Acoustic Stimulation ; Animals ; Corpus Striatum ; physiology ; Estrogens ; metabolism ; Estrus ; physiology ; Evoked Potentials, Auditory ; physiology ; Female ; Rats ; Rats, Sprague-Dawley

Generation 4 polyamidoamine (PAMAM) dendrimer was PEGylated with polyethylene glycol (PEG) at an average molecular weight 5 000 via amide bond. PAMAM and PEGylated PAMAM (PAMAM-PEG) dendrimer were used as drug nanocarriers. Methotrexate (MTX), an antineoplastic agent, was selected as a model drug. PAMAM/MTX and PAMAM-PEG/MTX complexes were prepared. The pharmacokinetic characters and anti-tumor activity of the PAMAM-PEG/MTX complex were studied as compared with MTX injection and PAMAM/MTX complex by intravenous injection in rats and S180 tumor bearing mice, separately. The plasma samples from normal rats were analyzed by HPLC method, and concentration-time data were analyzed using a non-compartmental analysis. Their anti-tumor effects in vivo were evaluated against S180 solid tumors in mice by measuring average tumor weight and calculating the inhibitory rate of tumor on day 17 after successive injections. The results showed that both plasma half-life and mean retention time (MRT) of the complexes were longer than that of MTX injection (P<0.01), while the area under the plasma concentration vs time curve (AUC) of PAMAM-PEG/MTX was the largest as compared with that of free drug and PAMAM/MTX complex (P<0.01). The inhibitory rate of tumor of PAMAM-PEG/MTX complex enhanced 2.1 and 1.8 times over that of free drug and PAMAM/MTX complex, respectively, indicating that PAMAM-PEG/MTX exhibited the highest antitumor activity. In summary, PEGylated PAMAM could be useful as a potential drug delivery carrier.
Animals ; Antimetabolites, Antineoplastic ; blood ; pharmacokinetics ; pharmacology ; Area Under Curve ; Cell Line, Tumor ; Dendrimers ; chemical synthesis ; pharmacokinetics ; Drug Carriers ; Female ; Male ; Methotrexate ; blood ; pharmacokinetics ; pharmacology ; Mice ; Neoplasm Transplantation ; Nylons ; chemical synthesis ; pharmacokinetics ; Polyethylene Glycols ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sarcoma 180 ; pathology ; Tumor Burden ; drug effects

There are sex differences in some brain areas in mammalians. Parkinson's disease (PD), caused by the mesencephalic substantia nigra (SN) dopaminergic neuronal loss, displays sexual difference, i.e., the incidence is higher and the symptoms are more intense in males than that in females. However, it has not been known whether sexual dimorphisms exist in the SN. Sixty adult Sprague-Dawley rats were randomly divided into 5 groups: (1) Female intact group (F-INT group); (2) Male intact group (M-INT group); (3) Ovariectomized group (OVX group); (4) Castration group (CAST group); (5) Ovariectomized + estrogen-replaced group (OVX-E(2) group): The rats received sequentially physiological dose of estrogen for 3 d from the 7th day after ovariectomization. P50 auditory evoked potential (P50) was recorded for 14 d from electrodes inserted in the rat right SN in quiet and awake state. After recording, the brain tissues were dissected and the tyrosine hydroxylase (TH)-expressing neurons in the compact zone of the SN were counted using immunohistochemical method. The results showed that the number of TH-positive (TH(+)) cells in the SN of normal male animals was less than that in normal female rats (P<0.05), and the T/C ratio of P50 in normal males was significantly less than that in normal females (P<0.01), indicating that there exists sexual difference in function and structure in the SN. No differences in the T/C ratio of P50 or the number of TH(+) cells were found between M-INT and CAST groups. The T/C ratio of P50 and the number of TH(+) cells in the SN in OVX group were reduced significantly compared with those in F-INT group (P<0.01). There was no significant difference in the T/C ratio of P50 and the number of TH(+) cells in the SN between OVX- E(2) and F-INT groups 15-20 d after estrogen replacement, suggesting that estrogen can promote the survival and functional recovery of dopaminergic neurons in the SN. The results suggest that there exist sex-specific differences in the dopaminergic neurons in the SN structurally and functionally. The difference of estrogen level in cerebra between male and female animals may account for the sexual differences. Endogenous estrogen plays an important role in maintaining the integrity and modulating the functional activity of dopamine system in the SN.
Animals ; Dopaminergic Neurons ; cytology ; Estrogens ; pharmacology ; Evoked Potentials, Auditory ; Female ; Male ; Orchiectomy ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Sex Characteristics ; Substantia Nigra ; cytology

OBJECTIVETo assess the in-stent lumen visibility and image quality of coronary stents by dual-source computed tomography (DSCT) coronary angiography, and the diagnostic accuracy of DSCT in the detection of coronary in-stent restenosis.

METHODSDSCT was performed at 147 stents in 78 patients at an interval of (21.8?22.2) months after coronary stent implantation. Axial multi-planar reconstruction of the stents and curved-planar reconstruction through the median of the stents were evaluated for stent image quality on a 5-point scale, and the stent lumen diameters were detected. Thirty out of these 78 patients underwent conventional coronary angiography within one month after CT angiography. The patency of 60 stents were independently evaluated by two blinded readers.

RESULTSImage quality was good to excellent on average score (1.6?0.6) . Stent image quality score was correlated with stent diameter, stent location, and heart rate. All stents were assessable in lumen visibility with an average visible lumen diameter percentage of (72.2?12.2) %. Visible lumen diameter percentage was correlated with stent diameter and stent location. For the stents with calcified plaques, the visible lumen diameter percentage at the calcified site was significantly lower than that at the non-calcified site (P<0.001) . Compared with the conventional coronary angiography, 12 out of 14 in-stent stenoses were correctly detected. The sensitivity, specificity, positive predictive value, and negative predictive value for the detection of in-stent stenosis was 85.7%, 95.7%, 85.7%, and 95.7%, respectively. For stents whose diameter >0.275cm, the sensitivity, specificity, positive predictive value, and negative predictive value were all 100%. The agreement between CT findings and coronary angiography results was 93.3%, and it was correlated with stent diameter and heart rate.

CONCLUSIONSUsing a DSCT, coronary stent lumen is partially visible and the image quality is high. Stent diameter and location can influence the stent lumen visibility and image quality. DSCT has a high diagnostic accuracy for the detection of in-stent restenosis and may be a valuable modality for the follow-up of coronary artery stent patency."

Aged ; Coronary Angiography ; methods ; Coronary Restenosis ; diagnostic imaging ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sensitivity and Specificity ; Stents ; Tomography, X-Ray Computed ; methods ; Vascular Patency

Objective To explore the effect of pedicled omental flap with Da Vinci surgical system for breast reconstruction,so as to facilitate the application of robot-assisted surgery in the breast reconstruction after breast cancer(BC) surgery.Methods A luminal BC patient,administrated in March 2017,received Da Vinci robot-assisted filling with pedicled omental flap after extensive lumpectomy and sentinel lymph node biopsy.Blood supply,wound healing,cosmetic outcome,complications and satisfaction of the patient were evaluated.Results The wound healed well,with no obvious scar.The reconstructed breast had desirable volume and looked symmetrical against the other side.The cosmetic outcome was excellent and the patient was satisfied with the surgery.No recurrence or metastasis was found during the twomonth post-operational follow-up.Conclusion Da Vinci robot-assisted breast reconstruction with pedicled omental flap has the advantages of clear vision,precise movement and flexible machinery joint.Breast conservation and autologous reconstruction with this technique substantially improves the psychological and social well-being of the patient.

OBJECTIVETo investigate the clinical value of 64-slice spiral CT (64SCT) in assessing global left ventricular function in patients with old myocardial infarction (OMI), with magnetic resonance imaging (MRI) as the reference standard.

METHODSA total of 28 patients (23 men and 5 women) with OMI underwent contrast-enhanced 64SCT with retrospective electrocardiographic-gating, MRI, and transthoracic echocardiography (Echo). The data sets of 64SCT and MRI were reconstructed at both end-diastole and end-systole to measure left ventricular end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), ejective fraction (EF), and myocardial mass at end-diastole (MM). The data acquired with 64SCT and Echo were compared with the results obtained on MRI as the standard of reference respectively.

RESULTSThe parameters of global left ventricular function obtained with 64SCT were significantly correlated with the MRI data (r = 0.788-0.976, P < 0.001). EDV, ESV, SV, EF, and MM were slightly overestimated by 64SCT compared with MRI; however, there was no significant difference among the measurements. 64SCT was in good agreement with MRI. For the EDV, ESV, SV, and EF, the limits of agreement with Echo were significantly larger than with 64SCT compared with MRI (P < 0.05). The sensitivity, specificity, and accuracy of 64SCT to identify EF value less than 50% were 84.6%, 100% and 92.9%, respectively, whereas those of Echo were 61.5%, 66.7% and 64.3%, respectively. The accuracy of 64SCT was significantly higher than that of Echo (P < 0.01). There was a significantly larger overestimation of EDV, ESV, and SV with Echo than with 64SCT compared with MRI (P < 0.05), whereas EF was not significantly different.

CONCLUSIONSA strong correlation between 64SCT and MRI is found for all parameters. 64SCT agrees well with MRI, and allows more reliable and accurate evaluation of global left ventricular function in patients with OMI than Echo compared with MRI.

Adult ; Aged ; Echocardiography ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Myocardial Infarction ; diagnostic imaging ; physiopathology ; Tomography, Spiral Computed ; methods ; Ventricular Function, Left ; physiology

OBJECTIVETo determine the accuracy of dual-source CT (DSCT) coronary angiography (CAG) for the diagnosis of coronary artery disease (CAD) that induces perfusion defects at myocardial perfusion imaging (MPI) with single photon emission computed tomography (SPECT), by using SPECT and conventional CAG as the reference standard.

METHODSThirty-five patients with suspected or known CAD underwent both DSCT coronary angiography and MPI (using exercise or adenosine stress-rest protocol) with technetium-99m sestamibi SPECT. All the patients were beta-blockers-naïve before DSCT scan. The DSCT CAG studies were classified as having no stenosis, non-obstructive (< 50% luminal narrowing) stenosis, or obstructive (> or = 50% luminal narrowing) stenosis. MPI examinations were classified as showing normal or abnormal (reversible and/or fixed defects). A comparative regional analysis of coronary stenosis on DSCT versus myocardial perfusion on SPECT was made. In a subset of 20 patients, CAG was performed as a reference standard for CT angiography.

RESULTSOn the basis of the DSCT data, 98.4% of coronary segments were assessable. Twenty-seven branches were classified as having no stenosis, among which 85% had normal MPI. Nine branches showed non-obstructive stenosis and 69 branches showed at least one obstructive lesion. Only 50 (64%) branches with an abnormal DSCT had abnormal MPI; even in branches with obstructive stenosis on DSCT, 23 (33%) still had a normal MPI. By receiver operating characteristic curve analysis, at the optimal cutoff value of 58% stenosis, the sensitivity and specificity of DSCT to detect myocardial perfusion defect as defined by SPECT were 85% and 65%. In the subgroup compared with CAG, the sensitivity and specificity of DSCT to identify obstructive stenosis were 93% and 96%.

CONCLUSIONSDSCT and SPECT provide mutually complementary information on CAD. CT angiography can help rule out functionally relevant CAD, but has poor capability in predicting ischemia. DSCT provides high-quality diagnostic image without heartbeat controlling and has a high accuracy in detecting obstructive stenosis.

Aged ; Coronary Angiography ; methods ; Coronary Artery Disease ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Myocardial Ischemia ; diagnostic imaging ; Technetium Tc 99m Sestamibi ; Tomography, Emission-Computed, Single-Photon ; Tomography, X-Ray Computed

OBJECTIVETo establish a protocol for culturing of human embryonic stem cells (HUES4) without any animal-derived feeder cells and to investigate the karyotype stabilities of HUES4 cells after long-term cultivation.

METHODSHUES4 cells were cultured on mitomycin C treated MEFs or human foreskin fibroblast feeder cells. The pluripotency of the ES cells was analyzed by immunocytochemistry staining to detect the expression of pluripotent marker, karyotype of the ES cells at passage 27, 34, 41, 44 and short tandem repeat (STR) at passage 27 were analyzed.

RESULTSThe HUES4 cells cultured on human feeder cells were positive for alkaline phosphatase activity, SSEA-4, TRA-1-60 and TRA-1-81 staining, but negative for SSEA-1. Analysis of karyotype at different passages suggested an abnormal karyotype 46, XY, t(9;15)(q22;q26) mosaicism occurred in HUES4, and the ratios of abnormal increased with passage.

CONCLUSIONHUES4 could be cultured without animal-derived feeder cells and the incidence of abnormal karyotype might be increased with long-term culture.

Cell Culture Techniques ; methods ; Cell Line ; Cell Proliferation ; Embryonic Stem Cells ; cytology ; metabolism ; Fibroblasts ; cytology ; metabolism ; Foreskin ; cytology ; Humans ; Immunohistochemistry ; Karyotyping ; methods ; Male ; Microsatellite Repeats ; genetics ; Polymorphism, Genetic

Substantial evidence strongly implies that sensory gating P50 (also called P50 auditory evoked potential, P50) and dopaminergic neurotransmitters are related. In animal experiment, P50 can be recorded in an awake and quiet state with freedom of movement. Until now there is lack of animal experimental data on the supportive effect of estrogen on function of dopaminergic neurons in substantia nigra (SN) in physiological state. In the present study, female Sprague-Dawley (SD) rats were used as subjects. The animals were divided randomly into four groups: (1) control group (normal animals); (2) Parkinson's disease (PD) model group: the right SN was lesioned with 6-hydroxydopamine (6-OHDA); (3) PD model with bilateral ovariectomized group (OVX-PD): bilateral ovariectomy was performed before administration with 6-OHDA; (4) estrogen + PD model with bilateral ovariectomized group (OVX-E(2)-PD): physiological dose of estrogen was given to the bilateral ovariectomy animals before administration with 6-OHDA. P50 induced by two brief acoustic stimuli were recorded in the right SN and the number of TH(+) dopaminergic neurons in the SN stained by immunohistochemistry was calculated after the determination of P50. The results showed that in the PD model group, the testing/conditioning (T/C) ratio of P50 decreased by 40.60% and the number of TH(+) cells in the right SN decreased by 64.74% as compared with that in the control group (P<0.01); In the OVX-PD group, the T/C ratio of P50 decreased by 45.88% and the number of TH(+) cells was reduced by 57.26% as compared with that in the PD group (P<0.01). Administration with 6-OHDA into the SN pars compacta of ovariectomized rats caused more decrease in the number of TH(+) cells as well as more damage to the function of sensory gating in SN. While in OVX-E(2)-PD group, intramuscular injection with estrogen at physiological dose 3 d before 6-OHDA administration decreased the degree of damage to the SN functionally and morphologically, and its degree of injury corresponded to PD group. These results indicate that the mechanism of protection of dopaminergic neurons in the SN provided by physiological level of estrogen is by promoting the resistibility of the neurons to harmful stimulation. If the gonads are resected resulting in a lack of estrogen, the degree of injury to the function and morphology of dopaminergic neurons in SN induced by 6-OHDA increases. Replacement of estrogen at physiological level on time is necessary. Sensory gating P50 in SN may reflect dynamically the protection of estrogen against dopaminergic neurons depletion in vivo.
Animals ; Disease Models, Animal ; Dopaminergic Neurons ; drug effects ; Estrogens ; pharmacology ; Evoked Potentials, Auditory ; Female ; Neuroprotective Agents ; pharmacology ; Ovariectomy ; Oxidopamine ; adverse effects ; Parkinson Disease ; physiopathology ; Parkinson Disease, Secondary ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; cytology ; drug effects

BACKGROUNDExperimental autoimmune myocarditis (EAM) in rats is a T-cell-mediated disorder. The initiation and maintenance of autoimmune responses in EAM depend on the maturation state of dendritic cells. IL-10 is a pleiotrophic immunomodulatory cytokine that functions at different levels of the immune response, so it has emerged as a promising therapeutic factor for the treatment of autoimmune/inflammatory diseases. This study was designed to test the hypothesis that IL-10 gene modified bone marrow-derived immature dendritic cells (iDCs) ameliorate EAM and to explore the underlying mechanisms.

METHODSEAM was induced using the methods of cardiac myosin immunization on day 0 and day 7. Immature and mature bone marrow-derived dendritic cells (BMDCs) were generated without or with the stimulation by lipopolysaccharide (LPS) and the phenotype was analyzed by flow cytometry. Some of the iDCs were transfected by pcDNA3-IL-10 plasmid. 2 x 10(6)/per rat mature DC (mDC), immature DC (iDC), pcDNA3 transfected iDC, pcDNA3-IL-10 transfected iDC or phosphate buffered saline (PBS) were injected intravenously for treatment 5 days after the first immunization. On day 21, HE staining was performed to detect the myocardial inflammation and T lymphocyte proliferation assay was used to determine the effects of IL-10 gene transfected iDC on autoreactive T cell proliferation. Expression of IkappaB, the inhibitor of NF-kappaB pathway, was determined by Western blot.

RESULTSBMDCs generated in a medium supplemented with granulocyte-macrophage-colony-stimulating factor (GM-CSF) were relatively immature, as determined by flow cytometry. However, stimulation with LPS induced these cells to become mature (m) DCs with higher levels of surface major histocompatibility complex (MHC)-II and costimulatory molecules. Intravenous administration of iDCs, especially pcDNA3-IL-10 transfected iDC, ameliorated the histopathological severity of the myosin induced-EAM, and the effect was lost after the DCs underwent maturation induced by in vitro exposure to LPS. IL-10 gene modified iDC inhibited the antigen specific T cell responses towards cardiac myosin. IkappaB protein was up-regulated significantly in the IL-10 gene modified iDC group.

CONCLUSIONSIL-10 gene modified iDC induced antigen-specific tolerance in EAM. The underlying mechanisms may be related to costimulatory molecules down-regulation and NF-kappaB pathway inhibition.

Animals ; Autoimmune Diseases ; immunology ; Dendritic Cells ; physiology ; Immune Tolerance ; Interleukin-10 ; genetics ; Lymphocyte Activation ; Male ; Myocarditis ; immunology ; Myosins ; immunology ; NF-kappa B ; physiology ; Rats ; Rats, Inbred Lew ; Signal Transduction ; Transfection