Based on the different permeability of DNA-intercalant dyes YO-PRO-1(YP) and propidium iodide (PI) to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4?mol/L YP and 4?g/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively. The correlation between YP fluorescence intensity and the apoptotic cell number was confirmed by fluorescence microscope and linear regression(r=0.999,P

OBJECTIVETo summarize clinical outcomes of locking compression plate (LCP) combined with minimally invasive percutaneous plate osteosynthesis (MIPPO) for the treatment of Pilon fracture.

METHODSFrom January 2009 to December 2012, Pilon fracture patients treated by LCP with MIPPO were retrospectively analyzed. All open fractures, pathologic fractures and those who had limb vascular disease or nerve injury were excluded. Thirty-eight patients were enrolled, including 29 males and 9 females aged from 21 to 78 years old with an average of 48 years old. According to AO classification, 20 cases were type B, 18 cases were type C. Operative time, blood loss, reduction quality, time of fracture healing complications and postoperative ankle joint function were applied for evaluating clinical outcomes, AOFAS scoring were used for assessing postoperative clinical effects.

RESULTSAll patients were followed up from 13 to 24 months (averaged 18 months). All patients obtained bone union without any plate failures or loss of fixation/reduction. One patient occurred superficial wound infection, and resolved with antibiotics and local wound care. Postoperative average AOFAS score was 81 (ranged 65 to 97).

CONCLUSIONLCP with MIPPO for Pilon fratcure has advantages of less invasion, fewer complications and satisfactory ankle function.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Retrospective Studies ; Tibial Fractures ; surgery

OBJECTIVETo explore the clinical effects of anterior decompression and fusion with a nano-hydroxyapatite/ polyamide 66 (n-HA/PA66) cage in treating lower cervical fracture and dislocation.

METHODSFrom January 2008 to December 2010, the clinical data of 42 patients with lower cervical fracture and dislocation were retrospectively analyzed. There were 29 males and 13 females aged from 20 to 65 years old. The mean age was 46.8 years. Five cases got injuried in C3, 14 cases in C4, 12 cases in C5, 7 cases in C6 and 4 cases in C7. According to Frankel grade, 4 cases were classified in grade A, 11 cases in grade B, 13 cases in grade C, 9 cases in grade D and 5 cases in grade F. Twenty-eight cases were treated with anterior corpectomy and fusion and 14 cases with anterior discectomy and fusion. Frankel grade was used to do neurologic assessment and visual analogue scale (VAS) was used to evaluate the improvement of clinical symptoms. Segmental height and sagittal lordosis were measured by radiographs and cage location. Cage appearance and fusion status were assessed by 3D-CT images.

RESULTSAll patients were followed up for 3 to 5.2 years with an average of 4.1 years. Frankel grade had obviously improved than preoperative (Z = -4.845, P < 0.001). There were 2, 3, 11, 8, 11 cases classified in grade A, grade B, grade C, grade D and grade E respectively. At the third day after operation and latest follow-up,VAS was (2.6 +/- 1.8),(1.3 +/- 1.0) scores respectively. Both had improved than preoperative (P < 0.05). Up to the latest follow-up, there was only one patient (2.4%) with slight cage translocation (less than 2 mm), however, no cage prolapsed, or collapse, or breakage were found. Both segmental height and lordosis improved significantly after surgery (P < 0.001). And there was not significant difference in both parameters between each postoperative time points (P > 0.05). The mean distance of cage subsidence was 1.5 mm and the rate of cage subsidence (> 3 mm) was 4.8%.

CONCLUSIONThe n-HA/PA66 cage can not only restore and maintain the fusion segmental height and radian, but also promote the osseous fusion and profit the radiographic assessment after operation. Thus, it was an ideal material for prop graft.

Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Decompression, Surgical ; methods ; Female ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fractures ; surgery ; Spinal Fusion ; methods

A novel genomic island (GI) in Streptococcus suis serotype 2(SS2) was identified, which resided in the highly virulent strains but not in the hypo-virulent strains or avirulent strains of SS2 of the Chinese isolates. This newly discovered GI strain was designated as SSGI4 and its whole length of genome was 11 269 bps, sharing the typical properties of pathogenicity islands, such as the distinct G+C content, a mosaic architecture characteristics and the specificity for virulent isolates. There were 11 genes within SSGI4, in which some genes were putative cell surface protein genes and others were amino acid-binding protein genes. Our finding sheds light on the investigation of horizontal gene transfer in SS2 and their influence on pathogenicity.

OBJECTIVETo observe the immune response after the transplantation of a deproteinized heterogeneous bone scaffold and provides the theoretic reference for clinical practice.

METHODSThe fresh pig bone and deproteinized bone were transplanted respectively to establish BABL/C thigh muscle pouches model of male mice and take the samples for detection at 1, 2, 4, 6 weeks after operation. Lymphocyte stimulation index, subset analysis, serum specific antibody IgG, cytokine detection and topographic histologic reaction after implantation were investigated.

RESULTSAfter the transplantation of deproteinized bone, lymphocyte stimulation index, CD(4)(+) and CD(8)(+) T-lymphocyte subsets, serum specific antibody IgG and cytokines in deproteinized bone group were significantly lower than those in fresh pig bone group at each time point (P<0.05). The histological examination found that in fresh bone group at each time point, a large quantity of inflammatory cells infiltrated in the surrounding of bone graft, and they were mainly lymphocytes, including macrophages and monocytes. In deproteinized bone group, there were few inflammatory cells infiltration around bone graft one week after operation. The lymphocytes were decreased as time went by. At 6 weeks, fibroblasts and fibrous tissue grew into the graft, and osteoclasts and osteoprogenitor cells appeared on the verge.

CONCLUSIONSThe established heterogeneous deproteinized bone has low immunogenicity and is a potentially ideal scaffold material for bone tissue engineering.

Animals ; Bone Transplantation ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds ; Transplantation, Heterologous

OBJECTIVETo assess the association between G894T (Glu298Asp) mutation in exon 7 of the endothelial nitric oxide synthase gene and premature coronary heart disease (P-CHD).

METHODSHospital-based case-control study was conducted. Newly-diagnosed CHD patients were recruited as study subjects. 132 CHD patients diagnosed at/before age 55 for males and 65 for females were assigned to P-CHD case group with other 172 CHD patients as the control group. Polymerase chain reaction with Ban II restriction enzyme digestion was performed to detect the G894T mutation.

RESULTSG894T mutant genotypes in P-CHD group (TT, GT and GG frequencies were 6.06%, 20.45% and 73.48%, respectively) were significant higher than those in control group (TT, GT and GG frequencies were 1.74%, 11.63% and 86.63%, respectively) (P = 0.01). Mutant T allele frequency in P-CHD group was also significantly higher than that in control group (16.29% versus 7.56%, P = 0.001, OR = 2.38, 95% CI: 1.38 - 4.16). Stepwise multiple logistic regression analysis at 0.05 significant level with sex, smoking, alcohol drinking, and overweight covariates indicated that G894T mutation also having significant effect on P-CHD (P = 0.01, OR = 2.25, 95% CI: 1.19 - 4.26).

CONCLUSIONThis study suggested that G894T mutation in endothelial nitric oxide synthase gene might serve as a major risk factor to the pathogenesis of P-CHD in this study population.

Age Factors ; Case-Control Studies ; Coronary Disease ; enzymology ; genetics ; Exons ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Middle Aged ; Nitric Oxide Synthase Type III ; genetics ; Point Mutation ; Risk Factors

BACKGROUNDThe p22phox is a critical component of the superoxide-generating vascular nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Several polymorphisms in p22phox gene are studied for their association with cardiovascular diseases. However, no publication is available to assess the relation of 549C > T polymorphism in p22phox gene to coronary artery disease (CAD) risk. This study was to investigate the effect of the p22phox gene 549C > T polymorphism on CAD risk.

METHODSHospital-based case-control study was conducted with 297 CAD patients and 343 healthy persons as the control group. Polymerase chain reaction and pyrosequencing using PSQ 96 MA Pyrosequencer (Biotage AB) were used to detect the polymorphisms. Multiple Logistic regression model was used to adjust the potential confounders and to estimate odds ratio (OR) with 95% confidence intervals (CIs).

RESULTSThe observed genotype frequencies of this polymorphism obeyed the Hardy-Weinberg equilibrium in both cases (P = 0.439) and controls (P = 0.668). The frequency of mutant genotypes (TT + CT) in cases (41.08%) was higher than that in controls (36.73%) with an OR = 1.20 (95%CI = 0.87-1.65). After the adjustment of the potential confounders, there was a significant association of the mutant genotypes with increased risk of CAD (OR = 1.57, 95%CI = 1.01-2.46, P = 0.047).

CONCLUSIONSThe mutant genotypes of the p22phox gene 549C > T polymorphism had a significant effect on the increased risk of CAD in this studied population.

Case-Control Studies ; Coronary Artery Disease ; etiology ; genetics ; Female ; Genotype ; Humans ; Logistic Models ; Male ; Middle Aged ; NADPH Oxidases ; genetics ; Polymorphism, Single Nucleotide

Objective:To determine the optimal condition for mouse embryonic stem cells (mESC) culture with stirred bioreactor,and to develop a method for mass production of embryoid bodies (EB). Methods:The different initial cell concentrations of mESC and the initial stirring speed of bioreactor were investigated to determine the optimal condition for EB formation. Induced by ascorbic acid,the differentiation of EBs formed in stirred bioreactor into cardiomyocytes was compared with EBs formed in Petri dish. Immunofluorescence staining and RT-PCR were used to identify the cardiomyocytes derived from mESC. Results:The formation of a large number of uniform relatively EBs was achieved in stirred bioreactor when mESC were seeded initially with 1?105~3?105 cells/ml and stirring speed was set to 15~30r/min. Most of cells in the EBs formed in bioreactor were viable. EBs produced in bioreactor differentiated into cardiomyocytes more efficiently compared with EBs from Petri dish. The cardiac specific genes were expressed in ESC-derived cardiomyocytes. Conclusions:Stirred bioreactor culture could enhance the efficiency of EB formation and differentiation into cardiomyocytes,which may be a more ideal culture system for EB formation.

OBJECTIVETo analyze the early clinical and radiographic outcomes of Hoffa fractures treated by a standard protocol of open reduction and internal fixation using headless compression screws combined with back buttress plate in a consecutive series of 8 Chinese patients.

METHODSOpen reduction and internal fixation was performed on all patients. The fractures were anatomically reduced and held temporarily by K-wire. If the ends of fractures were atrophic, autologous bone graft from the ipsilateral iliac crest was packed between the ends. Then the fracture fragments were fixed with AO 6.5 mm headless compression cannulated screws. At least two screws were used to provide rotational stability. One pre-contoured reconstruction plate was placed on the nonarticular surface posteromedially or posterolaterally as back buttress plate.

RESULTSAll the patients were followed up for at least 12 months (range 12-25 months). All fractures achieved anatomical reduction and healed clinically and radiographically. At recent follow-up, the mean flexion degree was 120.6° (range 110°-135°) and the mean extension degree was 2.5° (range 0°-5°). The average visual analogue scale score was 1.6 points (range 0-3). Six patients were assessed as excellent and 2 as good according to the hospital for special surgery knee score system. There were no superficial or deep infections, or hardware breakages. No patient had giving way or locking of the knee, though some had intermittent pain and swelling after strenuous exercise. Injury mechanism had significant influence on the functional outcome (P=0.046).

CONCLUSIONHeadless compression screws combined with back buttress plate and/or autologous bone grafting to treat old Hoffa fracture is one of effective measures. It would be conducive to not only fracture healing but also early exercise and functional recovery.

Adult ; Bone Plates ; Bone Screws ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate the effects of basic fibroblast growth factor (bFGF) on the expression of glial fibrillary acidic protein (GFAP) after tractive spinal cord injury in rats and to explore the recovery of spinal cord function.

METHODSThe rats were subjected to tractive spinal cord injury at T13-L2. Cortical somatosensory-evoked potential (CSEP) was closely monitored and when P1-N1 wave amplitude decreased to 70% of that before operation, a small-bore catheter was inserted below the injured plane through subarachnoid cavity. In the treatment groups, 20 microl of bFGF solution (containing 20 microg of bFGF) was injected through the catheter right after the operation and 1, 2, 3, 4, 8, 12 and 24 h postoperatively. In the control group, same volume of normal saline was injected and every four rats were killed at 1, 4, 7, 14 and 21 d after the operation. Combined behavior score (CBS) and electro-physiological examination were adopted to evaluate function recovery. Expression of GFAP was observed by immuno-histochemical staining and was analyzed quantitatively by computer image analysis.

RESULTSThere was statistically significant difference in GFAP-positive cells between bFGF treatment group and the control group (P<0.01). Similar tendency was indicated by the results of CBS and CSEP.

CONCLUSIONSbFGF can induce large expression of GFAP after tractive spinal cord injury in rats and promote spinal function recovery, which is highly important for spinal cord regeneration.

Animals ; Disease Models, Animal ; Evoked Potentials, Somatosensory ; drug effects ; Fibroblast Growth Factor 2 ; pharmacology ; Glial Fibrillary Acidic Protein ; drug effects ; metabolism ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; Reference Values ; Spinal Cord Injuries ; metabolism ; physiopathology ; Traction