To date, three techniques of total spondelyctomy were used for spinal tumor, including piecemeal technique, eggshell technique and total en bloc spondelyctomy, and each has specific features and indications. Due to limitations of technique, piecemeal technique and eggshell technique are gradually replaced by total en bloc spondelyctomy in treatment of spinal tumor. Moreover, with technical development, application of total en bloc spondelyctomy in treatment of spinal tumor becomes more rigorous and standard. However, it is controversial about its indications and surgical approach. Postoperative spinal stability reconstruction is achieved through various methods according to different conditions.

Dendritic Cells ; Humans ; Rhinitis, Allergic, Perennial ; Rhinitis, Allergic, Seasonal

BACKGROUND:Tissue engineering requires a lot of seed cells. Osteoblasts have become important seed cells in bone tissue engineering. However, it is difficult to culture the osteoblasts, and cellnumber, purity, proliferation and differentiation activity are different obtained by different culture methods. OBJECTIVE:To identify and compare three common primary osteoblat culture methods, and to explore a method for the primary culture of osteoblasts which is easy to operate, economical and effective, in order to provide basis for the further experimental research. METHODS:Calvarias were dissected from newborn Sprague Dawley rats in 72 hours, and osteoblasts were isolated with col agenase digestion method, sequential digestion method and bone tissue method respectively. The morphological observation and cytochemical staining were performed, the growth curve of the cells was drawn with cellCounting Kit-8 method, and the rate of living osteobalsts was counted with trypan blue staining. RESULTS AND CONCLUSION:The proliferation of the insolated and cultured osteoblasts was wel with typical characteristics of osteoblasts, cytochemical staining results were positive. Compared with the sequential col agenase digestion method, the col agenase digestion method presented higher production of osteoblasts and higher cellsurvival rate (P<0.05), and the col agenase digestion method was easier than the sequential col agenase digestion method and cost less than sequential col agenase digestion method. Bone tissue method was the easiest method with less damage to cells, but bone tissue method presented lower production of osteoblasts and cost much more time, which cannot be used in large-scale osteoblast culture. The col agenase digestion method is a simple, efficient and ideal method for isolation and culture of primary osteoblasts.

Objective To confirm the role of human parvovirus B 19 ( B19) infection in childhood immune thrombocytopenia ( ITP) patients.Methods A total of 416 cases of newly diagnosed childhood ITP patients from Jan .2011 to Dec.2013 had been summarized to be cases group , Then a total of 130 childhood patients with common respiratory tract infection were selected randomly as the control group . All patients had been divided in grouped by age as <1 years group(n=187), 1-3 years group(n=127), 3-7 years group(n=71), and 7-14 years group(n=31).The incidence rate of B19 infection in all age groups, the prognosis conditions in B19 infection positive and negative groups after the same therapy protocol were obserred .Results The B19 infection rate had no statistical significance in re-search groups and control groups .And no significant difference in the same age groups compared with control groups (all P>0.05) was found.All the ITP patients had not been given anti -B19 treatment.The PLT remission rate,respectively, after treated with the same pro-tocol including glucocorticoid and/or immunoglobulin had a declining trend as the ages increasing .The B19 infection groups of all ages al-so had no significant difference PLT remission rate had been confirmed in non -B19 infection patients in each age groups (all P>0.05). Conclusions B19 infection may not be a major cause in childhood newly ITP , and treatment protocol with no anti -B19 treatment had no influence on the clinical curative efficacy .

Objective To construct prokaryotic expression vector of XTP3 gene and induce the expression of fusion protein in E.coli,and prepare XTP3-specific rabbit polyclonal antibody,detect the specificity of the antibody in hepatic carcinoma tissue and normal liver tissue.Methods DNA fragment of XTP3 amplified by PCR was inserted into pET-32a(+) to construct prokaryotic expression vector pET-32a(+)-XTP3.After identified by sequencing,pET-32a(+)-XTP3 was transformed into E.coli BL21 and induced with IPTG.After analyzed by SDS-PAGE and Western blotting,the induced expression product was purified and renatured by Ni+ affinity column chromatography.The purified protein was used to immunize New Zealand rabbits to gain polyclonal antibody,and the polyclonal antibody was then detected by ELISA,immunohistochemistry and Western blotting.Results Prokaryotic expression vector pET-32a(+)-XTP3 was successfully constructed,and the XTP3 fusion protein of about 52kD was highly expressed in E.coli.DS-PAGE showed that the protein product was mainly in inclusion body.The purified protein and polyclonal antibody were obtained successfully.It was manifested by ELISA that the titer of polyclonal antibody was over 1∶128 000.Immunohistochemistry showed that XTP3 antibody presented membrane-positive in hepatic carcinomous tissue.Conclusions The recombinant XTP3 protein and polyclonal antibody have been obtained successfully.These results lay a foundation for studying the immuneogenicity and bionomics of XTP3 protein.

Objective To incestigate the fetures of cranial CT and MRI in the patients with hypertensive encephalopathy. Methods The CT and MRI findings of ten cases of hypertensive encephalopathy with the charge of CT,MRI appearance of FLAIR (fluid attenvated inversion-recovery), DWI(difussion weighted imaging),ADC(apparent diffusion coefficient) were analyzed retrospectively. Results Of ten patients,3 cases had abnormal finding in the cranial CT;10 cases had abnormal finding in the cranial MRI,the lesions were demonstrated as slightly hypointensity on T1WI and slightly hyperintensity on T2WI and remarkably hyperintensity on FLAIR,and iso or slightly hyperintensity on DWI,and remarkably hyperintensity on ADC.The lilateral parietal occipital lobes and cerebellar hemisphere and Brain Stem were the more common sites. Conclusions The only characteristric finding of hypertensive encephalopathy in MRI and CT imaging studies is vasogenic edema,especially in the subcortical white matter of the parietal and occipital lobes bilaterally,and cereballar hemisphere et al;especially FLAIR,DWI and ADC of MRI can be helpful for diagnosis and diffenential diagnosis,prognosis and curative effect of hypertensive encephalopathy.

OBJECTIVE To understand the distribution or change and drug resistance of the bacteria flora in nosocomial infection,in order to provide laboratory evidence for controlling nosocomial infection and to indicate clinical antimicrobial agents usage.METHODS All isolates were identified by routine procedure and VITEK microbe automatic system and API identified system.Drug susceptibility test was used K-B paper disk diffusion method in accordance with the CLSI/NCCLS standards.RESULTS There were 11 464 strains of bacteria which were obtained from 2002 to 2006.Sputum,secretion and middle urine were the major specimens.The major bacteria floras were Pseudomonas aeruginosa,Escherichia coli,Acinetobacter,Enterococcus,Staphylococcus and Candida albicans,whose isolating rates were 57.02% in 2002,64.46% in 2003,57.83% in 2004,57.49% in 2005,and 60.73% in 2006.E.coli and Klebsiella pneumoniae produced ESBLs rates were from 39.87% to 61.98% that drug resistance rates to cefoperazone/sulbactam and imipenem were 14.06% and 0.64%.The drug resistance rates of nonferment Gram-negtive bacteria to cefoperazone/sulbactam were below 33.22%,Gram-positive cocci to vancomycin was below 0.34%.CONCLUSIONS Now,the important bacteria flora is nonferment Gram-negative bacteria in nosocomial infection.The P.aeruginosa,E.coli,Acinetobacter baumannii,Enterococcus faecalis,C.albicans,and meticillin-resistant staphylococcus(MRS) were prevalent important bacteria in nosocomial infection.The situation of producing ESBLs in Enterobacteriaceae is very severe.Glycopeptides are the best choice to MRS.To zymogenic bacteria and the nonferment Gram-negtive bacteria that cause severe infection,combining-drug treatment can be chosen according to drug susceptibility test results.

Objective:To explore the relationship between YKL-40 and the proliferation of breast cancer and its mechanism.Methods:The expression of YKL-40 in breast cancer MCF-7 cells was detected by immunofluorescence assay.Fluorescence microscope was used to observe the conversion efficiency,and Real-time PCR was used to screen the most effective YKL-40 siRNA.Expression levels of PI3K,P-PI3K,AKT,P-AKT of the PI3K/AKT pathway associated proteins was test by Western blot.At the same time,MTT and flow cytometry were validated by YKL-40 siRNA treatment of human breast cancer MCF-7 ceils,the differences of 24h,48h and 72h groups of cell proliferation ability and cell cycle.Result:MCF-7 cell express YKL-40 protein,mainly located in the cytoplasm.Real-time PCR show that siRNA01,siRNA02,siRNA03 compared with NC group YKL-40 gene silencing effect is remarkable.Among of them the strongest silencing effect is siRNA02 (P＜0.01),Western blot show the experimental group than the control group,Total PI3K and AKT remain unchanged while P-PI3K,P-AKT expression decreased (P＜0.05).In the experimental group,the number of G1 cells in the control group was increased (P＜0.01),while the S phase cells decreased (P＜0.01).MTT results showed that the experimental group compared with the control group,the proliferation ability is decreased(P＜0.01).Conclusions This study suggests that YKL-40 can be used as the upstream regulatory factor of PI3K/AKT signaling pathway and affect the process of cell cycle in breast cancer,and then regulate the proliferation of breast cancer,YKL-40 may be a crucial target for the treatment of breast cancer.

objective To examine nuclear transIocation of peroxisome proliferator-activated receptor γ(PPARγ)in rats following focal cerebral ischemia/reperfusion(I/R),and to explore the significance of altered PPARγ,nuclear translocation in ischemic brain injury.Methods Healthy adult male SD rats underwent 60-min cerebral artery occlusion followed by reperfusion of 4,8,or 24 h,respectively.The cytoplasmic-to-nuclear shuttling of PPARγ was characterized by Western blot,immunohistochemical and immunofluoreseence staining.The effects of PPARγ agonist rosiglitazone (Ros) and antagonist GW9662 on I/R-induced PPARγ nuclear translocation were also examined in the present study. Furthermore,TTC staining war adopted to determine the change in cerebral infarction volume. Results (1)Western blot analysis revealed an increase of PPARγ in the nucleus and a simultaneous reduction in the cytosol following ischemia and reperfusion for 4 h(tcytosol=9.03,tmuclear=27.19,P=0.00).Prolonged the reperfusion further enhanced this I/R induced PPARγ translocation in a time-dependent manner.Using immunohistochemistry and immunofluorescence,nuclear PPAR γ positive staining increased from 48.3%in the sham control to 80.3% following ischemia and reperfusion for 24 h.(2)Western blot analysis revealed that PPARγ agonist Ros further increased I/R-induced nuclear enrichment of PPARγ,whereas PPARγ antagonist GW9662inhibited I/R-stimulated change in PPARγ.(3)When compared to the L/R group using TTC staining,Ros treatment significantly decreased the infarction volume by 48.40%(15.46±4.94 versus 29.96±3.39,t=5.93.P=0.00),whereas GW9662 increased by 58.95%(47.62±4.93 versus 29.96±3.39,t=7.23,P=0.00).Conclusions Cerebral I/R injury induces PPARγ translocation from the cytosol to the nucleus.This change may represent an intrinsic neuroprotective response against brain I/R injury.

BACKGROUND:Topping-off technique can be used for fixation treatment through the combination of fusion and interspinous dynamic device, in order to prevent or slow down the adjacent lumbar segment degeneration.
OBJECTIVE:To obverse the protective effect of Topping-off technique (posterior lumbar interbody fusion
procedure combined with the fixation of dynamic interspinous device Coflex) for the degenerative intervertebral disc. METHODS:A total of 32 patients with degenerative lumbar diseases who had been treated with Topping-off technique were included in this study. The Oswestry disability index, the Japanese Orthopaedic Association scores, range of motion for Coflex implanted segment and during the relative signal intensity of the Coflex implanted segment in MRI image were recorded and calculated preoperatively and the entire fol ow-up period.
RESULTS AND CONCLUSION:Al patients were fol owed-up for 20.6 months averagely. Up to the last fol ow-up, the Oswestry disability index and Japanese Orthopaedic Association scores were significantly improved when compared with those before treatment (P<0.001). There was no significant difference in the range of motion for Coflex implanted segment before and after treatment (P=0.19). The relative signal intensity of the Coflex implanted segment was significantly improved when compared with that before treatment (P<0.01). The clinical application of the Topping-off technique showed a protective effect on the intervertebral disc.