approaches and nailing directions should be taken into consideration in posterior bicondylar fractures according to their imaging features.

Objective To analyze the perioperative blood loss in the different surgical treatments of senile hip fractures (femoral neck and intertrochanteric fractures).Methods Enrolled in the study were 128 aged patients who had been treated in our department for hip fractures between December 2013 and May 2014.They were 53 men and 75 women,with a mean age of 80.5 years.They were divided into 2 groups according to fracture type,with 57 cases in group A (femoral intertrochanteric fracture) and 71 cases in group B (femoral neck fracture).Their perioperative blood loss was calculated according to the Gross equation.The preoperative hidden blood loss was compared between the 2 groups.The dominant blood loss,postoperative hidden blood loss,total perioperative hidden blood loss and total blood loss were compared between cannulated screw fixation,hemi-hip arthroplasty and total hip arthroplasty for the femoral neck fractures and intramedullary nailing for the intertrochanteric fractures.Results The preoperative hidden blood loss in group A (213.0 ±65.3 mL) was significantly greater than in group B (138.4 ±51.4 mL) (P ＜ 0.05).In comparison of dominant blood loss,total hip arthroplasty ＞ hemi-hip arthroplasty ＞ intramedullary nailing ＞ cannulated screw fixation.In comparisons of postoperative hidden blood loss,total perioperative hidden blood loss and total blood loss,total hip arthroplasty ＞ intramedullary nailing ＞ hemi-hip arthroplasty ＞ cannulated screw fixation.All the above differences were statistically significant (P ＜ 0.05) except the one regarding total perioperative hidden blood loss between total hip arthroplasty and intramedullary nailing (P ＞ 0.05).Conclusions The surgeons should have a comprehensive knowledge of the perioperative blood loss in the treatment of senile hip fractures.Enough attention should be paid to the preoperative and postoperative hidden blood loss.

Objective To evaluate the incidence and imaging features of tibial plateau fracture combined with dislocation.Methods A total of 298 patients with intact imaging data treated for tibial plateau fracture from December 2012 to June 2014 were analyzed retrospcctively.Tilt angle of medial fragment fracture line,ratio of width of medial fragment to total plateau,surface area percentage,fragment height,and major displacement were measured on CT images using picture archiving and communication system (PACS).Results Eighteen patients (6.0％) had tibial plateau fractures combined with dislocation,with tilt angle of medial or posteromedial fragment fracture line of (62.1 ± 14.1)°.Eleven patients (61％) had split of the medial tibial plateau with surface area percentage of (46.3 ± 2.5) ％,fragment height of (43.9 ± 6.2) mm,and major displacement of (5.3 ± 2.1) mm.Seven patients (39％) had posteromedial fragment fracture with surface area percentage of (25.1 ± 5.7) ％,fragment height of (40.0 ± 10.1) mm,and major displacement of (4.8 ± 6.45) mm.Conclusions Fracture line of fracture-dislocation tibial plateau fracture is much often through the eminence or located at the lateral eminence with large medial or posteromedial fragments.Surgical approaches and directions of nailing should be considered properly according to the morphological characteristics.

The objective of this study was to explore the therapeutic efficiency of hyaluronan tetrasaccharide (HA 4 ) treatment after spinal cord injury (SCI) in rats and to investigate the underlying mechanism. Locomotor functional and electrophysiological evaluations revealed that the behavioral function of rats in the HA 4 -treated group was significantly improved compared with the vehicle-treated group. The expression of brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), cluster determinant (CD44) and Toll-like receptor-4 (TLR-4) was obviously upregulated in the HA 4 -treated group than that in the sham and vehicle-treated group. Furthermore, HA 4 could induce BDNF and VEGF expression in the astrocytes in vitro. In addition, the high expression of BDNF and VEGF could be inhibited by blocking CD44 and TLR-4. These findings indicate that HA 4 could be useful as a promising therapeutic agent for SCI and might exert the effect by interaction with the CD44 and TLR-4.

Aim: To explore whether icaritin, a prenylflavonoid derivative of the Chinese tonic herb Epimedium, could suppress the proliferation of human osteosarcoma cells in vitro , and to elucidate the mechanisms of the action. Methods: Human osteosarcoma SaOS 2 cell line was used in the present study. The proliferation of the cells was examined using MTT assay and immunofluorescence DAPI staining. Cell motility was studied with the scratch assay. Cell apoptosis was determined by Annexin V-FITC and PI double staining using flow cytometry. Western blotting and RT-PCR were used to measure the expression of mRNAs and proteins in the cells. Results: Icaritin (5–15 μmol/L) suppressed the proliferation of SaOS 2 cells in vitro in a dose-dependent manner. Furthermore, the cell motility was significantly decreased after exposure to icaritin. Moreover, icaritin (5 μmol/L) time-dependently induced the apoptosis of SaOS 2 cells, markedly suppressed MMP-2 and MMP-9 expression, upregulated caspase-3 and caspase-9 expression, and increased the level of cleaved caspase-3 in the cells. Co-exposure to the caspase-3 inhibitor zVAD-fmk (10 μmol/L) compromised the icaritin-induced caspase-3 expression and apoptosis in SaOS 2 cells. Conclusion: Icaritin suppresses the proliferation of SaOS 2 human osteosarcoma cells by increasing apoptosis and downregulating MMP expression.

The objective of this study was to explore the correlation between the alteration in chondroitin sulfate proteoglycan (CSPG) expression at the epicenter of spinal cord and the loss of behavioral function in tiptoe walking Yoshimura mice. The tiptoe walking Yoshimura mice (twy) and Institute of Cancer Research (ICR) mice, aged 20 and 26 weeks, were used in the present study. The behavior assessment, micro-computed tomography and immunofluorescent staining were performed. The compressed spinal cord was histologically analyzed. The results showed that the expression of CSPG was statistically higher at the compressed spinal cord for twy mice compared with that at the normal spinal cord for ICR mice. At the 26th week, a large ossification block at the posterior longitudinal ligament of C1–3 was obviously observed at the micro-CT image We observed the BMS Score was significantly correlated with the expression of glial fibrillary acidic protein, CSPG and hyaluronan (P < 0.05). These findings suggest that compression injury induces the higher CSPG expression at the compressed spinal cord in the twy mice. Furthermore, the alteration in CSPG expression at the epicenter of spinal cord is associated with the loss of behavioral function in twy mice.

BACKGROUND:The objective of this study was to analyze the risk factors associated with the hemoglobin and hematocrit drops in the early postoperative period for intertrochanteric fracture patients with intramedullary nailing treatment. METHODS:From January 2003 to December 2013, 634 intertrochanteric fracture patients with complete information were recruited into the study. Their age, gender, operating time, medical diseases, blood routine examination at admission and postoperative first day, and the days between the trauma and operation were recorded. RESULTS:The hemoglobin (HGB) change of patients (<75 years) was significantly greater than that of patients (>75 years) (P = 0.039). Meanwhile, the change of hematocrit (HCT) level of patients (<75 years) was greater than that of patients (>75 years), but the difference was not significant (P = 0.062). The gender had no significant influence on HCT and HGB. The HGB and HCT change of patients with diabetes (ΔHCT, 8.47 ± 3.36 %; ΔHGB, 29.19 ± 13.10 g/l) were statistically greater than that of patients without diabetes (ΔHCT, 5.52 ± 3.84 %; ΔHGB, 19.81 ± 14.68 g/l) (P = 0.006, P = 0.022). The hypertension and coronary heart disease had no significant influence on the change of HCT and HGB levels. The operation time had a significant influence on the change of HCT and HGB. The ΔHCT and ΔHGB in the group for which the time was more than 48 h between the trauma and operation were greater than that in the group with less than 48 h between the trauma and operation but not significantly different (ΔHCT, P = 0.672; ΔHGB, P = 0.66). CONCLUSION:The factors of age, medical disease such as diabetes, operation time, and time between the trauma and operation may be associated with the change of perioperative hemoglobin and hematocrit levels for intertrochanteric fracture patients after intramedullary nailing treatment in the early postoperative period.

The aim of this study was to determine the therapeutic efficiency of bog bilberry anthocyanin extract (BBAE) treatment starting 1 d after spinal cord injury (SCI) in rats and to investigate the underlying mechanism. The BBAE contained cyanidin-3-glucoside, malvidin-3-galactoside and malvidin-3-glucoside. SCI models were induced using the weight-drop method in Sprague–Dawley rats and additionally with sham group (laminectomy only). The animals were divided into four groups: vehicle-treated group; 10 mg/kg BBAE-treated group; 20 mg/kg BBAE-treated group; sham group. BBAE-treated or vehicle-treated group was administered orally at one day after SCI and then daily for 8 weeks. Locomotor functional recovery was assessed during the 8 weeks post operation period by performing a Basso, Beattie, and Bresnahan (BBB) locomotor score test. At the end of study, the animals were killed, and 1.5 cm segments of spinal cord encompassing the injury site were removed for immunohistochemistry, histopathological and western blotting analysis. Immunohistochemistry for GFAP, aggrecan, neurocan and NeuN was used to assess the degree of astrocytic glial scar formation and neuron survival. Immunohistochemistry and western blotting analysis for TNF-α, IL-6, IL-1β was used to evaluate the anti-inflammation effect of BBAE. To evaluate its inhibition effect on the astrocytes, we performed the MTT assay and immunohistochemistry for Ki67 in vitro. Results show that the BBAE-treated animals showed significantly better locomotor functional recovery, neuron death and smaller glial scar formation after spinal cord injury in vivo. In addition, BBAE administration could inhibit astrocyte proliferation in vivo and vitro. Therefore, BBAE may be useful as a promising therapeutic agent for SCI.

We investigated whether 17 beta-estradiol (E2) treatment could prevent the apoptosis of neural cells after spinal cord injury (SCI) and cultured cortical cells through inhibition of JNK (c-Jun N-terminal kinase) phosphorylation. SCI-induced rats were randomly divided into three groups: control, E2-treated, and sham-treated. Five rats from each group were sacrificed at 2, 4, 6, 12, or 24 h postinjury. Apoptotic neural cells were assessed using the TUNEL method. JNK phosphorylation was detected with immunohistochemistry. Cultured cortical cells were pretreated with E2 and the specific JNK inhibitor SP600125 and then treated with glutamate-induced cytotoxicity in vitro. Neuron viability was determined with an methyl thiazolyl tetrazolium (MTT) assay, morphology of apoptotic cells was observed with 4',6-diamidino-2-phenylindole (DAPI) staining, and JNK phosphorylation was detected using Western blot analysis. Treatment with E2 reduced neuron apoptosis and inhibited JNK phosphorylation. Moreover, the number of apoptotic cells was correlated with JNK phosphorylation 24 h after the rats suffered the SCI. Pretreatment with E2 significantly maintained neural cell viability, attenuated apoptosis, and inhibited JNK phosphorylation induced by glutamate in vitro. These neuroprotective effects of E2 on neural cells were blocked by the co-administration of SP600125. Our results suggest that neuroprotection from E2 is partially mediated by the inhibition of JNK phosphorylation.

Background: Foreign studies have shown that hyaluronan can contribute to the improvement of motor function after spinal cord injury in rats, but its mechanism is unclear. Objective: To study the effect of hyaluronan on expression of neurotrophic factors in astrocytes. Methods: Cultured astrocytes were obtained from the brain of SD rats under sterile conditions. Passage 3 cells were identified by immunochemistry method. Passage 4 cells were treated with hyaluronan (Mr:125 000-175 000) for 24 hours, and control group was set. Results and Conclusion: The results of immunofluoresence showed that the positive rate of primary astrocytes was nearly 100%. The results of Western blotting and reverse transcription-PCR showed that expressions of brain-derived neurotrophic factor and vascular endothelial growth factor were significantly increased in 100 mg/L and 1 000 mg/L groups compared with the control, 1 mg/L and 10 mg/L groups (P < 0.05). Double-labeling immunofluoresence also showed that, under the effect of 1 000 mg/L hyaluronan, the expressions of brain-derived neurotrophic factor and vascular endothelial growth factor in astrocytes were strongly increased compared with the control group. It is confirmed that small molecular-weight hyaluronan can induce astrocytes to up-regulate expressions of brain-derived neurotrophic factor and vascular endothelial growth factor.

Objective:To explore proteins associated with non-traumatic osteonecrosis of the femoral head in adults by using high throughput proteomic approach.Methods:Eight necrotic bone tissue samples were harvested from the femoral head of patients with osteonercrosis of the femoral head,and 8 normal bone tissue samples were collected as the control group in the same patients.After bone tissues were demineralized by HCl solution,total bone proteins were extracted sequentially by using four different lysis buffers.Thereafter,multi-dimensional liquid chromatography-tandem mass spectrometry(MDLC-MS/MS) were applied to separate and identify the extracted proteins,and bioinformatics analysis was carried out then by the software tools such as TurboSEQUEST and GoMiner.Results:The in-depth proteome analysis of necrotic and normal bone tissue revealed 1233 and 999 high-confidence proteins respectively based on two peptides minimum,with the a false-positive rate of 0.9%.154 differential expressed proteins,the level of which were upregulated or downregulated at least 3 fold were identified by spectral counting,as compared with the normal samples.Enrichment analysis in Gene Ontology categories of the differentially expressed gene found that 34 differential expressed proteins were highly associated with osteonecrosis of the femoral head.Downregulated expression of GPCR26 and ChST2 was confirmed by Western Blot on the samples of ONFH.Conclusion:The pathogenesis of the non-traumatic osteonecrosis of the femoral head has been proved complicated and remains unknown.These identified differential expressed proteins might be promising biomarker candidates for early diagnosis of the devastating disease.The results of this study will affirmatively provide a new approach to understand the etiopathogenesis of the osteonecrosis of the femoral head.