Objective To investigate the feasibility of monitoring the tumor's hypoxic changes by 99Tcm-2-(2-methyl-5-nitro-1H-imidazol-1-yl) ethyl eihydrogen phosphate (MNLS) imaging after radiotherapy.Methods (1) H22 cells were cultured and mice model with liver cancer xenografte was made.The mice were imaged at 0.5,1,2,3,4,6 and 8 h (six mice in each group) after injected with 7.4 MBq 99TcmMNLS when the tumor size reach about 1 cm.Then the mice were sacrificed.The T/NT and ％ID/g of each time point was calculated.(2) The liver cancer bearing mice of radiotherapy group (25 Gy) and control group were imaged at 0,24,48 h,and then the technique of ROI was adopted to calculate the T/NT at each time point in the two groups.Immunohistochemical stain method was used to evaluate the expression level of HIF-1α in liver cancer.(3) One-way analysis of variance,the least significant difference t test,two-sample t test and Spearman correlation analysis were performed.Results (1) The uptake of 99TcmMNLS in the liver cancer bearing mice was significant at 2 h after injection and the ％ID/g was the highest.99Tcm-MNLS was excreted mainly through kidneys.(2) The T/NT and HIF-1α expression level in radiotherapy group at 24 h (2.65±0.27,(50.62±3.78)％) were lower than those at the instant (3.35±0.19,(85.32±0.94)％,t=5.640,6.701,both P＜0.05),but higher than those at 48 h (2.23±0.52,(21.69±0.75)％,t=7.674,4.911,both P＜0.05).The T/NT and HIF-1α expression in the radiotherapy group were significantly higher than those in the control group at the instant (2.74 ± 0.29,(28.26 ± 1.70) ％,t =4.235,3.473,both P＜0.05) but lower at 48 h (3.15±0.88,(67.64±3.55) ％,t =7.902,3.258,both P＜0.05).However,no significant difference was observed at 24 h between radiotherapy group and the control group (2.98±0.16,(58.45±0.98) ％,t =0.525,2.043,both P＞0.05).The change of T/NT closely correlated with the expression of HIF-1α in both the radiotherapy group and control group(r,=0.793,0.756,both P＜0.05).Conclusion 99Tcm-MNLS hypoxia imaging has potential to monitor changes of hypoxia in tumor after radiotherapy.

Purpose To explore the features of 99Tcm-AnnexinⅤscintigraphy of venous thrombus and its feasibility of discriminating fresh venous thrombus from old one. Materials and Methods The rabbits (n=15) were randomly divided into three groups (fresh thrombus group, old thrombus group and control group). The inferior vena cava thrombus models were developed in the rabbits of thrombus groups by inserting screw cooper wire into inferior vena cava. The rabbits of control group received sham operation. 99Tcm-AnnexinⅤwas injected in the rabbits of fresh thrombus group and control group one day after operation;the same was done in the rabbits of old thrombus group 14 days after operation. Planar anterior abdominal images were obtained at 30, 60, 90 and 120 min after 99Tcm-AnnexinⅤinjection in all groups respectively. The ratios of thrombus to background of the two thrombus groups and the ratios of the area correspondent to the thrombus groups to background of the control group were calculated by ROIs counts. Then rabbits were executed, and thrombus was used for pathology examination. Results 99Tcm-AnnexinⅤuptake in thrombi was clearly visualized in all rabbits of the fresh thrombus group;whilst negative images showed in all rabbits of the old thrombus group and control group. The thrombus to background ratios of the fresh thrombus group (4.06±0.49) were higher than that of the old thrombus group (2.46±0.38), and also higher than the inferior vena cava below inferior pole of right kidney level to background ratios (2.27±0.24) of the control group (t=5.746, 7.318;P<0.05). All the thrombi of the fresh thrombus group were confirmed as fresh mixed thrombi by HE staining, and those of the old thrombus group were confirmed as old mixed organized thrombi. Conclusion 99Tcm-AnnexinⅤmay become a new acute venous thrombus imaging tracer used to discriminate fresh venous thrombus from old one.

Objective To analyze clinical features and treatment results of primary non-Hodgkin lymphoma of bone (PLB) and further to investigate the rational treatment. Methods Clinical data of 26 patients with PLB were analyzed. Twenty-three (88.5 %) patients received radiotherapy in combination with chemotherapy, three received chemotherapy alone, and three patients also received surgical resection. Results The pathological types of lymphoma in the patients were diffused large B-cell iymphoma (DLBCL) in 15 patients (57.7 %), small B-cell lymphoma in 1 patient(3.8 %), B-cell lymphoma with unclassified subtypes in 4 patients (15.5 %), T-cell lymphoma in 5 patients (19.3 %,among which anaplastic large cell lymphoma in 3 patients), and unclassified lymphoma in one patient (3.8 %). Of the 26 cases of PLB, 15 were at stage Ⅰ, 3 at stage Ⅱ, 3 at stage Ⅲ and 5 at stage Ⅵ. The 3- and 5-year overall survival rates were 59.16 % and 31.37 %respectively. In the eleven patients who died of lymphoma, three had Iocol-regional relapse, and nine had systemically involved lymphoma. The radiation-induced bone fracture had not been observed after local radiotherapy with median dose of 50 Gy. Conclusion Pelvis maybe a common primary site of PLB, and DLBCL type are the most observed histological subtype. The optimal treatment for PLB is radiotherapy combined with chemotherapy. Local regional radiotherapy with median dose of 50 Gy can be safe and feasible.

Objective To express and detect the antigenicity of human laminin alpha4 LG3-4 module (hLN?4LG3-4) protein by gene engineering techniques. Methods The cDNA encoding hLN?4LG3-4 was amplified by RT-PCR from human placenta, then inserted into pMD-18T vector by T/A cloning and sequenced. Prokaryotic expression vector pET-28a-LG3-4 was constructed by recombinant DNA technique. The hLN?4LG3-4 fusion protein expressed in BL21(DE3)/pET system was identified by SDS-PAGE, purified by Ni-NTA resin, and assayed by Western blotting. Results The cDNA fragment of hLN?4LG3-4 was cloned successfully. While BL21 (DE3)/pET28a-LG3-4 bacterium was induced with IPTG, a new protein band with a relative molecular weight of 44 000 was shown on SDS-PAGE profile. hLN?4LG3-4 fusion protein of high purity (95%) was obtained and specific protein band was detected by Western blotting. Conclusion The hLN?4LG3-4 fusion protein was successfully expressed.

Objective To investigate the expressions of c-Fos,5-HT and spontaneous activities in single prolonged stress (SPS) stress rats,and to study the possible mechanism of posttraumatic stress disorder(PTSD).Methods forty-eight male SD rats were randomly divided into 6 groups:Individual living + SPS group (IS group),Individual living+control group (IC group),Enriched environment+SPS group (ES group),Enriched environment+control group (EC group),control+SPS group(CS group),control group(C group),8 rats in each group,spontaneous activities were measured before the experiment and post experiment at week 1,2.And the expressions of c-Fos,5-HT in the hippocampus were examined by immunohistochemical staining.Results (1) Spontaneous activity:there were no differences of crossing number and distance among 6 groups before experiment (P＞0.05).On SPS 14 days the crossing number and distance of IS group (12.12±9.64,(2.71 ± 1.99)m) decreased compared with the CS group(45.25±8.37,(6.37± 1.18) m,P＜0.05),and ES group (69.75± 10.05,(10.69± 1.50) m)showed significant increase compared with CS group (P＜0.05).(2)5-HT:the expression of 5-HT in the hippocampus of IS group(0.1125±0.0095) was significantly higher than that in CS group(0.6138±0.0059,P＜0.05),menwhile lower in ES group(0.0495±0.0074,P＜0.05).(3)c-Fos:compared with CS group(0.3108±0.0074),the expression of c-Fos in the hippocampus of IS group(0.3585±0.0150,P＜0.05) significantly increased,but decreased in ES group (0.2613±0.0063,P＜ 0.05).Conclusion Enriched environmental can improve the level of activities in PTSD rats,and to reduce the the expression of c-Fos and 5-HT in the hippocampal neurons.

ObjectiveTo determine the feasibility of using left ventricular volume-time curve in the evaluation of left ventricular diastolic function,and to analyze characteristics of left ventricular volume-time curve changcs in hypertrophic cardiomyopathy (HCM).MethodsSeventeen cases of HCM and 12 healthy volunteers received cardiac MRI (CMRI) examination,and left ventricular (LV) 2-chamber long and short axis cine imaging were performed,LV volume-time curves were reconstructed and platform time,different diastolic volume recovery (DVR) time and their corresponding filling velocity were calculated from LV volume-time curve off-line.The DVR time and their corresponding filling velocity were analyzed by using multiple linear regression analysis. Results Compared with the group of healthy volunteers,ventricular septal HCM group had delayed left ventricular 50％,70％ DVR time[ (8.9 ± 1.3) versus (7.7 ± 0.8 ) phase,F=6.787,P=0.016;(11.3 ±1.6) versus(9.7±1.8) phase,F=4.927,P=0.036] and shortened plateau time [ ( 1.8 ± 1.7) versus ( 4.1 ± 1.4 ) phase,t =6.787,P ＜ 0.01 ].Ventricular septal HCM group had reduced 30％,50％ DVR filling rates [ (0.22 ± 0.11 ) versus (0.40 ± 0.15 ) ml/ms,F =12.916,P ＜ 0.01 ; (0.20 ± 0.09 ) versus (0.30 ± 0.10) ml/ms,F =7.121,P =0.014 ] compared with those in the group of healthy volunteers.But 70％,80％,90％ DVR filling rates showed no statistically significant different in the two groups.In HCM patients,myocardial fibrosis caused 50％,70％,80％ DVR time delay [ (9.6 ± 1.0) versus ( 7.9 ± 1.5 ) phase,F =5.000,P =0.045 ; ( 12.3 ± 1.4 ) versus ( 9.6 ± 1.8)phase,F=8.039,P=0.015;(13.1 ±1.4 ) versus(10.9±1.9)phase,F=5.060,P=0.044],but no significant difference of DVR filling rate was found between the two groups. Conclusions Left ventricular volume curve analysis techniques can be used for detailed evaluation of left ventricular diastolic function.The left ventricular diastolic dysfunction of hypertrophic cardiomyopathy occurs mainly in early diastolic period,and accompanied by the shortening of the plateau time. Myocardial fibrosis can aggravate early left ventricular diastolic dysfunction of hypertrophic cardiomyopathy.

Objective To explore the influence of different injection techniques on the quality of bolus in 99mTc-DTPA renal dynamic imaging.Methods 395 patients accepted 99mTc-DTPA renal dynamic imaging were retrospectively analyzed.All patients were divided into three groups according to injection techniques:direct injection group (187 cases),intravenous route injection group (84 cases)and venous indwelling needle injection group (124 cases).The three groups were injected by each technique.Areas of interest (ROI) were drawn on abdominal aorta by Xeleris workstation in blood flow perfusion imaging.The time-radioactivity curves of ROI were got.The patients whose ROI curve formed a peak was successfully injected,and did not formed was unsuccessfully injected.The number of patients in three groups who were successfully or unsuccessfully injected was respectively calculated.The data of three groups was taken Chisquare test by SPSS17.0 software.Results 174 patients of the direct injection group,46 of the intravenous route injection group and 115 of the venous indwelling needle injection group were injected successfully.The successful rate respectively was 93.0％,54.8％ and 92.7％.The successful rate of the direct injection group and venous indwelling needle injection group were higher than intravenous route injection group.The difference had statistical significance.The successful rate of the direct injection group and venous indwelling needle injection group hadn't statistical significance.Conclusions The successful rates of the direct injection group and venous indwelling needle injection group were similar.The venous indwelling needle injection technique can be chosen.The successful rate of the intravenous route injection group was lower than the other two groups.The intravenous route injection technique should be chosen prudently.

Objective This study was designed to observe the impact of cigarette cessation on anxiety and depression in patients with coronary heart disease (CHD).Methods A total of 690 cigarette smoking patients with CHD identified by coronary angiography (CAG) were included and analyzed in the study.The mental state were scored with Hamilton anxiety (HAMA) and depression (HAMD) scale both on admission and at 6-month follow-up.The patients were divided into two groups based on the cigarette cessation.The score of mental state between the two groups were compared.The patients were treated with percutaneous coronary intervention (PCI),coronary artery bypass grafting (CABG),or medicine therapy (MT).Results The clinic data and score of mental state were similar at the time of admission (HAMA:10.66±5.53 vs 11.09 ±5.61,P =0.311;HAMD:29.81 ± 10.13 vs 28.94 ± 10.22,P =0.266 4) between the two groups.After 6 months,the proportions of subjects in smoking cession group with anxiety (24.2％ vs 32.3％,P ＜0.05),depression (18.0％ vs 27.5％,P ＜0.05),and anxiety and depression (7.0％ vs 16.2％,P ＜0.001) decreased significantly compared with those in smoking group irrespective of the treatment strategy.Both the HAMA and HAMD scores were lower in smoking cessation group (HAMA:9.83±3.40;HAMD:24.91 ±7.90) than in smoking group (HAMA:10.98 ±4.87;HAMD:27.70 ± 11.16) (all P ＜ 0.001).Conclusions Smoking cessation is good for the relief of anxiety and depression in CHD patient.

This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.

Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.