Objective To analyze the correlation between serum hs -CRP levels and MR Imaging in ischemic cerebrovascu-lar patients.To explore the hs -CRP serum levels in ischemic cerebrovascular patients incidence of early diagnosis and prognosis e-valuation application value.Methods 300 cases of ischemic cerebrovascular disease as the research object were selected from May 2013 to December 2014.The disease is classified into acute cerebral infarction group 100 cases, acute phase of cerebral infarction group 140 cases and transient ischemic attack 60 cases.Among 100 cases of patients with acute cerebral infarction group, in accord-ance with the MR Imaging data, they can be subdivided into three groups.Group A: lacunar infarction group 28 cases; Group B:small area of infarction group 50 cases and group C: large area of infarction group 22 cases.Another 40 healthy physical examination were selected as control group.The hs -CRP serum levels were made correlation analysis with MR Imaging.Results The acute cerebral infarction and transient ischemic attack group serum hs -CRP level difference compared with the control group with statisti-cal significance (p<0.05); The SPSS Spearman correlation analysis result shows that the hs -CRP serum levels and MR Imaging data shown in the infarction area and the body has a direct correlation .Conclusion The ischemic cerebrovascular disease patients se-rum hs-CRP level with MR Imaging have shown increased infarct size and raise.Damage area is bigger, and the corresponding serum hs-CRP levels changes more violently.

Aim To evaluate the effect of expression inhibition of spermine oxidase(SMO)on the actitumor activity of polyamine analogue CPENSpm (N~1-cyclopropylmethyl-N~(11)-ethylnorspermine).Methods siRNA technique was used to inhibit expression of SMO in human lung cancer line A549.QT-RT-PCR and enzyme activity assay was performed to determine the expression level of SMO.The cell proliferation was detected by MTT assay.The apoptosis of A549 cells were evaluated by DNA degradation and Sub-G_1/flow cytometry assay.Results The A549 cell line with silenced SMO expression was successfully obtained.Basic SMO mRNA and enzyme activity levels in the SMO-siRNA plasmid transfected cells were 0.53% and 14% lower than that in the control cells respectively. Treating A549 control cells by 10 μmol·L~(-1) CPENSpm for 24 hours resulted in a 10-folds up-regulation of SMO in mRNA level and 20-fold increase in enzyme activity,but this drug-induced SMO expression was obviously prevented in SMO-siRNA plasmid transfected cells.MTT assay demonstrated that SMO expression inhibition decreased the sensitivity of A549 cells to CPENSpm exposure(0～20 μmol·L~(-1)).DNA degradation and sub-G_1 assay proved a deceased ability of CPENSpm to induce apoptosis in SMO-siRNA plasmid transfected cells.Conclusion Up-regulation of SMO by CPENSpm is possibly one of the molecular basics for its antitumor activity.

Objective To investigate whether difluromethylornithine (DFMO) can be used in the treatment of human leukemia. Methods The cell proliferation was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] assay after treatment of human lymphocyte Jurkat cells by DFMO (0 to 10 mmol/L) for 24 to 72 h. Enzyme activity of spermine oxidase (SMO) and acetylpolyamine oxidase (PAO) was determined by chemiluminesence assay. DNA fragmentation assay was used to evaluate cell apoptosis. Fluorescent dye assay was performed to determine the changes in mitochondrial membrane potential. Western blotting was used to determine Bax content. Casepase-3 enzyme activity was measured by spectrophotometric method. Results DFMO treatment inhibited the proliferation of Jurkat cells significantly in a dosage- and time-dependent manner (P

Objective:To evaluate the effects of several kinds of surface treatment methods on the bond strength of zirconia prosthesis,and to provide references for improving the bond strength of zirconia prosthesis.Methods:Sixty zirconia blocks(10 mm×10 mm×2 mm) were divided into 6 groups according to the surface treatment methods:sandblasting,silica coating,Z-PRIME Plus,sandblasting+Z-PRIME Plus,sandblasting+silica coating,and silica coating+Z-PRIME Plus groups.The zirconia-resin specimens were fabricated using ScotchbondTM Universal;shear bond test was performed to detect the shear bond strength after treated with water storage(37℃) for 24 h by universal mechanical testing mechine;the fracture types were observed by stereoscopic microscope.One fracture specimen was randomly chosen from each group,and the morphology of the specimen was examined under scanning electron microscope.Results:The shear bond strength of the specimen in Z-PRIME Plus group was higher than those in the other groups(P<0.05).The adhesive failure was predominantly observed in sandblasting,silica coating,sandblasting+silica coating and silica coating+Z-PRIME Plus groups.There were 4 cases of mixed failure in Z-PRIME Plus group and there were 8 cases of mixed failure in sandblasting+Z-PRIME group.No cohesive failure was observed in all the groups.The scanning electron microscope results showed that the fracture surface occurred on the resin-zirconia interface of the specimens in sandblasting group,Z-PRIME Plus group,and sandblasting+Z-PRIME Plus group.The silica-zirconia fracture of specimens occurred in silica coating group and silica coating+Z-PRIME Plus group.The silica-resin interface fracture of specimens partially occurred in sandblasting+silica coating group.Conclusion:The application of Z-PRIME Plus can significantly improve the bond strength between zirconia and resin.

AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E. coli BL21 (DE_3). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificicity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO.

The genotypes of extended spectrum β-laetamases(ESBLs) and AmpC β-lactamases produced by Enterococcus gallinarum isolated from chloebia gouldiae were determined to elucidate the evolution mechanism of the resistant genes.The minimum inhibitory concentrations (MICs) of 18 antibacterial drugs against the Enterococcus gallinarum were detected with two dilution method,and the ESBLs and AmpC β-lactamases from the bacterium were amplified by PCR using the primers of TEM,SHV,CTX-M,ACC,CIT,DHA,EBC,FOX and MOX,respectively.The PCR products were cloned and then the cloned fragments were sequenced to identify their genotypes and subtypes.The bacterium was proved to be a ESBL-producing and AmpC β-lactamase-producing bacterium,showing severe resistant to the other drugs,except the third and forth cephalosporins,carbopenems and fosfomycin.Compared with that of AJ847364 (TEM-116),the sequence of the TEM-type was characterized by two nucleotide mutations (512T→A and 695A→C),which led to two mutations of amino acids(17111e→Lys and 232Lys→Thr),showing that the detected TEM-type was a new genotype,the sequence of the AmpC β-lactamase was similar to that of EF078894 (ACT-like type)with a 97% homology.The genotype of ESBLs of Enterococcus gallinarum was a new TEM-type derived from the TEM-type ESBLs of klebstella pneumoniae isolated from the same avian.The genotype of AmpC lactamase was ACT-type,which probably concerned with β-1actam antibiotics used.

Objective To study the correlation between non-alcoholic fatty liver disease (NAFLD)and glycosylated hemoglobin (HbA1 c) in middle-aged and aged population.Methods A total of 4 127 inservice workers and retirees aged 45 years old or above from one petrochemical enterprise in Ningbo were enrolled in our study.The waistline,body mass index,blood pressure,fasting blood-glucose,blood lipid profile,glutamyltranspeptidase,HbA1c and epigastrium B ultrasound were investigated.According to the quartile of HbA1c level,participants were divided into four groups,namely,Q1 group ≤5.2％,Q2 group ＞ 5.2％-5.4％,Q3 ＞ 5.4％-5.6％ and Q4 group ＞ 5.6％.The prevalence of NAFLD and clinical characteristics of each group were analyzed.Logistic regression was used to predict independent risk factors of NAFLD.Results The morbidity of NAFLD was 27.2％ with 31.9％ in male and 21％ in female,which was significantly higher in men.In Q1,Q2,Q3 and Q4 group,the prevalence of NAFLD were 18.5％ (178/961),22.8％ (185/812),25.6％ (280/1 095),38.1％ (480/1 259) respectively.With the increase of HbA1 c level,the morbidity of NAFLD increased synchronously.The age,systolic pressure,total cholesterol,low densitylipoprotein cholesterin and fasting blood-glucose were all elevated according to the increase of HbA1 c in 1 123 NAFLD patients.Multi-factor logistic regression analysis indicated that high HbAlc level was the risk factor of NAFLD (OR =1.67,95％ CI 1.15-2.43,P =0.007).Conclusion HbA1c is an independent risk factor of NAFLD and both of these are closely related to blood lipid metabolism disorder.

OBJECTIVETo investigate the effect of different length and diameters on the stability of mini implant and to select optimal length and diameter using continuous variation of parameters.

METHODSTo perform 3-dimensional finite element analysis, finite element models of a maxilla, and mini implants with length of 6-12 mm and diameters of 1.2-2.0 mm were generated. Load of two different forces were applied to the head of mini implant. One type was horizontal force (HF), the other was composite force (CF). The maximum equivalent stress (Max EQV) in maxilla and the maximum displacement (Max DM) of mini implant were evaluated.

RESULTSThe Max EQV in maxilla and Max DM of mini implant decreased as length and diameter increased. When length was more than 9 mm, the evaluation indexes were small and had a less change. Datas indicated that diameter played a more important role in reducing target, and was a more effective parameter in reducing Max EQV when CF was loaded.

CONCLUSIONFrom biomechanical point of view, the choice of the length should not be more than 9 mm. When CF is loaded using the mini implant, diameter exceeding 1.2 mm are optimal design for mini implant.

Objective: To explore the effects of endoplasmic reticulum stress-induced apoptosis in thyroid injury of rats caused by excessive fluoride intake. Methods: All 40 Wistar rats were randomly divided into four groups, control group, low fluoride group, medium fluoride group and high fluoride group. The rats in control group were fed with tap water (fluoride concentration=0.344 mg/L) and the experimental rats were fed with the water contaminated fluoride with the dose of 5, 10 and 20 mg/L. 10 rats (female: male=1∶1) in each group were sacrificed after 8 months of exposure through drinking water. The contents of urine fluoride were detected by fluorine ion selective electrode method. Morphology of thyroid was observed through light microscope and apoptosis in thyroid were detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. The mRNA and protein expressions of glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) were analyzed by RT-PCR and immunohistochemistry respectively, and results were compared among groups. Results: The contents of urine fluoride in all fluoride treated groups were separately (4.74±1.88), (7.70±2.82) and (10.50±2.92) mg/L, which were gradually higher than that of control group (2.23±0.54) mg/L (P<0.05). Morphological changes were found in thyroid tissues of fluoride treated groups, thyroid follicular hyperplasia or even no cavity cell clusters were observed. Apoptosis in thyroid were notably increased in fluoride treated groups. The mRNA expression levels of GRP78 in all fluoride treated groups were separately 1.30±0.42, 1.39±0.29 and 1.50±0.27, which were significantly higher than that of control group (0.93±0.24) (P<0.05). And the mRNA expression levels of CHOP in medium and high fluoride groups were separately 1.17±0.29 and 1.30±0.26, which were significantly higher than that of control group (0.91±0.20) (P<0.05). The protein expression levels of GRP78 and CHOP in medium and high fluoride groups were respectively 29.68±4.04, 29.90±3.74 and 4.05±1.62, 4.44±1.81, which were significantly higher than those in the control group (separately 23.80±6.36, 2.27±0.89) (P<0.05). Conclusion Excessive-fluoride intake can induce thyroid injury, and endoplasmic reticulum stress-induced apoptosis might be involved in the injury.

Objective To assess the efficacy of methylprednisolone (MP)pulse therapy in treatment of Hashimoto thyroiditis(HT) complicated with goiter.Methods 30 patients with HT complicated with goiter participated in the study and received MP pulse therapy.The patients had to be euthyroid for at least 3 months before the date of inclusion with plasma concentrations of thyroid hormones within their reference range.The goiter was still obvious and had no improvement.A dose of 250 mg MP was administered intravenously for seven consecutive days,and then treated with oral prednisone 10 mg,three times per day for 6 weeks with the dosage in each week gradually reduced at 5 mg to none.Ultrasonic was used to measure thyroid size before and after MP treatment.Results The treatment was successful at the end of the trial in all of the 30 patients receiving MP.The thyroid size from length,breadth,thickness to isthmus obviously decreased (P ＜ 0.05).The length of the left lobe was (57.42 ± 12.87) mm and (46.37 ± 7.67) mm (t =4.58) before and after treatment; The breadth of the left lobe was(26.68 ±7.71) mm and(22.21 ±6.09) mm(t =4.56) before and after treatment; The thickness of the left lobe was (27.18 ± 6.60) mm and (21.14 ± 5.67) mm(t =7.28) before and after treatment.The length of the right lobe was(58.17 ± 12.32)mm and(49.73 ±9.35) mm(t =3.84) before and after treatment; The breadth of the right lobe was (26.14 ± 7.37)mm and (23.00 ± 6.68) mm(t =3.29) before and after treatment ; The thickness of the right lobe was(27.57 ± 6.42)mm and(22.00 ±5.55)(t =5.88)before and after treatment.The isthmus before and after treatment was(9.94 ±4.15)mm and(6.19 ±2.57)mm(t =6.09).The recurrence rate was 17％ (5/30) after one year.Conclusions MP pulse therapy is an effective treatment for HT complicated with goiter.The recurrence rate is low.