AIM: To investigate the expression of peroxisome proliferator-activated receptors (PPARs) in human endothelial cells, and their effects on plasminogen activator inhibitor-1 transcription. METHODS: The expression of three types of PPARs in mRNA level were detected in human umbilical vein endothelial cells(HUVECs) by using RT-PCR. Cultured endothelial cells line-ECV304 were transfected with PAI-1 promoter controlling CAT reporter gene and co-transfected with varying doses (250, 500, 1 000 ng) of expression vectors PPAR? or PPAR?.The transcripton activity of PAI-1 promoter were detected with ELISA. RESULTS: There were all three types of PPARs mRNA expression in HUVECs, while the expression of PPAR? was less than that of PPAR?( P

Objective:To investigate the changes of default network functional connectivity (FC) in resting state of patients with insomnia disorder (ID).Methods:Fifty-six patients with insomnia disorder and fifty healthy controls were recruited. All subjects were assessed with Pittsburgh sleep quality index (PSQI), insomnia severity index (ISI), Epworth sleepiness score (ESS) and Hamilton depression scale (HAMD-17). All subjects were scanned with resting state functional magnetic resonance imaging (rs-fMRI). SPM12 and CONN18b were used to preprocess rs-fMRI data on MATLAB (R2013b).The medial superior frontal gyrus was defined as the seed point, and the differences in the functional connection strength of the two groups of subjects were compared. Pearson correlation analysis was performed between the FC value of the brain area with statistical significance and scores of PSQI and ISI.Results:(1)FC analysis showed that compared with the control group, patients with ID had abnormal default mode network (DMN) connection, as follows: FC of left medial superior frontal gyrus and left central anterior gyrus (MNI: x, y, z=-30, -15, 51) and anterior cingulate gyrus (MNI: x, y, z=-6, 24, 36), FC of right medial forehead between the last gyrus and the left central anterior gyrus (MNI: x, y, z=-48, -6, 48), FC of left anterior cuneiform and the left central back (MNI: x, y, z=-54, -18, 54) and left superior occipital gyrus (MNI: x, y, z=-21, -69, 30), FC of right precuneus and left central posterior gyrus (MNI: x, y, z=-60, -21, 48) was enhanced. The FC of left anterior cuneiform lobe and the right middle frontal gyrus (MNI: x, y, z=42, 33, 42) and the right angular gyrus (MNI: x, y, z=54, -51, 45), FC of right precuneus and left inferior temporal gyrus (MNI: x, y, z=-51, -12, -42) was weakened. (2)With the left medial superior frontal gyrus as the seed point, FC values in anterior cingulate gyrus ( r=0.451, 0.338, both P<0.01) and left anterior central gyrus ( r=0.324, 0.402, both P<0.05) were positively correlated with PSQI and ISI scores. With the right precuneus as the seed point, FC value of left posterior central gyrus was positively correlated with PSQI( r=0.333, P=0.013) and ISI scores( r=0.418, P=0.008), while FC value of left inferior temporal gyrus was negatively correlated with PSQI( r=-0.662, P=0.001) and ISI scores( r=-0.402, P<0.01).With the left precuneus as the seed point, FC value of left superior occipital gyrus was positively correlated with PSQI( r=0.438, P=0.001) and ISI scores( r=0.495, P=0.011). Conclusion:Patients with ID may have changes in the functional connection of multiple networks. The decrease of FC in DMN may be one of the potential causes of insomnia. The increase in FC between DMN and the visuospatial attention network may be the core of the mechanism of damage to the brain function network of insomnia link.

To investigate a new kind of tumor tracer 99mTc-YIGSR developed from a five amino structure (YIGSR) of the Laminin -chain, which can bind to the laminin receptors of tumor specifically, and radiolabeled with MAG3. (1) Preparation of the 99mTc-YIGSR probe: with S-Acetly-NH3-MAG3 as the chelator and with proper reductants YIGSR was labeled with 99mTc; (2) Cell culture and viability measurement: EAC was maintained in RPMI 1640 supplemented with calf serum; the trypan blue exclusion was applied to calculate the cell viability; (3) Study of the cell dynamic: The EAC's uptake of 99mTc-YIGSR and 99mTc-MIBI was observed at 37 degrees C and 22 degrees C, respectively. (1) The labeling efficiencies of 99mTc-YIGSR and 99mTc-MIBI were (62 +/- 3)% and (96 +/- 2)%, respectively; (2) The cell viability was declined with time of incubation; (3) At 37 degrees C, the EAC'S uptake of 99mTc-YIGSR and 99mTc-MIBI reached the peak of (43.16 +/- 2.4) % and (24.4 +/- 1.8) % at 60 min, respectively; and at 22 degrees C, the highest uptake was (26.5 +/- 2.1) % and (9.47 +/- 1.9) % at 60 min, respectively. The in vitro study suggests that 99mTc-YIGSR is superior to 99mTc-MIBI in cell uptake and has potential value in tumor imaging.
Carcinoma, Ehrlich Tumor/*radionuclide imaging ; Laminin ; Oligopeptides ; Radiopharmaceuticals/diagnostic use ; Radiopharmaceuticals/*pharmacokinetics ; Technetium Tc 99m Mertiatide/diagnostic use ; Technetium Tc 99m Mertiatide/*pharmacokinetics ; Technetium Tc 99m Sestamibi/diagnostic use ; Technetium Tc 99m Sestamibi/*pharmacokinetics ; Tissue Distribution

The validity of (99m)Tc-YIGSR, a novel receptor radio-tracer, in imaging the Ehrlich ascites tumor was evaluated. YIGSR, a pentapeptide of laminin, was labeled with (99m)Tc by using a bifunctional chelator S-Acetly-NH(3)-MAG(3). The MIBI was labeled with (99m)Tc by following the kit instruction. The mice of tumor group were intravenously injected 1-2 mCi of (99m)Tc-YIGSR or (99m)Tc-MIBI via caudal vein, immobilized and imaged under a Gamma camera. The same procedure was performed in mice of blockade group, in which the unlabeled YIGSR was previously injected to block the receptor-recognition sites, and inflammation group serving as control. The reverse-phase Sep-Pak C(18) chromatogram was found to have an essentially complete conjugation between YIGSR and S-Acetly-NH(3)-MAG(3). The conjugated YIGSR could be radio-labeled successfully with (99m)Tc at room temperature and neutral pH, with a radio-labeling yield of 62%. Without the chelator S-Acetly-NH(3)-MAG(3), the YIGSR was labeled with (99m)Tc at an efficiency of 4%. The imagological study revealed obvious tumor accumulation of (99m)Tc-YIGSR 15 min after the injection, and the uptake peaked after 3 h with a tumor-to-muscle ratio (T/M) of 11.36. The radio-tracer was slowly cleared up and resulted in a T/M of 3.01 at the 8th h after the injection. As for blocked group, the tumor uptake of radiotracer was significantly lower, with the highest T/M being 4.61 after 3 h and 0.89 after 8 h. The T/M was 3.72 at the 3rd h and 1.29 at the 8th h after the (99m)Tc-YIGSR injection in the inflammatory group. The T/M was significantly higher in tumor group than in inflammatory group or control group (P<0.001). In the 99mTc-MIBI group, the T/M was 1.40 at the 3rd h and 0.55 at the 8th h after the injection, which showed a significant difference as compared with (99m)Tc-YIGSR (P<0.001). It is concluded that YIGSR can be successfully radiolabelled by using S-Acetly-NH(3)-MAG(3). (99m)Tc-YIGSR has many advantages in tumor imaging, such as quick and clear visualization, high sensitivity and specificity, and satisfactory target/non-target ratio (N/NT). It promises to be tumor radio-tracer.
Carcinoma, Ehrlich Tumor/*radionuclide imaging ; Radioactive Tracers ; Radiopharmaceuticals/*diagnostic use ; Receptors, Laminin/*metabolism ; Technetium Tc 99m Mertiatide/*diagnostic use ; Technetium Tc 99m Sestamibi/*diagnostic use

Objective To evaluate the influences of FBP and OSEM (with different iteration numbers or subset numbers) on cardiac function parameters with MPI,and to identify a suitable clinical method.Methods The MPI of 66 patients (33 males,33 females,age range: 18-77 years) from January 2012 to December 2014 were retrospectively analyzed.The imaging data was reconstructed with FBP and 5 different OSEM (4-16 subsets and 2-4 iterations) in order to evaluate if there was any difference of cardiac function parameters calculated by QGS between the different image processing methods.In addition,the results were compared with those of cardiac ultrasonography.One-way analysis of variance was used for data analysis.Results The statistical differences of LVEF,EDV,and ESV(F values: 10.73,4.89 and 5.97,all P<0.05) were found among 6 reconstructed methods.The values of LVEF were (66.14±11.12)％,(75.05±12.10)％,(70.09±11.27)％,(66.88±10.38)％,(64.97±10.25)％,and (62.58±9.84)％.Those of EDV were (77.32±27.58),(67.97±27.56),(75.10±27.89),(81.03±28.11),(84.94±29.07),and (89.98±29.71) ml,and the ESV values were (28.71±10.04),(19.71±16.51),(25.13±17.66),(29.01±18.47),(32.10±19.63),and (35.83±20.41) ml respectively.The cardiac function parameters measured by OSEM (with 2 iterations,12 subsets) were much similar to that measured by cardiac ultrasonography.Conclusion Compared with other 5 processing methods of MPI,the OSEM (with 2 iterations,12 subsets) method may be more suitable for practical clinical application.

Objective To prepare a kind of 99Tcm-labeled estrogen receptor (ER) SPECT imaging agent,and evaluate its binding characteristics with ER by in vitro and in vivo studies.Methods EDL was prepared from estrone and then reacted with GAP to synthesize GAP-EDL.Then,GAP-EDL was labeled with 99Tcm to obtain 99Tcm-GAP-EDL.Cell uptake and blocking assays were performed in vitro on MCF-7and MDA-MB-231 cells.The biodistribution study of 99Tcm-GAP-EDL was performed on normal BALB/c mice at 30,60,120,180 and 240 min post injection.Nude mice bearing either MCF-7 or MDA-MB-231derived tumors were injected with 99Tcm-GAP-EDL,and subjected to γimaging at 1,2,4 h post injection.The mice injected with excess unlabeled GAP-EDL or estradiol were used as the blocking control.Two-sample t test was used for data analysis.Results The radiolabeling yield of 99Tcm-GAP-EDL was (98.8 ±0.5) ％ and the radiochemical purity after 24 h was over 90％.The cell uptake rate of MCF-7 cells at 240min was (4.84± 0.21) ％,which was significantly higher than that of MDA-MB-231 cells ((2.11±0.21) ％;t =15.96,P＜0.05).99 Tcm-GAP-EDL uptake rate of MCF-7 cells in blocking groups ((2.31 ± 0.28) ％ and (2.05±0.35) ％) decreased significantly compared to that of non-blocking group(t=11.52,11.16,both P＜0.05).Biodistribution studies showed that 99Tcm-GAP-EDL was mainly metabolized by the liver and kidneys,and exhibited quick blood clearance.Gamma imaging showed high uptake in MCF-7 tumors 1 h post injection and the uptake reached the highest at 4 h,while there was little 99Tcm-GAP-EDL uptake in MDA-MB231 tumors and blocked MCF-7 tumors.Conclusions 99Tcm-GAP-EDL may be prepared under mild conditions with high labeling purity and stability.The in vitro and in vivo characteristics of 99Tcm-GAP-EDL suggest that it may be a promising probe for ER positive tumor imaging.

In this study, a novel technique for the preparation of (125)I-5-trimethylstannyl-1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl) urail (FIAU) was developed, (125)I-FIAU biodistribution profile was detected in Kunming mice and the possibility of using FTAU radio-labeling for reporter gene imaging was explored. 5-trimethylstannyl-1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl) urail (FTAU) was labeled with radioiodine ((125)I). A rotary evaporation method was used to remove excess methanol. The reactant was purified through a Sep-Pak C18 reversal phase column. The radiochemical purity and in vivo stability were determined using silica gel thin layer chromatography (TLC). The biodistribution of (125)I-FIAU in Kunming mice was also detected. The results showed that (125)I-FIAU could be radiolabeled effectively with FTAU, with mean labeling rate being (81±0.38)% (n =5). The mean radiochemical purity of (98.01±0.40)% (n=5) was achieved after a reversal phase Sep-park column purification. (125)I-FIAU was stable when incubated in normal human serum or in saline at 37°C, with a radiochemical purity >96% during a 0.5-24 h time period. Biological experiments exhibited rapid clearance of (125)I-FIAU from the blood pool. (125)I-FIAU was mostly excreted by kidneys. (125)I-FIAU in myocardium dropped conspicuously after 8 h and there was hardly retention at 24 h. We were led to concluded that the new method of radioiodinization of FTAU for the preparation of (125)I-FIAU is easy, highly effective and stable in vivo. The biodistribution of (125)I-FIAU in Kunming mice showed it can serve as an imaging probe for myocardial reporter genes.

Objective To investigate factors related to chronic pain in those injured with fractures 27 months after the Sichuan earthquake.The correlation between intensity of pain and quality of life was also analyzed.Methods A total of 705 victims were investigated on site.Their residual pain was categorized using a visual analogue scale (VAS) score as no pain,mild pain,moderate pain or severe pain.The pain-related biological,psychological and social factors were analyzed using the Barthel Index ( BI ),Life Satisfaction Questionaire-11 ( LiSat-11 )and the SF-36 health questionnaire. Rehabilitation and surgical interventions,employment,income and emotional status were also investigated. Results The incidence of chronic pain was 88.5％ in this population,of which mild pain and moderate pain were 35.7％ and 33.3％,respectively.The percentage of the victims who had received fracture surgery was 65.8％ ; the percentage of those who had recovered was 96.9％.BI scores for the victims without pain,with mild,moderate and severe pain were 92.7 + 10.2,92.8 + 8.4,91.2 ± 9.9 and 90.4 + 14.7,respectively ; the differences between these groups were all statistically insignificant.The influence of pain intensity on life satisfaction showed a significant linear trend.The percentages of the victims with restricted occupational ability in the four groups were 38.3％,61.5％,75.7％ and 62.8％ respectively.The median of personal annual income were ￥ 3550,￥ 2500,￥ 2000 and ￥ 2500.The VAS scores were significantly related to abnormal emotions,life satisfaction,employment and annual income.The subjects with different levels of residual pain also showed significant differences in the physical functioning,role-physical,bodily pain,general health,vitality,social functioning,role-emotional and mental health sub-scales of the SF-36.The total SF-36 scores were highest among victims without pain (70.6 + 17.5) and declined significantly in those with mild (61.3 + 14.3 ),moderate (52.7 + 14.3 ) and severe pain (52.3 + 14.7 ).This negative correlation between pain intensity and SF-36 total score was statistically significant. Conclusions Chronic pain remains common among fracture victims 27 months after the earthquake.Its intensity is correlated with psychological and social factors as well as quality of life.

OBJECTIVETo investigate the effect of peroxisome proliferator-activated receptors (PPARs) activators on plasminogen activator inhibitor 1 (PAI-1) expression in human umbilical vein endothelial cells and elucidate a possible mechanism.

METHODSHuman umbilical vein endothelial cells (HUVECs) were obtained from normal fetus, and cultured conventionally. Then the HUVEC were exposed to fatty acids and prostaglandin J(2) in varying concentrations with fresh media. RT-PCR and ELISA were used to determine the expression of PPAR and PAI-1 in HUVECs. Transient co-transfection of PAI-1 promoter and PPARalpha gene or PPARgamma gene to ECV304 was performed.

RESULTSPPARalpha, PPARdelta and PPARgamma mRNA in HUVECs were detected by RT-PCR. Treatment of HUVECs with PPARalpha and PPARgamma activators-linolenic acid, linoleic acid, oleic acid and prostaglandin J(2), but not with stearic acid could augment PAI-I mRNA expression and protein secretion in a concentration-dependent manner. Proportional induction of PAI-1 promoter activity was observed through increasing amounts of PPARalpha DNA in HUVECs through a transient gene transfection assay, although the mRNA expression of the 3 subtypes of PPAR with their activators were not changed compared with controls.

CONCLUSIONSHUVECs express PPARs. PPARs activators may increase PAI-1 expression in endothelial cells (EC). Although PPARs expression was not enhanced after being stimulated by their activators in EC, the functionally active PPARalpha is probably involved in regulating PAI-1 expression in EC.

Cells, Cultured ; Fatty Acids ; pharmacology ; Gene Expression Regulation ; drug effects ; Humans ; Plasminogen Activator Inhibitor 1 ; genetics ; Prostaglandin D2 ; analogs & derivatives ; pharmacology ; RNA, Messenger ; analysis ; Receptors, Cytoplasmic and Nuclear ; genetics ; physiology ; Transcription Factors ; genetics ; physiology ; Transcription, Genetic ; drug effects

The validity of 99mTc-YIGSR, a novel receptor radio-tracer, in imaging the Ehrlich ascites tumor was evaluated. YIGSR, a pentapeptide of laminin, was labeled with 99mTc by using a bifunctional chelator S-Acetly-NH3-MAG3. The MIBI was labeled with 99mTc by following the kit instruction. The mice of tumor group were intravenously injected 1-2 mCi of 99mTc-YIGSR or 99mTc-MIBI via caudal vein, immobilized and imaged under a Gamma camera. The same procedure was performed in mice of blockade group, in which the unlabeled YIGSR was previously injected to block the receptor-recognition sites, and inflammation group serving as control. The reverse-phase Sep-Pak C18 chromatogram was found to have an essentially complete conjugation between YIGSR and S-Acetly-NH3-MAG3. The conjugated YIGSR could be radio-labeled successfully with 99mTc at room temperature and neutral pH, with a radio-labeling yield of 62%. Without the chelator S-Acetly-NH3-MAG3, the YIGSR was labeled with 99mTc at an efficiency of 4%. The imagological study revealed obvious tumor accumulation of 99mTc-YIGSR 15 min after the injection, and the uptake peaked after 3 h with a tumor-to-muscle ratio (T/M) of 11.36. The radio-tracer was slowly cleared up and resulted in a T/M of 3.01 at the 8th h after the injection. As for blocked group, the tumor uptake of radiotracer was significantly lower, with the highest T/M being 4.61 after 3 h and 0.89 after 8 h. The T/M was 3.72 at the 3rd h and 1.29 at the 8th h after the 99mTc-YIGSR injection in the inflammatory group. The T/M was significantly higher in tumor group than in inflammatory group or control group (P＜0.001). In the 99mTc-MIBI group, the T/M was 1.40 at the 3rd h and 0.55 at the 8th h after the injection, which showed a significant difference as compared with 99mTc-YIGSR (P＜0.001).It is concluded that YIGSR can be successfully radiolabelled by using S-Acetly-NH3-MAG3.99mTc-YIGSR has many advantages in tumor imaging, such as quick and clear visualization, high sensitivity and specificity, and satisfactory target/non-target ratio (N/NT). It promises to be tumor radio-tracer.