0.05).Under SEM,the BMSCs showed good adhesion to beta-TCP with obvious proliferation.Conclusion BMSCs can grow and proliferate well on the compound BMSCs/beta-TCP and beta-TCP has good biocompatibility with BMSCs in vitro,which may be used as a good scaffold material for injectable tissue engineering bone.

GRP78,as endoplasmic reticulum resident chaperone,is highly induced by a variety of tumor microenvironmental stresses,such as hypoxia and glucose deprivation.GRP78 is implicated in tumor cell proliferation,apoptosis,invasion,metastasis and angiogenesis.Recent evidence indicates that GRP78 levels is correlated with pathological grade,stage and prognosis for the majority of solid tumors.In addition,GRP78 plays an important role in drug tolerance and knockdown of GRP78 has been shown to sensitize malignant cells.GRP78 could not only be a good biomarker to predict cancer progression and chemo-responsiveness,but also an appealing target for the development of a more selective chemotherapy.

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 report our clinical outcomes of treating tibial defects combined with soft tissue defects using Ilizarov technique.Methods From May 2010 to February 2015,52 patients with combined bone and soft tissue defects of the tibia were treated at our department.They were 41 males and 11 females,aged from 19 to 65 years (average,37.7 years).By Gustilo classification,49 cases were type ⅢB and 3 type ⅢC.The areas of soft tissue defect ranged from 7 cm ×3 cm to 28 cm × 15 cm,and the tibial defects ranged from 5 cm to 15 cm in length (average,12.6 cm).The schemes of Ilizarov technique depended on the location and size of the tibial defects.Open wound dressing combined with bone transport was adopted in 21 cases,limb shortening followed by bone lengthening with compression at the fracture ends in 12 cases,and tissue flap transplantation combined with bone transport or lengthening in 19 cases.Results The follow-up time of the 52 patients ranged from 13 to 61 months (average,27.1 months).The distance of bone transport or lengthening ranged from 5.0 cm to 13.6 cm (average,10.8 cm);the bone transport speed averaged 0.81 mm/day.The tibiae united in all the 52 patients;the time for external fixation ranged from 13 to 21 months (average,15.3 months);the external fixation index was 2.3 months/cm.According to the Paley functional criteria,23 cases were excellent,19 good,9 fair,and one poor,yielding an excellent to good rate of 80.7％.Conclusion According to the location and size of the bone and soft tissue defects of the tibia,the 3 schemes of Ilizarov technique can be rationally chosen to obtain fine clinical outcomes.

Objective To investigate the effects of miR-135a on HOXA10 expression,proliferation and apoptosis of SKOV3 cells.Methods (1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined.(2) miR-135a mimics,miR-135a inhibitor and negative control were transfected into SKOV3 cells,respectively.Reverse transcription (RT)-PCR,western blot analysis were used to examine the expression levels of HOXA10 at different times (24,48 and 72 hours).(3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10.(4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium(MTT) assay [quantified by absorbance(A)].Western blot was used to examine the expression of apoptosis-associated protein bcl-2,bax and caspase-3 in SKOV3 cells after 48 hours transfection.Results (1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms.(2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24,48 and 72 hours) after miR-135a mimics transfectionin SKOV3 cells (0.94 ±0.04 vs 0.78 ±0.03 vs 0.70 ±0.03,P ＜0.05).While,the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ±0.03 vs 2.60 ±0.08,P ＜0.05).After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours,the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group,respectively (all P ＜ 0.01).Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected.(3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10,luciferase reporter assays showed that the luciferase activity reduced by 67.8％ (P ＜0.01).(4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05,0.67 ±0.05 vs 0.75 ± 0.06 ; respectively,all P ＜ 0.05).While,SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06,P ＜ 0.01).After miR-135a mimics transfection,the level of bcl-2 protein was significantly lower than that in control group (0.28 ±0.06 vs 0.76 ±0.09,P ＜0.01).The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1,P ＜ 0.01).While,there was no statistical difference of bax expression (P =0.142).However,after miR-135a inhibitor transfection,the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09,P =0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1,P ＜ 0.01).There was also no statistical difference of bax expression (P =0.066).Conclusion miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.

Objective To assess the efficacy and safety of intraoperative wake-up test during surgery for ossification of the ligamentum flavum(OLF). Methods Between June 2004 and June 2010,35 patients(23 males and 12 females,at age range of 32-71 years,mean 50.5 years) with OLF underwent posterior decompressive laminectomy and excision of OLF.The operation wag performed on the cervical vertebrae in four patients,on the thoracic vertebrae in 24 and on the lumbar spine in seven,with monosegmental OLF in 14 patients and multisegmental OLF in 21.The outcomes were evaluated using Japanese Orthopaedic Association (JOA) scale. Results The operation lasted for mean 190 minutes (120-260 minutes),with the blood loss for mean 350 ml (220-520 ml)and the mean 2.4(1-5) lamin receiving decompression.The follow-up period ranged from 6 to 72 months (mean 29.5 months).The JOA score was improved from 4.302±1.023 preoperatively to 8.327±1.032 at the final follow-up (P<0.01).The neurological symptoms was aggravated during operation in three patients,for whom the muscles force recovered gradually in two patients but one showed no change after special measures.No serious complication was found in the other patients postoperatively. Conclusions The wake-up test can serve as an effective monitoring during the procedure ofthe decompressive laninectomy in the patients with severe OLF and decrease the occurrence of neurologic deficit complications.

Objective To investigate the benefit of inferior vena cava filter on the treatment of deep vein thrombosis(DVT) and pulmonary embolism.Methods From January 2009 to September 2011,of the 115 patients with DVT,27 cases were treated by VCF under DSA,followed by thrombolysis and anticoagulation.Results All the cases were.successfully implanted with VCF and limb detumescence was achieved after thrombolysis and anticoagulation treatment without complications.Conclusion The implantation of inferior vena cava filter combined with thrombolysis and anticoagulation is favorable and safe to treat deep vein thrombosis and pulmonary embolism,and it is also effective to prevent PE in patients with deep venous thrombosis in the low extremity.

AIM:To investigate the construction of oligonucleotide microarray specialized for pancreatic adenocarcinoma-associated genes and its preliminary application of detecting differential expressed genes in pancreatic cancer.METHODS:Pancreatic cancer related genes were purposely selected,and oligonucleotide microarray was prepared by spotting oligonucleotide probes on glass slides coated with APS-PDC.Labeled cDNA targets for hybridizations were synthesized by reverse transcription from total RNA in the presence of Cy5-dCTP and Cy3-dCTP,respectively.Hybridized microarray was scanned by Agilent laser scanner,and the aquired image was analyzed by Imagene3.0 software.The intensity ratios of Cy3 and Cy5 were calculated.To confirm the expression profiles of these genes,quantitative reverse transcription-PCR (QRT-PCR) was carried out for CDC25B and TUSC3 genes,and β-actin gene was taken as internal control.The product of PCR was quantitated by comparative Ct method.RESULTS:The signal of microarray hybridization was clear,and the images had a lower background and higher signal-noise ratio.In comparison with normal pancreas,twenty -four differentially expressed genes were identified which included seventeen up-regulated and seven down-regulated genes.The results of QRT-PCR demonstrated that the expressions of CDC25B and TUSC3 in pancreatic cancer were up-regulated and down-regulated respectively,which is consistent with microarray results.CONCLUSION:The oligonucleotide microarray specialized for pancreatic cancer is desirable for its specialty and sensitivity,which can simultaneously and parallelly detect multiple pancreatic cancer-associated genes.

Studies on NaCl concentration, pH tolerance and selectivity to different concent rations of Na +, K +, Mg 2+ and Ca 2+ of 43 isolates from th e saline soils in XinJiang, HeBei and QingHai Provinces of China and 4 type stra ins were performed in this paper Results showed that halotolerant actinomycete s have extensiv e adaptability to Na +, K + and Mg 2+ and only a few of them can grow in low CaCl 2 concentration Halophilic actinomycetes have extensive adaptability to Na +, and for most halophilic actinomycetes, Na + can be substituted by K + , Mg 2+ , but not for Ca 2+ For some halophilic actinomycetes , it is necessary to have Na + for their growth It also showed that the growth of al l halophilic actinomycetes had se lectivity with different concentration of Na +, K +, Mg 2+ So it is pre sumed that only Kaliumphilic or Magnesiumphilic Actinomycetes maybe exist in hig h salt environments In addition, the growth pH range were 6 0～9 0 and the o ptimum pH were 7 0～8 0 not only for halophilic but also for halotolerant acti nomycetes The dis tribution of halophilic actinomycetes also have some relativity to isolation sit es

BACKGROUND: Many experiments indicate that the angiogenesis of tissue engineered bone graft plays a key role in the osteogenesis.OBJECTIVE: An experimental pattern was set up designed to prepare a kind of vascularized engineered-bone graft for repairing rhesus tibia defects and analyze the relation of angiogenesis and osteogenesis in vivo by rontgenographic and morphological approaches.DESIGN: Random controlled animal experiment.SETTING: Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University.MATERIALS: The composite graft was constructed by seeding the induced bone marrow stem cells (BMSCs) on to a beta-tricalcium phosphate(3-TCP) scaffold in vitro, a circular cylinder (20 mm × 8 mm diameter) with a slit (width 2 mm and length 3 mm ) open to both ends and slot. Porosity 60% and pore diameter 100-150 μm. Twenty-nine healthy rhesuses aged 4-5 years and weighted 3.5-5 kg were adopted without gender limitation.METHODS: The experiment was conducted in the Department of Orthopaedics and Traumatology, Nanfan Hospital, Southern Medical University from October 2003 to July 2005. ①Bone-periosteum defect of 20 mm was made in the middle part of right tibia of the 27 rhesuses, and randomly divided into 3 groups equally. ②The defect gaps in fascia-blood vessel group (A) were plugged with in vitro engineered composites constructed by bone marrow stem cells and 3-TCP scaffold, which were totally hugged by a sheet of pedicled deep fascia and additionally a corresponding portion of saphenous artery and veins. The gaps in fascia group (B) and control group(C), however, were inserted with fascia-coated tissue engineered bone and tissue engineered bone only, respectively. Furthermore, two rhesuses without filling materials on the defect were picked up as blanks fixed by steel pins. ③The angiogenesis and osteogenesis for each treatment was assessed by radioactive imaging, roentgenographic analyses, blocking density and vaso-area image analysis at time intervals of 4, 8 and 12 weeks postoperative.MAIN OUTCOME MEASURE: The score of radioactive imaging,roentgenographic, morphological and vaso-area image analyses RESULTS: Totally 29 rhesuses were involved in the result analysis.① General observation of samples: In group A, all the surfaces of the implanted material and the central part were wholly wrapped up or replaced by bonelike tissues which were hard and could not be broken. And 2/3 materials had been absorbed; In group B and C, partial materials of the medial surface and the front were not coated or replaced by bonelike tissues, which could be broken with force, and 1/3 material had been absorbed.②Histological observation of scaffolds: With time passing, the scaffold materials were absorbed to different degrees in group A, B and C, among which, group A was most significant; Under the microscope, the implanted materials at 12 weeks were completely coated with the bonelike tissues, while the blood vessels structures in the materials were mostly alveoli alike and multi-braches. In group B, most of the materials at 12 weeks were wrapped up by the new bone, and few blood vessels could be seen in the center of the materials. In group C, the implanted materials at 12 weeks were slightly absorbed. The new bone and the vascular structures were both increased a little, but still very few.③Analyses of vaso-area: The vaso-areas of both central and peripheral parts in group A were significantly bigger than those of group B and C (P ＜ 0.05). Furthermore, it tended to increase with the time.④X-rays observation: At 12 weeks, group A's images presented obviously decreased density which was lower than that of the normal bone in individual areas and the continual bony callus manifested. Whereas group B and C's images showed slightly decreased density and the continual bony callus appeared on the sections. ⑤The roentgenographic scores of bone defects: The results indicates that the scores of group A was better than those of group B and C at 4, 8 and 12 weeks, respectively (P ＜ 0.05).CONCLUSION: ①This study shows that a feasible and effective angiogenesis approach of tissue engineered bone can accelerate osteogenesis in vivo. ②The absorption level is positively related to local angiogenesis.