Objective　To discuss the development and the invasive transfer mechanism.Methods　Cell culture method, p53 protenin immunohistochemistry, PCR-SSCP examination, zymography, PCM and heterogenic inouclation test inside the nude mice were adopted. Results　p53 genetic mutation existed in human melanoma cells. The fluorescent positive rate of 67KD LN-R on the surface of the melanoma cell and the average flurescent intensity are repectively WM451＞WM983A＞WM1341B＞WM35.Early WM35 did not produce MMPs; WM1341B only produced 72KDa(MMP-2),not 92KDa(MMP-9);Both WM983A at progressive stage and distal transfering tumor WM45 produced 72KDa and 92KDa. WM983A and WM451 were also seen to form evident transfering tumor inside the naked mice.Conclusion　p53 genetic mutation, the production of MMPs and the high indication of 67KD LN-R on the celluar surface are the key factors for the human melanoma cell to obtain invasive transferring abililty.

Objective To explore the role of nucleotide-binding oligomerization domain receptor protein 3 (NLRP3) inflammasome in the pathogenesis of dementia. Methods 16 patients with with Alzheimer's disease (AD), 22 patients with vascular dementia (VD) and 20 healthy persons with similar age were detected the level of NLRP3 mRNA with RT-PCR in peripheral blood mononuclear cells January, 2014 to October, 2015. The serum level of interleukin (IL)-1βand IL-18 were detected with ELISA. The total number of white blood cells (WBC) and serum total calcium were collected. Single factor correlation analysis was performed between NLRP3mRNA and IL-1β, or IL-18, or WBC, or calcium concentration. Results The levels of NLRP3 mRNA ranged from high to low were as in AD group>VD group>the healthy group (q>11.48, P<0.05). The serum level of IL-1βwas higher in AD group than in the the healthy group (q=16.74, P<0.05), but not significantly different between VD group and the healthy group, nor between VD group and AD group (P<0.05). There was no signifi-cant difference about IL-18 among AD group, VD group and the healthy group (P<0.05). NLRP3 mRNA was positively correlated with the serum levels of IL-1β(r=0.64) and calcium (r=0.58) in AD group. There was no correlation between NLRP3 mRNA and IL-18 or WBC (P<0.05). Conclusion The expression of NLRP3 inflammasome may be related with the pathogenesis of AD, but little impact on the pathogene-sis of VD.

Objective To explore the relevance between nitrate transporters (Sialin) and ischemia-reperfusion injury (IRI) in free flaps.Methods Twenty rats were randomly divided into control group and experimental group,10 in each group.After preparatiing 4 cm×8 cm free tissue flap on left lower abdominal,the rats in experimental group were experienced ischemia in 6 hours,then reperfusion.The rats in control group were without ischemia treatment,and tissue flaps were sutured in situ.After postoperative observation for 7 days,the furthest edge of survival flaps were sampled.The expression of Sialin was detected by the Real-time PCR and Western blot.The change of nitrate content was analysed by chemiluminescence.Results Both of results of Real-time quantitative PCR and Western blot,it was found that expression levels of Sialin in the experimental group were significantly lower than the control group(0.945±0.0530)(P＜0.05).Applying Real-time quantitative PCR relative quantitative analysis,it was found that the experimental group Sialin mRNA expression level was 0.284±0.0486,significantly lower than that of control group (0.945±0.0530)(P＜0.05).Using Western blot technical analysis,experimental Sialin protein expression level was(0.1413±0.0446),significantly lower than the control group (0.3519±0.0368) (P＜0.05).The concentration of nitrate of test results found that the expression of nitrate transporters (Sialin) linearly related with the concentration of nitrate in free flaps,which means tbe Sialin expression level decreased,the nitrate content decreased(P＜0.05,R=0.81).Conclusion The concentration of nitrate was linearly related to the level of Sialin expression:when Sialin expression levels dropped,the concentration of nitrate decreased.Nitrate transporters(Sialin) may be correlated with IRI in free flaps.

Objective: To investigate the safety and effectiveness of three-dimensional (3-D) rotational angiography reconstruction of left atrial and pulmonary vein stereo image embedded with real-time X-ray fluoroscopy system for guiding radiofrequency catheter ablation (RFCA) in treating the patients with atrial ifbrillation (AF).
Methods: A total of 60 consecutive AF patients who received RFCA in our hospital from 2011-04 to 2013-04 were studied. The patients were randomly divided into 2 groups and 3-D Carto3 mapping system was applied for guiding RFCA in both groups. n=30 in each group. Treatment group, the patients received Siemens ARTIS Zeego digital subtraction system for left atrial and pulmonary vein rotational angiography, then, stereo images were reconstructed and embedded with real-time X-ray lfuoroscopy for RFCA guidance. Control group, conventional left and right pulmonary venography was conducted for RFCA guidance. The procedural and X-ray exposure times, rates of success and complications were recorded and compared between 2 groups.
Results: All 60 patients had successful RFCA, compared with Control group, the patients in Treatment group had obviously less procedural time and X-ray exposure time, while the success rate and complications were similar between 2 groups.
Conclusion: 3-D rotational angiography reconstruction of left atrial and pulmonary vein stereo image embedded with real-time X-ray lfuoroscopy system is safe and effective for guiding RFCA in treating the AF patients, which may reduce the procedural and X-ray exposure times.

Objective To evaluate the effectiveness and safety of multimodal postoperative analgesia of thoracotomy for malignant tumor excision.Method Sixty patients undergoing thoracotomy for malignant tumor excision were recuited.They were randomized into four groups:group A receiving single flurbiprofen axetil injection,group B using electronic analgesia pump for sufentanil PCIA,group C using electronic analgesia pump for dezocine PCIA and group D receiving sufentanil controlled by PCA-TCI system.Vital signs,artery lood gas analysis,PrinceHenry score,Ramsay score,remedial measure and untoward reactions were recorded.Results The falling down degree of Prince-Henry score of Group D was higher than those of Groups A,B and C (P ＜ 0.05).Reduction of Ramsay score of Group D was higher than those of Groups A,B and C (P ＜ 0.05).The number of the patients in Group D not undergoing Bolus remedial measure was larger than that of those in Groups B and C (P ＜ 0.05).Conclusion When referring to the effectiveness and safety of the models administrated the thoracotomy for malignant tumors excision,the analgesia model of PC A-TCI sufentanil (plasma target concentration:0.12～ 0.14 ng/ mL) used in Group D is superior to other models used in Groups A,B and C.Moreover,individual analgesia can be reached by the PCA-TCI sufentanil model.

Objective To evaluate dual-source dual-energy CT(DSCT) for the differentiation of urinary stone composition in vitro. Methods Ninety-seven urinary stones were obtained by endoscopic lithotripsy and scanned using dual-source dual-energy CT. The stones were divided into six groups according to infrared spectroscopy stone analysis: uric acid ( UA ) stones ( n = 10 ), cystine stones ( n = 5 ), struvite stones( n = 6), calcium oxalate ( CaOx ) stones ( n = 22 ), mixed UA stones ( n=7 ) and mixed calcium stones(n=47). Hounsfield units (HU) of each stone were recorded for the 80 kV and the 140 kV datasets by hand-drawing method. HU difference, HU ratio and dual energy index ( DEI ) were calculated and compared among the stone groups with one-way ANOVA. Using dual energy software to determine the composition of all stones, results were compared to infrared spectroscopy analysis. Results There were statistical differences in HU difference [(-17±13), (229±34),(309 ±45), (512 ±97), (201±64)and (530±71) HU respectively], in HU ratio (0.96±0.03, 1.34 ±0.04, 1.41 ±0.03, 1.47 ±0.03,1.30±0.07, and 1.49 ±0.03 respectively), and DEI( -0.006 ±0.004, 0.064 ±0.007, 0.080 ±0. 007, 0. 108±0.011 ,0. 055 ±0.014 and 0. 112 ±0.008 respectively ) among different stone groups(F=124. 894,407.028, 322. 864 respectively, P ＜0. 01 ). There were statistical differences in HU difference,HU ratio and DE1 between UA stones and the other groups( P ＜ 0. 01 ). There were statistical differences in HU difference, HU ratio and DEI between CaOx or mixed calcium stones and the other four groups (P＜0. 01 ). There was statistical difference in HU ratio between cystine and struvite stones ( P ＜ 0. 01 ). There were statistical differences in HU difference, HU ratio and DEI between struvite and mixed UA stones (P＜0. 05 ). Dual energy software correctly characterized 10 UA stones, 4 cystine stones, 22 CaOx stones and 6 mixed UA stones. Two struvite stones were considered to contain cystine. One cystine stone, 1 mixed UA stone, 4 struvite stones and 47 mixed calcium stones were considered to contain oxalate. Conclusions DSCT has the ability to differentiate urinary stone composition in vitro. With dual energy software, the UA, cystine and mixed UA stones can be differentiated from other types of stones.

A mouse-anti-human monoclonal antibody was produced by using the membrane proteins of human lung carcinoma cell line A549 as the immunogen to generate monoclonal antibodies against lung carcinoma with the use of hybridoma techniques. McAb4E7 was prepared successfully. To identify its antigen, proteomic technologies such as two-dimenstional electrophoresis, western blotting and mass spectrometry were employed. The targeting antigen of McAb4E7 expressed positive in human lung cancer cell lines A549 and human hepatocarcinoma cell line HepG2, moreover, the expression of the antigen was stronger in A549 cells. Finally, we obtained one positive protein in A549 cell line that has strong affinity and specificity for McAb4E7, which was identified to be ATP synthase beta subunit. We identified ATP synthase beta subunit as the targeting antigen of lung carcinoma special monoclonal antibody McAb4E7.
Animals ; Antibodies, Monoclonal ; biosynthesis ; chemistry ; immunology ; Antibodies, Neoplasm ; immunology ; Antibody Specificity ; Antigens, Neoplasm ; genetics ; immunology ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; immunology ; Membrane Proteins ; immunology ; Mice ; Mice, Inbred BALB C ; Mitochondrial Proton-Translocating ATPases ; immunology

Objective:To investigate the effects of naturally occurring mild hypothermia and artificial mild hypothermia on the expression of high mobility group box 1(HMGB1) in lung tissues of septic mice.Methods:One hundred and twenty BALB/C mice (SPF level) were randomly numbered.Twelve mice with integer multiples of 10 were used as the normal control (NC) group, and the remaining 108 mice were chosen as the septic group.The septic mouse model was established by intra abdominal injection of lipopolysaccharide (LPS) 10 mg/kg.The NC group was given the same dose of normal saline.Anal temperature of the septic mice were measured 1 hour after the model was established successfully, and then were divided into naturally occurring mild hypothermia group and non-mild hypothermia group according to T≤36℃ and T>36℃.In the naturally occurring mild hypothermia group, the mice with T<34℃ were eliminated, and the remaining septic mice were randomly divided into the naturally occurring mild hypothermia(NOMH) observation group and the keep normothermia (KN) group.NOMH group was not given preheating intervention, while KN group was placed in an incubator to maintain the anal temperature between 36.0℃ and 37.5℃.Septic mice in the non-mild hypothermia group were randomly divided into the nonhypothermia (NH) observation group and the artificial mild hypothermia (ATMH) group.The NH group was not treated with hypothermia, while the ATMH group was treated with physical hypothermia, so that the anal temperature of the mice were maintained at 34℃-36℃.Four mice in each group were randomly selected at 6 and 12 hours after modeling, and the concentrations of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) and HMGB1 in serum were detected by enzyme-linked immunosorbent assay(ELISA). At 12 hours, the survival rate of each group of mice was observed.Then 4 mice of each group were sacrificed and lung tissues were taken.The pathological changes of lung tissues were observed by hematoxylin-eosin (HE) staining, and the expression of HMGB1 in lung tissues was observed by immunohistochemical staining.Real time fluorescence quantitative PCR and Western blot were used to detect the relative expression of HMGB1 at mRNA and protein levels.Results:(1)Twelve hours after modeling, the survival number of NOMH group, ATMH group, KN group and NH group were 36(40), 6(11), 27(40), 4(11), respectively, and there were differences between the four groups (χ 2=32.286, P=0.002). Compared with the other three groups of septic mice, the survival rate was highest in the NOMH group (compared with ATMH group: χ 2=5.222, P=0.022; compared with the KN group: χ 2=6.050, P=0.013; and the NH group: χ 2=11.672, P=0.001), but the differences between the other two groups were not statistically significant (all P>0.05). (2)Compared with the NC group, the concentrations of serum TNF-α, IL-6 and HMGB1 of septic mice in each group were significantly increased at 6 h and 12 h (all P<0.05). Compared with NOMH group, the concentrations of TNF-α, IL-6 and HMGB1 in ATMH group, KN group and NH group were significantly increased at 6 h and 12 h(all P<0.05), and the concentrations of TNF-α, IL-6 and HMGB1 in NH group were the highest at all time points (all P<0.05). The concentrations of TNF-α at 12 h decreased compared with 6 h (all P<0.05), while the concentrations of IL-6 and HMGB1 at 12 h increased compared with 6 h (all P<0.05). (3)HE staining showed that the lung tissue damage were minimal in NOMH group, followed by ATMH group.(4)Immunohistochemical staining showed that the expression of HMGB1 protein was in order of NOMH group, ATMH group, KN group and NH group; (5)The relative expressions of HMGB1 protein in lung tissues of septic mice in NOMH group, ATMH group, KN group, and NH group was 0.280±0.013, 0.320±0.016, 0.340±0.018, and 0.380±0.014, respectively, and the relative expression level of HMGB1 mRNA was 4.86±0.22, 6.02±0.18, 6.26±0.20, and 7.98±0.28, respectively, compared with NC group (HMGB1 protein content was 0.240±0.013, and the relative expression level of HMGB1 mRNA was 2.21±0.12) significantly increased (all P<0.05). Cmpared with NOMH group, the relative expression levels of HMGB1 protein and HMGB1 mRNA in the lung tissues of the ATMH group, KN group and NH group were significantly increased(all P<0.05), with the highest expression level in the NH group(all P<0.05). Conclusion:Mild hypothermia may reduce lung tissue damage by down-regulating the expression of HMGB1 in lung tissues of septic mice, and the improvement of spontaneous mild hypothermia was more significant.

ObjectiveTo evaluate the feasibility of determining the chemical composition of kidney stones using gemstone spectral imaging ( GSI ).Methods One hundred and sixty eight extracted human kidney stones immersed in a 10 cm deep water tank underwent CT (Discovery CT750 HD) scans with GSI mode and conventional polychromatic imaging ( CPI,120 kVp) mode.All GSI data were transferred to Workstation AW 4.4 to acquire monochromatic images of 50 keY,effective atomic number (Zeff) mapping images,water (calcium)-based images and calcium (Water)-based images with GSI Viewer.CT numbers of stones were measured and compared at 50 keV monochromatic images and 120 kVp polychromatic images,the mean Zeff,calcium density and water density were measured at Zeff mapping images,Calcium (Water) -based images and Water (Calcium)-based images,respectively.The mean Zeff,spectral HU curve slope and calcium water ratio (CWR) were compared with ANOVA and Wilcoxon test.The composition of kidney stones was determined by infrared spectrometer after CT examination.According to the result of stone composition determined by infrared spectroscopy,108 pure kidney stones were divided into five groups:Uric acid stones ( UA,n = 13 ),struvite stones ( STR,n = 24),cystine stones ( CYS,n = 14),calcium phosphate stones ( CaP,n = 18),and calcium oxalate stones ( COX,n = 39).ResultsThe mean Zeff,CWR,the mean CT numbers at 50 keV images,120 kVp images and spectral HU curve slope of each group were listed as the following:UA [ 7.4 ± 0.4,0.0085 ± 0.0021,( 503 ± 168 ) HU,(495 ± 106 ) HU and - 0.77 ] ; STR [ 11.8 ± 0.9,0.1743 ± 0.0677,( 1056 ± 290 ) HU,( 799 ± 165 ) HU and 18.72 ] ; CYS [ 11.2 ± 0.6,0.1253 ± 0.0297,( 740 ± 172 ) HU,( 565 ± 129 ) HU and 12.79 ] ; CaP [ 16.0 ± 0.4,0.6781 ± 0.0952,( 2567 ±178 ) HU,( 1602 ± 200 ) HU and 37.14 ] ; COX [ 15.4 ± 0.4,0.5683 ± 0.0759,( 2267 ± 385 ) HU,( 1489 ±284) HU and 36.36 ],there were significant differences among groups ( P ＜ 0.01 ).The differences in the mean Zeff,CRW,spectral HU curve slope were statistically significant among the five groups ( P ＜ 0.05 ).Conclusion Spectral CT imaging provides a new method to characterize the kidney stones with the information orovided by mean Zeff,CRW and the CT numbers at 50 keV.

Objective To evaluate the potential cerebral cortical volume alterations in type 2 diabetes mellitus (T2DM) patients with mild cognitive impairment (MCI) compared with T2DM patients and healthy controls,and to observe the correlations with the scores of neuropsychological scales.Methods Cortical volume based on high-resolution MR T1WI data from 30 healthy controls (HC),30 T2DM patients and 30 T2DM with MCI patients were evaluated with FreeSurfer software and compared with variance analysis.The correlations between cerebral cortical volume which had statistical difference and the scores of neuropsychological scales were analyzed.Results There were significant differences in auditory verbal learning test (AVLT) scores,complex figure test-delayed recall (20 min) scores,digit symbol-coding subtest scores,MoCA scores and higher trail-making test-A scores,as well as trail-making test-B scores between T2DM and T2DM with MCI patients (all P＜0.05).Compared with T2DM patients,cortical volume of left entorhinal cortex,left lateral orbitofrontal gyrus,left posterior cingulate gyrus and the right lateral orbitofrontal gyrus,right pars orbitalis,right insula reduced in T2DM with MCI patients (all P＜0.05).In T2DM with MCI patients,AVLT scores were positively correlated with volume of the left entorhinal cortex (r=0.452,P=0.018).Conclusion Several cortical volume reductions are exhibited in T2DM patients with MCI.The volume of the left entorhinal cortex may be a potential biomarker to diagnose and evaluate MCI in T2DM.