ABSTRACT
Objective: To compare the patient-reported outcomes and short-term clinical outcomes between robotic-assisted and laparoscopic-assisted radical gastrectomy for locally advanced gastric cancer. Methods: This single-center prospective randomized controlled trial was conducted in the Department of Gastrointestinal Surgery,Affiliated Hospital of Qingdao University from October 2020 to August 2022. Patients with locally advanced gastric cancer who were to undergo radical gastrectomy were selected and randomly divided into two groups according to 1â¶1, and received robotic surgery and laparoscopic surgery, respectively. Patient-reported outcomes and short-term clinical outcomes (including postoperative complications, surgical quality and postoperative short-term recovery) were compared between the two groups by t test, Mann-Whitney U test, repeated ANOVA, generalized estimating equation, χ2 test and Fisher's exact test. Results: A total of 237 patients were enrolled for modified intention-to-treat analysis (120 patients in the robotic group, 117 patients in the laparoscopic group). There were 180 males and 59 females, aged (63.0±10.2) years (range: 30 to 85 years). The incidence of postoperative complications was similar between the robotic group and laparoscopic group (16.7% (20/120) vs. 15.4% (18/117), χ2=0.072, P=0.788). The robotic group had higher patient-reported outcomes scores in general health status, emotional, and social domains compared to the laparoscopic group, differences in time effect, intervention effect, and interaction effect were statistically significant (general health status: χ2 value were 275.68, 3.91, 6.38, P value were <0.01, 0.048, 0.041; emotional: χ2 value were 77.79, 6.04, 6.15, P value were <0.01, 0.014, 0.046; social: χ2 value were 148.00, 7.57, 5.98, P value were <0.01, 0.006, 0.048). However, the financial burden of the robotic group was higher, the differences in time effect, intervention effect and interaction effect were statistically significant (χ2 value were 156.24, 4.08, 36.56, P value were<0.01, 0.043,<0.01). Conclusion: Compared to the laparoscopic group, the robotic group could more effectively relieve postoperative negative emotions and improve recovery of social function in patients.
ABSTRACT
OBJECTIVE: Peripheral nerve block can provide effective postoperative analgesia to patients undergoing total hip arthroplasty (THA). This study aimed to compare ultrasound-guided pericapsular nerve group (PENG) block against anterior quadratus lumborum (AQL) block for pain management in primary THA. PATIENTS AND METHODS: In this prospective, double-blind, randomized controlled trial, 90 patients undergoing primary THA under general anesthesia were randomly allocated to receive ultrasound-guided PENG block + sham AQL block ("PENG group") or ultrasound-guided AQL block + sham PENG block ("AQL" group). The primary outcome was the highest pain score on a visual analogue scale while the patient was in the recovery room. Secondary outcomes included pain scores after transfer out of the recovery room, morphine consumption, quadricep strength, duration of hospitalization, pain level one year after surgery, and incidence of complications. RESULTS: Patients in the PENG group reported significantly lower maximum pain scores in the recovery room (31.3±9.1 vs. 37.3±7.4, p=0.001), as well as significantly lower pain scores at rest at 3 h after surgery and during motion at 3 and 6 h after surgery. The two groups did not differ significantly in postoperative morphine consumption, length of hospitalization, pain level at one year after surgery, or incidence of complications. Neither block significantly weakened the quadriceps. CONCLUSIONS: PENG block may provide slightly more effective postoperative analgesia than AQL block during the early recovery period after primary THA.
Subject(s)
Arthroplasty, Replacement, Hip , Humans , Femoral Nerve , Prospective Studies , Morphine/therapeutic use , Ultrasonography, Interventional , PainABSTRACT
OBJECTIVE: The aim of our study was to determine whether abnormal hyperplasia of chondrocytes occurs in glucocorticoid-induced osteonecrosis of the femoral head (GC-ONFH) using a well-established rat model. MATERIALS AND METHODS: Rats were injected with lipopolysaccharide and methylprednisolone to induce GC-ONFH, while control animals were injected with saline (12 animals per group). Establishment of the disease model was confirmed using micro-computed tomography and hematoxylin-eosin (HE) staining of femoral head tissue sections. Chondrocyte hyperplasia was detected using HE staining and semi-quantitated using toluidine blue and saffron O staining. Expression of the autophagy marker LC3B was assessed in cartilage tissues of femoral head using immunohistochemistry. RESULTS: GC-ONFH animals showed significantly greater area of abnormal chondrocyte hyperplasia in femoral head tissue sections than control animals. They also showed significantly higher expression of LC3B in articular cartilage of the femoral head. CONCLUSIONS: GC-ONFH may be associated with abnormal chondrocyte hyperplasia in articular surface cartilage, which may be related to glucocorticoid-induced overactivation of autophagy.
Subject(s)
Femur Head Necrosis , Femur Head , Animals , Chondrocytes , Eosine Yellowish-(YS)/adverse effects , Femur Head/pathology , Femur Head Necrosis/chemically induced , Femur Head Necrosis/pathology , Glucocorticoids , Hematoxylin , Hyperplasia/chemically induced , Lipopolysaccharides/adverse effects , Methylprednisolone/adverse effects , Rats , Tolonium Chloride/adverse effects , X-Ray MicrotomographySubject(s)
Chromothripsis , Multiple Myeloma , Humans , Multiple Myeloma/genetics , Chromosome AberrationsSubject(s)
Coronavirus Infections , Pneumonia, Viral , COVID-19 , Child , China , Humans , Infant , Respiratory Tract InfectionsABSTRACT
Objective: To investigate the clinical and laboratorial characteristics of patients with myelodysplastic syndrome (MDS) and erythroid hyperplasia. Methods: MDS patients whose bone marrow was hypercellular with erythroid lineage more than 50% and blasts account for less than 20% of non-erythroid cells were enrolled in this study. The ratio of mature erythrocytes to nucleated erythrocytes was no more than 20, namely MDS patients with erythroid hyperplasia(MDS-E). The retrospective analysis comprised 102 patients with MDS-E from the First Affiliated Hospital of Suzhou University. Clinical characteristics, karyotype, and the prognostic significance of erythroid hyperplasia were evaluated. Results: A total of 48 MDS-E patients (47.1%) presented a variety of cytogenetic abnormalities. The most frequently involved chromosomes were chromosome 8 (39.5% of all abnormal karyotypes), chromosome 7 (22.9%), followed by chromosome 5 (18.8%), chromosome 1 (16.7%) and chromosome 20 (16.7%). Hemoglobin (Hb) level affected the prognosis by survival analysis. The overall survival (OS) of MDS-E patients with Hb equal or more than 70 g/L was longer than that of patients less than 70 g/L (P<0.001). Allogeneic hematopoietic stem cell transplantation (HSCT) significantly improved the OS compared with best supportive care (P<0.001) and chemotherapy (P<0.001). The extent of erythroid hyperplasia in bone marrow did not impact on prognosis (P=0.187). Conclusions: Compared with previous reports of MDS patients, MDS-E patients have higher level of erythroid hyperplasia, more common erythroid dyshematopoiesis, more frequent 8 and 1 chromosome abnormalities. The degree of erythroid hyperplasia is not correlated with prognosis. Allogeneic hematopoietic stem cell transplantation improves the prognosis.
Subject(s)
Chromosome Aberrations , Hematopoietic Stem Cell Transplantation , Hyperplasia/pathology , Myelodysplastic Syndromes/genetics , Myelodysplastic Syndromes/pathology , Adult , Aged , Bone Marrow/pathology , Female , Humans , Karyotype , Karyotyping , Male , Middle Aged , Myelodysplastic Syndromes/surgery , Prognosis , Retrospective Studies , Survival Analysis , Treatment OutcomeABSTRACT
Objective: To investigate EVI1 expression and its associated clinical and cytogenetic characteristics in 447 acute myeloid leukemia (AML) patients. Methods: EVI1 expressions were measured in 447 AML cases from Jan. 2007 to Apr. 2015 to couple with clinical, cytogenetic and mutations' characteristics to summarize the features of AMLs with high EVI1 expression. Results: 17.9% of AML were high EVI1 expression (EVI1 +), and the remainder low EVI1 expression (EVI1-). No significant differences between the two groups in terms of age, sex, hemoglobin level, white blood cell count and platelet count were observed. More M0, M5 and M6 subtypes were observed in EVI1+ group (P= 0.027, 0.004 and 0.011, respectively). Cytogenetic abnormalities of 11q15, 11q23/MLL, 3q26, -7/7q- and t (9;11) were observed more frequently in EVI1 + group (P<0.001, <0.001, <0.001, <0.001, =0.014, respectively). Normal karyotype, inv (16), t (8;21) were observed more frequent in EVI1- group (P=0.001, 0.009, 0.002, respectively). EVI1 + was more observed in high risk cytogenetics. Mutation of NPM1 was more observed in EVI1- group (P <0.001). Remission rate in EVI1 + group was significantly lower than EVI1- group (P<0.001). Leukemia-free survival was improved in EVI1 + AML patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). Conclusions: High EVI1 expression was more observed in FAB subgroup M5, harbored more cytogenetic abnormalities of 11p15, 11q23/MLL, 3q26 rearrangement, -7/7q- and t (9;11). Remission rate of high EVI1 expression AML was lower, which could be improved by allo-HSCT.
Subject(s)
Chromosome Aberrations , DNA-Binding Proteins , Leukemia, Myeloid, Acute/genetics , MDS1 and EVI1 Complex Locus Protein/metabolism , Proto-Oncogenes , Adolescent , Adult , Chromosome Disorders , Cytogenetics , Female , Humans , Leukemia, Monocytic, Acute , Leukemia, Myeloid, Acute/pathology , Male , Middle Aged , Mutation , Nucleophosmin , Prognosis , RNA, Messenger , Transcription FactorsABSTRACT
The complete mitochondrial genome of the Southern catfish (Silurus meridionalis) and the Chinese catfish (S. asotus), was determined using the long and accurate polymerase chain reaction (LA-PCR) method. The mitochondrial DNA nucleotide sequences of S. meridionalis and S. asotus were compared with those of 47 other catfish species in the same order. The total length of mitochondrial DNA for S. meridionalis and S. asotus was 16,526 and 16,525 bp, respectively, and included 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and a non-coding control region. This mitochondrial gene arrangement is identical to that observed in other Siluriformes. To determine the relative phylogenetic positions of S. meridionalis and S. asotus, and to discover phylogenetic relationships among 24 families of Siluriformes, analyses were conducted, based on mitochondrial DNA, 12S ribosomal RNA, 16S ribosomal RNA, and 13 protein-coding gene sequence data sets. Phylogenetic analyses were congruent with a basal split of the order into Clupeiformes, Characiformes, Cypriniformes, and Siluriformes, and supported a closer relationship of the Southern catfish (family Siluridae) and the Chinese catfish (family Siluridae) to Pimelodidae than to Bagridae. We concluded that these two species are part of a molecular clade that is different from that proposed in recent studies, in which Amblycipitidae appears as a sister group. Our results showed Amblycipitidae appearing as the most basal extant, and Bagridae appearing as a sister group of Cranoglanididae and Pangasiidae. The Siluriformes showed close phylogenetic relationship to the Characiformes.
Subject(s)
Catfishes/genetics , Genome, Mitochondrial , Phylogeny , Sequence Analysis, DNA , Animals , Molecular Sequence Annotation , RNA, Ribosomal/genetics , RNA, Ribosomal, 16S/geneticsABSTRACT
PURPOSE: Cantharidin, a natural toxin, is the active substance of mylabris and has antitumor effects in man. Norcantharidin, the demethylated analogue of cantharidin, has been used in the treatment of patients with primary hepatoma and those with leukopenia in China. The present study was designed to investigate whether norcantharidin exerts cytotoxic activity against colorectal cancer cells by inducing apoptosis and to examine the possible mechanism in the phenomenon. METHODS: Inhibition of proliferation of norcantharidin on Colo205, HT-29, and SW480 colorectal cancer cells was determined by the trypan blue dye exclusion test. Apoptosis of norcantharidin-treated cells was determined by morphological analysis, agarose gel DNA electrophoresis, and quantitated by flow cytometry after staining with propidium iodide. Cell cycle and the cell surface expression of the CD95/CD95 ligand were evaluated by flow cytometry. Caspase 8-like protease and protein phosphatase 1 and 2A activities were also analyzed. RESULTS: Treatment with norcantharidin of colorectal cancer cells not only inhibited cell proliferation, but also induced apoptosis. Norcantharidin induced apoptosis mainly in two phases: rapid apoptosis in S-phase cells and delayed apoptosis in G2/M arrested cells. Treatment with norcantharidin resulted in an upregulation of the CD95 receptor and CD95 ligand on the cell surface. Furthermore, stimulation with anti-CD95 monoclonal antibody (mAb) resulted in further induction of apoptosis after treatment with norcantharidin. In addition, the apoptosis-inducing effect of norcantharidin was almost completely inhibited by anti-CD95 ligand mAb. Norcantharidin-treated cells showed the activation of caspase 8. Both zVAD-FMK (a broad range caspase inhibitor) and IETD-FMK (a caspase-8 inhibitor) showed apparent inhibition of the apoptosis-inducing effect. Norcantharidin did not show an inhibitory effect on protein phosphatase. CONCLUSIONS: These results suggest that norcantharidin triggers apoptosis in colorectal cancer cell lines via the activation of the CD95 receptor/ligand system, and that this agent may be useful for developing new therapeutic regimens for the treatment of colorectal carcinoma.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Bridged Bicyclo Compounds, Heterocyclic/pharmacology , Colorectal Neoplasms/pathology , Membrane Glycoproteins/biosynthesis , Antibodies, Monoclonal , Caspase 8 , Caspase 9 , Caspases/metabolism , Fas Ligand Protein , Humans , Membrane Glycoproteins/physiology , Tumor Cells, Cultured , Up-RegulationABSTRACT
AIM: To study the effects of cimetidine (Cim) on the production of interferon gamma (IFN gamma) and tumor necrosis factor alpha (TNF alpha) by splenocytes in immune-derived aplastic anemic (AA) mice. METHODS: Aplastic anemic mice model was constructed first, and then the splenocytes were induced to secrete IFN gamma and TNF alpha. Concentration of IFN gamma was assayed using sandwich ELISA, while that of TNF alpha was measured with L929 cytotoxicity methods. RESULTS: (1) Concentrations of IFN gamma and TNF alpha secreted by splenocytes from AA mice were (137 +/- 36) ng/L and (6 +/- 3) microg/L, respectively, much more than the irradiated and the control mice. (2) Treatment with Cim 10 micromol/L reduced the concentrations of IFN gamma and TNF alpha to (14 +/- 8) ng/L and (2.7 +/- 0.6) microg/L, respectively. CONCLUSION: Cim could effectively reduce the production of IFN gamma and TNF alpha from splenocytes of AA mice.
Subject(s)
Anemia, Aplastic/metabolism , Cimetidine/pharmacology , Histamine H2 Antagonists/pharmacology , Interferon-gamma/biosynthesis , Lymphocytes/metabolism , Tumor Necrosis Factor-alpha/biosynthesis , Anemia, Aplastic/pathology , Animals , Cells, Cultured , Female , Male , Mice , Mice, Inbred BALB C , Mice, Inbred DBA , Spleen/cytologyABSTRACT
Overcoming immune tolerance of the growth factors associated with tumor growth should be a useful approach to cancer therapy by active immunity. We used vascular endothelial growth factor (VEGF) as a model antigen to explore the feasibility of the immunogene tumor therapy with a vaccine based on a single xenogeneic homologous gene, targeting the growth factors associated with angiogenesis. To test this concept, we constructed a plasmid DNA encoding Xenopus homologous VEGF (XVEGF-p) and control vectors. We found that immunogene tumor therapy with a vaccine based on XVEGF was effective at both protective and therapeutic antitumor immunity in several tumor models in mice. VEGF-specific autoantibodies in sera of mice immunized with XVEGF-p could be found in Western blotting analysis and ELISA assay. The purified immunoglobulins were effective at the inhibition of VEGF-mediated endothelial cell proliferation in vitro, and at antitumor activity and the inhibition of angiogenesis by adoptive transfer in vivo. The elevation of VEGF in the sera of the tumor-bearing mice could be abrogated with XVEGF-p immunization. The antitumor activity and production of VEGF-specific autoantibodies, significantly elevated IgG1 and IgG2b, could be abrogated by the depletion of CD4(+) T lymphocytes. The observations may provide a vaccine strategy for cancer therapy through the induction of autoimmunity against the growth factors associated with tumor growth in a cross reaction with single xenogeneic homologous gene and may be of importance in the further exploration of the applications of other xenogeneic homologous genes identified in human and other animal genome sequence projects in cancer therapy.
Subject(s)
Endothelial Growth Factors/immunology , Lymphocyte Subsets/immunology , Lymphokines/immunology , Animals , Antibodies, Monoclonal/immunology , Endothelial Growth Factors/genetics , Enzyme-Linked Immunosorbent Assay , Female , Fibrosarcoma/immunology , Gene Library , Immunohistochemistry , Immunotherapy , Killer Cells, Natural/immunology , Liver Neoplasms, Experimental/immunology , Lymphocyte Depletion , Lymphokines/genetics , Mammary Neoplasms, Experimental/immunology , Mice , Models, Immunological , T-Lymphocytes/immunology , Tumor Cells, Cultured , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factors , Xenopus laevisABSTRACT
The title compound, 2,2'-(2,4,8,10-tetrathiaspiro[5.5]undecane-3,9-diylidene)bis(propanedinitrile), C(13)H(8)N(4)S(4), has been designed and synthesized for use as a potential new organic molecular electronic material. The spiro-annulated structure has twofold symmetry and is formed by two equal push-pull ethylene units, with the cycloalkylthio groups as electron donors and the cyano groups as electron acceptors. The intermolecular S.N non-bonded separation within a layer in the lattice is 3.296 (6) A, indicating a strong intermolecular interaction between the cyano groups and the S atoms, while the S atoms in two neighbouring molecules have a shortest S.S contact of 3.449 (3) A. In addition, attractive C-H.N and C-H.S interactions bridge adjacent molecules either within a layer or between layers. In short, these four types of intermolecular interactions combine to form an extended three-dimensional network in the lattice, resulting in a highly ordered array of molecular packing.
ABSTRACT
Bone marrow endothelial cells are the essential component of the bone marrow microenvironment. They produce many kinds of cytokines, including stimulators and inhibitors. Many researchers have suggested that in the presence of endothelial cell layer, CD34+CD38- cells are capable of expansion. The ability of the endothelial cell layer to protect hematopoietic stem cells from extensive differentiation may be related to the inhibitors derived from endothelial cells. The aim of the present study was to determine whether the inhibitors thymosin beta4 and AcSDKP are elaborated by murine bone marrow endothelial cells. Murine bone marrow endothelial cells (mBMECs) were cultured in serum-free conditioned medium. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to analyze the differential expression of the thymosin-beta gene, and reverse phase high-performance chromatography (HPLC) and mass spectroscopy were used to determine the concentration of thymosin beta4 (Tbeta4) and AcSDKP in EC lysate and in the medium (mBMEC-CM). Colony-forming unit granulocyte-macrophage (CFU-GM) colony assays were used to examine the effect of components (mw 3-10 kD, <3 kD) of mBMEC-CM, thymosin beta4, and AcSDKP on the proliferation of hematopoietic cells.mBMECs expressed Tbeta4 mRNA. In EC lysate and mBMEC-CM, Tbeta4 and AcSDKP were detected. After adding protease inhibitors, the concentration of Tbeta4 in EC lysate increased significantly, while the concentration of AcSDKP decreased. mBMEC-CM (mw 3-10 kD) had no effect on the formation of CFU-GM. However, mBMEC-CM (mw <3 kD) could inhibit the growth of CFU-GM. Tbeta4 (10(-11) approximately 10(-7)mol/L) and AcSDKP (10(-11) approximately 10(-5)mol/L) had dose-dependent inhibitory effects on the growth of CFU-GM. Angiotensin converting enzyme (ACE), the enzyme degrading AcSDKP, could partially eliminate the inhibitory effect of mBMEC-CM (mw <3 kD) on CFU-GM.BMECs express and secrete Tbeta4 and AcSDKP. Tbeta4 exists in the 3-10 kD component of mBMEC-CM, while AcSDKP exists in the <3 kD component of ECCM. Both components exert inhibitory effects on the proliferation of hematopoietic progenitors.
Subject(s)
Bone Marrow Cells/metabolism , Hematopoiesis/physiology , Oligopeptides/metabolism , Thymosin/metabolism , Animals , Cells, Cultured , Chromatography, High Pressure Liquid , Colony-Forming Units Assay , Culture Media, Conditioned/pharmacology , Culture Media, Serum-Free , Endothelium/cytology , Endothelium/metabolism , Gene Expression Regulation/drug effects , Mice , Peptidyl-Dipeptidase A/pharmacology , Protease Inhibitors/pharmacology , RNA, Messenger/biosynthesis , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
In the present study, the effects of murine bone marrow endothelial cell conditioned medium (ECM) combined with flt3 ligand (FL) or/and thrombopoietin (TPO) on the proliferation of HPP-CFC and CFU-GM were investigated. Both ECM and the concentrated retentate of ECM (MW>10 kD) promoted the growth of CFU-GM and HPP-CFC, and this promoting effect was further enhanced by addition of FL or TPO. Using reverse transcriptase-polymerase chain reaction (RT-PCR) technique, the expression of FL and TPO mRNA was not found in murine bone marrow endothelial cells.
Subject(s)
Bone Marrow Cells/cytology , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Hematopoietic Stem Cells/cytology , Membrane Proteins/pharmacology , Thrombopoietin/pharmacology , Animals , Cell Division/drug effects , Cells, Cultured , Colony-Forming Units Assay , Culture Media, Conditioned , Culture Media, Serum-Free , Endothelial Cells/cytology , Hematopoiesis/drug effects , Mice , RNA, MessengerABSTRACT
The breaking of immune tolerance against autologous angiogenic endothelial cells should be a useful approach for cancer therapy. Here we show that immunotherapy of tumors using fixed xenogeneic whole endothelial cells as a vaccine was effective in affording protection from tumor growth, inducing regression of established tumors and prolonging survival of tumor-bearing mice. Furthermore, autoreactive immunity targeting to microvessels in solid tumors was induced and was probably responsible for the anti-tumor activity. These observations may provide a new vaccine strategy for cancer therapy through the induction of an autoimmune response against the tumor endothelium in a cross-reaction.
Subject(s)
Cancer Vaccines/pharmacology , Endothelium/cytology , Endothelium/immunology , Immunotherapy/methods , Neoplasms, Experimental/therapy , Amino Acid Sequence , Animals , Antigens, CD/immunology , Autoantibodies/immunology , CD4-Positive T-Lymphocytes/immunology , Cattle , Cells, Cultured , Cross Reactions , Endothelium, Vascular/cytology , Endothelium, Vascular/immunology , Humans , Integrin alphaV , Mice , Molecular Sequence Data , Neovascularization, Pathologic/drug therapy , Neovascularization, Pathologic/immunology , Peptides/immunology , Receptor Protein-Tyrosine Kinases/immunology , Receptors, Growth Factor/immunology , Receptors, Vascular Endothelial Growth FactorABSTRACT
Recently, cytokines and interleukins such as SCF, GM-CSF, G-CSF, TGF-beta, IL-6, IL-7, IL-8, IL-11 have been reported to be elaborated by endothelial cells. For further study, serum free bone marrow endothelial cell conditioned medium (BMEC-CM) was collected and ultrafiltrated by using a centriprep 10. The concentrated retentate (R-BMEC-CM) contained some substances whose molecular weight was more than 10 000 daltons. The filtrate (F-BMEC-CM) contained some substances whose molecular weight was less than 10 000 daltons. The effects of R-BMEC-CM and F-BMEC-CM on the growth of haematopoietic progenitors and the expression of cytokine and interleukin mRNAs of BMEC were investigated. The results showed that R-BMEC-CM stimulated the growth of CFU-GM, HPP-CFC, BFU-E, CFU-E, and CFU-Meg; while F-BMEC-CM inhibited the growth of these progenitors. Using the method of hybridizing to the Atlas cDNA Array, we were able to detect the presence of mRNAs of cytokines and interleukins in bone marrow endothelial cells. Our finding of the existence of mRNAs of SCF, GM-CSF, IL-6, TGF-beta, IL-1, and IL-11 in these cells was in agreement with the data reported previously. Furthermore, we detected mRNAs of MIP-2, Thymosion-beta4, PDGF, MSP-1, IFN-gamma, IL-13 and inhibin, which are related to haematopoiesis. Among these cytokines and interleukins, SCF, GM-CSF, IL-6, IL-1, and IL-11 are haematopoietic stimulators which may be responsible for the stimulative effects on the growth of haematopoietic progenitors. One of our new findings, the thymosin-beta4, is a small molecular haematopoietic inhibitor. It may be responsible for the inhibitory effect of F-BMEC-CM on haematopoietic progenitors. The presence of mRNAs of BMP, MSP-1, MIP-2, PDGF and IL-13 suggests that bone marrow endothelial cells might elaborate these substances. Their influence on haematopoietic progenitors needs further study.
Subject(s)
Bone Marrow Cells/metabolism , Cytokines/biosynthesis , Hematopoiesis/physiology , Animals , Bone Marrow Cells/cytology , Cell Division/drug effects , Cells, Cultured , Culture Media, Conditioned/pharmacology , Endothelium/cytology , Endothelium/metabolism , Endothelium, Vascular/cytology , Endothelium, Vascular/metabolism , Gene Expression , Hematopoietic Stem Cells/drug effects , Mice , RNA, Messenger , S PhaseABSTRACT
The effects of > 10 kD component of serum-free conditioned medium from murine bone marrow-derived endothelial cell line on the formation of CFU-GM and HPP-CFC were investigated. We found that > 10 kD component alone could stimulate CFU-GM colony formation in dose-dependent. Above- 10 kD component supplemented with GM-CSF not only resulted in significant increase of CFU-GM as compared with > 10 kD or GM-CSF alone, but also could stimulate HPP-CFC formation. The effect of > 10 kD component on the formation of HPP-CFC was also in dose-dependent. The component above- 10 kD combined with IL-3, IL-6, EPO, SCF or GM-CSF further enhanced HPP-CFC colony formation. The results suggest that > 10 kD component may be used as the substitutes of hematopoietic growth factors to stimulate the formation of CFU-GM and HPP-CFC.
Subject(s)
Bone Marrow Cells/cytology , Culture Media, Conditioned/pharmacology , Endothelium/cytology , Hematopoietic Stem Cells/cytology , Animals , Cell Line , Colony-Forming Units Assay , Female , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Male , MiceABSTRACT
In this study, the effects of bone marrow endothelial cell conditioned medium (BMEC-CM) on hematopoietic progenitors and the expression of cytokines mRNA of BMEC were investigated. BMEC-CM was ultrafiltrated in Centriprep-10, an ultrafiltering concentrator. The retentate (MW>10 kD) and filtrate (MW<10 kD) were obtained. The effects of different components of BMEC-CM on the murine hematopoietic progenitors were examined. The results showed that the retentate of BMEC-CM (MW>10 kD) significantly promoted the growth of CFU-GM, CFU-E, BFU-E, CFU-Meg and HPP-CFC, while the growth of CFU-GM, CFU-E, BFU-E, CFU-Meg and HPP-CFC were markedly inhibited by the filtrate of BMEC-CM (MW<10 kD). The effects of BMEC-CM were dose-dependent. The presence of mRNAs for GM-CSF, TGF-beta, bone morphogenetic protein 2A, basic fibroblast growth factor receptor, endothelin-2, macrophage-stimulating 1 (MSP-1), thymosin-beta10, connective tissue growth factor, macrophage inflammatory protein 2 alpha (MIP-2alpha), IL-6, IL-11, IL-13 and IFN-gamma mRNA were successfully detected by Atlas Arrays.
Subject(s)
Cytokinins/biosynthesis , Endothelium/metabolism , Hematopoiesis/physiology , Thymosin/biosynthesis , Animals , Bone Marrow Cells/metabolism , Cell Division , Cells, Cultured , Chemokine CXCL2 , Culture Media, Conditioned , Endothelium/cytology , Interleukin-11/biosynthesis , Interleukin-11/genetics , Mice , Monokines/biosynthesis , Monokines/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Thymosin/geneticsABSTRACT
By using CFU-GM/CFU-L colony assay, NBT/MTT reductant test and DNA fragmentation analysis, we studied the effects of Sophora flavescens (SF) on CFU-GM proliferative ratio in human normal bone marrow/umbilical cord blood and on proliferation, differentiation and apoptosis in human acute myelogenous leukemia HL-60 cells. The results showed that 5, 10, 15, 20 micrograms.microliter-1 of SF significantly inhibited proliferation and induced apoptosis in the HL-60 cells in a dose-dependent fashion. Fifteen micrograms.microliter-1 of SF also induced differentiation in HL-60 cells. Furthermore, cytotoxic activity of SF(5-15 micrograms.microliter-1) was not apparent on human normal hematopoietic progenitors(CFU-GM). The results indicate that an appropriate concentration of SF has a selective antileukemic effect. Thus, these are important impetuses for further research of SF as an anticancer agent.