Objective Multimeric cyclic RGD (Arg-Gly-Asp) peptides are capable of improving the integrin αvβ3-binding affinity due to the polyvalence effect.In this study,the authors prepare 99Tcm-la-bearing cyclic RGD tetramer E{E[c(RGDfK)]2}2,and evaluate its biodistribution and imaging in nude mice beating U87 MG human glioma xenografts with integrinαvβ3-positive.Methods 99Tcm-hydrazino-nictinamide (HYNIC)-E{E[c(RGDfK)]2}2 was prepared by two-step method,while HYNIC wag chosen as bifunctional chelator,and tricine and trisodium triphenylphosphine-3,3,3-trisuifonate (TPPTS) as coligands.The af-finity of c (RGDyK) monomer,HYNIC-E[c(RGDfK)]2 dimer and HYNIC-E{E[c(RGDfK)]2}2 tetramer to integrin αvβ3 was compared by in vitro competitive assay against binding of 125I-c(RGDyK)to integrin αvβ3.positive U87 MG human glioma cells.The biodistribution [the percentage of injection dose per gram of tissue(%ID/g)] and imaging were performed in nude mice bearing UB7MG human glioma xenografts.Re-suits The labeling yield of 99Tcm-HYNIC-E{E[c(RGDfK)2}2 was over 95%,and the radiochemical purity was more than 99%after purification with Sop-Pak C18 cartridge.The 50%inhibiting concentration (IC30) val-ues of c(RGDyk),HYNIC-E[c(RGDfK)]2 and HYNIC-E{E[c(RGDfK)]2}2 were 85.9,9.5 and 4.5 nmol/L, respectively.The result indicated that RGD tetramer possessed a significantly higher affinity to in-tegrinαvβ3.The biodistribution data showed that 99Tcm-HYNIC-E{E[c(RGDfK)]2}2 was excreted mainly through kidneys.The tumor uptake of 99Tcm-HYNIC-E{E[c(RGDfK)]2}2 was two times higher than 99Tcm- HYNIC-E[c(RGDfK)]2,at 1h postinjection,with the uptake of(10.32±0.07)%ID/g and(5.15±0.52)%ID/g,respectively,which was consistent with the in vitro competitive binding data.The tumor up-tale of 99Tcm-HYNIC.E{E[c(RGDfK)]2}2 was still as higher as(9.35±1.35)%ID/g at 4 h postinjec-tion, which demonstrated that the retention time of radiotracer in tumor was long enough.The imaging showed that tumor was clearly visualized at 1h postinjection,and the image at 4 h postinjection Was better.Conclusion The higher tumor uptake and longer retention time in tumor make 99Tem-HYNIC-E{E[c(RG-DfK)J 2}2 a promising radiotracer for integrinαvβ3-positive tumors imaging,furthermore,suggest that radi-onuelides(such as 90Y).1abeled RGD tetramer is more suitable for the therapy of integrin αvβ3-positive tumors.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome ; Young Adult

Objective:To observe the effect of electroacupuncture (EA) on the expressions of acetylcholine (ACh) and mucin 5AC (MUC5AC) in the lungs of rats with chronic obstructive pulmonary disease (COPD),and explore the mechanism of EA in treating COPD.Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a control group,a COPD group,and an EA group,with 10 rats in each group.The control group was a group of normal rats.The COPD rat model was induced by cigarette smoke combined with lipopolysaccharide (LPS).The COPD rats were treated with EA at bilateral Feishu (BL 13) and Zusanli (ST 36) in the EA group,30 min each time,once a day,successively for 14 d.The lung function was tested.The contents of ACh and MUC5AC in lungs and bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay (ELISA).Pearson method was used to analyze the correlation between pulmonary function and the content of MUC5AC in lungs.The mRNA and protein expressions of MUC5AC in lung tissues were detected by real-time polymerase chain reaction (RT-PCR) and Western blot (WB),respectively.The immune response of MUC5AC was observed by immunohistochemistry.Results:Eight rats were left in each group,and the other two died.Compared with the control group,the total airway resistance (Raw) increased significantly and dynamic compliance (Cdyn) decreased significantly in the COPD group (P＜0.01);compared with the COPD group,the Raw level declined significantly and Cdyn increased significantly in the EA group (P＜0.01).The contents of ACh and MUC5AC in the lungs and BALF were remarkably higher in the COPD group compared with those in the control group (P＜0.01,P＜0.001);compared with the COPD group,the contents of ACh and MUC5AC were significantly lower in the EA group (P＜0.05,P＜0.001).There was a negative correlation between MUC5AC content and lung function (P＜0.001).The mRNA and protein expressions of MUC5AC in the lungs were significantly higher in the COPD group than in the control group (P＜0.001);compared with the COPD group,the expressions were significantly lower in the EA group (P＜0.01).Compared with the control group,the immune response of MUC5AC in the airway epithelium significantly increased in the COPD group (P＜0.001);the immune response of MUC5AC was significantly lower in the EA group compared with that in the COPD group (P＜0.001).Conclusion:EA treatment can improve the lung function of COPD rats,which may be related to its effect in the down-regulation of ACh and MUC5AC contents in the lungs as well as the inhibition of mucus hypersecretion.

OBJECTIVETo explore the clinical effect of low dose pituitrin in children with septic shock.

METHODSA total of 48 pediatric cases with septic shock, in whom 6 hours, conventional treatment could not reverse shock from January 2008 to December 2010, were selected for this study. The patients were divided into two groups randomly (completely random design) (control group 24, remedial group 24). The conventional treatment included antibiotics/fluid resuscitation/correcting acid-base imbalance, glucocorticoid, organ (heart/lung) support, dopamine 1 - 15 µg/(kg·min) and norepinephrine 0.5 - 2 µg/(kg·min) pumped in continuously in the control group. In initial 6 hours the same treatment was given to the remedial group, while low dose pituitrin (0.01 - 0.03 U/min) was pumped additionally during the rest of time. The therapeutic effect on correcting shock was evaluated in both groups.

RESULTSThe total effective rate was 76.2% in the remedial group and 40.0% in the control group; the mortality was 33.3% and 60% respectively. The difference between both groups was significant (P = 0.025).

CONCLUSIONLow dose pituitrin could improve the clinical effect significantly in children with septic shock in whom 6 hours conventional treatment failed to correct shock, shorten the total periods of treatment, and decrease mortality.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Norepinephrine ; therapeutic use ; Pituitary Hormones, Posterior ; administration & dosage ; therapeutic use ; Shock, Septic ; drug therapy ; Treatment Outcome ; Vasoconstrictor Agents ; therapeutic use

OBJECTIVETo study the effect of methylene blue on the growth and acid production of Streptococcus mutans, and the effect of methylene blue on acid production metabolism in plaque glycolysis model (i-PGM) in vitro, and investigate the practicability of methylene blue as a new kind of dental caries prevention agent.

METHODSNephelometer method was used to measure OD value of Streptococcus mutans culture fluid in the different incubation conditions. The kinds and quantities of acid produced by Streptococcus mutans in the different incubation conditions were measured with gas chromatography. pH values of glycolysis buffer media of i-PGM in the different treatment conditions were measured by ORION electrode.

RESULTS(1) The OD value of Streptococcus mutans treated by methylene blue was lower than that by normal saline, and there was significant statistical difference between them. (2) The kinds of acid in three different culture fluid were same, but the total quantities of acid were significantly different among three different culture fluid, in which the total quantities of acid of culture fluids treated by glucose was the greatest, and treated by methylene blue was the lest. (3) The pH value of i-PGM treated by methylene blue was significantly different compared with negative control group, but was not significantly different compared with positive control group.

CONCLUSIONMethylene blue can inhibit the growth and acid production metabolism of Streptococcus matans and acid production metabolism of i-PGM.

Dental Caries ; Dental Plaque ; Glycolysis ; Methylene Blue ; Streptococcus mutans

Embryonic hematopoiesis in mammals is characterized by successive temporal and spatial changes. Previous investigations indicate that in vitro differentiation of embryonic stem cells (ES cells) derived from 129 mice can mimic embryonic hematopoiesis to some extent. To investigate the in vitro hematopoietic differentiation capacity of ES cells derived from C57BL/6 mice, the authors initially established the murine ES cell line with standard identification methods employed. Next, two-step culture system was utilized for embryoid bodies formation and the appearance of different hematopoietic precursors was confirmed by CFC assay, cellular chemical staining as well as RT-PCR. The results demonstrated that the ES cell line MES-1 fulfilled the criteria of ES cell line and its progeny after in vitro differentiation included primitive and definitive erythrocyte precursors, mixed colony-forming cells and granulocyte/macrophage colony-forming cells. RT-PCR analysis revealed the molecular consistence of transcription factors and hematopoietic markers with cellular event. In conclusion, MES-1 established from C57BL/6 mice was able to differentiate in vitro to a variety of hematopoietic precursors, thus could partly recapitulate embryonic hematopoiesis.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; genetics ; Cell Line ; Colony-Forming Units Assay ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; cytology ; Erythroblasts ; cytology ; metabolism ; Erythroid Precursor Cells ; cytology ; metabolism ; Erythroid-Specific DNA-Binding Factors ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics

OBJECTIVETo observe the effects of Stragalus membranaceus injection on nitric oxide and endothelin levels of intestinal mucosa in reperfusion injury after hemorrhage shock.

METHOD32 SD rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with Astragalus membranaceus 10 g x kg(-1)); high dosage group (treated with Astragalus membranaceus 20 g x kg(-1)). Models of hemorrhagic shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology was observed, and the concentration of lactic acid (LD), nitric oxide (NO), endothelin (ET) of intestinal mucosa were detected.

RESULTThe intestinal pathology showed that intestinal mucosa epithelial cells damage in model group was severe, in low dosage group was medium, in high dosage group was slight, and no obvious damage was found in normal group. The concentration of LD and NO of small intestine mucous membrane in model group and low dosage group were significantly higher than those in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05). The concentration of ET of small intestine mucous membrane in model group was the highest of the four groups (P < 0.05). The concentration of ET in low dosage group was significantly higher than that in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05).

CONCLUSIONStragalus membranaceus injection can reduce small intestine mucous damage by protecting endothelium function in injury after hemorrhage shock-reperfusion.

Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endothelins ; metabolism ; Ileum ; metabolism ; pathology ; Injections, Intravenous ; Intestinal Mucosa ; metabolism ; pathology ; Lactic Acid ; metabolism ; Male ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; pathology ; Shock, Hemorrhagic ; complications

This study was aimed to investigate the function defect of partial homing receptor on cord blood hematopoietic stem cells (CBHSC) and explore efficacy and feasibility of intervention in vitro. The expression and activity of active groups in P, E-selectin ligands on CD34+ cells from cord blood, bone marrow and peripheral blood were detected by flow cytometry; meanwhile the expression of active groups in selectin ligands on CD34+ cells treated by fucosyl transferase in vitro was determined by flow cytometry. The results indicated that the expression levels of CD26 on the surface of stem/progenitor cells (CD34+) from cord blood, bone marrow and peripheral blood were (7.62+/-0.63)%, (6.35+/-0.89)% and (6.18+/-0.91)% (p>0.05) respectively. And the activities of CD26 of the three sources of stem cells were 67.15 U/1000 cells (1 U=1 pmol/min), 26.85 U/1000 cells and 20.95 U/1000 cells respectively, in which the activity of CD26 on surface of CD34+ from cord blood was significantly higher than that from other both sources (p<0.01). The expression levels of P-selectin ligand on the stem/progenitor cells three kinds were (83.46+/-6.33)%, (15.65+/-0.89)% and (80.17+/-6.85)%, and the expression levels of E-selectin ligand on stem/progenitor cells of three kinds were (25.31+/-1.03)%, (26.34+/-0.89)% and (29.79+/-1.78)% respectively. The expression of E-selectin ligand on the surface of cord blood stem/progenitor cell CD34+ increased from (25.31+/-1.03)% to (63.23+/-1.08)% after glycosylation engineering. It is concluded that there is no significant difference of the expression of CD26 between the three sources of stem/progenitor cells, but the activity of CD26 in cord blood was obviously higher than that in bone marrow and peripheral blood. The expression of P-selectin ligand on bone marrow stem/progenitor cell was lower than that on stem cells of cord blood and peripheral blood. Glycosylation engineering can promote and elevate the expression of E-selectin ligand on the surface of CD34+ cells from cord blood.
Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; metabolism ; Cells, Cultured ; Dipeptidyl Peptidase 4 ; metabolism ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Receptors, Fibroblast Growth Factor ; metabolism ; Sialoglycoproteins ; metabolism ; Stem Cells ; cytology ; metabolism

This study was aimed to retrospectively analyze and compare the clinical curative efficacy of patients with hematologic malignancies after G-CSF-mobilized sibling HLA-matched (sm) peripheral blood hematopoietic stem cell transplantation (sm-allo-PBHSCT) and sm-allo-PBHSCT combined with bone marrow transplantation (BMT). 100 patients received sm-allo-HSCT in a single center from October 2001 to October to 2010, included 38 patients received sm-allo-PBHSCT and 62 patients received sm-allo-PBHSCT combined with BMT. The myeloablative or reduced intensity conditioning regimens were chosen according to the condition of patients. All patients received standard cyclosporine (CsA) and mycophenolate mofetil (MMF) as prophylaxis for GVHD. The results showed that the rapid hematopoietic reconstitution was observed in all patients. The median time of ANC ≥ 0.5 × 10(9)/L in both groups were 12 days, the median time of platelet count ≥ 20 × 10(9)/L was 15 days in sm-allo-PBHSCT group and 16 days in sm-allo-PBHSCT + BMT group. The incidence of acute GVHD, acute GVHD of III-IV grade and chronic GVHD in sm-allo-PBHSCT and sm-allo-PBHSCT + BMT groups were 37.1% and 34.2%, 7.89% and 8.06%, 36.11% and 41.38% respectively, there were no statistical differences. The relapse rates were similar in two groups (sm-allo-PBHSCT 13.16% vs sm-allo-PBHSCT + BMT 12.9%). The 3-year disease-free survivals in sm-allo-PBHSC and sm-allo-PBHSCT + BMT groups were 57.1 ± 8.7% and 61.3 ± 6.4% respectively (p = 0.852). The 2-year overall survival of high-risk patients was 41.4 ± 12.8% in sm-allo-PBHSCT group, while 60.9 ± 9.6% in sm-allo-PBHSCT + BMT group (p = 0.071). It is concluded that the rhG-CSF mobilized sibling matched allo-PBHSCT + BMT is superior to the rhG-CSF mobilized sibling matched allo-PBHSCT in increasing the overall survival of high-risk hematologic malignancies.
Adolescent ; Adult ; Aged ; Bone Marrow Transplantation ; Child ; Child, Preschool ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; HLA Antigens ; immunology ; Hematologic Diseases ; immunology ; therapy ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Retrospective Studies ; Siblings ; Tissue Donors ; Young Adult

This study was to investigate the reconstitution of NK cells and their receptors after unrelated cord blood stem cell transplantation (UCBT) and its clinical importance. 11 cases of acute leukemia underwent UCBT were enrolled in this study. The reconstitution of NK cells and their surface receptors as well as the the recovery of T and B cells within 90 days after clinical engraftment following UCBT were measured and analysed by flow cytometry. The results indicated that the recovery of NK cells appears to be relatively early. CD3(-)56(+) NK cell count was (35.12 +/- 18.66)% of peripheral blood (PB) lymphocytes on the day of clinical engraftment and higher than that in normal. The peak of the NK cells reached to (37.8 +/- 17.52)% of lymphocyte at 30 days after clinical engraftment. NK count was (30.4 +/- 19.14)% at 60 days after clinical engraftment when the absolute NK cell count reached to the peak (up to 544 cells/microl) in PB. The activated receptor NKG2D was reconstituted fast and high expressed [(79.58 +/- 8.71)%] at the time of clinical engraftment with a tendency of gradual elevation, which reached to peak value (82.55 +/- 9.10)% at day 60. Another activated receptor NKp46 also reconstituted fast, and maintained at a high level even at 90 days after clinical engraftment. The expression of NKG2A was lower than that of the activated receptor of NK cells, which tendency lasted for at least 90 days after clinical engraftment. The reconstitution of T cells in PB after UCBT was relatively slow with lower expression rate. It is concluded that the reconstitution of NK cells in patients with acute leukemia is earlier following UCBT. The earlier recovery of activated receptor of NK cells, especially NKG2D, suggests that the activation of NK cells may play a role in graft versus leukemia (GVL) effect in the early period after UCBT.
Adolescent ; Adult ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Killer Cells, Natural ; Leukemia ; immunology ; surgery ; Lymphocyte Count ; Male ; Postoperative Period ; Receptors, Natural Killer Cell ; Young Adult