Objective To compare patient radiation dose of two techniques of spine X-ray panoramic imaging under the same image quality.Methods An anthropomorphic phantom was used.The reasonable imaging parameters were found for spine panoramic radiography using Sonialvision safire17 Slot scan with HQ mode.And the panoramic films were obtained with different parameters using GE XR650 DR system.The panoramic images were scored by three experienced radiologists.The imaging parameters with the same score in groups were input into PCXMC 2.0 software to get effective dose(E) and organ dose.Results The reasonable imaging parameters of Slot scan were high quality(HQ) mode, SID 150 cm, 100 kVp, and 2 mAs;and the corresponding parameters of XR650 were SID 200 cm, 100 kVp, and 3.2 mAs.The E of the Slot scan with HQ mode, XR650 with manual mode and XR650 with AEC mode was(0.118 7±0.001 4),(0.0847±0.0008), and (0.1580±0.001 5) mSvrespectively, while the E of XR650 with manual mode was lower than the others(F =3 007.293, P ＜0.05).The organ dose using XR650 DR with manual mode were lower than that using Slot scan with HQ mode in all samples except breasts(P ＜0.05);the organ dose using XR650 DR with AEC mode were higher than that using XR650 DR with manual mode and Slot scan with HQ mode for all samples except for thyroid, oesophagus and lungs (P ＜0.05).Conclusions The radiation doses of both spine X-ray panoramic imaging with manual mode are low, and low dose spine X-ray panoramic imaging can be achieved if reasonable parameters are used.

Objective To determine the functional motif of the recombinant Ricin B(rRicin B) in Vibrio vulnificus cytolysin (VVC) and understand its molecule pathogenic mechanism.Methods The motif of VVC was predicted through bioinformatics analysis and cloned into a procaryotic expression vector pET28a-rRicin B.The recombinant plasmid was transformed into E.coli BL21 (DE3) and induced by IPTG to express rRicin B.The expressed protein was further analyzed by SDS-PAGE and purified by Ni2+-NTA agarose.Renaturation of the rRicin B were also carried out for further analysis.ELISA assay and confocal microscope was applied to identify the activity of the rRicin B on human Hela cells.Results Ricin B motif located in the 336-465 amino acids of Vibrio vulnificus cytolysin with a relative molecular weight of 20×103.The result of ELISA showed that the antigenicity of rRicin B was 28.71 U/L after renaturation.FITC labeled rRicin B could bind to the cell membrane and enter the cytoplasm of human Hela cells.Conclusion The Ricin B motif in Vibrio vulnificus cytolysin bearing the similar ability with the natural Ricin B can bind to the cell membrane and enter the cytoplasm.This feature may play an important role in the activity of pore-forming and the cytotoxicity of Vibrio vulnificus cytolysin.

Objective Exploration of TSH, FT3, FT4 changes in critical condition prior to death.Methods 84 patients that were anti-tumor therapy acceptable and not associated with the treatment of thyroid disease in patients with terminal tumors. Age from 24 to 85-year-old, male 56, female 28. The use of SELDI technology (surface-enhanced laser description / ionization time-of-flight mass spectrometry protein chip technology) checks to M/Z for the 11 100 + H～11 900 + H peak of the cluster, divided into the positive group for critically ill patients with early warning protein (LGT) and the negative group for it. All patients were given TSH, FT3, FT4 inspection. The results were statistically analyzed. Results TSH, FT3, FT4 in the LGT positive group was significantly lower than in the negative LGT group. There was no correlation between TSH,FT3 and FT4 in the LGT positive group;In the negative LGT group FT3 and FT4 had a positive correlation,and FT3 and TSH no relevance. Conclusion Thyroid function is normal in the stage of cancer patients stable condition. Possible pituitary-hypothyroidism may occur as the disease advances to the end or before death.This may be an important factor causing the acceleration to the death of the patients. However, the cause and effect between this phenomenon and the early warning of protein fingerprint group(LGT) is still not clear. In critical condition checking thyroid function and render supplementary amount of thyroid hormone in accordance with the checking results may be beneficial.

Objective: To analyze the aberrant methylation of death-asscciated protein kinase (DAPK) gene in blood from non-small cell lung cancer (NSCLC) patients and to evaluate its clinical significance. Methods: A methylation-specific PCR was performed for the detection of promoter hypermethylation of DAPK gene in blood DNA from 65 cases of NSCLC patients,and the relation of the aberrant methylation of DAPK gene and the clinical pathological data was analyzed. Results: 30.8% (20/65) of the blood from 65 cases of NSCLC patients showed hypermethylation in DAPK promoter, whereas no methylated DAPK gene promoter was found in blood from normal control or patients with benign lung diseases. DAPK gene promoter methylation in blood did not strongly correlate with the pathological class, stage, metastasis and differentiation of NSCLC. Conclusion: Detection of the aberrant methylation of DAPK gene in blood DNA from NSCLC patients might offer an effective means for the earlier auxiliary diagnosis of the malignancy.

Objective To identify drug resistance status of Mycobacterium tuberculosis (MTB) strains by the GenoType MTBDRplus line-probe assay (LPA),compare its performance with traditional drug susceptibility testing (DST),and to assess its predictive value for the prognosis of patients with drug resistance tuberculosis.Methods Pulmonary tuberculosis patients who visited Zhuji People's Hospital,Zhejiang Province during February 2011 and January 2012 with a positive result of sputum smear at baseline were all recruited.A total of 275 culture positive specimens were collected,then isolated and cultured for Mycobacterium tuberculosis in the laboratory.DST were performed,meanwhile,GenoType MTBDRplus were also applied to detect resistance to isoniazid (INH) and rifampin (RMP).All the tuberculosis patients who were recruited were followed,including sputum culture and chest radiography.Results There were 192 strains showing drug resistance both by DST and MTBDRplus LPA.Fourteen multidrug resistant (MDR),21 INH mono-resistant and 2 RMP mono-resistant strains were detected by DST.As for GenoType MTBDRplus LPA,MDR,INH mono-resistant and RMP mono-resistant strains were 14,18 and 2,respectively.Taken DST as the gold standard,LPA was more accurate in the detection of resistance to RMP,while it failed to detect 23.8％ (5/21) of the INH-resistant strains.We analyzed the prognosis of patients with drug resistance by GenoType MTBDRplus LPA,the rates of treatment success were 84 ％ (110/131),9/15,3/11 in patients infected with susceptible,INH mono-resistant and MDR strains,respectively.For the 2 cases of RMP mono-resistanee,one was cured and the other failed.The predictive value of molecular drug resistance test for treatment failure in INH mono-resistant patients was 40.0 ％,while that was 83.5 ％ for treatment success in INH susceptible patients.The predictive value for treatment failure in RMP mono-resistant patients was 50.0％,while that was 81.5％ for treatment success in RMP susceptible patients.The predictive value for treatment failure in MDR patients was 72.7％,while that was 81.1％ for treatment success in patients without MDR.Conclusion The GenoType MTBDRplus LPA assay is a rapid and reliable diagnostic test for resistance of MTB,which can be used to predict the prognosis of drug resistant tuberculosis in the clinical practice.

Objective To inVestigate the Potential mechanism and ProtectiVe effects of osthole on rats with focal cerebral ischemia_rePerfusion injury. Methods Middle cerebral artery occlusion was induced by the suture_occluded method. After 2 h of cerebral ischemia and 24 h of rePerfusion,scores of neurological deficits and infarct Volume were eValuated. The leVels of SOD,MDA,GSH and ATPase were also determined. Results Scores of neurological deficits and infarct Volume were lower in the osthole grouPs than in the model grouP. Meanwhile,the actiVity of SOD and GSH content were increased,while the MDA content was decreased in osthole grouPs. Conclusion Osthole exerts ProtectiVe effects on rats with focal cerebral ischemia_rePerfusion injury.

BACKGROUND: Inhibiting the degradation of extracellular matrix in the intervertebral disc can delay the degenerative process of intervertebral disc. Matrix metalloproteinases-3 (MMP3) is considered as a key enzyme for degradation of extracelular matrix components such as type II collagen and aggrecan.
OBJECTIVE:To construct the short hairpin RNA lentiviral vector targeting human MMP3 gene and to detect its efficiency of gene silence by infecting human degenerative nucleus pulposus cells.
METHODS:According to the human MMP3 mRNA (NM_002422.4) sequence, four groups of the short hairpin RNA gene sequences targeting MMP3 were designed, synthesized and annealed to form double stranded DNA fragments, which were connected with the LV3 vectors digested by BamHI andEcoRI enzymes, and then transfected into the competent cels. The positive clones were identified by PCR, and analyzed by sequencing. The packaging and titer of lentivirus were determined after transfecting 293T cells. Human degenerative nucleus pulposus cels were infected with lentivirus vector, and the transfection efficiency of each group was observed under inverted fluorescence microscope. The interfering efficiency was detected by real time-PCR and western blot at 72 and 96 hours.
RESULTS AND CONCLUSION:The ds-oligo DNA was successfully inserted into the lentiviral vector as confirmed by electrophoresis and sequence analysis. The recombinant lentivirus was harvested from 293T cels with a viral titer of 1-5 ×108 TU/mL. RNA interference targeting the GCC AGG CTT TCC CAA GCA AAT sequences with the highest interfering efficiency in MMP3 gene at 72 and 96 hours resulted in suppression of MMP3 mRNA expression by 98% and 72%, respectively; and at 96 hours, the interfering efficiency of protein expression was 57.2%. The recombinant lentivirus vector containing RNA interference targeting MMP3 gene is successfuly constructed, which lays a foundation for further studies on the MMP3 function and gene therapy.

Objective To compare the differences of pulmonary nodule detection rates between iterative reconstruction (sinogram affirmed iterative reconstruction,SAFIRE)algorithm and filtered back projection (FBP) algorithm in chest CT, and to evaluate the detection accuracy.Methods Three groups of tube voltage values of 80,100,and 120 kV were defaulted on the new dual-source CT,with automatic mAs care dose 4D technology, the chest phantom with simulated pulmonary nodules was scanned, then the images were reconstructed with FBP and SAFIRE (grade 1-5 ),respectively. The detection rates of simulated pulmonary nodules in the chest CT images reconstructed of SAFIRE (grade 1-5 )and FBP were compared, and their diameters and CT values were measured.Results With the same tube voltage, no significant difference was found in the detection rate of simulated pulmonary nodules between SAFIRE (grade 1-5 )and FBP (P>0.05 ), the diameter deviation of simulated nodules of SAFIRE (grade 3 )was less than FBP, and the difference in the average CT value of the simulated nodules between SAFIRE (grade 3)and FBP was not statistically significant (P>0.05);the simulated nodule detection rate of 100 kV was equivalent to the detection rate of 120 kV,the simulated nodule (-800 HU and 3 mm )detection rate of 80 kV was less than that of 120 kV;as the tube voltage reduced,or simulated nodule diameter decreased,or the density of simulated nodule reduced,the nodule’s diameter deviation was increased. Conclusion Compared with FBP,the capabilities of SAFIRE in pulmonary nodule detection in different densities and different sizes are same,and SAFIRE algorithm is helpful for accurate displaying of pulmonary nodules,and it can be used for low-dose CT lung cancer screening program.

Objective To evaluate the effect of NGX6 combined with cisplatin on the inhibition rate of A549 cells and NCI-H1975 cells and antitumor effects in vivo.Methods The NGX6 was loaded in the LPD, and prepare Liposome protamine DNA complexes.A549 cells and NCI-H1975 cells were seperately divided into NGX6 group ( 30 μg/mL NGX6 concentration ) , cisplatin group, NGX6 +cisplatin group, PBS as negative control group.The effect of cytotoxicity of four group on A549 cells and NCI-H1975 cell were evaluated by MTT assay.the clone forming rate and the inhibition rate were determined by Cell colony count.lung transplantation tumor model were successfully established, then nude mice were divided into four groups as abeve, each group of 10, tumor size and survival period were determined tumor cell apoptosis were observed.Results The cell viability of A549 cells and NCI-H1975 cells of (NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The cloning efficiency of A549 cells and NCI-H1975 cells of ( NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The tumor inhibitory rate was in vivo for (NGX6 +cisplatin) was higher than other group(P<0.01).The median survival of nude mice in (NGX6 +cisplatin), NGX6, cisplatin and saline group were 43,31,29 and 15 days.Conclusion NGX6 combination with cisplatin can inhibit the cell proliferate of lung cancer cells and inhibit the tumor growth and the combination of NGX6 and cisplatin may be a potentially effective treatment for lung cancer.

Objective To observe the morphological structures of WHBE rabbit brain in vivo based on 3.0 T magnetic resonance imaging system (MRI), accumulate the basic biological data of WHBE rabbit brain imaging, and provide a background information to further expand the WHBE rabbit application.Methods Nine healthy adult male WHBE rabbits were intravenously anesthetized with 3% pentobarbital sodium.3.0 T MRI plus rabbit brain dedicated coil was used to perform routine transverse and sagittal scans, and the size of brain structures were measured.Results MRI scanning can be successfully performed to obtain sagittal and transverse T2WI or T1WI images of WHBE rabbit brain in vivo, and can be clearly observed the basic structures of WHBE rabbit brains in vivo, such as olfactory bulb, cerebrum, cerebellum and pituitary gland.In addition, high signal was found in the hippocampus of the left and right temporal lobes in 4 rabbits with T2WI, but also low signal appeared in the corresponding regions in T1WI, and the others were not abnormal.Meanwhile, the reference data of frontal lobe, hippocampus, cerebrum, lateral ventricles, pituitary gland and other related anatomical structures were also obtained.Conclusions Using the 3.0 T magnetic resonance imaging system and rabbit brain coil,the morphological and anatomical structures of rabbit brain can be clearly observed, and the basic imaging data of WHBE rabbits brain have been established preliminarily.