PurposeTo evaluate the motion correction algorithm (snapshot freeze, SSF) and multi-cycle reconstruction on the imaging quality and interpretation of coronary CT angiography (CCTA).Materials and Methods Sixty patients with suspected coronary heart disease were divided into two groups according to the heart rate. Twenty-ifve patients with heart rate ≤65 beats per minute underwent prospectively ECG-gated CCTA on gemstone CT. Image quality and interpretability of standard (STD) and SSF reconstructions at 75% R-R interval were compared. Thirty-five patients with heart rate>65 beats per minute underwent retrospective ECG-gated CCTA. Image quality and interpretability of STD and SSF reconstructions at 45% and 75% R-R interval were compared. The image qualities and interpretability were assessed based on coronary artery segment, coronary artery bifurcation and overall coronary artery.Results For 25 patients with prospectively ECG gated CCTA, SSF reconstructions showed higher interpretability than STD reconstructions on per-segment level [96.3% (341/354)vs 90.7% (321/354),P<0.01]. Image qualities with SSF were higher than that with STD reconstructions for left anterior descending (LAD), left circumflex (LCX) and right coronary artery (RCA). There was no statistic significance for left main coronary artery (P>0.05). For 35 patients with retrospective ECG-gate CCTA, SSF reconstructions at 45% R-R interval showed higher interpretability (P<0.01,P<0.001). But there were no signiifcant difference for 75% R-R interval images (P>0.05). Image qualities with SSF were higher than that with single and double sector STD reconstructions (P<0.001). The image qualities were signiifcant different among SSF, single and double sector STD reconstructions for 45% R-R interval images of LM (P<0.01), but it was similar for 75% R-R interval images (P>0.05). There were signiifcant different for LAD, LCX and RCA on both 45% and 75% R-R interval images (P<0.01,P<0.001). Conclusion SSF may improve the image quality and interpretability of CCTA images using prospectively and retrospective ECG-gated techniques, especially for improving the image quality.

Human CD3AK cells were prepared from peripheral blood mononuclear cells by culturing with recombinant IL-2 and antiCD3AK McAb. The mechanism and regulation of CD3AK cytotoxic activity with cytokines (rhIFN-?, rhIFN-?, TNF) and chernotherapeutic agents (CDDP or ADM) were observed by LDH-release assay, ABC-CELISA and the flow cytometric assay. The results showed: (1) Adhesion molecules ICAM-l/LFA-1 participated in CD3AK-mediated killing of tumor cells, hrlFN-? and TNF enhanced cytotoxicity of CD3AK through this pathway. (2) CD3AK could indirectly kill tumor cells by releasing soluable cytotoxic factors. (3) The membrane-associated TNF may be involved in CD3AK-mediated cytotoxicity. (4) CD3AK cells could induce the apoptosis of tumor cells. (5) Pretreatment of tumor cells with CDDP or ADM resulted in the increased vulnerability of tumor cells to CD3AK-mediated killing, the enhancement of CD3AK-mediated cytotoxicity by CDDP was relative to the increased expression of ICAM-1, HLA-ABC on tumor cell membrane.

BACKGROUND:CTLA-4Ig as a tolerance-induction agent is a potential strategy in graft-versus-host disease prevention. OBJECTIVE:To investigate the efficacy of CTLA4Ig-gene-modified bone marrow stromal cels mediated by adenovirus to induce T-cel tolerance of haploidentical donors. METHODS: The bone marrow stromal cels isolated culture from the bone marrow of HLA haploidentical donors were transfected by recombinant adenovirus encoding CTLA4IgcDNA (AdCTLA4Ig) at a multiplicity of infection=50 for 72 hours. The expression rate and the location of CTLA4Ig in the transfected cels were detected by fluorescence microscope after immunofluorescence staining. CTLA4Ig-modified bone marrow stromal cels (2×104, 4×104and 8×104) were respectively co-cultured with 105 T cels from the peripheral blood of HLA haploidentical donors and 105 peripheral blood mononuclear cels from recipients. The proliferative inhibition rate was determined by MTT assay, and the level of interleukin-2 in the supernatant was detected by ELISA. The bone marrow mononuclear cels (1×105/wel) were co-cultured with CTLA4Ig-modified bone marrow stromal cel layers constructed in 6-wel plates. The number of bone marrow mononuclear cels and colony-forming unit-granulocyte macrophages were calculated after 5-day culture. RESULTS AND CONCLUSION: The expression rate of CTLA4Ig at the multiplicity of infection=50 was as high as 85%, and the immunofluorescence signals of CTLA4Ig were distributed unevenly in the cytoplasm. The inhibition rates of 2×104, 4×104, and 8×104 CTLA4Ig-modified bone marrow stromal cels on proliferation of T cels were higher than that of untransfected cels. The levels of interluekin-2 in the corresponding cel groups were significantly lower than that in the untransfected cels (P < 0.05). At 5 days of culture, there was no significant difference in the number of bone marrow mononuclear cels and colony-forming unit-granulocyte macrophages between the transfected and untransfected cel groups (P > 0.05). These findings indicate that CTLA4Ig-modified bone marrow stromal cels mediated by adenovirus can induce immune tolerance of T-lymphocyte from HLA haploidentical donors in vitro.

Objective To assess the early prognosis of 117 patients after carduopulmonary resuscitation (CPR) in ICU by using the markers of inflammation,Glasgow Coma Scale (GCS) and Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) scores.Methods A total of 117 CPR patients admitted between 2010 January to 2012 December were enrolled for study.Within 24 h after admission,inflammatory markers,blood routine items,hepatorenal function,electrolytes of blood were measured.The GCS and APACHE Ⅱ scores were recorded.Arterial blood gas analyses were performed at 0,12,and 24 h after hospitalization,and the 12-h and 24-h lactate clearance rates were calculated.Seven days after treatment,according to the outcomes,the patients were divided into survival group and death group,and the clinical data of two groups were analyzed.Results (1) Of them,73 patients died and 44 survived.Factor analysis showed that age,time elapsed from resuscitation to ICU admission,D-dimer,arterial oxygenation index (FiO2),arterial blood pH,arterial blood lactate concentration upon ICU admission,GCS score and APACHE Ⅱ score were significantly different between the two groups (P ＜ 0.05or P ＜ 0.01); (2) Two classification logistic regression analysis showed that D-Dimer,GCS score and APACHE Ⅱ score significantly correlated with the mortality risk of the patients in the wake of CPR with relative odds ratios of 1.000,2.091,and 0.531,respectively (P ＜ 0.05 or P ＜ 0.01) ; (3) Receiver operating characteristic curve analysis indicated that the area under the curve of GCS (0.821) and APACHE Ⅱ (0.869) had higher predictive value than D-dimer (0.655).The highest accuracy (84.6％) in predicting patient survival was achieved when the GCS score was 6.5.Meanwhile,the highest accuracy (82.1％) in predicting patient death was achieved when the APACHE Ⅱ score was 17.5.Conclusions Both GCS score and APACHE Ⅱ score has obvious correlation with the prognosis of the critically ill patients after CPR and could be used to predict prognosis at early stage.

Objective To investigate the expression of long non - coding RNA(lncRNA)in neonatal rats with hypoxic - ischemic brain damage(HIBD). Methods SD rats of 10 postnatal days were divided into the sham -operated control and the hypoxic - ischemic(HI)group. At 24 h after HI,the animals were sacrificed. HE staining was used to assess histopathological damage. Microarray was used to detect the expression of lncRNA and mRNA in hypoxic -ischemic and sham control brain. Real - time PCR was used to verify the microarray result. The differentially expressed mRNA was analyzed by gene ontology(GO),pathway and coding - noncoding RNA co - expression(CNC)network analysis. Results HE staining showed that cells in HI brains became swollen and disordered with ambiguous cell struc-ture. Microarray data demonstrated that 322 lncRNAs and 375 mRNAs were significantly altered in the neonatal brains following hypoxic - ischemic injury compared with sham control(P ﹤ 0. 05). The real - time PCR results agreed with those of the microarray. GO analysis showed that the most enriched biological process associated with the upregulated mRNA had response to wounding,whereas the biological process mostly enriched among the downregulated mRNA was so-matic stem cell division. Pathway analysis indicated that upregulated mRNA was primarily corresponded with cytokine -cytokine receptor interaction pathway and that downregulated mRNA mainly correlated to axon guidance pathway. CNC network analysis demonstrated that 177 lncRNAs were correlated to the expression of mRNA involved in inflammation and cell death(P ﹤0. 05). Conclusions HI injury significantly influences cerebral lncRNA and mRNA expression profiles in the neonatal rat brains. Deregulated lncRNAs might contribute to the pathogenesis of HIBD via interacting with mRNA.

High expression of Fightless I (FLII) is associated to multiple tumors. Based on our previous study that FLII might be involved in the nuclear export, we assessed the possible interaction of FLII with the nuclear envelop associating proteins Importin β and Nup88. We first constructed GST-FLII, GST-LRR recombinant plasmids and transformed them into the Rosetta strain to produce GST-FLII, GST-LRR fusion protein. After purification of these proteins, GST-pull down, as well as co-immunoprecipitation, were used to test the interaction of FLII with Importin β and Nup88. FLII interacted with Importin β and Nup88, and FLII LRR domain is responsible for these interactions. Thus, FLII may play a role in nuclear export through interaction with Importin β and Nup88.
Humans ; Microfilament Proteins ; metabolism ; Nuclear Pore Complex Proteins ; metabolism ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Recombinant Fusion Proteins ; metabolism ; beta Karyopherins ; metabolism

Objective To investigate the effectiveness and safety in patients with largeartery occlusive acute cerebral infarction who received multi-interventional modes mainly with mechanical thrombectomy and its related factors affecting prognosis. Methods The clinical data of 56 patients with large artery occlusive acute cerebral infarction were analyzed retrospectively. The clinical characteristics (gender,age,and underlying diseases),timing of treatment (time from ictus to puncture,time from puncture to recanalization), multi-interventional mode therapies (intra-arterial thrombolysis,thrombectomy,balloon dilation,and stenting, etc. ),and distribution of offending vessels were observed. The modified Thrombolysis in Cerebral Ischemia Scale (mTICI)grade was used to evaluate revascularization. The National Institute of Health Stroke Scale (NIHSS)score was used to observe the neurological function at 24 h before and after procedures. The modified Rankin scale (mRS)was used to evaluate the prognosis at 3 months after procedure. The safety of the treatment was evaluated with operative complications (mainly symptomatic intracranial hemorrhage)and mortality. The patients were divided into either a good prognosis group (n = 34;mRS≤2)or a poor prognosis group (n =22;mRS≥3)according to the prognosis at 3 months after procedure. They were analyzed with univariate analysis. The factors influencing the prognosis were further analyzed with multivariate logistic regression analysis. Results (1)The recanalization rate in 56 patients was 78. 6%(n = 44),in which basilar artery was the highest,reaching 93. 8% (15 / 16),middle cerebral artery was 87. 0% (20 / 23). The NIHSS score at 24 hours was 10 ± 7,it was lower than 16 ± 6 on admission. There was significant difference (t =6. 401,P <0. 01). At 3 months,34 patients (60. 7%)had good prognosis,4 (7. 1%)died,and 8 (14. 3%) had symptomatic intracranial hemorrhage. (2)Multiple factor analysis showed that the high level of recanalization was a protective factor for good prognosis (OR,0. 465,95% CI 0. 267 -0. 809,P =0. 007). Diabetes was an independent risk factor for poor prognosis (OR,5. 535,95% CI 1. 101 -27. 835, P = 0. 038). Conclusion Acute large artery occlusive cerebral infarction treated with the intra-arterial multi-interventional modes may quickly and effectively restore intracranial blood flow. It has the characteris-tics of high recanalization rate and good prognosis,and the higher the level of recanalization,the better the prognosis. Diabetes is an independent risk factor for poor prognosis.

Objective To evaluate the role of Toll-like receptor 4 (TLR4)-p38 mitogen-assoliated protein kinase (p38MAPK)-nuclear factor kappa B (NF-κB) signaling pathway in sevoflurane-induced decrease in cognitive function of aged rats.Methods Sixty SPF healthy male Sprague-Dawley rats,aged 20 months,weighing 550-750 g,were divided into 5 groups (n =12 each) using a random number table method:control group (C group),sevoflurane group (S group),TAK242 plus sevoflurane group (TS group),SB202190 plus sevoflurane group (SS group),and PDTC plus sevoflurane group (PS group).All the rats were intubated after anesthesia and connected to an animal ventilator.TAK242,SB202190 and PDTC 10 μl were injected into the lateral cerebral ventricle in TS,SS and PS groups,respectively,and normal saline containing the equal volume of DMSO was given in C and S groups.Starting from 10 min after lateral cerebral ventricle injection,4％ sevoflurane was inhaled for 6 h via the tracheal tube,with the inhaled oxygen concentration 30％ and oxygen flow rate 2 L/min.The mixture of air and oxygen was inhaled in C group.The learning and memory ability was assessed by Morris water maze test at 7 days after the end of sevoflurane anesthesia,and the escape latency and swimming distance were recorded.Animals were sacrificed after the end of Morris water maze test,and brains were removed and hippocampi were isolated for determination of neural apoptosis (by TUNEL),contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in hippocampal tissues (by enzyme-linked immunosorbent assay),and expression of caspase-3,phosphorylated p38MAPK (p-p38MAPK),total p38MAPK (t-p38MAPK) and NF-κB in nucleus (by Western blot).The apoptosis rate and p-p38MAPK/t-p38MAPK ratio were calculated.Results Compared with C group,the escape latency and swimming distance were significantly prolonged at each time point,the apoptosis rate and contents of TNF-oα and IL-1β were increased,the expression of caspase-3,p-p38MAPK and NF-κB was up-regulated,and p-p38MAPK/t-p38MAPK ratio was increased in the other four groups (P＜0.05).Compared with S group,the escape latency and swimming distance were significantly shortened at each time point,the apoptosis rate and contents of TNF-α and IL-1β were decreased,and the expression of caspase-3 was down-regulated in TS,SS and PS groups,the expression of NF-κB was significantly down-regulated in TS and SS groups,and the expression of p-p38MAPK was significantly down-regulated,and p-p38MAPK/t-p38MAPK ratio was decreased in TS group (P＜0.05).Compared with TS group,the escape latency and swimming distance were significantly prolonged at each time point,the apoptosis rate and contents of TNF-α and IL-1β were increased,the expression of caspase-3 and p-p38MAPK was up-regulated,and p-p38MAPK/t-p38MAPK ratio was increased in SS and PS groups,and the expression of NF-κB was significantly up-regulated in PS group (P＜0.05).The expression of NF-κB was significantly up-regulated in PS group when compared with SS group (P＜0.05).Conclusion TLR4-p38MAPKNF-κB signaling pathway is involved in sevoflurane-induced decrease in cognitive function of aged rats.

Objective To explore the effect of erythropoietin (EPO) attenuating apoptosis in old rat hippocampal neuronal cells exposed to sevoflurane and the role of toll like receptor 4.Methods Twenty months old SD rats,male,550-750 g,in accordance with the random number table,were divided into 3 groups (n =9):control group (group C),sevoflurane treatment (group S),and sevoflurane plus EPO treatment (group ES).The rats in group S and ES were subjected to inhale 4％ sevoflurane for 6 h,but the rats in group C were inhaled air-oxygen only.The rats in group ES were injected with EPO into caudal vein at 24 h,48 h,and 72 h after sevoflurane exposure.The cognitive ability was assessed by Morris water maze test;the effects of hippocampal apoptosis were assessed by TUNEL assays;the expressions of TLR4 mRNA was measured by RT-PCR assay;mitochondrial membrane potential (MMP) was assessed by JC-1 fluorescence;the expressions of APP and Aβ were assessed by western blot.Results Compared with group C,there were significant increases of escape latency period,neuronal apoptosis,TLR4 mRNA,and APP and Aβ expression,but a decrease of MMP in group S (P＜0.05).Compared with group S,there were significant decreases of escape latency period,neuronal apoptosis,TLR4 mRNA,and APP and Aβ expression,but a increase of MMP in group ES (P＜0.05).Conclusion The attenuation of rat hippocampal neuronal apoptosis induced by EPO could be associated with inhibition of TLR4,improvement of MMP,as well as inhibition of APP and Aβ activity.

Objective To evaluate the role of Toll-like receptor 4 (TLR4) signaling pathway in sevoflurane anesthesia-induced cognitive dysfunction in aged rats.Methods Twenty-seven SPF healthy male Sprague-Dawley rats,aged 20 months,weighing 550-750 g,were divided into 3 groups (n =9 each) using a random number table:control group (C group),sevoflurane anesthesia group (S group) and TLR4 antagonist plus sevoflurane anesthesia group (TS group).TLR4 monoclonal antibody 30 μl was injected into the lateral cerebral ventricle in group TS,and the equal volume of serum containing no antibody was injected into the lateral cerebral ventricle in C and S groups.At 10 min after completion of injection,S and TS groups inhaled the mixture of 4％ sevoflurane and 30％ oxygen for 6 h,and group C only inhaled the mixture of air and oxygen.Morris water maze test was performed at 24 h after the end of sevoflurane anesthesia.The animals were sacrificed after completion of Morris water maze test,brains were removed and hippocampi were isolated for determination of nerve cell apoptosis (using TUNEL) and expression of activated caspase-3 (using immunofluorescent staining).Nerve cell apoptosis rate was calculated.The expression of high-mobility group box 1 protein (HMGB1) mRNA in hippocampi was measured by Northern blot assay at 6 h after the end of sevoflurane anesthesia.The expression of amyloid precursor protein (APP) and amyloid beta protein (Aβ) in hippocampi was assessed by Western blot at 24 h after the end of sevoflurane anesthesia.Results Compared with C group,the escape latency was significantly prolonged,nerve cell apoptosis rate was increased,the expression of activated caspase-3,HMGB1 mRNA,APP and Aβ was up-regulated in group S,and nerve cell apoptosis rate was increased,the expression of activated caspase-3,HMGB1 mRNA,APP and Aβ was up-regulated (P＜0.05),and no significant change was found in the escape latency in group TS (P＞0.05).Compared with S group,the escape latency was significantly shortened,nerve cell apoptosis rate was decreased,and the expression of activated caspase-3,HMGB1 mRNA,APP and Aβ was down-regulated in group TS (P＜0.05).Conclusion Activation of TLR4 signaling pathway is involved in the mechanism of sevoflurane anesthesia-induced cognitive dysfunction in aged rats.