BACKGROUND:To improve the survival rate of transplanted tissue, most scholars focus on cel therapy, particularly cel-assisted fat grafting.
OBJECTIVE:To analyze the effect of recombinant adenovirus-mediated hypoxia inducible factor 1alpha (HIF1α) mutant on survival rate of transplanted fat particles through transfection of adipose-derived mesenchymal stem cels.
METHODS:Recombinant adenovirus-mediatedtriple-mutantHIF1αwas inserted into an adenovirus pAdEasy-1 system, folowed by viral packaging and titer determination. Human adipose-derived mesenchymal stem cels were cultured, passaged and identified, and subsequently transfected with three kinds of viruses and blank vector (experimental group with transfection of the triple mutantof HIF1α;positive control group; negative control group; blank control group). Transfection efficiency was determined using enhanced green fluorescent protein labeling. Additionaly, MTT assay was used to detect cel proliferation.
RESULTS AND CONCLUSION:The recombinant adenovirus was successfuly constructed and packaged in line with transfection requirements. Moreover, adipose-derived mesenchymal stem cels were successfuly identified by adipogenic, osteogenic and chondrogenic induction and could be used asseed cels for subsequent experiments. RT-PCR results showed that HIF1α mRNA expression in the experimental group and positive control group was significantly higher than that in the other two groups (P< 0.05). Western blot analysis showed that the relative absorbance value in the experimental group was significantly higher than that in the other three groups (P< 0.05). A significant increase in the cel proliferation was found in the experimental group, significantly different from the other three groups (P<0.05). Therefore, our findings indicate that transfection of adenovirus-mediated triple-mutant HIF1α not only can sustain the expression of target protein in transfected adipose-derived mesenchymal stem cels under normoxic conditions, but also can promote the proliferation of transfected adipose-derived mesenchymal stem cels.

Objective To evaluate and compare the presence of the nerve root sedimentation sign in patients with lumbar spondylolisthesis (LS) and lumbar disc herniation(LDH). Methods One hundred and fifty-one patients with degen?erative lumbar spinal disease treated by surgery from July 2012 to March 2014 were reviewed retrospectively in Tianjin Hos?pital. All the patients were divided into two groups:LS group (48 cases) and LDH group (103 cases). The clinical outcomes were evaluated by Japanese Orthopedic Association (JOA). The probability of positive sedimentation sign was compared be?tween LS group and LDH group. Correlation between the JOA score and nerve root sedimentation sign were analysed in two groups. Results There was no statistical difference in JOA score between LS group and LDH group (15.83±3.57 vs 16.76± 3.10, t=1.624, P=0.107). A positive sedimentation sign was identified in 39 patients in the LS group (81.25%) but in 65 pa?tients in the LDH group (63.11%). The difference between LS group and LDH group was statistically significant(χ2=5.028, P=0.031). In LS group, JOA scores of patients with positive nerve root sedimentation sign were lower than those of patients with negative nerve root sedimentation sign(15.33±3.50 vs 18.00±3.20, t=2.092,P=0.042);but in LDH group, there was no statistical difference in JOA scores of patients between positive nerve root sedimentation sign and negative nerve root sedi?mentation sign (16.40±3.13 vs 17.37±2.99, t=1.539, P=0.127). Conclusion A positive sedimentation sign also occurs in pa?tients with LDH, but positive nerve root sedimentation sign are more likely to appear in patients with lumbar spondylolisthe?sis who have worse clinical symptoms.

Objective We investigated the value of endoscopic evacuation and craniotomy of the hypertensive in?tracerebral hemorrhage to determine which methods are more suitable for the patients. Methods One hundred twenty pa?tients with hypertensive intracerebral hemorrhage participated this study. They were divided into classic surgical evalua?tion group (n=60) and endoscopic surgical evaluation group (n=60) according to their corresponding surgery strategies. Each patient was assessed by the preoperative Glasgow Coma Scale (GCS), the mean rate and time of hematoma evacua?tion from onset to operation, the postoperative GCS, the mean time of admission in neuro-intensive care unit (NICU) and Glasgow Outcome Score (GOS) at 3 month after surgery. Results The continuous (≥3 months) follow-up surveys were all completed by 120 patients. There was no statistical difference in clinical data before operation between two groups (P>0.05). However, clearance of hematoma was much faster and more efficient in endoscopic surgical group than in classic surgical evaluation group (1.5 ± 0.4 vs.3.9 ± 0.6 h, P<0.01; 95.84 ± 2.72% vs.87.48 ± 7.84%, P<0.01). The GCS scores were 10(6,12),12(8,13) and 13(10,13) in endoscopic surgical group whereas were 6(5,9),7(5,11).8(5, 12) in craniotomy group at 1,3 and 7 d followed operation. GCS scores were higher in surgical group than in craniotomy group at all time points (P<0.01). In addition, patients receiving endoscopic treatment showed a shorter NICU admission time than those receiving craniotomy (3.55±4.21d vs. 9.10±4.72d, P<0.01). The intracranial infection and hypostatic pneumonia were sig?nificantly lower in endoscopic than in craniotomy surgery group (0 vs.6 cases; 5 vs. 41 cases, P<0.05). The endoscopic treatment significantly improved the GOS score compared with craniotomy [3(3, 4)vs. 2(2, 3)] (P<0.01). Conclusion Endoscopic evacuation of hematoma for hypertensive intracerebral hemorrhage is efficient and minimally invasive, which is superior to craniotomy.

This article introduces the general concepts and methods of sample size estimation and testing power analysis. It focuses on parametric methods of sample size estimation, including sample size estimation of estimating the population mean and the population probability. It also provides estimation formulas and introduces how to realize sample size estimation manually and by SAS software.

ABSTRACT:Objective To investigate changes in the neutrophils in rats with D-galactosamine (D-GalN)-induced acute liver failure (ALF)and to explore the therapeutic effect of interventions treatment of neutrophils on ALF.Methods Liver function,the expressions of inflammatory cytokines TNF-αand IL-1β,and the changes of neutrophils in the peripheral blood and the liver were observed in rats with D-GalN (intraperitoneal injection)-induced ALF.SD rats were randomly divided into three groups when treated with intervention of neutrophils:control group,ALF group (intraperitoneal injection of D-GalN),and treatment group (intravenous injection of anti-PMN serum via tail vein 24 h before modeling).Biochemical analysis was used to detect serum ALT,AST, TBIL and blood ammonia.Hematology analyzer was applied to analyze the number and percentage of peripheral blood neutrophils.The number of neutrophils in the liver was evaluated by immunohistochemistry.Liver RT-PCR was adopted to detect the mRNA expression of inflammatory cytokines TNF-αand IL-1β.Results We found that 6 h after D-GalN injection,serum ALT,AST,TBIL and blood ammonia in ALF rats were significantly increased (P <0.05).The mRNA expression levels of inflammatory cytokines TNF-αand IL-1βin the liver reached the peak at 6 h after modeling (P <0.001),and it was still notably higher at 24 h than before modeling (P <0.001 ).The number and percentage of peripheral blood neutrophils and the number of neutrophils in the liver were all markedly increased 12 h after modeling (P <0.001 ),and the increase continued at least until 24 h (P <0.001 ).24 h after intravenous injection of anti-PMN serum via tail vein,ALF rats had a distinct decrease in the number of peripheral blood neutrophils and neutrophils in the liver 24 h after modeling (P <0.001).Meanwhile,serum ALT,AST,TBIL and blood ammonia were all greatly decreased compared with those in ALF group (P <0.05);a significant reduction of hepatocyte apoptosis was observed.Also,the expressions of TNF-α and IL-1β in the liver were remarkably decreased after treatment (P <0.05).Conclusion Neutrophils accumulated in peripheral blood and liver of rats with D-GalN-induced ALF.The treatment of anti-PMN serum may have a therapeutic effect on liver function and immune microenvironment in ALF rats.

Objective To explore the relationship of the radiological characteristics and clinical symptoms of ossification of the posteriorlongitudinal ligament (OPLL) of cervical spine. Methods 200 OPLL patients were recruited into this study. The data of JOA scores of theOPLL patients were collected. The stenotic rate of the cervical spine canal was calculated by the sagittal plane CT. The sorts and ossificatedcervicle were also identified by the images. Results The stenotic rate of the cervical spine canal was 19.8% to 60.3% with average 42.9%,the JOA scores was 4 to 15, averaged 10. Linear regression showed the stenotic rate of the cervical spine canal was negatively correlatedwith the JOA scores (P<0.01). There were 94 cases for segmental-type, 46 for mixed-type, 23 for continuous-type and 37 for focal-type. Thethird and fourth cervical vertebrae were susceptible to ossify. Conclusion The higher the OPLL occupation ratio, the worse the clinicalOPLL symptoms. The OPLL patients following clinical symptoms with spinal cord oppression are more likely the segmental-type.

Objective To explore the relationship between the somatosensory evoked potential (SEP) diversification during operation and the short term outcome of ossification of posterior longitudinal ligament (OPLL) of cervical spine. Methods 70 OPLL inpatients from February 2008 to February 2011were included in this study. All patients received anterior cervical operation. According to the diversification of the SEP during surgery, the patients were divided into 2 groups. The preoperative and postoperative JOA scores of the OPLL patients were collected. Results There were 35 patients in the SEP no change group and 30 in SEP improved group. There was no difference in the preoperative JOA scores between them (P>0.05), but was significantly different in the postoperative JOA scores (P>0.05). Conclusion There is some relationship between the SEP diversification during operation and the short term outcome of OPLL of cervical spine.

ObjectiveTo investigate the proliferation of rat's osteoblasts promoted by mechanical strain via the protein kinases C ξ(PKCξ) protein path-way.MethodsOsteoblasts were retrieved from SD rats' skulls in the sterilized environment.BioDynamic testing instrument was used to exert 2％ and 0％ mechanical strain on rats' osteoblasts for 180 min on the each group(2％ mechanical strain was experimental group and 0％ mechanical strain was control group); and the same method was applied on the rats' osteoblasts which pretreated with the classical PKC ξ inhibitor with 2％ mechanical strain (inhibitor group).The cell cycles of rats' osteoblasts were measured by flow cytometry; and Western blotting and immunofluorescence were used to detect expression of protein PKC ξ and phosphorylationed PKC ξ in the rats' osteoblasts.ResultsThe mechanical strain obviously increased the ratio of S period in the cell cycles.Compared to the control group and inhibitor group,a significant increase of the expression of protein PKC ξ and phosphorylationed PKC ξ in the osteoblasts was detected in the mechanical strain experimental group.ConclusionPKCξ can respond to the stimulus of the mechanical strain,which promotes the proliferation of osteoblasts through PKCξ pathway.And it plays an important role in the process of signal conduction.

ObjectiveTo measure the displacement parameters of the femoral head in space through CT three-dimensional reconstruction so as to re-understand impacted femoral neck fractures.MethodsFifty patients with impacted femoral neck fractures were included in the study.There were 17 males and 33 females.Bilateral proximal femurs of each patient were scanned by spiral CT preoperatively.Then these primitive CT data were reconstructed by Mimics 10.01 software.Registered the normal femur and fracture mirror model,the registered mirror model was generated a new mask in the CT cross-sectional images,marked key points in the masks and measured the displacement parameters of femoral head with three-dimensional measurement technology.ResultsIn the impacted femoral neck fractures,the femoral head displacement angle was 17.17°±10.40°,70％(35/50) of the patients had femoral head rotation angle was at 10° to 35°,and 30％(15/50) of the patients had femoral head space displacement more than 20°.The average displacement distance of femoral head center and the deepest point of fovea capitis were(6.49±3.60) mm and(10.42±5.92) mm,respectively.A high proportion of the impacted femoral neck fractures had a large of displacement in the three-dimensional space.It is not scientific to equal impacted femoral neck fractures to undisplaced fractures.ConclusionThree-dimensional reconstruction and digital measurement is a precise,efficient and convenient method for the measurement of femoral head displacement parameters in femoral neck fracture patients.The classic Garden classification for impacted femoral neck fractures has certain defects and limitations.Clinicians should re-understand impacted femoral neck fractures and choose more reasonable treatment.

Objective To investigate the effects of trichostatin A(TSA)on the mice model of collagen induced arthritis(CIA).Methods Mice model of rheumatoid arthritis(RA)was induced in DBA/1 mice with type Ⅱ collagen.Paws were scored for histological severity of arthritis.The severity of inflammation of mouse joint was evaluated by histological examination.Real-time PGR was used to determine the cytokine mRNA expression.Cytokine production was measured by ELISA from serum,spleen cell culture or dendritic cell and T cell co-culture supematant.T cell proliferation was examined by MTT method.Results TSA can significantly suppress the severity of the arthritis in CIA.IFN-γ was elevated in CIA mice,but was inhibited significantly by TSA introduced either at the same time with immunization or at the onset of manifestation of arthritis.Collagen specific T cell proliferation was significantly suppressed by introduction of TSA.Increased level of IL-4 by T cells was observed in TSA treated group compared to that of control group.Conclusion IL-4 level was increased and played a critical role in the protective effects of TSA in CIA.TSA suppresses the progress of CIA by regulates the balance of Th1/Th2 differentiation.