Actins ; metabolism ; Animals ; Antigens, CD ; metabolism ; Cadherins ; metabolism ; Carotid Artery Injuries ; metabolism ; pathology ; Endothelium, Vascular ; metabolism ; ultrastructure ; Fluorescent Antibody Technique ; Microscopy, Confocal ; Rats ; Troponin ; metabolism

Electrocardiography ; Humans ; Takotsubo Cardiomyopathy ; Ventricular Dysfunction, Left

Drug-Eluting Stents ; adverse effects ; Humans ; Middle Aged ; Myocardial Infarction ; etiology ; Sirolimus ; administration & dosage ; Thrombosis ; etiology

Objective To optimize methods of culturing smooth muscle progenitor cells(SPCs) from mononuclear cells(MNCs) of peripheral blood. Methods Human MNCs isolated from buffy coat were seeded on M-199 with bovine pituitary extraction.On the eighth day outgrowth cells were stimulated with platelet-derived growth factor-BB(PDGF-BB).Fifteen days later,immunofluorescence,Western blot or RT-PCR was used to analyzed the expression of smooth muscle cell specific ?-actin(?-SMA),smooth muscle myosin heavy chain(SM MHC),Calponin,CD34,Tie-2 and Flk-1,and fluorescence activated cell sorter was employed to examine ?-SMA positive cells ratio. Results The cells stimulated by PDGF-BB for 15 d were positive for ?-SMA,SM MHC,Calponin,CD34 and Flk-1,but negative for Tie-2.The ?-SMA positive cells ratio was(90.57?5.63)%,significantly different from that of the control(P

Alanine Transaminase ; metabolism ; Animals ; Liver ; drug effects ; Male ; Mice ; Plant Extracts ; pharmacology

OBJECTIVETo study the blood pressure (BP) changes in the liver transplant recipients.

METHODSA total of 206 patients without preoperation hypertension received liver transplantation in our hospital from February 2001 to July 2005. The BP level and serum immunosuppressant concentration at preoperation and various time points post operation were determined.

RESULTSCompared with the preoperation, the average systolic and diastolic pressure was significantly increased at the 2 week, 1, 2, 4 and 6 months post operation. The mobility of hypertension increased significantly after liver transplantation, with the highest mobility (46.49%) at the 1st month post operation. There was no linear correlation between the immunosuppressant (FK506) concentration and the BP level at any time point.

CONCLUSIONThere was a high hypertension incidence after liver transplantation. Although the use of immunosuppressive drugs accompanied with the BP increase, there was no linear correlation between the immunosuppressant concentration and the BP level post operation.

Blood Pressure ; Female ; Follow-Up Studies ; Humans ; Hypertension ; etiology ; Immunosuppressive Agents ; adverse effects ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Postoperative Complications ; physiopathology ; Retrospective Studies

Secretory anti-gp96 scFv fragment was expressed in Pichia pastoris to obtain a small molecule antibody that specifically recognizes heat shock protein gp96. The gp96-scFv fragment gene was synthesized and cloned to Pichia pastoris expression plasmid pPICZa-A. Pichia pastoris X33 was electroporated with the linearized recombinant expression vector, and expression of gp96-scFv fragment was induced by methanol. The His-tagged recombinant protein was then purified by affinity chromatography and analyzed with SDS-PAGE and Western blotting assays. The biological activities of recombinant gp96-scFv fragment were determined by Western blotting, Immunofluorescence, ELISA and FACS assays. The gp96-scFv fragment was expressed successfully in Pichia pastoris. About 50 mg of recombinant protein could be purified from 1 liter of the Pichia pastoris culture supernatant. Its molecular weight was about 15 kDa. The gp96-scFv fragment could specifically bind to gp96 protein by Western blotting, immunofluorescence, ELISA and FACS analyses. Pichia pastoris-expressed gp96-scFv fragment specifically recognizes gp96 protein, which could be used for Western blotting, Immunofluorescence, ELISA and FACS analyses.
Blotting, Western ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Membrane Glycoproteins ; immunology ; Pichia ; metabolism ; Plasmids ; Recombinant Proteins ; biosynthesis ; Single-Chain Antibodies ; biosynthesis

Objective To investigate the effects of hypertonic saline/hetastarch 40 injection (HSH40) on intracranial pressure and cerebral oxygenation in patients undergoing neurosurgery for supratentorial glioma. Methods Thirty adult ASA grade Ⅰ or Ⅱ patients undergoing excision of supratentorial glioma were randomly divided into 2 groups: HSH40 4 mL ·kg~(-1) (trial group, 15 cases) or 20% mannitol 1 g·kg~(-1) (control group, 15 cases) was given intravenously over 15 min when hemodynam-ics was stable. Mean arterial pressure (MAP) , cerebrospinal fluid pres-sure(CSFP) and urine output were measured and recorded at different time-points. Cerebral arterial to jugular venous oxygen content differ-ences (Da-jvO_2) and cerebral oxygen extraction rate (CERO_2) were calculated. Results As compared to their baseline, CSFP significantly decreased from 15 min to 120 min after infusion in the 2 groups, and the reduction at 15 min in trial group was more significant than that in control group. Urine output was significantly lower in trial group than that in control group. Da-jvO_2and CERO_2 significantly decreased and internal jugular venous oxygen saturation(SjvO_2) markedly increased at 60 min, 120 min as compared to their baseline in the 2 groups. Conclusion HSH40 is more effective than 20% mannitol in reducing CSFP and can improve cerebral oxygenation during excision of intracranial tumour.

OBJECTIVEThe purpose of this study was to observe histopathological changes post cryoablation in canine myocardium, to characterize the specific ablation lesion post cryoablation.

METHODSCryothermal ablation was applied on myocardium (both epicardium and endocardium) of 14 mongrel dogs with different ablation parameters (-25 degrees C x 4 min, -50 degrees C x 4 min, -75 degrees C x 4 min, -75 degrees C x 2 min, -75 degrees C x 6 min, -75 degrees C x 8 min). Lesion dimensions and histopathologic changes were observed.

RESULTSThe discrete, sharply delimited lesions were detected in cryoablated myocardium. Histologically, cryoablation in all temperatures studied induced heterogeneous necrosis of the myocardium. Lesion dimensions are related to freezing time and temperature.

CONCLUSIONCryoablation is a feasible and preferably choice for clinical application due to its controllable myocardium lesions.

Animals ; Catheter Ablation ; Cryosurgery ; Dogs ; Endocardium ; pathology ; Myocardium ; pathology

OBJECTIVEThe purpose of this study was to observe the endothelial progenitor cells (EPCs) related gene expression changes before and early after revascularization in patients with acute myocardial infarction.

METHODSPeripheral blood samples were taken from patients with acute anterior myocardial infarction 6 hours and 7 days after PCI and stenting. Mononuclear cells (MNCs) were isolated by Ficoll-density centrifugation and cultured in M-199 medium. After 14 days culture, attaching cells incorporated DiI-acetylated low-density lipoprotein (EPCs) were collected and RNA was isolated by Trizol for microarray analysis on 24 genes associated with permissibility/vessel tone (angiotensin system: ACE, AGTR-1, AGTR-2; NO system: eNOS; prostacyclin system: COX-2; endothelin system: ET-1, ETA, ETB; superoxide anions system: SOD-1), angiogenesis (adhesion molecule: CDH5; growth factors and receptors: VEGFR1, VEGFR2, VEGF) and endothelial cell activation (adhesion molecules expression: ICAM1, ICAM2, ICAM3, PECAM-1, E-Selectin, L-Selection, VCAM1; change phenotype from antithrombotic to prothrombotic: tPA, uPA, PAI, vWF). VEGFR2, PECAM-1 and VE-cadherin positive cells were identified by flow cytometry.

RESULTEight gene expressions (AGTR-1, AGTR-2, COX-2, eNOS, ET-1, ETA, VEGF) were significantly downregulated 7 days post PCI compared to pre-PCI (P < 0.05). Flow cytometry results showed that VEGFR2 positive cells were also significantly reduced post PCI than that of before PCI (P < 0.05).

CONCLUSIONPCI down-regulated endothelial progenitor cells related gene expressions in patients with acute myocardial infarction.

Aged ; Aged, 80 and over ; Centrifugation, Density Gradient ; Endothelial Cells ; cytology ; Endothelium, Vascular ; cytology ; Female ; Gene Expression ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; metabolism ; therapy ; Myocardial Reperfusion ; Myocardial Revascularization ; Oligonucleotide Array Sequence Analysis ; Receptor, Angiotensin, Type 1 ; biosynthesis ; genetics ; Receptor, Endothelin A ; biosynthesis ; genetics ; Stem Cells ; cytology ; Superoxide Dismutase ; biosynthesis ; genetics