Objective To constitute a higher expression system and to obtain the breast cancer cell strains in which BSP is stably expressed. Methods hBSP gene was subcloned from pB-hBSP vector by PCR. The PCR fragment was inserted into the eukaryon expression vector, pIRES2-EGFP, which allow exogenous protein and EGFP to express respectively. The recombinant vector, pIRES2-hBSP-EGFP, was transfected into human breast cancer cells with Lipofectamine TM 2000. The expression of symbol protein EGFP, could be conveniently observed with fluorescent microscope. Results The recombinant pIRES2-hBSP-EGFP plasmid was constituted and successfully transfected into breast cancer cells. In the breast cancer cell strain hBSP and EGFP were expressed. Conclusion The successful constitution and transfection of hBSP and EGFP nonfusion expression vector laid a foundation for the further study on the effect of BSP on breast cancer metastasizing to bone in vivo or in vitro.

Objective To explore the manifestation and diagnostic value of CT in retinoblastoma. Methods Twentythree cases of retinoblastomas proven by operation and pathologic examination were collected. Their CT images, clinical symptoms and pathologic results were retrospectively analyzed. Results Twentythree patients of retinoblastomas aged from 4 months to 9 years (mean 2.4 years). CT images showed soft tissue mass (shaped as node, crescentshaped or occupied the entire space of the eyeball) was in the globe of all cases. There was calcification in 19 cases (82.6%), including “small pinch” in 12 cases or “patches” in 7 cases. Five cases were found the optic nerve thickened (21.7%), and 1 case of intracranial invasion through the optic nerve canal (1/23), and 6 cases of retina detachment (6/23). The lesion showed mild or moderate enhancement after intravenous contrast medium were performed. Conclusion CT has an advantage on showing the size, shapes, intrastructures and invasive degree of the lesion, and is meaningful to the diagnosis and differential diagnosis of retinoblastoma.

Objective To study the neuroprotective effects of neuregulin-1?(NRG-1?) on the nervous behavioral function,cerebral infarction volume,brain water content(BWC),neuronal apoptosis and aquaporin-4(AQP-4) expression in astrocytes after cerebral ischemic reperfusion and the related mechanism in mice.Methods Intraluminal thread methods were applied to establish the middle cerebral artery occlusion reperfusion models in the mice.Neuregulin-1?(2?g / kg) was injected into the internal carotid artery for treatment.The nervous behavioral function was evaluated with Bederson's test.The cerebral infarction volume was observed with tetrazolium chloride staining.The BWC was measured by dry-wet weight comparing.The apoptosis positive cells were counted by immunofluorescence assay.The expression of AQP-4 was determined by immunohistochemical assay.Results Nervous behavioral malfunction appeared in all the mice with left middle cerebral artery occlusion and/or reperfusion.The infarction focus showed in the ischemic hemisphere after the injury.The BWC,the number of neuronal apoptosis cells and AQP-4 expression in astrocytes were higher than those in the sham group. In the NRG-1? treatment group,the nervous behavioral function was improved 24 hours after ischemia,the number of apoptosis positive cells reduced and the infarction volume decreased significantly compared with the control group(P0.05).In the groups of reperfusion for 22,46 and 70 hours,the five indexes mentioned above were significantly different from those in the corresponding control groups(P

Aim To investigate the method of culturing vascular smooth muscle cells (VSMC) of coronary arteries derived from experimental diabetic rats and to establish the model of VSMC for the study of chronic cardiovascular diseases of diabetes. Method After the establishment of the model of diabetic rats, the coronary arteries of rats were separated carefully to culture VSMC using the method of explanting culture. Results The primary culture of VSMC was performed successfully by the method mentioned above. The cells could be isolated by different kinds of digestive enzyme in three steps. 7 ~10 days afterward, the cells overlapped in layers and displayed the typical ‘hill and valley’ pattern and positive immunochemical staining for smooth muscle actin. Conclusion VSMC from experimental diabetes proliferates faster than normal VSMC and its cultivating condition is strictly limited. The morphology of diabetic VSMC is as same as that of normal rats.

Objective:To study the methods of early qualitative diagnosis of laryngeal displasia and earlycarcinoma. Methods:We investigated displasia (34 cases),squamous cell carcimoa (19 cases) and vocalcord polyps (17 cases) of larynx which were embeded in paraffin by PCR－SSCP analysis of p53 gene,im－munochemi stain of p53 protein and Ⅳ collagen,cytometer (determining fraction of cell cycle) and AgNORstain. Results :Eleven of 34 cases of displasia were all positive in the indexes. Seven of 11 cases were alsopositive in pathology,but others were negative. Five of 34 cases of displasia were all negative in the index-es,2 of 5 cases were positive and 1 of 5 cases were negative,other two cases did not have following-up re-sults in pathology. Conclusion :The results suggest that it is the combination of the morphological indexeswith biological indexe that is more scientific and more accurate in quatitative diagnosis of premalignantchange.

Objective To analyze the ultrasonic imaging characteristics of primary breast lymphoma and their significance for diagnosis.Methods Data of ultrasonograph from 21 cases of primary breast lymphoma were reviewed, and all cases had been certificated by pathology.Results All of 21 cases with primary breast lymphoma were solid mass.The contours of 16 cases were regular, and the boundaries of 9 cases were clear.20 cases were hypoechoic masses, besides 3 cases with lower echo like liquefactive degeneration.The internal echoes of 19 cases were disorderly, part of them with hyperechoic trabs.The rear echoes of 17 cases were neither enhanced nor fading.The surrounding tissues of 19 cases were not changed.13 cases had more than 3 blood vessels on color Doppler imaging.Conclusion There are some ultrasound characteristics in primary breast lymphoma, which would be helpful in the diagnosis of primary breast lymphoma.

Objective Nutrition becomes a focus to professionals as an important factor during the development of pressure ulcers.However, there is no evidence for the effectiveness of eternal nutrition support for pressure ulcer prevention .This research aimed to evaluate the effectiveness of eternal nutrition support for pressure ulcer prevention and provide evidence for patients with the risk of pressure ulcer . Methods Key words such as pressure ulcer prevention,eternal nutritional supportwere used to search information in the Cochrane Library , JBI Database of Systematic Reviews , MEDLINE, Web of Science, EMbase, CBMdisc, CNKI and Wanfang Database to collect randomized controlled trials (RCTs).The quality of RCTs was critically appraised and data were extracted by 3 reviewers independently .Meta-analyses were conducted for the eligible RCTs using RevMan 5.0 software, and Stata12.0 software was applied to detect bias or sensitivity analysis to determine the source of heterogeneity if necessary . Results 7 RCTs were includ-ed, among which 1 RCT was excluded by sensitivity analysis .6 RCTs ( n=1553 ) were finally included for meta analysis ( n=728 in trail group, n=825 in control group), and all the patients were given oral nutrition support for 7days to 26 weeks.The results showed that the ulcer incidence in the intervention group reduced by 16%in comparison to the control group (P=0.006).The protein and energy intake in intervention group were higher than those in control group , and the length of stay in hospital was shortened by 16 days. No significant difference was found in the albumin values . Conclusion The enteric nutrition support can reduce the incidence of pressure ulcer obviously with the highest level of evidence , IA .

Objective To explore the effect of different ages on behavioral dimensions of anxiety animal models in Kunming mice.Methods Young/Old,male,Kunming mice were recorded twice,for five minutes,with a one-week inter-trial interval in open field test (OFT),elevated plus maze (EPM) and light/dark box (LDB).The following parameters were evaluated:percentage of time spent in the OFT central area (Ctime％),percentage of number of squares visited in the OFT central area (Ccross％),the total number of horizontal squares in OFT (Cross),the total number of rears in OFT (Rear),percentage of time spent in the EPM open arms (Otime％),percentage of number of entries into the EPM open arms (Oentries％),the total number of entries into the EPM open and closed arms (Entries),percentage of time exploring in the LDB light area (Ltime％),percentage of number of squares crossing in the LDB light area (Lcross％),percentage of rears in the LDB light area (Lrear％),the total number of transition in LDB (Transition),the total number of horizontal squares in LDB (Cross) and the total number of rears in LDB (Rear).Results OFT-Ctime％,OFT-Ccross％,EPM-Otime％,EPM-Oentries％,LDB-Ltime％,LDB-Lcross％ and LDB-Lrear％ had significant loadings on anxiety factor; OFTCross,EPM-Entries,LDB-Transition and LDB-Cross loaded on locomotion-exploration factor.Old mice had higher anxiety emotion (Ctime％,t =3.12,P＜ 0.01; Ccross％,t =2.37,P＜ 0.01; Otime％,t =1.31,P＜ 0.05; Oentries％,t=1.57,P＜0.05; Ltime％,t=4.15,P＜0.01 ; Lcross％,t=3.28,P＜0.01 ; Lrear％,t=2.73,P＜0.01) and lower locomotion and exploratory activity (Cross,t =2.05,P ＜ 0.01 ; Entries,t =2.09,P ＜ 0.05 ;Transition,t=2.51,P＜0.01; Cross,t=3.02,P＜0.01),compared with Young.Conclusion Ages have no effect on structural dimensions of anxiety animal models.With the growth of ages,old mice have higher anxiety emotion but lower locomotion and exploratory activity.

A sequential extraction procedure has been proposed for the evaluation of the speciation of heavy metals including Cd,Cu,Pb and Zn in sediments from mariculture area,and the speciation of heavy metals was separated and defined as acid soluble fraction,reducible fraction,fraction bound organic matter,fraction bound sulfides and residual fraction. Matrix effects of high salinity on the determination of heavy metals in sediments were eliminated by matrix matching and internal standard methods when inductively couple plasma optical emission spectroscopy (ICP-OES) and mass spectroscopy (ICP-MS) were used,respectively. The results showed that the measured values of marine sediment reference materials were consistent with the standard values when the digestion solutions were determined after dilution. The extraction results of the prepositional extraction procedure and European Community Bureau of Reference Program (BCR) procedure were compared and the selectivity of extractants was investigated. The preliminary studies indicated that this sequential extraction procedure was applicable for evaluating the speciation of heavy metals in sediment with organic substances pollution and eutrophication,especially for fraction bound organic matter and fraction bound sulfides.

BACKGROUND: Most animal experiments of transgene are derived from mice; therefore, it is necessary to establish a focal cerebral ischemiareperfusion model and significant to prevent and cure ischemic cerebrovascular diseases.OBJECTIVE: To establish a convenient and reliable model with middle cerebral artery occlusion reperfusion (MCAO/R) in mice.DESIGN: Randomized controlled animal study.SETTING: Institute of Cerebrovascular disease, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: Twenty healthy BALB/c mice, of both genders, weighing 25-30 g, of SPF grade, were divided into sham operation group (n=5), ischemia group (n=10) and 22-hour reperfusion group (n=5) on the basis of digital table. In addition, according to digital table, 130 healthy male Kunming mice were divided into sham operation group (n=10), 24-hour ischemia group (n=30), 2-hour ischemia/22-hour, 46-hour and 70-hour reperfusion groups with 30 in each group; meanwhile, 30 female mice were divided into sham operation group, 24-hour ischemia group and 2-hour ischemia/22-hour reperfusion group with 10 in each group. All Kunming mice were weighing 25-30 g and of SPF grade.METHODS: The experiment was carried out in the Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University from July 2005 to March 2006. The 6-0 suture with silica gel covered at an end was sent from the left external carotid artery (ECA) into internal carotid artery (ICA) till arriving at the initiation of middle cerebral artery (MCA) to block the blood stream in it, then drawing the suture from ICA 2 hours after occlusion to accomplish reperfusion. Mice were cut off their heads in sham operation group at 24 hours after operation, in ischemia group at 24 hours after blocking blood stream and in reperfusion group at 24, 48 and 72 hours after operation. Reliability of models was evaluated with neurology score and tetrazolium chloride stain. Longa standard scores: neurology score ≥ 1 point was regarded as successful models;coronal sections of brain tissue were stained with tetrazolium chloride, and the white region was regarded as infarcted volume.MAIN OUTCOME MEASURES: Neurology score and infarcted volume after staining of triphenyltetrazolium chloride in brain tissue.RESULTS: All mice were involved in the final analysis. ① Successful rate was 20% of BALB/c mice, 66.7%-73.3% of male Kunming mice and 40%-50% of female Kunming mice. ② Brain sections of BALB/c mice in sham operation group were orange at both sides of cortex and infarction focus was not observed. A big infarcted volume was observed on brain sections of mice in ischemia group, and infarcted volume counted for 50%-70% as homonymy hemisphere on optochiasmatic coronal sections. The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%. In addition, condition in ischemiareperfusion group was similar to that in ischemia group. A big infarcted volume was observed on brain sections, and infarcted volume counted for 50%-75% as homonymy hemisphere on optochiasmatic coronal sections.The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%.CONCLUSION: The model with MCAO/R in mice characterizes by relatively smaller trauma, and the ischemic region is stable; therefore, it can be used to accurate timing control of ischemia/reperfusion. This model is an ideal one for researching pathophysiological changes, prognosis and therapy in cerebrovascular disease.