Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production...Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.展开更多
Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to t...Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assayand Western blot. Cell growth kinetics analyses throughgrowth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth testwere performed to identify cells proliferation potential.Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormalappearance of M-CSF in nucleus could enhance cellproliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.展开更多
AIM： Recently it has been reported that granulocyte colony stimulating factor （G-CSF） can induce hypercoagulability in healthy bone marrow donors. It is conceivable that the induction of a prothrombotic state in a ...AIM： Recently it has been reported that granulocyte colony stimulating factor （G-CSF） can induce hypercoagulability in healthy bone marrow donors. It is conceivable that the induction of a prothrombotic state in a recipient of an organ graft with already impaired perfusion might cause further deterioration in the transplanted organ. This study evaluated whether G-CSF treatment worsens liver perfusion following liver transplantation in the rat model. METHODS： A non-arterialized rat liver transplantation model was employed to evaluate the effect of G-CSF treatment on the liver in a syngeneic and allogeneic strain combination. Study outcomes included survival time and liver damage as investigated by liver enzymes and liver histology. Observation times were 1 d, 1 wk and 12 wk. RESULTS： Rats treated with G-CSF had increased incidence and severity of biliary damage following liver transplantation. In these animals, hepatocellular necrosis was accentuated in the centrilobular region. These lesions are indicative of impaired perfusion in G-CSF treated animals. CONCLUSION： G-CSF should be used with caution in recipients of liver transplantation, as treatment might enhance preexisting, undetected perfusion problems and ultimately lead to ischemia induced biliary complications .展开更多
Background and Aim: Despite the fact that acute appendicitis is the most common surgical emergency all around the world, its diagnosis is still based on clinical evaluation and accuracy of the diagnosis depending on e...Background and Aim: Despite the fact that acute appendicitis is the most common surgical emergency all around the world, its diagnosis is still based on clinical evaluation and accuracy of the diagnosis depending on experience. The aim of this study is to evaluate the role of inflammatory markers in diagnosis of acute appendicitis. Material and Method: The study includes 77 cases with histopathologically proven acute appendicitis and 17 control cases. Blood samples were obtained from all cases and C-reactive protein (CRP), Granulocyte Colony Stimulating Factor (G-CSF) and Total Antioxidant Capacity (TAC) were measured. Findings: In cases with acute appendicitis, CRP and G-CSF levels were found to be related to acute appendicitis;however, TAC was not affected by the disease process. Moreover, CRP and G-CSF levels were correlated with the disease severity. Conclusion: Both CRP and G-CSF can be used in diagnosis of acute appendicitis. Furthermore, increased CRP level can be a marker to show advanced cases. However, G-CSF is not an effective marker to show disease severity.展开更多
Objective: To verify the antigen association of MAF-J6-1 receptor with M-CSFR and to further study the role of M-CSF and its receptor mediated juxtacrine in promoting leukemic cell proliferation. Methods: Monoclonal a...Objective: To verify the antigen association of MAF-J6-1 receptor with M-CSFR and to further study the role of M-CSF and its receptor mediated juxtacrine in promoting leukemic cell proliferation. Methods: Monoclonal antibody (McAb) of MAF-J6-1R RE2 and polyclonal antibody (PolyAb) of rhM-CSFR were prepared. The specificity of McAb RE2 to M-CSFR was confirmed by indirect ELISA, cross-neutralizing assay with J6-1 cell colony formation and neutralization test by ELISA. Results: the reactive activity of purified RE2 to M-CSFR was over 1: 16000. The inhibitory activity of M-CSFR and MAF-J6-1R could be blocked by RE2 and anti-M-CSFR antibody. The reactivity of RE2 to M-CSFR could be reduced by M-CSFR. Conclusion: The specificity of RE2 to M-CSFR was confirmed and the antigen association of MAF-J6-1R with M-CSFR was proved. It suggests that M-CSF and its receptor mediated auto-juxtacrine stimulation could be an operative mechanism in either leukemia or nonhematological malignancies.展开更多
The dysfunction of vascular endothelial cells plays a key role in startingand facilitating restenosis. The acceleration of intima repair and the recovery of endothelialfunction would reduce the restenosis rate. This s...The dysfunction of vascular endothelial cells plays a key role in startingand facilitating restenosis. The acceleration of intima repair and the recovery of endothelialfunction would reduce the restenosis rate. This study was undertaken to assess the effect ofgranulocyte-macrophage colony stimulating factor ( GM-CSF) on the repair of damaged iliac arteries.Twenty-four male New Zealand white rabbits undergoing primary iliac artery deendothelialization wererandomly divided into two groups ( GM-CSF group and control group) . The GM-CSF group received asubcutaneous injection of GM-CSF (10 μg ? kg^(-1) ? d^(-1) ) , and the control groupwas given a subcutaneous injection of equivalent saline. The iliac arteries of all animals weredamaged by balloon after 7 days. The levels of nitric oxide ( NO) were detected before, 1 week, 2weeks and 4 weeks after angioplasty. The repair and hyperplasia of the intima were observedmicroscopically and the indices of stenosis were evaluated by computerized planimetry after 4 weeksof angioplasty. The NO levels of the GM-CSF group were higher than those of the control group 2weeks and 4 weeks after angioplasty [91.92 +-11.57) μmol/L vs. (81. 67 +- 12. 18) μmol/L; (97. 67+- 10. 13 ) ( μmol/L vs. (83. 16 +-12. 64) μmol/L]. Four weeks after balloon damage,histological examination showed that neointima formation, vascular smooth muscle cells and fibroustissue of the GM-CSF group were less than those of the control group. The endothelium of the GM-CSFgroup was more integrated, and stenosis of lumen was slighter than that of the control group.Morphometry showed the lumen area of the GM-CSF group was larger than that of the control group[(1.27 +-0. 31) mm^2 vs. (0. 92 +- 0. 24) mm^2 ] , the neointimal area and percent of intimahyperplasia were significantly smaller than those of the control group [ (0. 85 +-0. 34) mm vs. (1.18 +-0. 38) mm^2; (40 +- 7)% vs. (55 +- 6)%]. GM-CSF could facilitate the repair of the intima,reduce neointima formation, better the function of the endothelium, and decrease the rate ofrestenosis.展开更多
Objective To analyze efficacy and safety of CLAG regimen in patients with refractory or relapsed acute myeloid leukemia(AML).Methods Efficacy and adverse events of patients with refractory or relapsed AML who were tre...Objective To analyze efficacy and safety of CLAG regimen in patients with refractory or relapsed acute myeloid leukemia(AML).Methods Efficacy and adverse events of patients with refractory or relapsed AML who were treated with one course of CLAG from April 1st,2014 through December 9th,2015 in our hospital were retrospectively reviewed.Results Thirty-three展开更多
Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy.However,it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting.Herein we observed ...Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy.However,it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting.Herein we observed that two cycles of fasting treatment significantly inhibited breast tumor growth and lung tissue metastasis,as well as prolonged overall survival in mice bearing 4T1 and 4T07 breast cancer.During this process,both the immunosuppressive monocytic-(M-)and granulocytic-(G-)myeloid-derived suppressor cell(MDSC)decreased,accompanied by an increase in interleukin(IL)7R^(+)and granzyme B^(+)T cells in the tumor microenvironment.Interestingly,we observed that Ly6G^(low)G-MDSC sharply decreased after fasting treatment,and the cell surface markers and protein mass spectrometry data showed potential therapeutic targets.Mechanistic investigation revealed that glucose metabolism restriction suppressed the splenic granulocytemonocyte progenitor and the generation of colony-stimulating factors and IL-6,which both contributed to the accumulation of G-MDSC.On the other hand,glucose metabolism restriction can directly induce the apoptosis of Ly6G^(low)G-MDSC,but not Ly6G^(high)subsets.In summary,these results suggest that glucose metabolism restriction induced by fasting treatment attenuates the immune-suppressive milieu and enhances the activation of CD3^(+)T cells,providing potential solutions for enhancing immune-based cancer interventions.展开更多
Objective To evaluate the effect of recombinant human granulocyte colon y stimulating factor (rhGCSF) on accelerating neutrophil recovery and decrease fatal infections for childhood acute myeloid leukemia (AML) Meth...Objective To evaluate the effect of recombinant human granulocyte colon y stimulating factor (rhGCSF) on accelerating neutrophil recovery and decrease fatal infections for childhood acute myeloid leukemia (AML) Methods From November 1992 to March 1997, 45 patients wer e enrolled into our study and 15 were newly diagnosed All were treated with hi gh dose chemotherapy combined with rhGCSF Results Of 15 newly diagnosed patients, 13 achieved complete remission (CR) after one course of therapy and 2 achieved CR after two courses of therapy For newly diagnosed patients, the durations of absolute neutrophil counts (ANC ) <05109/L were 5 days and 10 days in rhGCSF group and control group res p ectively ( P <005) The incidences of infection of these two groups w ere 40% and 60% respectively ( P <005) As for patients who receive d intensive therapy, the durations of ANC <05109/L were 5 days and 8 days i n rhGCSF group and control group, respectively ( P <005), and the i ncidences of infection were 25% and 444% respectively ( P <005) Conclusions The application of rhGCSF in children with AML after chem otherapy may hasten the hematopoietic recovery The duration of neutropenia wa s shortened by 3-4 days, and the incidence of fatal infection was reduced rhG CSF does not stimulate AML growth in vivo展开更多
Objective To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte macrophage colony stimulating factor (GM CSF) or hepatitis B i...Objective To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte macrophage colony stimulating factor (GM CSF) or hepatitis B immunoglobulin (HBIG) plus recombinant hepatitis B vaccine (rHBvac) Methods A total of 27 chronic HBV infected children, who were born to HBV carrier mothers and received hepatitis B immunoprophylaxis at birth, were randomized into 2 groups: one receiving a combined therapy of 50 μg of GM CSF plus 10 μg of rHBvac injected intramuscularly at the same location (GM CSF group, 14 children) or 200 IU HBIG and 10 μg rHBvac in different muscles (HBIG group, 13 children) on a monthly four dose schedule HBV DNA quantification and other HBV serological markers were tested before and after the four dose therapy Results Twelve children in each group completed the study Of them, 3 children in the GM CSF group and 4 in the HBIG group had elevated serum alanine transaminase (ALT) before the trial, and then 2 in each group became ALT normal after the treatment Before the therapy, hepatitis B e antigen (HBeAg) positivity was found in nine children in the GM CSF group and 10 in the HBIG group One from each group had an HBeAg/anti HBe seroconversion after the treatment The quantity of HBV DNA was significantly lower after the treatment ( P =0 023) in GM CSF group, but was not significantly reduced in HBIG group No subjects were found to be negative for hepatitis B surface antigen (HBsAg) after the treatment, and no serious adverse events occurred in either group Conclusion Combined GM CSF and rHBvac therapy inhibit HBV replication in carrier children who were not protected after treatment with immunoprophylaxis展开更多
Objective To explore the mechanism of leukocytosis Methods Enzyme linked immunosorbent assay (ELISA) method was used for detecting levels of serum granulocyte colony stimulating factor (G CSF) in 47 cases of acu...Objective To explore the mechanism of leukocytosis Methods Enzyme linked immunosorbent assay (ELISA) method was used for detecting levels of serum granulocyte colony stimulating factor (G CSF) in 47 cases of acute promyelocytic leukemia (APL) during the treatment with all trans retinoic acid (ATRA) Results The peak of increased serum G CSF level occurred on the 9th day, and WBC number was the highest on the 11th day After ATRA treatment, both serum G CSF level and WBC number increased in 68 1% of the cases In 19 2% of the cases treated, serum G CSF level was increased but without obvious change in WBC number, and the reverse was true in 12 7% of the cases Conclusion Serum G CSF level was statistically correlated to the number of WBC, promyelocytes and its late stage by Spearman's rank order correlation coefficient展开更多
The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated wit...The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated with cyclophosphamide (CY) 300 mg/kg once alone was 13.3 ± 7.2d(n=7) however,average survival time of mice treated with CY 300 mg/kg in com-bination with PAP-1 10 mg/kg,3 times/wk was 36.7± 16.4d(n=7,P<0.01).PAP-1,ip had benefi-cial effect on the recovery of the CY induced decrease of peripheral leukocyte number,and the nu-cleated bone marrow cell(BMC)number and[3￣H]TdR uptaken by BMC induced by rmGM-CSF in S180 bearing mice treated with CY,In mice,after the first ip treatment with CY 100 mg/kg on d7,the peripheral leukocyte number decreased on d9 and recovered to normal level about d13 to d15. Such recovery was accelerated by administrating PAP-1,10mg/kg, 3 times/wk.A significant in-crease of the activity to form colony in spleen(colony-forming unit in spleen, CFU-S_8, CUF-S12) in mice irradiated with 550 rad 6O￣Co γ-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with PAP-1,10mg/kg,3 times/wk, ip were observed.After PAP-1,10 mg/kg,ip once,a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor(CSF) were also confirmed.The types of CSF in serum were M-CSF and other hematopoietic growth factors,which were confirmed by using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of PAP-1 on hematopoiesis may be related to its activityinducing CSFs and other hematopoietic growth factors and warrant further evaluation of its use-fulness.展开更多
Objective： The aim of the study was to investigate whether dendritic cell （DC） precursors, recruited by injection of chemokine ligand 3 （CCL3）, induce enhanced anti-tumor immunity after granulocyte-macrophage col...Objective： The aim of the study was to investigate whether dendritic cell （DC） precursors, recruited by injection of chemokine ligand 3 （CCL3）, induce enhanced anti-tumor immunity after granulocyte-macrophage colony stimulating factor （GM-CSF） transfection in mice ex vivo. Methods： The 615 mice were injected with CCL3 via the tail vein. Freshly isolated B220–CD11c＋ cells were cultured with cytokines. For adenoviral （Ad）-mediated gene transduction, DCs were transferred AdGM-CSF gene at different ratios of multiplicity of infection （MOI） to determine the optimal gene transfection conditions, and detecting the expression of GM-CSF after transfection. The variation of GM-CSF gene-modified DCs were analyzed by morphological observation, phenotype analysis, and mixed lymphocyte reaction （MLR）. DCs were loaded with gastric cancer antigen obtained by frozen and thawed method. The stimulated DCs vaccination induced T lymphocytes, and the killing effect of T cells to gastric cancer cells was assayed by MTT. INF-γ production was determined with the INF-γ ELISA kit. Results： B220–CD11c＋ cells numbers increased after CCL3 injection. ELISA results showed that after GM-CSF gene modification, DC could produce high level of GM-CSF. When DCs were transferred AdGM-CSF gene at MOI equal to 1：100, GM-CSF level in culture supernatants reached saturation [（130.00 ± 12.61） pg/mL]. After GM-CSF gene-modification, DCs tended to more maturated through morphological observation and were phenotypically identical to typical DC and gained the capacity to stimulate allogeneic T cells. T lymphocytes stimulated with DC transduced with GM-CSF gene showed the specific killing effect on gastric carcinoma cells and produced high level of INF-γ [（1245.00 ± 13.75） pg/mL]. Conclusion： CCL3-recruited DCs modified by adenovirus-transducted GM-CSF could produce high level of GM-CSF, which tended to more maturated, and the capacity of activating allogeneic T lymphocytes proliferation was enhanced greatly. Moreover, they could stimulate specific cytotoxic T lymphocyte （CTL） to gastric cancer ex vivo.展开更多
AIM:To evaluate the efficacy of granulocyte colony stimulating factors(G-CSF)in liver transplanted patients with hepatitis C(HCV)recurrence and Pegylated-IFN α-2b induced neutropenia,and to evaluate the impact of G-C...AIM:To evaluate the efficacy of granulocyte colony stimulating factors(G-CSF)in liver transplanted patients with hepatitis C(HCV)recurrence and Pegylated-IFN α-2b induced neutropenia,and to evaluate the impact of G-CSF administration on virological response. METHODS:Sixty-eight patients undergoing antiviral treatment for post-liver transplantation(OLT)HCV recurrence were enrolled.All patients developing neutropenia received G-CSF. RESULTS:Twenty three(34%)received G-CSF.Mean neutrophil count at the onset of neutropenia was 700/mmc(range 400-750/mmc);after 1 mo of G-CSF it increased to 1210/mmc(range 300-5590/mmc) (P<0.0001).Three patients did not respond to G-CSF. Treatment duration was similar in neutropenic and non-neutropenic patients.No differences in the rate of discontinuation,infections or virological response were observed between the two groups.G-CSF was protective for the onset of de novo autoimmune hepatitis(P<0.003). CONCLUSION:G-CSF administration is effective in the case of Peg-IFN induced neutropenia increasingneutrophil count,prolonging treatment and leading to sustained virological response(SVR)rates comparable to non-neutropenic patients.It prevents the occurrence of de novo autoimmune hepatitis.展开更多
Objective: To recombinant the nearly natural human granulocyte-macrophage-colony stimulating factor (GM-CSF) for supplying more safe and steady expressed cytokine in clinic. Method: The eukaryotic recombinant pcDNA3.1...Objective: To recombinant the nearly natural human granulocyte-macrophage-colony stimulating factor (GM-CSF) for supplying more safe and steady expressed cytokine in clinic. Method: The eukaryotic recombinant pcDNA3.1-GM-CSF plasmid which was controlled by the CMV promoter was transferred into CHO cell by lipofectamine, selected by G418 and the positive clones was got. The recombinant vector which was rejoined into the groups of DNA of CHO was identified by PCR. Results: The results showed that the protein of rhGM-CSF was about 28 KD by using ELISA, SDS-PAGE and Western blot. Conclusion: rhGM-CSF was expressed steadily and highly. The rhGM-CSF will be of more use value.展开更多
Objective To investigate the induction cytotoxic T cells (CTLs) with antitumor activity and therapeutic efficacy after dendritic cells(DCs) acquired antigen from apoptotic cholangiocarcinoma cells caused by y-irradiat...Objective To investigate the induction cytotoxic T cells (CTLs) with antitumor activity and therapeutic efficacy after dendritic cells(DCs) acquired antigen from apoptotic cholangiocarcinoma cells caused by y-irradiation.Methods DCs from peripheral blood mononuclear cells (PBMC) that maintain the antigen capturing and processing capacity charateristicof immature cells have been established in vitro, using granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then, in cholangiocarcinoma cells apoptosis was induced by J-irradiation. The experimental groups were as follows: (1) coculture ofDCs and apoptotic cancer cells and T cells; (2) coculture of DCs and necrotic cancer cells and T cells; (3) coculture of DCs, culturedcancer cell and T cells. They are cocultured for 7 days. DCs and T cells were riched, isolated and their antitumor response was tested.Results The cells had typical dendritic morphology, expressed high levels of GDI a and B7, acquired antigen from apoptotic cells causedby y-irradiation and induced an increased T cell stimulatory capacity in mixed lymphocyte reactions (MLR) .Conclusion DCs obtained from PBMCs using GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by y-irradiation and efficiently induce T cells. This strategy, therefore, may present an effective approach to transduce DCs with antigen.展开更多
This word was supported by grant from Military Medical Research Foundation of china (96z032). ** To whom correspondence and requests for reprints should be addressed. This is one of papers of the special ...This word was supported by grant from Military Medical Research Foundation of china (96z032). ** To whom correspondence and requests for reprints should be addressed. This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 1997). In order to investigate the antitumor effects of the in vivo G CSF gene therapy mediated by liposome and its mechanisms, human G CSF gene was encapsulated into liposome and was directly injected into tumor mass of C 26 colon adenocarcinoma bearing mice. After direct intratumoral injection of liposome encapsulated G CSF DNA, the subcutaneous tumor growth was dramatically inhibited and the survival time was prolonged signifi cantly. Tumor regression could be observed in about 30% of C 26 bearing mice. By the analysis of the antitumor mechanisms, we found that anti G 418 (600ug/ml) clone could be selected from the tumor cells freshly separated from the treated C 26 tumor mass, and secretion of G CSF in the supernatant could be detected. Northern blot also confirmed the expression of hG CSF by the tumor cells. Higher expressions of MHC class I(H 2k d) molecule and ICAM 1 on the tumor cells could be observed. The results demonstrated that liposome can effectively transfect G CSF gene into tumor cells in situ , and then increase the immunogenicity of the tumor cells which may contribute to the activation of the local antitumor immune responses effectively.展开更多
The full-length cDNA encoding human Granulocyte macrophage colony stimulating factor (GM CSF) was cloned by RT PCR, placed under the control of CMV promoter, and inserted into adenovirus vector of E1 substitution...The full-length cDNA encoding human Granulocyte macrophage colony stimulating factor (GM CSF) was cloned by RT PCR, placed under the control of CMV promoter, and inserted into adenovirus vector of E1 substitution type, pAx1cw. Subsequently, the cassette cosmid was cotransfected into 293 cells together with EcoT22I digested Ad5 TPC, and the replication deficient recombinant adenoviruses(Ad) of human GM CSF were generated efficiently by homologous recombination, with the titers of 1.51×10 9pfu/ml. 48 hours after infection with prepared human GM CSF recombinant adenoviruses in vitro, HeLa cells and primary human skin fibroblasts expressed high levels of human GM CSF (80～400ng/ 10 6cells/24hr). These suggest that the recombinant Ad of human GM CSF prepared by COS/TPC method is effective in mediating GM CSF gene transfer and might be used in cancer gene therapy.展开更多
Background Recent studies have shown that interleukin-3 receptor α （CD123） is highly expressed on leukemia stem cells of patients with acute myeloid leukemia, and is correlated with tumor load and poor prognosis.Th...Background Recent studies have shown that interleukin-3 receptor α （CD123） is highly expressed on leukemia stem cells of patients with acute myeloid leukemia, and is correlated with tumor load and poor prognosis.The expression of CD123 may also be high in patients with myelodysplastic syndrome （MDS）.In this study, the expression and clinical significance of CD123 and granulocyte colony stimulating factor （G-CSF） receptor （CD114） on the bone marrow cells of patients with MDS were investigated to explore the molecular marker of the malignant clone of MDS.Methods Forty-two patients with MDS, who were diagnosed in the Hematological Department of General Hospital of Tianjin Medical University from 2008 to 2009, and twelve normal controls were enrolled in this study.Fluorescence activiated cell sorter （FACS） was used to measure the expression of CD123 on CD34＋CD38- cells and CD114 on CD34＋cells of the bone marrow of these patients and controls and the clinical significance was analyzed.The expression of CD114 on CD123＋CD34＋CD38- cells was further measured to explore the molecular marker of the malignant clone in MDS.Results MDS patients displayed significantly higher proportion of CD34＋CD38-/CD34＋ （（14.03±5.27）%） than normal controls （（7.70±4.36）%, P 〈0.05）.The expression rate of CD123＋CD34＋CD38-/CD34＋CD38- was significantly higher in MDS patients （（48.39±28.15）%） than that in normal controls （（8.75±11.71）%, P 〈0.01）.The expression level of CD123 was significantly correlated with the proportion of bone marrow blasts （r=0.457, P 〈0.05）.The expression rate of CD114＋CD34＋/CD34＋ was lower in MDS patients （（33.05±21.71）%） than that in normal controls （（38.99±19.07）%） but was not statistically significant （P 〉0.05）.The expression of CD114 on CD123＋CD34＋CD38- cells （（34.82±29.58）%） was significantly lower than that on CD123-CD34＋CD38- cells （（53.48±27.41）%） of M DS patients （P 〈0.05）.Conclusions MDS patients displayed higher proportion of CD34＋CD38-/CD34＋ than normal controls.CD123 was highly expressed in the bone marrow of the patients with MDS, significantly correlated with the proportion of bone marrow blasts, and thus might be the marker of MDS malignant clone.CD123＋CD34＋CD38- cells exhibited lower expression of G-CSF receptors, which might partly explain why MDS clone responds worse to G-CSF in vitro and in vivo.展开更多
Background Lipopolysaccharide （LPS） forms outer membrane of the wall of Gram-negative cells. LPS can directly cause damage to epithelia of respiratory tract and is the major factor responsible for the chronic inflam...Background Lipopolysaccharide （LPS） forms outer membrane of the wall of Gram-negative cells. LPS can directly cause damage to epithelia of respiratory tract and is the major factor responsible for the chronic inflammation of respiratory passage. The mitogen-activated protein kinase （MAPK） signal transduction pathway of the airway epithelia is intimately associated with the action of LPS. The chronic inflammation of respiratory tract and smoking are interrelated and entwined in the development and progression of chronic lung diseases. This study was designed to examine the effects of cigarette smoke extract （CSE） and LPS on MAPK signal transduction pathway in order to further understand the roles CSE and LPS play in chronic lung inflammation. Methods Cultured primary human epithelial cells of airway were divided into four groups according to the stimulants used： blank control group, LPS-stimulation group, CSE-stimulation group and CSE plus LPS group. Western blotting was employed for the detection of phosphorylation level of extracellular-signal-regulated-kinase （ERK1/2）, p38 MAPK and c-Jun N-terminal kinase （JNK）. The expression of cytokines of MAPK transduction pathway （granulocyte-macrophage colony stimulating factor （GM-CSF） and mRNA of IL-8） in the primary epithelial cells of respiratory tract was also determined. Results Western blotting revealed that the phosphorylation levels of ERK1/2, p38 MAPK and JNK were low and 2 hours after the LPS stimulation, the phosphorylation of ERK1/2, p38 MAPK and JNK were all increased. There was a significant difference in the phosphorylation between the LPS-stimulation group and blank control group （P〈0.05）; no significant difference was found between CSE-stimulation group and blank control group （P〉0.05）; there was a significant difference between CSE ＋ LPS group and blank control group and between CSE ＋ LPS group and LPS group （P〈0.05）. The phosphorylation of CSE-LPS group was higher than that of blank control group but lower than that of LPS group. In blank control group, the expression of IL-8 and GM-CSF mRNA was low in the epithelial cells of airway and the release of IL-8 and GM-CSF was also at a low level. One hour after LPS stimulation, the level of IL-8 mRNA increased （P〈0.05） and reached a peak after 2 hours. On the other hand, GM-CSF mRNA level increased 2 hours after the stimulation （P〈0.05） and reached the highest level 4 hours after the stimulation. Two hours after LPS stimulation, IL-8 and GM-CSF protein level began to rise （P〈0.05）, and the level was the highest 8 hours after the stimulation （P〈0.01）. Stimulation with CSE alone had no effect on the release of IL-8 and GM-CSF and expression of IL-8 mRNA （P〉0.05）, but pre-treatment with CSE could delay the LPS-induced release of IL-8 and GM-CSF and the expression of IL-8 mRNA and its peak was lower. Conclusions LPS stimulation can significantly increase the phosphorylation of ERK1/2, p38 MAPK and JNK in the epithelial cells of airway and activate the MAPK transduction pathway, thereby can activate the downstream signal transduction pathway, and can ultimately result in the release of cytokines by the epithelial cells of airway. CSE can partially abolish the LPS-induced activation of MAPK signal transduction pathway and the expression of cytokines of the pathway, which might contribute to the development and progression of the inflammatory reactions in COPD patients.展开更多
文摘Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.
基金This work was supported by a grant from Tianjin Science and Technology Development Project (No. 003119311).
文摘Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assayand Western blot. Cell growth kinetics analyses throughgrowth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth testwere performed to identify cells proliferation potential.Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormalappearance of M-CSF in nucleus could enhance cellproliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.
基金Supported by the Deutsche Forschungsgemeinschaft (KFO 117/1) and the IFORES Research Program, University Hospital Essen
文摘AIM： Recently it has been reported that granulocyte colony stimulating factor （G-CSF） can induce hypercoagulability in healthy bone marrow donors. It is conceivable that the induction of a prothrombotic state in a recipient of an organ graft with already impaired perfusion might cause further deterioration in the transplanted organ. This study evaluated whether G-CSF treatment worsens liver perfusion following liver transplantation in the rat model. METHODS： A non-arterialized rat liver transplantation model was employed to evaluate the effect of G-CSF treatment on the liver in a syngeneic and allogeneic strain combination. Study outcomes included survival time and liver damage as investigated by liver enzymes and liver histology. Observation times were 1 d, 1 wk and 12 wk. RESULTS： Rats treated with G-CSF had increased incidence and severity of biliary damage following liver transplantation. In these animals, hepatocellular necrosis was accentuated in the centrilobular region. These lesions are indicative of impaired perfusion in G-CSF treated animals. CONCLUSION： G-CSF should be used with caution in recipients of liver transplantation, as treatment might enhance preexisting, undetected perfusion problems and ultimately lead to ischemia induced biliary complications .
文摘Background and Aim: Despite the fact that acute appendicitis is the most common surgical emergency all around the world, its diagnosis is still based on clinical evaluation and accuracy of the diagnosis depending on experience. The aim of this study is to evaluate the role of inflammatory markers in diagnosis of acute appendicitis. Material and Method: The study includes 77 cases with histopathologically proven acute appendicitis and 17 control cases. Blood samples were obtained from all cases and C-reactive protein (CRP), Granulocyte Colony Stimulating Factor (G-CSF) and Total Antioxidant Capacity (TAC) were measured. Findings: In cases with acute appendicitis, CRP and G-CSF levels were found to be related to acute appendicitis;however, TAC was not affected by the disease process. Moreover, CRP and G-CSF levels were correlated with the disease severity. Conclusion: Both CRP and G-CSF can be used in diagnosis of acute appendicitis. Furthermore, increased CRP level can be a marker to show advanced cases. However, G-CSF is not an effective marker to show disease severity.
文摘Objective: To verify the antigen association of MAF-J6-1 receptor with M-CSFR and to further study the role of M-CSF and its receptor mediated juxtacrine in promoting leukemic cell proliferation. Methods: Monoclonal antibody (McAb) of MAF-J6-1R RE2 and polyclonal antibody (PolyAb) of rhM-CSFR were prepared. The specificity of McAb RE2 to M-CSFR was confirmed by indirect ELISA, cross-neutralizing assay with J6-1 cell colony formation and neutralization test by ELISA. Results: the reactive activity of purified RE2 to M-CSFR was over 1: 16000. The inhibitory activity of M-CSFR and MAF-J6-1R could be blocked by RE2 and anti-M-CSFR antibody. The reactivity of RE2 to M-CSFR could be reduced by M-CSFR. Conclusion: The specificity of RE2 to M-CSFR was confirmed and the antigen association of MAF-J6-1R with M-CSFR was proved. It suggests that M-CSF and its receptor mediated auto-juxtacrine stimulation could be an operative mechanism in either leukemia or nonhematological malignancies.
文摘The dysfunction of vascular endothelial cells plays a key role in startingand facilitating restenosis. The acceleration of intima repair and the recovery of endothelialfunction would reduce the restenosis rate. This study was undertaken to assess the effect ofgranulocyte-macrophage colony stimulating factor ( GM-CSF) on the repair of damaged iliac arteries.Twenty-four male New Zealand white rabbits undergoing primary iliac artery deendothelialization wererandomly divided into two groups ( GM-CSF group and control group) . The GM-CSF group received asubcutaneous injection of GM-CSF (10 μg ? kg^(-1) ? d^(-1) ) , and the control groupwas given a subcutaneous injection of equivalent saline. The iliac arteries of all animals weredamaged by balloon after 7 days. The levels of nitric oxide ( NO) were detected before, 1 week, 2weeks and 4 weeks after angioplasty. The repair and hyperplasia of the intima were observedmicroscopically and the indices of stenosis were evaluated by computerized planimetry after 4 weeksof angioplasty. The NO levels of the GM-CSF group were higher than those of the control group 2weeks and 4 weeks after angioplasty [91.92 +-11.57) μmol/L vs. (81. 67 +- 12. 18) μmol/L; (97. 67+- 10. 13 ) ( μmol/L vs. (83. 16 +-12. 64) μmol/L]. Four weeks after balloon damage,histological examination showed that neointima formation, vascular smooth muscle cells and fibroustissue of the GM-CSF group were less than those of the control group. The endothelium of the GM-CSFgroup was more integrated, and stenosis of lumen was slighter than that of the control group.Morphometry showed the lumen area of the GM-CSF group was larger than that of the control group[(1.27 +-0. 31) mm^2 vs. (0. 92 +- 0. 24) mm^2 ] , the neointimal area and percent of intimahyperplasia were significantly smaller than those of the control group [ (0. 85 +-0. 34) mm vs. (1.18 +-0. 38) mm^2; (40 +- 7)% vs. (55 +- 6)%]. GM-CSF could facilitate the repair of the intima,reduce neointima formation, better the function of the endothelium, and decrease the rate ofrestenosis.
文摘Objective To analyze efficacy and safety of CLAG regimen in patients with refractory or relapsed acute myeloid leukemia(AML).Methods Efficacy and adverse events of patients with refractory or relapsed AML who were treated with one course of CLAG from April 1st,2014 through December 9th,2015 in our hospital were retrospectively reviewed.Results Thirty-three
基金supported by the Postdoctoral Research Funds of Hebei Medical University(30705010016-3759)Natural Science Foundation of China(32272328)+4 种基金Natural Science Foundation of Hebei Province(B2022321001)National Key Research Project of Hebei Province(20375502D)Postdoctoral Research Project of Hebei Province(B2022003031)Science and Technology Research Program of Hebei Provincial Colleges(QN2023229)Hebei Provincial Key Laboratory of Nutrition and Health(2023YDYY-KF05)。
文摘Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy.However,it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting.Herein we observed that two cycles of fasting treatment significantly inhibited breast tumor growth and lung tissue metastasis,as well as prolonged overall survival in mice bearing 4T1 and 4T07 breast cancer.During this process,both the immunosuppressive monocytic-(M-)and granulocytic-(G-)myeloid-derived suppressor cell(MDSC)decreased,accompanied by an increase in interleukin(IL)7R^(+)and granzyme B^(+)T cells in the tumor microenvironment.Interestingly,we observed that Ly6G^(low)G-MDSC sharply decreased after fasting treatment,and the cell surface markers and protein mass spectrometry data showed potential therapeutic targets.Mechanistic investigation revealed that glucose metabolism restriction suppressed the splenic granulocytemonocyte progenitor and the generation of colony-stimulating factors and IL-6,which both contributed to the accumulation of G-MDSC.On the other hand,glucose metabolism restriction can directly induce the apoptosis of Ly6G^(low)G-MDSC,but not Ly6G^(high)subsets.In summary,these results suggest that glucose metabolism restriction induced by fasting treatment attenuates the immune-suppressive milieu and enhances the activation of CD3^(+)T cells,providing potential solutions for enhancing immune-based cancer interventions.
文摘Objective To evaluate the effect of recombinant human granulocyte colon y stimulating factor (rhGCSF) on accelerating neutrophil recovery and decrease fatal infections for childhood acute myeloid leukemia (AML) Methods From November 1992 to March 1997, 45 patients wer e enrolled into our study and 15 were newly diagnosed All were treated with hi gh dose chemotherapy combined with rhGCSF Results Of 15 newly diagnosed patients, 13 achieved complete remission (CR) after one course of therapy and 2 achieved CR after two courses of therapy For newly diagnosed patients, the durations of absolute neutrophil counts (ANC ) <05109/L were 5 days and 10 days in rhGCSF group and control group res p ectively ( P <005) The incidences of infection of these two groups w ere 40% and 60% respectively ( P <005) As for patients who receive d intensive therapy, the durations of ANC <05109/L were 5 days and 8 days i n rhGCSF group and control group, respectively ( P <005), and the i ncidences of infection were 25% and 444% respectively ( P <005) Conclusions The application of rhGCSF in children with AML after chem otherapy may hasten the hematopoietic recovery The duration of neutropenia wa s shortened by 3-4 days, and the incidence of fatal infection was reduced rhG CSF does not stimulate AML growth in vivo
基金ThisstudywassupportedbyagrantfromtheNationalPublicHealthMinistry (No97030223)andagrantfromtheNationalNaturalScienceFoundationofChina (No 39670 667)
文摘Objective To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte macrophage colony stimulating factor (GM CSF) or hepatitis B immunoglobulin (HBIG) plus recombinant hepatitis B vaccine (rHBvac) Methods A total of 27 chronic HBV infected children, who were born to HBV carrier mothers and received hepatitis B immunoprophylaxis at birth, were randomized into 2 groups: one receiving a combined therapy of 50 μg of GM CSF plus 10 μg of rHBvac injected intramuscularly at the same location (GM CSF group, 14 children) or 200 IU HBIG and 10 μg rHBvac in different muscles (HBIG group, 13 children) on a monthly four dose schedule HBV DNA quantification and other HBV serological markers were tested before and after the four dose therapy Results Twelve children in each group completed the study Of them, 3 children in the GM CSF group and 4 in the HBIG group had elevated serum alanine transaminase (ALT) before the trial, and then 2 in each group became ALT normal after the treatment Before the therapy, hepatitis B e antigen (HBeAg) positivity was found in nine children in the GM CSF group and 10 in the HBIG group One from each group had an HBeAg/anti HBe seroconversion after the treatment The quantity of HBV DNA was significantly lower after the treatment ( P =0 023) in GM CSF group, but was not significantly reduced in HBIG group No subjects were found to be negative for hepatitis B surface antigen (HBsAg) after the treatment, and no serious adverse events occurred in either group Conclusion Combined GM CSF and rHBvac therapy inhibit HBV replication in carrier children who were not protected after treatment with immunoprophylaxis
文摘Objective To explore the mechanism of leukocytosis Methods Enzyme linked immunosorbent assay (ELISA) method was used for detecting levels of serum granulocyte colony stimulating factor (G CSF) in 47 cases of acute promyelocytic leukemia (APL) during the treatment with all trans retinoic acid (ATRA) Results The peak of increased serum G CSF level occurred on the 9th day, and WBC number was the highest on the 11th day After ATRA treatment, both serum G CSF level and WBC number increased in 68 1% of the cases In 19 2% of the cases treated, serum G CSF level was increased but without obvious change in WBC number, and the reverse was true in 12 7% of the cases Conclusion Serum G CSF level was statistically correlated to the number of WBC, promyelocytes and its late stage by Spearman's rank order correlation coefficient
文摘The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated with cyclophosphamide (CY) 300 mg/kg once alone was 13.3 ± 7.2d(n=7) however,average survival time of mice treated with CY 300 mg/kg in com-bination with PAP-1 10 mg/kg,3 times/wk was 36.7± 16.4d(n=7,P<0.01).PAP-1,ip had benefi-cial effect on the recovery of the CY induced decrease of peripheral leukocyte number,and the nu-cleated bone marrow cell(BMC)number and[3￣H]TdR uptaken by BMC induced by rmGM-CSF in S180 bearing mice treated with CY,In mice,after the first ip treatment with CY 100 mg/kg on d7,the peripheral leukocyte number decreased on d9 and recovered to normal level about d13 to d15. Such recovery was accelerated by administrating PAP-1,10mg/kg, 3 times/wk.A significant in-crease of the activity to form colony in spleen(colony-forming unit in spleen, CFU-S_8, CUF-S12) in mice irradiated with 550 rad 6O￣Co γ-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with PAP-1,10mg/kg,3 times/wk, ip were observed.After PAP-1,10 mg/kg,ip once,a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor(CSF) were also confirmed.The types of CSF in serum were M-CSF and other hematopoietic growth factors,which were confirmed by using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of PAP-1 on hematopoiesis may be related to its activityinducing CSFs and other hematopoietic growth factors and warrant further evaluation of its use-fulness.
基金Supported by grants of Medical Science and Technology Development Foundation, Jiangsu Province Department of Health (No. H201013)the Program for Postgraduate Research Innovation in University of Jiangsu Province (No. CX10B_054Z)the Project of Youth Foundation in Science and Education of Department of Public Health of Suzhou (2010, No. 4)
文摘Objective： The aim of the study was to investigate whether dendritic cell （DC） precursors, recruited by injection of chemokine ligand 3 （CCL3）, induce enhanced anti-tumor immunity after granulocyte-macrophage colony stimulating factor （GM-CSF） transfection in mice ex vivo. Methods： The 615 mice were injected with CCL3 via the tail vein. Freshly isolated B220–CD11c＋ cells were cultured with cytokines. For adenoviral （Ad）-mediated gene transduction, DCs were transferred AdGM-CSF gene at different ratios of multiplicity of infection （MOI） to determine the optimal gene transfection conditions, and detecting the expression of GM-CSF after transfection. The variation of GM-CSF gene-modified DCs were analyzed by morphological observation, phenotype analysis, and mixed lymphocyte reaction （MLR）. DCs were loaded with gastric cancer antigen obtained by frozen and thawed method. The stimulated DCs vaccination induced T lymphocytes, and the killing effect of T cells to gastric cancer cells was assayed by MTT. INF-γ production was determined with the INF-γ ELISA kit. Results： B220–CD11c＋ cells numbers increased after CCL3 injection. ELISA results showed that after GM-CSF gene modification, DC could produce high level of GM-CSF. When DCs were transferred AdGM-CSF gene at MOI equal to 1：100, GM-CSF level in culture supernatants reached saturation [（130.00 ± 12.61） pg/mL]. After GM-CSF gene-modification, DCs tended to more maturated through morphological observation and were phenotypically identical to typical DC and gained the capacity to stimulate allogeneic T cells. T lymphocytes stimulated with DC transduced with GM-CSF gene showed the specific killing effect on gastric carcinoma cells and produced high level of INF-γ [（1245.00 ± 13.75） pg/mL]. Conclusion： CCL3-recruited DCs modified by adenovirus-transducted GM-CSF could produce high level of GM-CSF, which tended to more maturated, and the capacity of activating allogeneic T lymphocytes proliferation was enhanced greatly. Moreover, they could stimulate specific cytotoxic T lymphocyte （CTL） to gastric cancer ex vivo.
文摘AIM:To evaluate the efficacy of granulocyte colony stimulating factors(G-CSF)in liver transplanted patients with hepatitis C(HCV)recurrence and Pegylated-IFN α-2b induced neutropenia,and to evaluate the impact of G-CSF administration on virological response. METHODS:Sixty-eight patients undergoing antiviral treatment for post-liver transplantation(OLT)HCV recurrence were enrolled.All patients developing neutropenia received G-CSF. RESULTS:Twenty three(34%)received G-CSF.Mean neutrophil count at the onset of neutropenia was 700/mmc(range 400-750/mmc);after 1 mo of G-CSF it increased to 1210/mmc(range 300-5590/mmc) (P<0.0001).Three patients did not respond to G-CSF. Treatment duration was similar in neutropenic and non-neutropenic patients.No differences in the rate of discontinuation,infections or virological response were observed between the two groups.G-CSF was protective for the onset of de novo autoimmune hepatitis(P<0.003). CONCLUSION:G-CSF administration is effective in the case of Peg-IFN induced neutropenia increasingneutrophil count,prolonging treatment and leading to sustained virological response(SVR)rates comparable to non-neutropenic patients.It prevents the occurrence of de novo autoimmune hepatitis.
基金This work was supported by the Natural Science foundation of Fujian Province, China (No. C97067).
文摘Objective: To recombinant the nearly natural human granulocyte-macrophage-colony stimulating factor (GM-CSF) for supplying more safe and steady expressed cytokine in clinic. Method: The eukaryotic recombinant pcDNA3.1-GM-CSF plasmid which was controlled by the CMV promoter was transferred into CHO cell by lipofectamine, selected by G418 and the positive clones was got. The recombinant vector which was rejoined into the groups of DNA of CHO was identified by PCR. Results: The results showed that the protein of rhGM-CSF was about 28 KD by using ELISA, SDS-PAGE and Western blot. Conclusion: rhGM-CSF was expressed steadily and highly. The rhGM-CSF will be of more use value.
文摘Objective To investigate the induction cytotoxic T cells (CTLs) with antitumor activity and therapeutic efficacy after dendritic cells(DCs) acquired antigen from apoptotic cholangiocarcinoma cells caused by y-irradiation.Methods DCs from peripheral blood mononuclear cells (PBMC) that maintain the antigen capturing and processing capacity charateristicof immature cells have been established in vitro, using granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then, in cholangiocarcinoma cells apoptosis was induced by J-irradiation. The experimental groups were as follows: (1) coculture ofDCs and apoptotic cancer cells and T cells; (2) coculture of DCs and necrotic cancer cells and T cells; (3) coculture of DCs, culturedcancer cell and T cells. They are cocultured for 7 days. DCs and T cells were riched, isolated and their antitumor response was tested.Results The cells had typical dendritic morphology, expressed high levels of GDI a and B7, acquired antigen from apoptotic cells causedby y-irradiation and induced an increased T cell stimulatory capacity in mixed lymphocyte reactions (MLR) .Conclusion DCs obtained from PBMCs using GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by y-irradiation and efficiently induce T cells. This strategy, therefore, may present an effective approach to transduce DCs with antigen.
文摘This word was supported by grant from Military Medical Research Foundation of china (96z032). ** To whom correspondence and requests for reprints should be addressed. This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 1997). In order to investigate the antitumor effects of the in vivo G CSF gene therapy mediated by liposome and its mechanisms, human G CSF gene was encapsulated into liposome and was directly injected into tumor mass of C 26 colon adenocarcinoma bearing mice. After direct intratumoral injection of liposome encapsulated G CSF DNA, the subcutaneous tumor growth was dramatically inhibited and the survival time was prolonged signifi cantly. Tumor regression could be observed in about 30% of C 26 bearing mice. By the analysis of the antitumor mechanisms, we found that anti G 418 (600ug/ml) clone could be selected from the tumor cells freshly separated from the treated C 26 tumor mass, and secretion of G CSF in the supernatant could be detected. Northern blot also confirmed the expression of hG CSF by the tumor cells. Higher expressions of MHC class I(H 2k d) molecule and ICAM 1 on the tumor cells could be observed. The results demonstrated that liposome can effectively transfect G CSF gene into tumor cells in situ , and then increase the immunogenicity of the tumor cells which may contribute to the activation of the local antitumor immune responses effectively.
文摘The full-length cDNA encoding human Granulocyte macrophage colony stimulating factor (GM CSF) was cloned by RT PCR, placed under the control of CMV promoter, and inserted into adenovirus vector of E1 substitution type, pAx1cw. Subsequently, the cassette cosmid was cotransfected into 293 cells together with EcoT22I digested Ad5 TPC, and the replication deficient recombinant adenoviruses(Ad) of human GM CSF were generated efficiently by homologous recombination, with the titers of 1.51×10 9pfu/ml. 48 hours after infection with prepared human GM CSF recombinant adenoviruses in vitro, HeLa cells and primary human skin fibroblasts expressed high levels of human GM CSF (80～400ng/ 10 6cells/24hr). These suggest that the recombinant Ad of human GM CSF prepared by COS/TPC method is effective in mediating GM CSF gene transfer and might be used in cancer gene therapy.
文摘Background Recent studies have shown that interleukin-3 receptor α （CD123） is highly expressed on leukemia stem cells of patients with acute myeloid leukemia, and is correlated with tumor load and poor prognosis.The expression of CD123 may also be high in patients with myelodysplastic syndrome （MDS）.In this study, the expression and clinical significance of CD123 and granulocyte colony stimulating factor （G-CSF） receptor （CD114） on the bone marrow cells of patients with MDS were investigated to explore the molecular marker of the malignant clone of MDS.Methods Forty-two patients with MDS, who were diagnosed in the Hematological Department of General Hospital of Tianjin Medical University from 2008 to 2009, and twelve normal controls were enrolled in this study.Fluorescence activiated cell sorter （FACS） was used to measure the expression of CD123 on CD34＋CD38- cells and CD114 on CD34＋cells of the bone marrow of these patients and controls and the clinical significance was analyzed.The expression of CD114 on CD123＋CD34＋CD38- cells was further measured to explore the molecular marker of the malignant clone in MDS.Results MDS patients displayed significantly higher proportion of CD34＋CD38-/CD34＋ （（14.03±5.27）%） than normal controls （（7.70±4.36）%, P 〈0.05）.The expression rate of CD123＋CD34＋CD38-/CD34＋CD38- was significantly higher in MDS patients （（48.39±28.15）%） than that in normal controls （（8.75±11.71）%, P 〈0.01）.The expression level of CD123 was significantly correlated with the proportion of bone marrow blasts （r=0.457, P 〈0.05）.The expression rate of CD114＋CD34＋/CD34＋ was lower in MDS patients （（33.05±21.71）%） than that in normal controls （（38.99±19.07）%） but was not statistically significant （P 〉0.05）.The expression of CD114 on CD123＋CD34＋CD38- cells （（34.82±29.58）%） was significantly lower than that on CD123-CD34＋CD38- cells （（53.48±27.41）%） of M DS patients （P 〈0.05）.Conclusions MDS patients displayed higher proportion of CD34＋CD38-/CD34＋ than normal controls.CD123 was highly expressed in the bone marrow of the patients with MDS, significantly correlated with the proportion of bone marrow blasts, and thus might be the marker of MDS malignant clone.CD123＋CD34＋CD38- cells exhibited lower expression of G-CSF receptors, which might partly explain why MDS clone responds worse to G-CSF in vitro and in vivo.
文摘Background Lipopolysaccharide （LPS） forms outer membrane of the wall of Gram-negative cells. LPS can directly cause damage to epithelia of respiratory tract and is the major factor responsible for the chronic inflammation of respiratory passage. The mitogen-activated protein kinase （MAPK） signal transduction pathway of the airway epithelia is intimately associated with the action of LPS. The chronic inflammation of respiratory tract and smoking are interrelated and entwined in the development and progression of chronic lung diseases. This study was designed to examine the effects of cigarette smoke extract （CSE） and LPS on MAPK signal transduction pathway in order to further understand the roles CSE and LPS play in chronic lung inflammation. Methods Cultured primary human epithelial cells of airway were divided into four groups according to the stimulants used： blank control group, LPS-stimulation group, CSE-stimulation group and CSE plus LPS group. Western blotting was employed for the detection of phosphorylation level of extracellular-signal-regulated-kinase （ERK1/2）, p38 MAPK and c-Jun N-terminal kinase （JNK）. The expression of cytokines of MAPK transduction pathway （granulocyte-macrophage colony stimulating factor （GM-CSF） and mRNA of IL-8） in the primary epithelial cells of respiratory tract was also determined. Results Western blotting revealed that the phosphorylation levels of ERK1/2, p38 MAPK and JNK were low and 2 hours after the LPS stimulation, the phosphorylation of ERK1/2, p38 MAPK and JNK were all increased. There was a significant difference in the phosphorylation between the LPS-stimulation group and blank control group （P〈0.05）; no significant difference was found between CSE-stimulation group and blank control group （P〉0.05）; there was a significant difference between CSE ＋ LPS group and blank control group and between CSE ＋ LPS group and LPS group （P〈0.05）. The phosphorylation of CSE-LPS group was higher than that of blank control group but lower than that of LPS group. In blank control group, the expression of IL-8 and GM-CSF mRNA was low in the epithelial cells of airway and the release of IL-8 and GM-CSF was also at a low level. One hour after LPS stimulation, the level of IL-8 mRNA increased （P〈0.05） and reached a peak after 2 hours. On the other hand, GM-CSF mRNA level increased 2 hours after the stimulation （P〈0.05） and reached the highest level 4 hours after the stimulation. Two hours after LPS stimulation, IL-8 and GM-CSF protein level began to rise （P〈0.05）, and the level was the highest 8 hours after the stimulation （P〈0.01）. Stimulation with CSE alone had no effect on the release of IL-8 and GM-CSF and expression of IL-8 mRNA （P〉0.05）, but pre-treatment with CSE could delay the LPS-induced release of IL-8 and GM-CSF and the expression of IL-8 mRNA and its peak was lower. Conclusions LPS stimulation can significantly increase the phosphorylation of ERK1/2, p38 MAPK and JNK in the epithelial cells of airway and activate the MAPK transduction pathway, thereby can activate the downstream signal transduction pathway, and can ultimately result in the release of cytokines by the epithelial cells of airway. CSE can partially abolish the LPS-induced activation of MAPK signal transduction pathway and the expression of cytokines of the pathway, which might contribute to the development and progression of the inflammatory reactions in COPD patients.
