Objective To investigate the effect of peroxisome proliferatoractivated receptor-γ coactivator-1α(PGC-1α) on early ischemic preconditioning (IPC) which may act as an important role in early IPC. Methods Building isolated working rat heart Langendorff model, thirty Wistar rats were divided randomly into three groups. Control group(CON group,n=10): a 120-min perfusion was performed without any intervension; ischemia and reperfusion group(I/R group,n=10): a 30-min equilibration period perfusion, a 30-min ischemia and a 60-min reperfusion were performed.; IPC group (n=10): a 10-min equilibration period perfusion was performed, then was elicited by two cycles of 5-min of ischemia interspersed with 5-min reperfusion prior to 30-min ischemia and a 60-min reperfusion. Frozen sections of myocardium at cardiac apex were made and immunohistochemical staining was used to detect expression and the intergrated optical density average (IODA) of PGC-1α. Ultrathin sections were made and the mitochondria under each specimen was evaluated according to Flameng score. Results PGC-1α expression in IPC group (IODA 10.94±5.23) was significantly higher than that in I/R group (IODA 3.88±1.72) and that in CON group (IODA 3.39±2.46; P=0.009, 0.007). The mitochondria changes in I/R group were significant edema and severe damage; but there were not so severe in CON group and IPC group.Flameng score of IPC group (0.44±0.13) and CON group (0.88±0.22) were lower than that in I/R group(1.78±0.14;P=0.003, 0.014) respectively. Conclusion IPC can protect myocytes mitochondria from ischemia and reperfusion.The cardioprotection may be related with the activation and the high expression of PGC-1α, which may act as one of the most important endogenous defence factors of the heart.
Objective To investigate the effect of combined therapy of granulocyte colony stimulating factor (G-CSF) and bone marrow mesenchymal stem cells (BMSCs) carrying hepatocyte growth factor (HGF) gene on the angiogenesis of myocardial infarction (MI) in rats and the mechanisms of the synergistic effect. Methods BMSCs were aspirated from the femur and tibia of 3-week-old Sprague Dawley (SD) male rats. The third generation of BMSCs were harvested and transfectedwith Ad-HGF. The MI models were establ ished in 44 SD male rats (weighing 200-250 g) by l igating the left coronary artery. At 4 weeks after l igation, the shorting fraction (FS) of the left ventricle being below 30% was used as a criteria of model success. The BMSCs (5 × 107/ mL) transfected with Ad-HGF were transplanted into the infarct zone of 12 SD rats, and the expression of HGF protein was detected by Western blot method at 2, 7, and 14 days after transplantation. At 4 weeks, the other 32 SD rats were randomly divided into 4 groups (n=8). The 0.1 mL normal sal ine was injected into the infarct zone in control group; 0.1 mL normal sal ine was injected combined with intraperitoneal injection G-CSF [100 μg/ (kg•d)] for 5 days in G-CSF group; 0.1 mL BMSCs (5 × 107/ mL) transfected with Ad-HGF was injected into the infarct zone in HGF group; 0.1 mL BMSCs (5 × 107/ mL) transfected with Ad-HGF was injected combined with intraperitoneal injection G-CSF [100 μg/ (kg•d)] for 5 days in combined therapy group. At 2 weeks after transplantation, heart function was detected by cardiac ultrasound and hemodynamic analysis, and then myocardial tissue was harvested to analyse the angiogenesis of the infarct zone, and the expression of VEGF protein by immunofluorescence staining. Results The expression of HGF protein in vivo was detected at 2 days and 7 days of BMSCs transfected with Ad-HGF transplantation. There was no significant difference in left ventricular systol ic pressure (LVSP), left ventricular end-diastol ic pressure (LVEDP), dP/dtmax, and FS between G-CSF group and control group (P gt; 0.05). When compared with the control group, LVEDP decreased significantly; LVSP, FS, and dP/dtmax increased significantly (P lt; 0.05) in HGF group and combined therapy group. When compared with HGF group, FS and dP/dtmax increased significantly in combined therapy group (P lt; 0.05). Immunofluorescence staining showed that the vascular endothel ial cells were observed in myocardial infarction border zone. The vascular density and the expression of VEGF protein were significantly higher in combined therapygroup than in other 3 groups (P lt; 0.05). Conclusion The combined therapy of G-CSF and BMSCs carrying HGF gene has a synergistic effect and can enhance infarct zone angiogenesis through inducing the expression of VEGF protein.
Objective To investigate the mechanisms of local application of granulocyte macrophage- colony stimulating factor (GM-CSF) on healing of colonic anastomoses impaired by intraperitoneal oxaliplatin in rats. Methods Sixty 10-week-old male Wistar rats were made the colonic anastomosis model and randomized into 3 groups, 20 rats in each. The rats received intraperitoneal injection of 5% dextrose in group A, and intraperitoneal injection of 5% dextrose and 10 mL oxaliplatin (25 mg/kg) in group B at 1 day; and 50 μg GM-CSF was injected into the perianastomotic area immediately after operation and 10 mL intraperitoneal oxaliplatin (25 mg/kg) was given at 1 day. The general situation of rats was observed after operation. Anastomotic healing was tested by measuring the bursting pressure in vivo at 2, 3, 5, 7 days. Anastomotic healing score was evaluated by histological staining. Immunohistochemical staining of the anastomotic site was used to determine the amount of collagen type I content. Results All animals survived to the experiment end. There was no significant difference in the bursting pressure among 3 groups at 2 and 3 days (P gt; 0.05); the bursting pressure of group B was significantly lower than that of groups A and C (P lt; 0.05). There was no significant difference in mononuclear cells infiltration, mucosal epithelialization, submucosa-muscle layer connection degree, and granulation tissue formation between groups A and C at different time points (P gt; 0.05); groups A and C were significantly better than group B in mucosal epithelialization and granulation tissue formation (P lt; 0.05). Groups A and C were significantly better than group B in mononuclear cells infiltration at 2 and 3 days, and in submucosa-muscle layer connection degree at 5 and 7 days (P lt; 0.05). There was no significant difference in collagen type I content among 3 groups at 2 and 3 days (P gt; 0.05); the content of collagen type I in groups A and C were significantly higher than that in group B (P lt; 0.05) at 5 and 7 days. Conclusion Local administration of GM-CSF may enhance colonic anastomotic healing by early stimulating infiltration of macrophages and increasing collagen deposition.
Objective To assess the clinical effectiveness and safety of granulocyte colony stimulating factor (G-CSF) for patients with acute lymphoblastic leukemia (ALL). Methods We searched the Cochrane Library, PubMed, EMbase, CNKI, and VIP databases from January 2000 to October 2009. Randomized controlled trials (RCTs) about G-CSF for patients with ALL were retrieved. The methodological quality of the included studies was assessed and the data was extracted according the Cochrane Reviewer’s Handbook. Meta-analyses for overall survival, complete remission, quality of life, infections, relapse rate, and adverse events were performed using RevMan 5.0 software. Results Six RCTs involving 620 patients with ALL were included. The results of meta-analyses showed that the G-CFS group was superior to the control group in the overall survival of adult ALL patients (RR=2.24, 95%CI 1.28 to 3.90, P=0.004). Conclusion G-CSF can improve the overall survival of adult ALL patients. However, it is not demonstrated that G-CSF could improve complete remission rate and quality of life, and reduce infections and relapse rate. More high-quality and large scale RCTs are required.
Objective To observe the changes in the number and function of bone marrow-derived endothel ial progenitor cells (EPCs) after bone-marrow stimulation, and to investigate the possible mechanism of improving ischemicl imb disease after bone-marrow stimulation through autologue bone-marrow stem cell implantation. Methods Twelvemale Lewis rats, weighing 200-250 g, were classified into the bone marrow stimulation group (n=6) and the control group(n=6). In the stimulation group, the bone marrow of each rat was stimulated by injection of recombinant human granulocytemacrophage colony-stimulatory factor. Mononuclear cells were harvested from bone marrow and cultured in EBM-2 medium. After 7-day culture, EPCs were stained by 1, 1-dioctadecyl-3, 3, 3, 3-tetramethyl indocarbocyanine-labbled acetylated low density l ipoprotein/fluorescein isothiocyanate-ulex europaeus agglutinin 1, and the double positive cells were counted by the fluorescent microscope. The adhesive abil ity of EPCs was determined by counting the number of re-cultured EPCs. The unilateral ischemia hindl imb model was made with 12 Lewis rats. Three days later, EPCs were transplanted into the ischemic tissues. According to different sources of EPCs, the 12 rats were divided into 2 groups: the stimulation group (n=6) and the control group (n=6). At 3 weeks after EPCs transplantation, the quantity of the collateral vascular was observed by digital subtraction angiography (DSA). Results After 7-day culture, the number of EPCs in the stimulation and control groups was (145.2 ± 37.0)/HP and (95.2 ± 39.4)/HP, respectively, and there was significant difference between the two groups (P lt; 0.05). Meanwhile, the number of adhesive EPCs in the stimulation and control groups was (21.8 ± 4.3)/HP and (15.0 ± 5.2)/HP, respectively, and the difference between the two groups was significant (P lt; 0.05). At 3 weeks after the EPCs implantation, the number of the collateral vascular was significantly larger in the stimulation group (4.2 ± 1.2) compared with the control group (2.7 ± 0.8), (P lt; 0.05). Conclusion Bone marrow stimulation increases the number of EPCs and improves the function concurrently, which may be the reason why autologue bone-marrow stem cell implantation improves the curative effect of ischemic l imb diseases after bone-marrow stimulation.
Objective To investigate the effectiveness and mechanism of recombinant human granulocyte-macrophage colony-stimulating factor (rhGMCSF) gel on wound debridement and healing of deep II thickness burn. Methods Between December 2008 and December 2010, 58 patients with deep II thickness burn, accorded with the inclusive criteria, were collected. There were 36 males and 22 females with an average age of 32.4 years (range, 12-67 years). The causes were hot liquid in 38 cases and fire in 20 cases. The time from injury to treatment was 1-3 days (mean, 2.1 days). In this randomized, double-blind, and self-control study, all patients were randomly divided into 2 groups, wounds were treated with rhGMCSF gel (test group) or gel matrix (control group). There was no significant difference in wound area between 2 groups (P gt; 0.05). The time of completed removal eschar and the percentage of removal-area of eschar were recorded at 2, 6, 10, 14, and 18 days during healing process. The time of wound healing was also recorded. Results Compared with control group, the necrotic tissues on the burn wound got soft in test group at 4 days after treatment. At 6 days, they loosened and eventually sloughed off. The wound bed presented in red and white, followed by rapidly growing granulation tissues. Except 18 days after treatment, there were significant differences in the percentage of removal-area of eschar between 2 groups (P lt; 0.05). The time of completed removal eschar in test group [(7.71 ± 2.76) days] was significantly shorter than that in control group [(14.71 ± 3.63) days] (t=13.726, P=0.000). The time of wound healing in test group was (18.41 ± 2.47) days, which was significantly shorter than that in control group [(23.58 ± 3.35) days] (t=15.763, P=0.000). Conclusion Compared with the gel matrix, the rhGMCSF gel may promote wound debridement and healing in deep II thickness burn.
Objective To identify the best therapy regimen for a patient with rare chronic leg ulcer in necrobiosis lipoidica diabeticorum (NLD). Methods We searched the Cochrane Library (Issue 3, 2006), PubMed (1966-July 2006), EMbase (1974-July 2006) and CBM (1978-July 2006) to identify relevant evidence. The quality of the retrieved studies was critically assessed. Results A total of 153 records were retrieved. No clinical guidelines, systematic reviews or clinical randomized studies were identified. Thirty treatment-related studies involving 17 interventions showed that, fumaric acid esters, oral pentoxifylline, topical use of growth factors and surgical skin flap transplant were relatively more effective and safer than conventional therapies. Conclusion After treatment with tropical granulocyte colony-stimulating factor, the patient with chronic leg ulcers was healed.
ObjectiveTo find out an effective method for amplification and purification of dendritic cells(DC) from peripheral blood of patients with pancreatic carcinoma. MethodsPeripheral blood mononuclear cells were purified from peripheral blood of health volunteers(control group,10 cases) and patients with pancreatic carcinoma (experimental group,12 cases) with incubation of granulocyte/macrophage colonystimulating factor(GMCSF) and interleukin4(IL4).The quality of DC were detected by immumofluorescence method and the expression levels of HLADR and B72 on DC were detected by flow cytometer after and before DC incubation with GMCSF and the IL4. ResultsThe expression level of HLADR and B72 of DC in experimental group were significantly less than those in control group(P＜0.01).DC in experimental group was significantly proliferated in the presence of GMCSF and IL4(P＜0.01).On day 7,the expression level of HLADR and B72 of DC in experimental group were significantly increased(P＜0.01) and there was no difference versus control group(Pgt;0.05).ConclusionIt is suggested that combination of GMCSF and IL4 can selectively and effectively enhance proliferation and immune function of DC from peripheral blood of patient with pancreatic carcinoma.
Objective To evaluate therapeutic efficacy and safety of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) inhalation in patients with recurrent pulmonarv alveolar proteinosis (PAP). Methods Three cases of recurrent PAP were treated by GM-CSF inhalation after whole lung lavage. The clinical data of the pulmonary function and SpO 2, the clinical symptoms and pulmonary lesions were compared before and after treatment. Results The pulmonary function and manifestations were improved obviously after GM-CSF inhalation. Also the ground-glass opacity was improved in high-resolution CT. The pulmonary function and SpO 2 increased obviously after received GM-CSF inhalation. There were no any adverse reactions in 3 cases. Conclusion GM-CSF inhalation therapy is effective and safe in recurrent PAP, but the long-term effect remains to be seen.