Résumé
<p><b>OBJECTIVE</b>To investigate the risk stratification of aggressive B cell lymphoma using the immune microenvironment and clinical factors.</p><p><b>METHODS</b>A total of 127 patients with aggressive B cell lymphoma between 2014 and 2015 were enrolled in this study. CD4, Foxp3, CD8, CD68, CD163, PD-1, and PD-L1 expression levels were evaluated in paraffin-embedded lymphoma tissues to identify their roles in the risk stratification. Eleven factors were identified for further evaluation using analysis of variance, chi-square, and multinomial logistic regression analysis.</p><p><b>RESULTS</b>Significant differences in 11 factors (age, Ann Arbor stage, B symptom, ECOG performance status, infiltrating CD8+ T cells, PD-L1 expression, absolute blood monocyte count, serum lactate dehydrogenase, serum iron, serum albumin, and serum β2-microglobulin) were observed among patient groups stratified by at least two risk stratification methods [International Prognostic Index (IPI), revised IPI, and NCCN-IPI models] (P < 0.05). Concordance rates were high (81.4%-100.0%) when these factors were used for the risk stratification. No difference in the risk stratification results was observed with or without the Ann Arbor stage data.</p><p><b>CONCLUSION</b>We developed a convenient and inexpensive tool for use in risk stratification of aggressive B cell lymphomas, although further studies on the role of immune microenvironmental factors are needed.</p>
Sujets)
Adulte , Humains , Marqueurs biologiques tumoraux , Régulation de l'expression des gènes tumoraux , Allergie et immunologie , Lymphome B , Classification , Anatomopathologie , Pronostic , Études rétrospectives , Analyse de survieRésumé
Burkitt lymphoma is a highly aggressive B-cell neoplasm. New therapeutic methods are needed to overcome the adverse effect of intensive chemotherapy regimens. Valproic acid and (-)-gossypol are two kinds of chemical compounds used as new anti-tumor drugs in recent years. To investigate the anti-tumor effect of valproic acid and (-)-gossypol, Burkitt lymphoma Namalwa cells were cultured and treated with valproic acid and (-)-gossypol at different concentrations. The proliferation of Namalwa cells was dramatically suppressed after the combination treatment with 2 mmol/L valproic acid and 5 μmol/L (-)-gossypol. The combined treatment also enhanced intrinsic apoptosis by down-regulating anti-apoptotic protein Mcl-1. Moreover, the autophagy flux significantly increased in Namalwa cells after combined treatment. However, the enhanced autophagy showed little effect on cell survival with present regimen. The results confirmed that combination of valproic acid and (-)-gossypol had synergistic anti-tumor effect to Burkitt lymphoma Namalwa cells. The related mechanisms might include the down-regulation of anti-apoptotic protein Mcl-1 and avianized pro-survival role of autophagy.
Sujets)
Humains , Antinéoplasiques , Pharmacocinétique , Utilisations thérapeutiques , Apoptose , Lymphome de Burkitt , Traitement médicamenteux , Lignée cellulaire tumorale , Contraceptifs masculins , Pharmacocinétique , Utilisations thérapeutiques , Synergie des médicaments , Antienzymes , Pharmacocinétique , Utilisations thérapeutiques , Régulation de l'expression des gènes tumoraux , Gossypol , Pharmacocinétique , Utilisations thérapeutiques , Acide valproïque , Pharmacocinétique , Utilisations thérapeutiquesRésumé
Hypoxia inducible factor-1α (HIF-1α) is an important transcription factor, which plays a critical role in the formation of solid tumor and its microenviroment. The objective of the present study was to evaluate the expression and function of HIF-1α in human leukemia bone marrow stromal cells (BMSCs) and to identify the downstream targets of HIF-1α. HIF-1α expression was detected at both the RNA and protein levels using real-time PCR and immunohistochemistry, respectively. Vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1α (SDF-1α) were detected in stromal cells by enzyme-linked immunosorbent assay. HIF-1α was blocked by constructing the lentiviral RNAi vector system and infecting the BMSCs. The Jurkat cell/BMSC co-cultured system was constructed by putting the two cells into the same suitable cultured media and conditions. Cell adhesion and secretion functions of stromal cells were evaluated after transfection with the lentiviral RNAi vector of HIF-1α. Increased HIF-1α mRNA and protein was detected in the nucleus of the acute myeloblastic and acute lymphoblastic leukemia compared with normal BMSCs. The lentiviral RANi vector for HIF-1α was successfully constructed and was applied to block the expression of HIF-1α. When HIF-1α of BMSCs was blocked, the expression of VEGF and SDF-1 secreted by stromal cells were decreased. When HIF-1α was blocked, the co-cultured Jurkat cell’s adhesion and migration functions were also decreased. Taken together, these results suggest that HIF-1α acts as an important transcription factor and can significantly affect the secretion and adhesion functions of leukemia BMSCs.
Sujets)
Humains , Sous-unité alpha du facteur-1 induit par l'hypoxie/métabolisme , Leucémie à cellules T/métabolisme , Cellules souches mésenchymateuses/métabolisme , Adhérence cellulaire , Test ELISA , Immunohistochimie , Cellules Jurkat , Réaction de polymérisation en chaine en temps réelRésumé
<p><b>OBJECTIVE</b>To investigate the effect of compound Danshen injection combined with prostaglandin E1 low molecular weight heparin calcium to dextran-40 preventing hepatic veno-occlusive disease (HVOD) after hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>A total of 520 patients who received HSCT in the authors' hospital from May 1998 and December 2009 were subjected, among whom 231 patients received autologous peripheral blood stem cell transplantation, 125 received HLA-identical sibling HSCT, 49 received HLA-identical/mismatched unrelated HSCT and 115 received HLA-haplotype HSCT. All patients were treated by intravenous dripping of CSI 40-60 mL, dextran-40 250-500 mL, prostaglandin-E1 40-60 microg, and subcutaneous injection of low molecular weight heparin calcium 3 000-5 000 IU every day, the preventive effect on HVOD after HSCT was observed.</p><p><b>RESULTS</b>HVOD occurred and caused death only in 1 case of the 520 patients observed, the incidence was 0.19%. Neither obvious adverse reaction nor coagulation disorder was found.</p><p><b>CONCLUSION</b>Compound Danshen Injection combined with prostaglandin E1, low molecular weight heparin calcium and dextran-40 is a safe and effective protocol for the prevention of HVOD after HSCT.</p>
Sujets)
Adolescent , Adulte , Enfant , Enfant d'âge préscolaire , Femelle , Humains , Mâle , Adulte d'âge moyen , Alprostadil , Utilisations thérapeutiques , Association thérapeutique , Dextrane , Utilisations thérapeutiques , Médicaments issus de plantes chinoises , Utilisations thérapeutiques , Transplantation de cellules souches hématopoïétiques , Héparine bas poids moléculaire , Utilisations thérapeutiques , Maladie veno-occlusive hépatique , Phénanthrolines , Utilisations thérapeutiquesRésumé
<p><b>OBJECTIVE</b>To observe the therapeutic effect of acpuncture treatment for chronic fatigue syndrome (CFS).</p><p><b>METHODS</b>Nighty cases of CFS were randomly divided into an observation group and a control group, 45 cases in each group. The observation group was treated with acupunture at Renying (ST 9), Fengfu (GV 16), Baihui (GV 20); the control group was treated with 250 mL 5% Glucose injectio combined with 20 mL Shenmai injectio. Fatigue Scale (FS) was used to compare the scores between the two groups after treatment.</p><p><b>RESULTS</b>The total scores in the observation group were 9.37 +/- 2.33 and 5.41 +/- 1.96 before and after treatment respectively, and in the control group, they were 9.08 +/- 2.27 and 7.34 +/- 2.03 respectively. FS brainwork integral, physical fatigue integral, and total integral all decreased after treatment in two groups (all P < 0.001), and it decreased much more obviously in the observation group (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Both of the acpuncture treatment and Shenmai injectio are able to decrease fatigue scale score, improve the fatigue symptoms of CFS patients, and the effect of acupucture treatment is obviously superior to that of Shenmai injectio.</p>
Sujets)
Adulte , Femelle , Humains , Mâle , Adulte d'âge moyen , Thérapie par acupuncture , Syndrome de fatigue chronique , Thérapeutique , Résultat thérapeutiqueRésumé
<p><b>OBJECTIVE</b>To summarize the clinical characteristics of secondary coagulation disorders caused by exposure to poison (raticide) in children and to investigate the diagnosis and corresponding treatment.</p><p><b>METHOD</b>The process of diagnosis, clinical characteristics, response to treatment and the prognosis were analyzed.</p><p><b>RESULTS</b>The main clinical manifestation was mucosal bleeding (66.6%), including epistaxis, gingival bleeding, hematomas and so on. All these children were previously well and had no history of bleeding. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged, factor II was undetectable and the levels of factors VII, IX, and X were lower. The fibrinogen was normal. A raticide was detected in blood and urine of 13 children although 12 of the patients had no definite history of raticide ingestion. Prothrombin complex, fresh frozen plasma and vitamin K(1) were effective in these cases. However, 2 - 3 weeks later, 6 patients presented with recurrent bleeding.</p><p><b>CONCLUSION</b>For children with secondary coagulation disorders of unknown cause, intoxication of raticide should be considered. The administration of blood coagulation factors and vitamin K(1) are effective in early treatment, and the treatment period should be more than 2 months. The PT and APTT should be followed up. Vitamin K(1) should be stopped when PT and APTT are normal.</p>
Sujets)
Femelle , Humains , Nourrisson , Nouveau-né , Mâle , Troubles de l'hémostase et de la coagulation , Diagnostic , Thérapeutique , Raticides , Intoxication , Phytoménadione , Utilisations thérapeutiquesRésumé
<p><b>OBJECTIVE</b>To provide reliable evidence of "J in three-needle therapy" for treatment of stroke.</p><p><b>METHODS</b>Multi-central randomized controlled trials were adopted, 180 hemiplegia patients of ischemic stroke were randomly divided into a fin three-needle group (90 cases) and a routine acupuncture group (90 cases). Two groups were both treated with basic neurology therapies, and J in three-needle group was treated with J in three-needle therapy, three acupoints of tempora, hand and foot etc. were selected; the routine acupuncture group was treated with traditional acupuncture, Quchi (LI 11), Huantiao (GB 30), Futu (ST 32) etc. were selected. Both groups were treated with acupuncture for 5 weeks. The cognitive function score of functional comprehensive assessment scale (FCA), the scores of mini-mental state examination scale (MMSE) and modified Barthel index (BI) were compared before and after treatment between two groups. Results After treatment, the scores of FCA, MMSE and BI in both groups were significantly improved compared to those before treatment (P < 0.01, P < 0.05); the improvement of FCA score, MMSE score and BI score in the J in three-needle group were superior to those of the routine acupuncture group after treatment (P < 0.01, P < 0.05). The total effective rate of 85.4% in the J in three-needle group was superior to tohat of 70.0% in the routine acupuncture group (P < 0.05).</p><p><b>CONCLUSION</b>J in three-needle acupuncture treatment can obviously improve the cognitive function and activity ability of daily life of hemiplegia patients after stroke, and the therapeutic effect of J in three-needle therapy is superior to that of traditional acupuncture treatment.</p>
Sujets)
Adulte , Sujet âgé , Femelle , Humains , Mâle , Adulte d'âge moyen , Activités de la vie quotidienne , Thérapie par acupuncture , Cognition , Hémiplégie , Psychologie , Réadaptation , Thérapeutique , Accident vasculaire cérébralRésumé
<p><b>OBJECTIVE</b>To observe the therapeutic effect of muscular needling combined with scarring moxibustion on active stage of rheumatoid arthritis (RA).</p><p><b>METHODS</b>Sixty cases of RA were randomly divided into a muscular needling group and a medication group, 30 cases in each group. The muscular needling group was treated by muscular needling on Quchi (LI 11), Sanyinjiao (SP 6), etc. combined with scarring moxibustion on Dazhui (GV 14), Zusanli (ST 36) etc., while the medication group was treated by oral administration of Diclofenac sodium and intramuscular injection of Methotrexate. The therapeutic effects, main symptoms and signs, erythrocyte sedimentation rate (ESR) and rheumatoid factor were observed in two groups before and after treatment.</p><p><b>RESULTS</b>The total effective rate of muscular needling group was 76.7%, and that of medication group was 73.3%, there was no significant difference between two groups (P > 0.05). The clinical symptoms, signs, and E8R of two groups were improved obviously compared with those before treatment (P < 0.01, P < 0.05), however there were no significant differences between the two groups after treatment (all P > 0.05). The adverse reactions of medication group were more eminent compared to the muscular needling group.</p><p><b>CONCLUSION</b>Muscular needling can obviously relieve the symptoms and signs of active stage rheumatoid arthritis and the effect is equivalent to oral administration of western medicine, the incidence of adverse reactions in the muscular needling group is obviously lower than that of western medication. Muscular needling is a safe and effective method for treatment of RA.</p>
Sujets)
Adulte , Sujet âgé , Femelle , Humains , Mâle , Adulte d'âge moyen , Jeune adulte , Points d'acupuncture , Thérapie par acupuncture , Polyarthrite rhumatoïde , Métabolisme , Thérapeutique , Association thérapeutique , Moxibustion , Facteur rhumatoïde , Métabolisme , Résultat thérapeutiqueRésumé
<p><b>OBJECTIVE</b>To sum up the clinical experience of the diagnosis and treatment of intracerebral infiltration by monoclonal plasmacytoid cells in Waldenstrom's macroglobulinemia(Bing-Neel syndrome).</p><p><b>METHODS</b>The clinical data of the diagnosis and treatment of a case of Bing-Neel syndrome was analyzed.</p><p><b>RESULTS</b>A 56-year-old male was diagnosed as Waldenstrom's macroglobulinemia one year ago, and presented with persistent headache during the treatment period. Magnetic resonance imaging showed a high intensity area on T2-weighed images in the right frontal lobe which was well enhanced by gadolinium-diethylenetriaminepenta-acetic acid. Infiltration of neoplastic cells was confirmed by biopsy. Immunohistochemical examination showed that mature plasmacytoid cells in the cerebral parenchyma were immunoglobulin M positive.</p><p><b>CONCLUSION</b>Infiltration in CNS (Bing-Neel syndrome) is uncommon in Waldenstrom's macroglobulinemia. As there is no effective therapy for this Bing-Neel syndrome, combination of radiation and chemotherapy should be considered for this situation.</p>
Sujets)
Humains , Mâle , Adulte d'âge moyen , Encéphale , Anatomopathologie , Invasion tumorale , Macroglobulinémie de Waldenström , AnatomopathologieRésumé
In order to investigate the effect of stromal cell derived factor-1 (SDF-1)/CXCR4 on the proliferation of megakaryocytic line-HEL cells co-cultured with human umbilical cord blood-derived stromal cells (hUCBSCs) and to further elucidate the mechanism of SDF-1/CXCR4-mediated functions, the HEL cells were co-cultured with hUCBSCs or human bone marrow stromal cells (hBMSCs), the suspended HEL was used as control. The concentrations of SDF-1 in supernatant of hUCBSCs and hBMSCs were detected by ELISA assay. The expression of CXCR4 membrane-bound protein of HEL cells was detected by laser confocal scanning microscopy and flow cytometry, and the expression of CXCR4 mRNA was detected by RT-PCR. The result showed that the concentrations of SDF-1 in different groups were the same at the early stage of culturing. But at 6 days after seeding, the concentrations of SDF-1 increased significantly in the hUCBSCs group, even though the passage was done. By means of laser confocal microscopy, the expression of CXCR4 protein and also red dots of fluorescence could be detected in cytoplasm of HEL cells co-cultured with hUCBSCs. However, there was no significant differences of the CXCR4 mRNA level between different groups (p > 0.05). It is concluded that hUCBSCs may play important roles in secreting high level of SDF-1 and regulating megakaryocyte expression of CXCR4.
Sujets)
Humains , Prolifération cellulaire , Cellules cultivées , Chimiokine CXCL12 , Génétique , Métabolisme , Techniques de coculture , Sang foetal , Biologie cellulaire , Métabolisme , Cytométrie en flux , Mégacaryocytes , Biologie cellulaire , Métabolisme , Monocytes , Biologie cellulaire , ARN messager , Génétique , Récepteurs CXCR4 , Génétique , Métabolisme , Cellules stromales , Biologie cellulaireRésumé
In order to study the influence of different gelatin concentrations, and lymphocyte isolation liquid on primary culture of umbilical cord blood-derived adhesive cells (hCBACs), the red blood cells of umbilical cord blood was separated by 3% and 6 % gelatin for detecting the effectiveness of sedimentation, then the adhesion rate at 48 hours, the day of initial expansion and the rate of culture success were detected for hCBACs cultured with CD34(+) cells after the mononuclear cells were separated by 6% gelatin followed by Ficoll and Percoll, and the morphological characteristics and growth status were observed by invert microscopy. Cytochemistry stain for nonspecific esterase stain (NSE), peroxidase (POX), periodic acid Schiff reaction (PAS) and alkali phosphatase (ALP) and immunocytochemistry labeling for CD31, CD45, CD68 and fibronectin (Fn) were detected. The results showed that 6 % gelatin was better than that 3% gelatin for red blood sedimentation. The Percoll was predominant over Ficoll in adhesion rate at 48 hours, the day of initial expansion, the time of initial formation of adhesive cell colony units, the time of maximal numbers of adhesive cell colony units, the the cell fusion time and ratio of culture success. 60% fibroblast-liked cells, 36% macrophage liked cells and 4% small-round cells were observed in cells isolated by both isolated methods. The cytochemistry stain for NSE, POX, PAS and ALP was similar in two groups, the difference was not statistically significant between these two groups. The immunocytochemistry labeling for CD31, CD45, CD68 and Fn was also similar in both groups and the difference was also not statistically significant between these two groups. It is concluded that the combination of 6% gelatin with Percoll is an ideal separation method for primary culture of hCBACs, which provides basic information for clinical application.
Sujets)
Humains , Séparation cellulaire , Méthodes , Cellules cultivées , Sang foetal , Biologie cellulaire , Gélatine , Pharmacologie , Lymphocytes , Biologie cellulaireRésumé
This study was aimed to investigate the effect of vcam-1 gene-modified human umbilical cord blood derived stromal cells (CBDSCs) on hematopoietic regulation so as to establish the experimental foundation for further study. The target gene vcam-1 was cloned into the shuttle plasmid with the report gene GFP. The recombinant shuttle plasmid was transformed into BJ5183 bacteria to recombine with backbone vector pAdeasy-l, and the recombinant adenoviral vector ad-vcam-1-gfp was confirmed after transfection with CBDSCs. The results indicated that two fragments of about 9 kb and 2 kb were obtained after digestion of recombinant plasmid pAdTrack-vcam-1 with NotIand XhoI, and single fragment of 600 bp was obtained after amplification with PCR; two fragments of about 31 kb and 4 kb were obtained after digestion of recombinant plasmid pad-vcam-1-gfp with PacI, which suggested a successful homologous recombination. The expression of vcam-1 gene in ad-vcam-1-gfp transfected CBDSCs could be detected by immunocytochemistry, RT-PCR and fluorescent microscopy. It is concluded that the recombinant adenoviral vector ad-vcam-1-gfp has been constructed successfully, and the expression of vcam-1 is up-regulated in CBDSCs transfected by gene ad-vcam-1-gfp.
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Humains , Adenoviridae , Génétique , Métabolisme , Sang foetal , Biologie cellulaire , Vecteurs génétiques , Génétique , Protéines à fluorescence verte , Génétique , Métabolisme , ARN messager , Génétique , Métabolisme , Protéines recombinantes , Génétique , Métabolisme , Cellules stromales , Biologie cellulaire , Transfection , Molécule-1 d'adhérence des cellules vasculaires , Génétique , MétabolismeRésumé
This study was aimed to investigate the expression of SDF-1 mRNA in SDF-1 cDNA-modified human bone marrow mesenchymal stem cells (hBMSCs) before and after transfection. The hBMSCs were isolated, cultured and identified, the SDF-1-pIRES2-EGFP eukaryotic expressing vector was constructed, and then the hBMSCs were transfected with the vector encapsulated by lipofectamine 2000. The transfection efficiency was measured by observing the expression of green fluorescence protein and detecting the mRNA by RT-PCR. The results indicated that the expression of SDF-1 mRNA increased by about 20% after hBMSCs were transfected instantaneously by SDF-1-pIRES2-EGFP. It is concluded that SDF-1 cDNA eukaryotic expression vector can be instantly transfected into hBMSCs by lipofectamine 2000, but the efficiency was too low to obtain enough steady transferred hBMSCs. Other procedures should be trialed to improve the transfection efficiency.
Sujets)
Humains , Cellules de la moelle osseuse , Biologie cellulaire , Chimiokine CXCL12 , Génétique , Métabolisme , ADN complémentaire , Génétique , Vecteurs génétiques , Cellules souches mésenchymateuses , Biologie cellulaire , ARN messager , Génétique , Métabolisme , TransfectionRésumé
<p><b>OBJECTIVE</b>To observe effects of acupuncture under guidance of qi street theory on endocrine function in the patient of epilepsy.</p><p><b>METHODS</b>Sixty cases of epilepsy were randomly divided into an acupuncture-medicine group and a medication group, 30 cases in each group. The medication group were treated with Valpromide, and the acupuncture-medicine group were treated with Valpromide and acupuncture at main points Renying (ST 9), Fengfu (GV 16), Tianzhu (BL 10). Epilepsy scores and plasma total cortisol (CORT), estradiol (E2), pregnendione (P), and prolactin (PRL) levels before and after treatment were observed. Results The total effective rates were 96.7% in the two groups with no significant difference between the two groups (P>0.05); the epilepsy score before treatment was (13.58+/-4.01) points and after treatment was (8.30+/-4.52) points in the acupuncture-medicine group, and (12.97+/-3.91) points and (7.86+/-4.90) points in the medication group, respectively, with significant improvement after treatment in the score in the two groups (P<0.05). After treatment, plasma CORT, E2, P and PRL levels in the acupuncture-medicine group and plasma CORT, E2, P levels in the medication group had significant changes (P<0.05 or P<0.01). There was a significant difference between the two groups in CORT level after treatment (P<0.05).</p><p><b>CONCLUSION</b>Acupuncture under guidance of qi street theory is an effective method for improving endocrine function in the patient of epilepsy.</p>
Sujets)
Adulte , Femelle , Humains , Mâle , Points d'acupuncture , Thérapie par acupuncture , Glandes endocrines , Épilepsie , Thérapeutique , Médecine traditionnelle chinoise , Guides de bonnes pratiques cliniques comme sujet , QiRésumé
This study was purposed to investigate the connexin 43 (Cx43) expression level in acute leukemia bone marrow stromal cells (ABMSCs) and normal bone marrow stromal cells (NBMSCs), and to explore the difference in communicating functions between these cells. The Cx43 expression levels of ABMSCs and NBMSCs were detected by using immunohistochemistry and computer gray scale assay, and the difference of gap junction intercellular communication (GJIC) was examined through dry transfer technique. The results showed that expression level of Cx43 in ABMSCs was lower than that in NBMSCs and its function of GJIC in ABMSCs was also weaker than that in NBMSCs. It is concluded that cell-cell communication function is lowered in ABMSCs.
Sujets)
Humains , Maladie aigüe , Cellules de la moelle osseuse , Biologie cellulaire , Métabolisme , Anatomopathologie , Communication cellulaire , Physiologie , Connexine 43 , Métabolisme , Jonctions communicantes , Métabolisme , Leucémies , Métabolisme , Cellules stromales , Biologie cellulaire , Métabolisme , Cellules cancéreuses en cultureRésumé
Objective To investigate the effects and safety of the treatment of continuous veno-venous hemofiltration (CVVH)in refractoriness heart failure with renal insufficiency patients.Methods 35 hospitalized patients of refractori- ness heart failure with renal insufficiency were chosen from 2001 to 2006,25 patients were treated by CVVH,and other 10 patients were treated by regular hemodialysis.The serum BUN,Cr,left ventricle ejection fraction(LVEF),were observed before and after the therapy in each group.Results The clinical symptom were obviously improved in Group HF,and LVEF,cardiac output(CO) were both increased by continuous hemofiltration treatment(P
Résumé
The aim of this study was to evaluate the effect of all-trans retinoic acid (ATRA) on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells (BMSC) in patients after conditioning treatment for peripheral blood stem cell transplantation (PBSCT). BMSC of 27 patients before and after conditioning treatment for PBSCT were cultured in vitro. After treated with ATRA at 0.01, 0.1, or 1 micromol/L, expression of intercellular adhesion molecule-1 (ICAM-1) protein and vascular adhesion molecule-1 (VCAM-1) protein were detected by flow cytometry, and soluble ICAM-1 (sICAM-1) protein was determined by using radioimmunoassay. Then BMSC was co-cultured with CD34+ cells, and adhesion rate of BMSC to CD34+ cells was measured. The results showed that after pretreatment with conditioning regimen for PBSCT, the expressions of ICAM-1 and VCAM-1 proteins in BMSC and the expression level of sICAM-1 protein in supernatant of BMSC culture were down-regulated, and the adhesion rate of BMSC to CD34+ cells was decreased, after administration of ATRA, the expression of ICAM-1 protein in BMSC, sICAM-1 protein in culture medium and adhesion rate of BMSC to CD34+ cells all increased significantly, but expression of VCAM-1 protein changed no significantly. It is concluded that the ATRA can partly restore adhesion function of BMSC injured by pretreatment for PBSCT and contribute to hematopoietic reconstitution.
Sujets)
Adolescent , Adulte , Enfant , Humains , Adulte d'âge moyen , Antigènes CD34 , Antinéoplasiques , Pharmacologie , Cellules de la moelle osseuse , Métabolisme , Anatomopathologie , Adhérence cellulaire , Molécules d'adhérence cellulaire , Génétique , Techniques de coculture , Tumeurs hématologiques , Métabolisme , Anatomopathologie , Thérapeutique , Transplantation de cellules souches de sang périphérique , Cellules stromales , Métabolisme , Anatomopathologie , Trétinoïne , Pharmacologie , Cellules cancéreuses en culture , Molécule-1 d'adhérence des cellules vasculaires , GénétiqueRésumé
The aim of this study was to evaluate the effect of dimethyl amiloride (DMA), a specific inhibitor of Na(+)/H(+) exchanger-1 (NHE-1), on intracellular pH value (pHi), proliferation and apoptosis of HL-60/ADM cells in vitro. After treatment with DMA at different doses, pHi of HL-60 and HL-60/ADM cell lines were determined by using pH-sensitive fluorescence dye BECEF-AM; the rate of growth inhibition of cells was detected with MTT assay; cell cycle was detected by flow cytometric DNA analysis; cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The results showed that pHi in HL-60/ADM cells was higher than that in HL-60 cells. After treatment with DMA at different doses, pHi decreased, the rate of growth inhibition and the rate of apoptotic cells in HL-60/ADM cells were all higher than those in HL-60 cells. Meanwhile, after treatment with DMA during 100 micromol/L to 150 micromol/L, the increase amplitude of G(0)/G(1) phase cells and the decrease amplitude of S + G(2)/M cells in HL-60/ADM cells were higher than those in HL-60 cells. It is concluded that by causing intracellular acidification, the NHE-1-specific inhibitor DMA inhibits proliferation of HL-60/ADM cells and induces apoptosis of HL-60/ADM cells, and the degree of this growth inhibition of HL-60/ADM cells is higher than that of HL-60 cells.
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Humains , Amiloride , Pharmacologie , Apoptose , Cycle cellulaire , Prolifération cellulaire , Doxorubicine , Pharmacologie , Résistance aux médicaments antinéoplasiques , Cellules HL-60 , Concentration en ions d'hydrogène , Antiport des ions sodium-hydrogèneRésumé
<p><b>OBJECTIVE</b>To investigate the inhibition effect of leukemic bone marrow stromal cells (BMSCs) on daunorubicin (DNR) induced apoptosis of human Jurkat cell line, and analyze the differentially expressed genes between Jurkat cells cocultured with leukemic BMSCs or without.</p><p><b>METHODS</b>Suppression subtractive hybridization (SSH) was employed to establish subtracted cDNA library of differentially expressed genes in Jurkat cells cocultured with leukemic BMSCs and DNR. The cDNA fragments were sequenced and analyzed.</p><p><b>RESULTS</b>The differentially expressed gene cDNA library was successfully developed. Primary screening was done by reverse Northern hybridization. Thirty up-regulated and 22 down-regulated cDNA fragments were isolated and sequenced. Analysis and comparison were performed in GenBank using BLAST. These genes are related to cell cycle regulation, cell apoptosis and energy metabolism.</p><p><b>CONCLUSION</b>Leukemic BMSCs influence gene expression of Jurkat cells. The resulting differentially expressed genes might be associated with the protection of leukemic cells by BMSCs from injury.</p>
Sujets)
Humains , Apoptose , Cellules de la moelle osseuse , Métabolisme , Anatomopathologie , Cellules cultivées , Techniques de coculture , Daunorubicine , Pharmacologie , Analyse de profil d'expression de gènes , Régulation de l'expression des gènes dans la leucémie , Banque de gènes , Cellules Jurkat , Cellules stromales , Métabolisme , AnatomopathologieRésumé
<p><b>OBJECTIVE</b>To construct a multiple myeloma (MM)-specific APE1siRNA expression vector, and detect the specific knock-down effect of the siRNA on expression of APE1 protein.</p><p><b>METHODS</b>APE1siRNA cDNA sequence was designed, synthesized and inserted into pSilencer 2.0-U6 linear expression vector. pSilencer APE1siRNA was digested by enzyme EcoRI and BamHI, then linear vector and IgP fragments were conjugated by T4 DNA ligase. pSilencer IgP-APE1siRNA and pSilencer IE-IgP-APE1siRNA were digested by enzyme EcoRI or XhoI. Linear vector and IE or Kappa fragments were conjugated by T4 DNA ligase. Then a MM specific pSilencer K-IE-IgP-APE1siRNA was cloned. The recombinant products were identified by DNA sequencing and enzyme digestions at each step. pSilencer K-IE-IgP-APE1siRNA plasmid was transfected to KM3, HOS, MDA-231 cells by liposome. APE1 gene silence induced by RNAi was analysed by Western blot.</p><p><b>RESULTS</b>APE1 protein in KM3 cells could be knocked down effectively and specifically by pSilencer K-IE-IgP-APE1siRNA vector. After 2 days, the level of APE1 protein in KM3 cells transfected with siRNA was 0.118 +/- 0.047, while that transfected with plasmid only was 0.988 +/- 0.029. The efficiency of gene silence was 90%.</p><p><b>CONCLUSION</b>A MM specific APE1siRNA expression vector was successfully constructed.</p>