<p>OBJECTIVETo observe the expression of P-selectin in the penile vascular epithelial cells and the morphological changes in the ultrastructure of the penile cavernous tissues of smoking rats, and to explore the pathogenesis of smoking-induced erectile dysfunction.p><p>METHODSFifty healthy Wistar rats were randomly divided into a normal control, a long-term heavy smoking group, a long-term light smoking, a short-term heavy smoking and a smoking cessation group. Their erectile function was tested by subcutaneous injection of apomorphine (APO), the P-selectin expression in the penile vascular epithelial cells detected by ELISA and the morphological changes in the ultrastructure of the penile cavernous tissues observed under the transmission electron microscope (TEM).p><p>RESULTSThe levels of P-selectin were 10.78 +/- 1.71 ng/L, 62.62 +/- 5.95 ng/L, 40.06 +/- 3.97 ng/L, 41.37 +/- 4.06 ng/L and 22.80 +/- 3.15 ng/L respectively in the normal control, long-term heavy smoking, long-term light smoking, short-term heavy smoking and smoking cessation groups, with significant differences between the control group and the other four (P < 0.05). Electron microscopy showed abnormal arrangement of endothelia, penile cavernous sinuses and smooth muscle cells, disrupted continuity of endothelia, damaged ultrastructure of endothelial and smooth muscle cells in the penile cavernous tissue, and obvious proliferation and fibrosis of interstitial tissues in the smoking rats.p><p>CONCLUSIONSmoking increases the P-selectin expression in the penile vascular epithelial cells and damages the ultrastructure of the penile cavernous tissue, which may be the main contributors to smoking-induced erectile dysfunction.p>
Animals ; Epithelium ; metabolism ; Male ; P-Selectin ; biosynthesis ; Penile Erection ; Penis ; blood supply ; metabolism ; ultrastructure ; Rats ; Rats, Wistar ; Smoking ; adverse effects

Cell adhesive molecular P-selectin was cloned, expressed and anchored on CHO cell membrane through GPI for selection specific antibodies. Total human platelet RNA was extracted and the functional segment of P-selectin gene was cloned by RT-PCR. The P-selectin functional segment gene was cloned into a eukaryotic expression vector pMCEw2-GPI containing an attenuated neo gene together with a downstream GPI, which was synthesized by overlapping PCR. The recombinant plasmid pMCEw2-GPI-P-selectin was then transfected to CHO(dhfr-) cells and screened with G418. ELISA, western-blot and immunofluorescence were carried out to detect the stability of P-selection expression on cell membrane. These results provided a necessary basis for the following study of selection the antibodies targeting P-selectin.
Animals ; Antibodies ; metabolism ; CHO Cells ; Cell Membrane ; metabolism ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Epidermal Growth Factor ; metabolism ; Glycosylphosphatidylinositols ; metabolism ; Lectins ; metabolism ; P-Selectin ; biosynthesis ; genetics ; immunology ; RNA ; metabolism ; Transfection

The study was purposed to develop a novel cryopreserved agent (CPA) for platelets, to investigate the morphology of cryopreserved platelets in different CPA and the CD62P expression on membrane of platelets after stimulating by thrombin, as well as to compare the effect of adding UDP-Gal on preserved efficiency of preservation solutions. A novel cryopreserved agent consisting of 2% DMSO, thrombosol and UDP-Gal was developed on basis of using higher concentration of DMSO. The morphology of chilled platelets was observed by transmission electron microscope and compared with fresh platelets. The expression of CD62P on the membrane of platelets was detected at 0, l, 3 months. The results indicated that the significant effect of cryopreservation on morphology of platelets was found according to percentages of round, dendritic and irregular shapes of cryopreserved platelets. The protective effects of 2% DMSO + thrombosol and 2% DMSO + thrombosol + UDP-Gal were better than that of 5% DMSO. Compared with fresh platelets, the expression of CD62P on platelet membrane decreased obviously after cryopreservation, but not observed difference at preservation for 1 month and 3 months, as well as among 3 kinds of different CPA. It is concluded that the protective effects of 2% DMSO + thrombosol and 2% DMSO + thrombosol + UDP-Gal on morphology of platelets are similar, but better than that 5% DMSO. The reaction of cryopreserved platelets to thrombin decreases, while the significant difference is not found among these 3 kinds of CPA. The addition of UDP-Gal to cryopreserved agents not show the protective effect on platelets.
Blood Platelets ; Blood Preservation ; methods ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dimethyl Sulfoxide ; pharmacology ; Humans ; P-Selectin ; biosynthesis ; genetics ; Uridine Diphosphate Galactose ; pharmacology

It is generally accepted that tissue factor plays an important role in coagulation and intravascular thrombus formation. Tissue factor is not only found primarily on the surface of certain cells that are located outside the vasculature, but also found in circulating cells. Monocyte express tissue factor induced by endotoxin. Recently, many researches indicate that P-selectin, CD40 ligand and GPIIb/IIIa receptor of platelet can also affect expression of tissue factor by monocyters. In addition, a lot of studies showed that tissue factor exist in the circulation including contained platelet. Tissue factor in the platelet releases under certain condition, and initiates coagulation. In this review the relation between platelet and tissue factor was elaborated.
Blood Platelets ; drug effects ; metabolism ; physiology ; CD40 Ligand ; physiology ; Humans ; Monocytes ; drug effects ; metabolism ; P-Selectin ; physiology ; Peptides ; pharmacology ; Platelet Glycoprotein GPIIb-IIIa Complex ; physiology ; Thromboplastin ; biosynthesis ; drug effects ; physiology

<p>OBJECTIVETo explore the relationship between expression thange of P-selectin after brain injury and secondary brain damage.p><p>METHODSSixty SD rats were randomized into 3 equal groups, namely the control group, mild injury group and severe injury group and animal models of brain injury were established in SD rats according to the method of Feeney. P-selectin expression in the brain tissues were determined at 6 h and l, 3, and 7 days following brain injury (n=5 for each time point). Imaging analysis was performed using computerized imaging technique.p><p>RESULTSP-selectin expression and neutrophil infiltration in the brain tissues increased significantly 6 h after brain injury (P<0.05), reaching the peak level at postoperative 24 h and then gradually decreased.p><p>CONCLUSIONP-selectin expression and neutrophil infiltration increase significantly following brain injury, and the time course and distribution of P-selectin expression are consistent with the secondary damage of the brain, strongly suggesting the involvement of P-selectin upregulation in the secondary insult after brain injury.p>
Animals ; Brain Chemistry ; Brain Injuries ; etiology ; metabolism ; pathology ; Female ; Immunohistochemistry ; P-Selectin ; biosynthesis ; Random Allocation ; Rats ; Rats, Sprague-Dawley

<p>OBJECTIVETo investigate the clinical effect of Erigeron injection (El) on positive expression rate of CD62p in platelet and content of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in serum of patients with acute cerebral infarction (ACI).p><p>METHODSSixty-eight patients with ACI were randomly divided into the treated group (n = 35) and the control group (n = 33). Conventional treatment were given to both groups, and EI 40 ml/d were given additionally to the treated group, the treatment course for both groups was 15 days. The positive expression of platelet CD62p and the serum TNF-alpha and IL-6 in patients before and after treatment were determined with flow cytometric (FCM) and electrochemical-luminescence (ECL) techniques respectively.p><p>RESULTSThe total curative effect in the treated group were significantly higher than that in the control group (P < 0.05). Levels of platelet CD62p and serum TNF-alpha and IL-6 in ACI patients before and after treatment were significant higher than those in the healthy group (P < 0.05), all the three parameters were significantly decreased after treatment, and the lowering in the treated group was more significant than that in the control group (P < 0.05).p><p>CONCLUSIONThe effect of El on ACI patients may relate to its action in down-regulating the expression of platelet CD62p, alleviating the immune response and inflammatory injury of central nervous system induced by cytokines.p>
Aged ; Aged, 80 and over ; Asteraceae ; chemistry ; Blood Platelets ; metabolism ; Cerebral Infarction ; immunology ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Injections ; Interleukin-6 ; blood ; Male ; Middle Aged ; P-Selectin ; biosynthesis ; blood ; Phytotherapy ; Tumor Necrosis Factor-alpha ; metabolism

To investigate the changes of platelet activated state and platelet activated function by trace whole blood flow cytometry (FCM), and to explore the mechanism of hemorrhage and infiltration in adults with acute leukemia, the expression percentage and changes of these expressions of CD62p and PAC-1 on platelet surface were determined by FCM of trace whole blood after platelet activated by ADP in patients with new diagnosed AL (group I), complete remission (CR, group II) and continuously complete remission (CCR, group III). Healthy adults were used as control group. The result showed that the expression of CD62p in group I and II was higher than that in control group, before and after platelet activated by ADP (P < 0.01). The expression of PAC-1 in group I was higher than that in control group (P < 0.01), the expression of PAC-1 in group II was lower than that in control group (P > 0.01), There was no significant difference in expression of CD62p and PAC-1 between group III and control group (P > 0.01), and no significant difference was found between AL group with megakaryocyte malignant pathological changes and AL group without megakaryocyte malignant pathological changes before platelet activated by ADP (P > 0.01). After platelet activated by ADP, the expression of PAC-1 in the former was lower than that in the latter (P < 0.01). It is concluded that (1) high level activated platelet in peripheral blood of AL patients show that interaction between activated platelet and leukemia cells can be one of reason resulting in widespread hemorrhage and infiltration AL patiens; (2) the decrease of number and activted function of platelet at the first stage of AL patients may be caused by malignant hyperplasia of leukemia cells and damage of megakaryopoiesis in bone marrow.
Acute Disease ; Adenosine Diphosphate ; pharmacology ; Adolescent ; Adult ; Aged ; Blood Platelets ; cytology ; metabolism ; Cell Membrane ; drug effects ; metabolism ; Female ; Flow Cytometry ; Humans ; Leukemia ; blood ; pathology ; Male ; Middle Aged ; P-Selectin ; biosynthesis ; Platelet Activation ; drug effects ; physiology ; Platelet Glycoprotein GPIIb-IIIa Complex ; biosynthesis

<p>OBJECTIVETo evaluate the effects of Yi-Shen-Huo-Xue Fang on expression of GMP-140 and cleaning out the oxygenic free radicle on rabbits blood stasis model.p><p>METHODThirty rabbits were divided randomly into five groups as the normal group, model group, large dose of "Yi-Shen-Huo-Xue Fang" group, small dose of "Yi-Shen-Huo-Xue Fang" group and "Xue-Shuan-Xin-Mai-Ning" group. After being treated respectively, granule membrane protein 140(GMP-140), erythrocyte sueroxide dismutase (E-SOD), erythrocyte lipid peroxide(E-LPO), plasma lipid peroxide(P-LPO) were checked up.p><p>RESULTThe GMP-140, E-SOD, E-LPO, P-LPO in normal control were compared with those in model groups, With the difference(P < 0.01), model control group was compared with large dose group and small dose group (P < 0.01), with "Xue-Shuan-Xin-Mai-Ning" group(P < 0.05), large dose group was compared with "Xue-Shuan-Xin-Mai-Ning" group(P < 0.05), and large dose group were compared with small dose group (P > 0.05).p><p>CONCLUSIONThe model was made successfully. Large dose group, small dose group and "xue-shuan-xin-mai-ning" group can inhibit expression of GMP-140, enhence SOD activity and decrease LPO content on blood stasis rabbit model. Large dose group and small dose group have stronger effect than "xue-shuan-xin-mai-ning" group.p>
Animals ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythrocytes ; metabolism ; Female ; Free Radical Scavengers ; pharmacology ; Lipid Peroxides ; blood ; Male ; Medicine, Chinese Traditional ; P-Selectin ; biosynthesis ; Plants, Medicinal ; chemistry ; Rabbits ; Random Allocation ; Superoxide Dismutase ; blood

In order to determine the tolerance ability of platelet to change of osmotic pressure in solution, the isotonic fresh platelets were exposed to a series of crystal salt solutions with osmotic pressure range from 47 to 611 mOsm for 15 minutes. Then the platelets were returned to isotonic condition and kept for 15 minutes. The expressions of phosphatidylserine and CD62p were assayed in platelets. The results showed that the phosphatidylserine and CD62p expressions were increased when the osmotic pressure of solution was below 238 mOsm, but no significant rise was detected when the platelets were exposed to 611 mOsm solution. No increases of positive rate of CD62p and phosphatidylserine were detected in platelets returned to isotonic condition. It is concluded that platelets are sensitive to hypoosmotic solution and tolerated to hyperosmotic solution. Exceeding the platelet safe volume limitation may lead to injure of platelet osmosis in crystal salt solution.
Blood Platelets ; drug effects ; metabolism ; Humans ; Hypotonic Solutions ; pharmacology ; Isotonic Solutions ; pharmacology ; Osmotic Pressure ; P-Selectin ; biosynthesis ; Phosphatidylserines ; biosynthesis ; Saline Solution, Hypertonic ; pharmacology ; Sodium Chloride ; pharmacology

Human platelets have distinct characters when preserved by different methods. A efficient flow cytometric assay for different preserved platelets expression of CD62p and phosphatidylserine(PS) is in dire need. Efficient flow cytometric assay for CD62p and PS expressed by preserved platelets was established and the major conditions were optimized. The platelets need not to be washed to wipe off plasma and can be labelled diredtly during the sample preparation. It is efficient for flow cytometric analysis when fresh platelet riched plasma (FPRP) was set as negative control, thrombin actived FPRP, and liquid nitrogen treated FPRP were set as positive control respectively. Gly-Pro-Arg-Pro acetate salt (GPRP) was applied to prevent platelets aggregation and fibrin formation, stabilize platelets and minimize the artificial platelets activation. This is also the key to conquer difficulty of flow cytometric quantitive analysis when platelet, Ca(2+) and plasma coexist. This flow cytometric method is specially suitable for the multi-parameter assay including PS expression for cryopreserved platelets. Minimal sample manipulation, no fixation, and GPRP application resulted in minor artifacts and good sample stability. Results suggested, this flow cytometric assay for preserved platelets is simple and efficient. In addition, the author prepared four different methods treated platelets that can be easily distinguished through this flow cytometric assay. It not only makes sure the practicability of this flow cytometric assay, but also suggests the value of the treated platelets applied in preserved platelets flow cytometric ass
Blood Platelets ; metabolism ; Flow Cytometry ; methods ; Humans ; P-Selectin ; biosynthesis ; Phosphatidylserines ; analysis ; Reproducibility of Results ; Tissue Preservation