Objective To study the characteristics of cerebral glucose metabolism in adults with hypoxic-ischemic encephalopathy (HIE) using positron emission tomography (PET) and statistical parameter mapping (SPM), and evaluate the degree of the cerebral glucose metabolism damaged. Methods 26 HIE patients and 20 healthy controls received 18F-FDG PET imaging. The scope and degree of the brain radioactivity decrease were observed with visual analysis. The three-dimensional projection images and the KE value were obtained by SPM analysis. Results The glucose metabolic decrease in HIE was primarily bilaterally. The bilateral basal ganglia and thalamus metabolism decreased most obviously. The brain cortical lobes varied degrees of metabolic decrease according to the order from high to low was the frontal, occipital, parietal and temporal lobes. The basal ganglia and thalamus were taken as the important target area of the evaluation of damage degree, the degree of damage of HIE was divided into Level Ⅰ(mild), Level Ⅱ(moderate) and Level Ⅲ (severe) combined with cortex damage. Conclusion The basal ganglia and thalamus are the target areas of metabolic damage, the classification combined with brain cortex damage degree and scope can be used to guide the clinical treatment and prognosis evaluation.

Objective To evaluate the therapeutic effect of limb ischemic preconditioning (LIPC) by observing the changes of brain glucose metabolism using positron emission tomography (PET) and statistical parametric mapping (SPM). Methods 40 patients with severe stenotic or occlusion cerebral artery lesions were enrolled and randomized into LIPC group (n=20) and control group (n=20). Brain lesions and cerebral hemorrhagic lesions were excluded after magnetic resonance imaging. The glucose metabolism of patients was analyzed before and after treatment in two groups, respectively, using the methods of radioactivity ratio and SPM. Results There were 5 patients drop-out in the control group. Comparison of the glucose metabolism ratio of the impaired area to the opposite area: LIPC group improved better than the control group (P<0.01) while the control group aggravated heavier than LIPC group (P<0.05). Comparing the glucose metabolism of patients before and after treatment in two groups, respectively, by paired-t test, 1) Setting the glucose metabolism of patients increased after therapy: There were 9 areas activated in LIPC group, including frontal, parietal, temporal, occipital lobes, basal ganglia and thalamus, and the KE=927, while there were only 3 areas activated in the control group, including frontal, parietal and occipital lobes, and the KE=289. 2)Setting the glucose metabolism of patients decreased after therapy: There was no area activated in LIPC group, while there were 2 areas activated in the control group, including parietal and temporal lobes, and the KE=115. Conclusion The improvement of glucose metabolism was observed in cerebral cortex, basal ganglia and thalamus of the patients with severe stenotic or occlusion cerebral artery lesions after LIPC by PET and SPM.

Objective To establish a flow-cytometric method to detect microvesicles (MVs) in rat peripheral blood, and to detect platelets-derived MVs (PMVs) and endothelial cell-derived MVs (EMVs) in blood from ischemic precondition-ing (IPC) treated rats. Methods Blood was withdrawn from rat abdominal aorta and anticoagulated with sodium citrate. Platelets-free plasma (PFP) was isolated through two centrifugations at room temperature. PFP was incubated with FITC-conjugated mouse anti-rat CD61 or PE-conjugated mouse anti-rat CD144. Standard beads in diameter of 1 and 2μm were used for calibration and absolute counting, respectively. Analysis was performed on flow cytometer. Results When 3.5%so-dium citrate was mixed with blood at volume ratio of 1∶4, clear supernatant was collected after centrifugation. Signals of parti-cles smaller than 1μm accounted for more than 99%of overall signals. PMVs and EMVs were CD61 positive and CD144 positive, respectively. Their diameters were both smaller than 1 μm. The concentration of PMVs and EMVs in peripheral blood from IPC treated rats was (4 053±1 987)/μL and (4 870±825)/μL, respectively. Conclusion The method for MVs de-tection by flow cytometry was successfully established and optimized, and verified through detecting PMVs and EMVs in pe-ripheral blood from IPC treated rats.

To gain a better understanding of the genetic diversity and evolution of PRRSV in the Ningxia Hui Nationality Autonomous Region (Ningxia) of China, the nsp2 genes from a series of PRRSV strains collected from the region in 2007 were partially sequenced. These sequences were then analyzed along with the classical strain (ch-la) and two other epidemic strains SD (3) and SD2006. Comparison of the nucleotide sequence with ch-la indicated that nsp2 genes of seventeen Ningxia isolates (NX strain) have deletions of 87 nucleotides. Sequence analysis indicated that homology between the Ningxia strain and ch-la was 60.3%-79.9% in the nucleotide sequence, and homology between the NX strains and SD strains was 80.3%-98.8% in the nucleotide sequence. The nsp2 genes of the seventeen isolates had 74.9%-100% nucleotide sequence identities with each other. This study was undertaken to assess the regional variation of prevalent PRRSV and to establish a sequence database for PRRSV molecular epidemiological studies.

Objective To analyze the complications of coronary rotational atherectomy and to evaluate the safety of the procedure. Methods We evaluated the procedural and angiographic outcomes of 248 consecutive procedures to rotational atherectomy between January 2000 and October 2016. Results 27 cases(10.9％)were found to have rotational atherectomy related complications. Among these 27 cases,coronary spasm occurred in 2 cases(0.8％),no reflow in 8 cases(3.2％), coronary dissection in 5 cases(2.0％),burr entrapment in 6 cases(2.4％),wire breakage in 3 cases(1.2％), and coronary perforation in 3 cases(1.2％). There were no death,acute myocardial infarction and emergent coronary bypass graft.14 case(5.6％)had PCI-related myocardial infarction during hospital stay.In-hospital major adverse cardiaccerebral event(MACCE)rate was 5.6％. Conclusions Coronary rotational atherectomy can be performed with high success rates and procedure-related complications are rare.

OBJECTIVETo investigate the effects of endothelial microvesicles (EMVs) induced by calcium ionophore A23187 on H9c2 cardiomyocytes.

METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with 10 micromol/L A23187 for 30 min. EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. EMVs were characterized using 1 and 2 microm latex beads and anti-PE-CD144 antibody by flow cytometry. For functional research, EMVs at different concentrations were cocultured with H9c2 cardiomyocytes for 6 h. Cell viability of H9c2 cells and the activity of LDH leaked from H9c2 cells were tested by colorimetry. Moreover, apoptosis of H9c2 cells was observed through Hoechst 33258 staining and tested by FITC-Annexin V/PI double staining.

RESULTSEMVs were induced by A23187 on HUVECs, and isolated by ultracentrifugation. We identified the membrane vesicles (< 1 microm) induced by A23187 were CD144 positive. In addition, the EMVs could significantly reduce the viability of H9c2 cells, and increase LDH leakage from H9c2 cells in a dose dependent manner (P < 0.05). Condensed nuclei could be observed with the increasing concentrations of EMVs through Hoechst 33258 staining. Furthermore, increased apoptosis rates of H9c2 cells could be assessed through FITC-Annexin V/PI double staining by flow cytometry.

CONCLUSIONMicrovesicles could be released from HUVECs after induced by A23187 through calcium influx, and these EMVs exerted a pro-apoptotic effect on H9c2 cells by induction of apoptosis.

Annexin A5 ; Apoptosis ; Calcimycin ; pharmacology ; Calcium ; metabolism ; Cell Line ; Cell Membrane ; drug effects ; Coculture Techniques ; Flow Cytometry ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Human Umbilical Vein Endothelial Cells ; Humans ; Myocytes, Cardiac ; drug effects ; Staining and Labeling

OBJECTIVETo investigate CD4+ CD25+ regulatory T cell frequencies in the peripheral blood of poor or non-responsiveness to Hepatitis B vaccine, and try to understand the relationship between CD4+ CD25+ regulatory T cell and poor or non-responsiveness to Hepatitis B vaccine.

METHODSFlow cytometric analysis was employed for CD4+ CD25+ regulatory T cell frequencies in the peripheral blood of 25 cases of non-responsiveness, 30 cases of poor-responsiveness, and collected 20 cases of responsiveness as control.

RESULTSCD4+ CD25+ regulatory T cell frequencies of responsiveness was (4.32 +/- 1.21)%, poor-responsiveness was (7.01 +/- 1.06)% and non-responsiveness was (12.75 +/- 2.01)%. It was found that non and poor-responsiveness showed a high percentage of CD4+ CD25+ regulatory T cell as compared with responsiveness (t = 8.426, t = 3.289, P<0.01).

CONCLUSIONThe poor and non-responsiveness should be related with the increase of CD4+ CD25+ regulatory T cell and this might be considered as an important cause of poor and non-responsiveness.

Adolescent ; Adult ; Blood ; immunology ; CD4 Antigens ; immunology ; Female ; Flow Cytometry ; Hepatitis B Vaccines ; immunology ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Young Adult

The off-label drugs use is a common practice in clinical medicine,which has both medical ethical legitimacy and ethical confusion,as well as hidden medical and legal risks.Appropriate the off-label drugs use can alleviate patients'pain,otherwise endangering their safety.The new Physician Law establishes legal norms for the off-label drugs use.The medical ethics and health law knowledge learned by resident physicians during their undergraduate studies all are basic knowledge and general principles,while the legal and ethical knowledge about the off-label drugs use is insufficient.A questionnaire survey on the ethical and legal issues of the off-label drugs use for resident physicians found that resident physicians urgently need to learn ethical and legal knowledge,as well as receive clinical application teaching and training in the aspect of the off-label drugs use.Therefore,in the standardized training of resident physicians,ethical and legal education on the off-label drugs use has been added.The core of this training is to conduct theoretical lectures focusing on the six ethical and legal principles of the off-label drugs use,as well as providing practical teaching drawing on the"clinic-style legal teaching"model.This paper explored a method for conducting ethical and legal education on the off-label drugs use in the future during the resident training stage.

OBJECTIVETo investigate the effects of microvesicles (MVs) derived from hypoxia/reoxygenation (H/R)-treated human umbilical vein endothelial cells (HUVECs) on endothelium-dependent relaxation of rat thoracic aortic rings.

METHODSH/R injury model was established to induce HUVECs to release H/R-EMVs. H/R-EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. H/R-EMVs were characterized using 1 μm latex beads and anti-PE-CD144 by flow cytometry. Thoracic aortic rings of rats were incubated with 2.5, 5, 10, 20 μg/ml H/R-EMVs derived from H/R-treated HUVECs for 4 hours, and their endothelium-dependent relaxation in response to acetylcholine (ACh) or endothelium-independent relaxation in response to sodium nitroprusside (SNP) was recorded in vitro. The nitric oxide (NO) production of ACh-treated thoracic aortic rings of rats was measured using Griess reagent. The expression of endothelial NO synthase (eNOS) and phosphorylated eNOS (p-eNOS, Ser-1177) in the thoracic aortic rings of rats was detected by Western blotting. Furthermore, the levels of SOD and MDA in H/R-EMVs-treated thoracic aortic rings of rats were measured using SOD and MDA kit.

RESULTSH/R-EMVs were induced by H/R-treated HUVECs and isolated by ultracentrifugation. The membrane vesicles (< 1 μm) induced by H/R were CD144 positive. ACh-induced relaxation and NO production of rat thoracic aortic rings were impaired by H/R-EMVs treatment in a concentration-dependent manner (P < 0.05, P < 0.01). The expression of total eNOS (t-eNOS) was not affected by H/R-EMVs. However, the expression of p-eNOS decreased after treated with H/R-EMVs. The activity of SOD decreased and the level of MDA increased in H/R-EMVs treated rat thoracic aortic rings (P < 0.01).

CONCLUSIONACh induced endothelium-dependent relaxation of thoracic aortic rings of rats was impaired by H/R-EMVs in a concentration-dependent manner. The mechanisms included a decrease in NO production, p-eNOS expression and an increase in oxidative stress.

Acetylcholine ; pharmacology ; Animals ; Aorta, Thoracic ; physiology ; Cell Hypoxia ; Endothelium, Vascular ; physiology ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; In Vitro Techniques ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Nitroprusside ; pharmacology ; Oxidative Stress ; Rats

OBJECTIVETo establish a flow cytometric method to detect the alteration of phenotypes and concentration of circulating microvesicles (MVs) from myocardial ischemic preconditioning (IPC) treated rats (IPC-MVs), and to investigate the effects of IPC-MVs on ischemia/reperfusion (I/R) injury in rats.

METHODSMyocardial IPC was elicited by three.cycles of 5-min ischemia and 5-min reperfusion of the left anterior descending (LAD) coronary artery. Platelet-free plasma (PFP) was isolated through two steps of centrifugation at room temperature from the peripheral blood, and IPC-MVs were isolated by ultracentrifugation from PFR PFP was incubated with anti-CD61, anti-CD144, anti-CD45 and anti-Erythroid Cells, and added 1, 2 µm latex beads to calibrate and absolutely count by flow cytometry. For functional research, I/R injury was induced by 30-min ischemia and 120-min reperfusion of LAD. IPC-MVs 7 mg/kg were infused via the femoral vein in myocardial I/R injured rats. Mean arterial blood pressure (MAP), heart rate (HR) and ST-segment of electro-cardiogram (ECG) were monitored throughout the experiment. Changes of myocardial morphology were observed after hematoxylin-eosin (HE) staining. The activity of plasma lactate dehydrogenase (LDH) was tested by Microplate Reader. Myocardial infarct size was measured by TTC staining.

RESULTSTotal IPC-MVs and different phenotypes, including platelet-derived MVs (PMVs), endothelial cell-derived MVs (EMVs), leucocyte-derived MVs (LMVs) and erythrocyte-derived MVs (RMVs) were all isolated which were identified membrane vesicles (<1 Vm) with corresponding antibody positive. The numbers of PMVs, EMVs and RMVs were significantly increased in circulation of IPC treated rats (P<0.05, respectively). In addition, at the end of 120-min reperfusion in I/R injured rats, IPC-MVs markedly increased HR (P<0.01), decreased ST-segment and LDH activity (P < 0.05, P < 0.01). The damage of myocardium was obviously alleviated and myocardial infarct size was significantly lowered after IPC-MVs treatment (P < 0.01).

CONCLUSIONThe method of flow cytometry was successfully established to detect the phenotypes and concentration alteration of IPC-MVs, including PMVs, EMVs, LMVs and RMVs. Furthermore, circulating IPC-MVs protected myocardium against I/R injury in rats.

Animals ; Cell-Derived Microparticles ; metabolism ; Coronary Vessels ; pathology ; Flow Cytometry ; Heart Rate ; Ischemic Preconditioning, Myocardial ; Myocardial Infarction ; physiopathology ; Myocardial Reperfusion Injury ; physiopathology ; Myocardium ; pathology ; Phenotype ; Rats