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1.
Article in Chinese | WPRIM | ID: wpr-689588

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of arsenic trioxide (AsO) on Cdc20 and Mad2 in process of AML HL-60 cell proliferation.</p><p><b>METHODS</b>The proliferation of HL-60 cells was detected by CCK-8 method at different concentrations of arsenic trioxide for 24, 48 and 72 hours. The cell morphological changes were observed by inverted microscopy. The expressions of Mad2 and Cdc20 mRNA and protein in HL-60 cells treated with AsO for 48 h were detected by real-time PCR and Western blot respectively.</p><p><b>RESULTS</b>Arsenic trioxide significantly inhibited the HL-60 cell proliferation and displayed a good time-dose correlation. RT-PCR and Western blot showed that the expression of Mad2 was up-regulated and the expression of Cdc20 was down-regulated in HL-60 cells treated with arsenic trioxide of different concentration (4,8,10 µmol/L).</p><p><b>CONCLUSION</b>Arsenic trioxide can inhibit the human acute myeloid leukemia HL-60 cell proliferation, and its mechanism may be related with up-regulation of Mad2 expression and down-regulation of Cdc20 expression.</p>


Subject(s)
Antineoplastic Agents , Apoptosis , Arsenic Trioxide , Arsenicals , Cdc20 Proteins , HL-60 Cells , Humans , Leukemia, Myeloid, Acute , Oxides
2.
Article in Chinese | WPRIM | ID: wpr-271969

ABSTRACT

<p><b>OBJECTIVE</b>To detect the expression levels of MRE11 and Ku80 mRNA, and telomere length in bone marrow mononuclear cells of aplastic anemia(AA) patients, and to explore their correlation with pathogenesis of aplastic anemia.</p><p><b>METHODS</b>Bone marrow mononuclear cells were collected from 40 cases of AA and 20 normal controls for detecting mRNA expression of MRE11 and Ku80 and telomere length by using real-time quantitative polymerase chain reaction (qPCR), then MRE11, Ku80 and telomere length were analyzed for their correlation.</p><p><b>RESULTS</b>As compared with controls, the expression levels of MRE11 and Ku80 in patients with AA were significantly reduced, and the telomere length in patients with AA was obviously shortened, respectively (P<0. 05). The telomere length was significantly shorter in the persons aged ≥45 years in comparison with the AA patients and normal control younger than 45 years old (P<0.05). For the AA patients older than or equal to 45 years and less than 45 years in comparison with the controls at the same age, the telomere length was significantly shorter(P<0.05). The expression levels of MRE11 and Ku80 didn't correlate with telomere length (P>0.05). The mRNA expression level of MRE11 correlated positively and significantly with that of Ku80 (r=0.863, P<0.05).</p><p><b>CONCLUSION</b>The change of telomere length may play an important role in the pathogenesis and progression of aplastic anemia. The lower expression of MRE11 and Ku80 may be involved in the pathogenesis of aplastic anemia.</p>

3.
Article in Chinese | WPRIM | ID: wpr-357271

ABSTRACT

<p><b>OBJECTIVE</b>To detect the mRNA expression levels of TERT and TIN2 in peripheral blood mononuclear cells of acquired aplastic anemia(AAA) patients, and to explore their correlation with pathogenesis of acquired aplastic anemia.</p><p><b>METHODS</b>Peripheral blood mononuclear cells of 40 cases of AAA including 33 cases of non-severe aplastic anemia(NSAA), 7 cases of severe aplastic anemia (SAA) and 20 subjects as control group were collected to detect mRNA expression of TERT and TIN2 by using real-time quantitative polymerase chain reaction(RT-qPCR), the correlation of TERT and TIN2 mRNA expression levels with classification of peripheral blood cells were analyzed.</p><p><b>RESULTS</b>The expression levels of TERT and TIN2 mRNA in patients with AAA were lower significantly than those in control group (P<0.05), and the SAA (P<0.01). The expression levels of TERT and TIN2 mRNA in patients with SAA were all lower significantly than those in patients with NSAA (P<0.05). The expression levels of TIN2 mRNA in patients with NSAA were lower significantly than those in control group (P<0.01). There were no significant difference in the expression level of TERT mRNA between patients with NSAA and control group (P=0.082). There was significant correlation between the expression level of TERT mRNA and red blood cells count (r=0.437, P=0.029), and hemoglobin level (r=0.522, P=0.007). There was significant correlation between the expression levels of TIN2 mRNA and the lymphocyte percentage (r=-0.404, P=0.045).</p><p><b>CONCLUSION</b>The expression level of TERT mRNA may be associated with the red blood cells and hemoglobin level. The expression level of TIN2 mRNA may be associated with the lymphocyte percentage.</p>


Subject(s)
Anemia, Aplastic , Gene Expression Regulation , Humans , Leukocytes, Mononuclear , RNA, Messenger , Telomerase
4.
Article in Chinese | WPRIM | ID: wpr-302406

ABSTRACT

This study was aimed to sort the side population (SP) cells from human multiple myeloma cell lines, then detect the biological characteristics of those SP cells. After Hoechst33342 staining, intracellular Hoechst33342 fluorescence staining differences of myeloma cell lines observed by the fluorescence microscopy. The fluorescence-activated cell sorting (FACS) technology was used to isolate SP cells and main population (MP) cells; proliferative capacity in vitro was determined by cell growth curve; the cell colony forming ability was compared by colony forming test. The CD138 expression was detected by flow cytometry. The expression of ABCG2 mRNA was detected by reverse transcription PCR; CCK-8 assay and colony forming test were used to evaluate the effect of bortezomib on the cell proliferation, vitality and colony forming ability of the two populations. The results showed that the myeloma cell lines had a small proportion of SP cells, especially, RPMI 8226 cells accounted for the highest proportion of SP cells (7.10 ± 2.69)%, which have also been confirmed under the fluorescence microscope; the proliferative activity and cell colony forming ability of SP cells were significantly higher than those of MP cells (P < 0.05). The expression levels of CD138 in SP and MP cells were not significantly different (P > 0.05). RT-PCR results showed that SP cells expressed the drug-resistance gene ABCG2, but MP cells hardly express these genes. The inhibition rate of bortezomib on SP cells was significantly lower than that on MP cells (P < 0.05), however, the difference was not significant (P > 0.05) at bortezomib 40 nmol/L. Bortezomib could reduce colony formation in the both two cell populations, but more severe reduction appeared in the MP cells. It is concluded that the myeloma cell line contain a small amount of SP cells with the cancer stem cell characteristics.


Subject(s)
Cell Line, Tumor , Cytological Techniques , Methods , Humans , Multiple Myeloma , Neoplastic Stem Cells , Cell Biology , Side-Population Cells , Cell Biology
5.
Chinese Journal of Hematology ; (12): 941-945, 2013.
Article in Chinese | WPRIM | ID: wpr-295767

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the down-regulated TRAF6 gene expression and its effects on proliferation and apoptosis in multiple myeloma (MM) cells.</p><p><b>METHODS</b>Detection of TRAF6 expression were conducted by RT-PCR and Western blot in MM cell lines of KM3, U266, RPMI8226 and primary cells from patients. RPMI8226 cell lines were transfected with siRNA of TRAF6. The efficiency of transfection was identified by using of fluorescence microscope, RT-PCR, and Western blot. The levels of proliferation were analyzed by CCK-8 method under the different concentrations of siRNA. Apoptosis rate were detected with Hoechst33258/PI double staining by flow cytometry. Apoptosis related proteins Bcl-2, BAX, and NF-κB signal pathway were observed before and after siRNA transfection by Western blot.</p><p><b>RESULTS</b>The levels of TRAF6 mRNA and protein in MM cell lines, especially in primary myeloma cells, were significantly higher than those in controls. After transfected with 50 nmol/L siRNA in RPMI8226 cells, the relative level of TRAF6 mRNA (0.49±0.24) was significantly lower than that in non-transfected group (1.87±0.23) and idling group (1.74±0.35). The proliferation rate of siRNA transfected cells decreased with dose dependence (P<0.01). The apoptosis rates increased from 11.20% (before transfection) to 51.82% (after transfection), accompanied by down-regulated Bcl-2 protein, NF-κB signal pathway (p-p65 and p52), and up-regulated BAX protein.</p><p><b>CONCLUSION</b>TRAF6 expression was high in myeloma cells. TRAF6 siRNA could inhibit proliferation of myeloma cells and induce apoptosis mediated by NF-κB classical and alternative pathway in myeloma cells.</p>


Subject(s)
Case-Control Studies , Cell Proliferation , Down-Regulation , Female , Gene Expression , Humans , Male , Multiple Myeloma , Metabolism , Pathology , TNF Receptor-Associated Factor 6 , Genetics , Metabolism , Tumor Cells, Cultured
6.
Journal of Experimental Hematology ; (6): 1158-1161, 2012.
Article in Chinese | WPRIM | ID: wpr-278415

ABSTRACT

This study was aimed to investigate the expression levels of TRF1, TRF2 and RAP1 mRNA in peripheral blood mononuclear cells of patients with acquired aplastic anemia, and to explore their relation with onset of acquired aplastic anemia. Peripheral blood mononuclear cells of 40 patients with acquired aplastic anemia and 20 normal subjects as control were collected to detect mRNA expression of TRF1, TRF2 and RAP1 by using real-time quantitative polymerase chain reaction. The results showed that the expression levels of TRF1 and RAP1 in peripheral blood mononuclear cells of patients with acquired aplastic anemia were significantly higher than that in normal controls (P < 0.05), while the expression level of TRF2 was lower than that in normal controls (P < 0.01). There was significant correlation between TRF2 and RAP1 expressions level (r = 0.522, P = 0.001). It is concluded that the changes in expression levels of TRF1, TRF2 and RAP1 may play a role in the pathogenesis of acquired aplastic anemia.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Anemia, Aplastic , Blood , Case-Control Studies , Child , Female , Humans , Leukocytes, Mononuclear , Metabolism , Male , Middle Aged , RNA, Messenger , Genetics , Telomeric Repeat Binding Protein 1 , Metabolism , Telomeric Repeat Binding Protein 2 , Metabolism , Young Adult , rap1 GTP-Binding Proteins , Metabolism
7.
Article in Chinese | WPRIM | ID: wpr-252488

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the correlation between Chinese medical syndrome type (CMST) and the ID4 gene promoter methylation state in the bone marrow cells of acute myeloid leukemia (AML) patients, and to discuss the correlation between ID4 gene methylation and the occurrence, development of AML.</p><p><b>METHODS</b>Thirty-five inpatients or outpatients from the Department of Hematology, the Affiliated Hospital of Shandong University of Traditional Chinese Medicine were recruited as the test group, while 10 healthy volunteers from the health medical center of the Affiliated Hospital, Shandong University of Traditional Chinese Medicine were recruited as the control group. The ID4 gene promoter methylation states were detected using methylation specific polymerase chain reaction (MS-PCR) in the two groups. Inter-group comparison was performed and CMSTs were compared to analyze the correlation between CMSTs and the gene promoter methylation.</p><p><b>RESULTS</b>Twenty-seven AML patients (77.1%) had methylation of ID4 gene, showing statistical difference when compared with the control group (P < 0.01). The ID4 methylation positive rate of ID4 gene promoter methylation was sequenced from low to high as qi and yin deficiency syndrome < inter-accumulation of blood stasis and phlegm syndrome < toxic heat inflaming syndrome, showing statistical difference when compared with the control group (P < 0.05). The peripheral white blood cells and the bone marrow blast cells were higher in ID4 methylation positive patients than in the ID4 methylation negative control patients with statistical difference (P < 0.05).</p><p><b>CONCLUSIONS</b>Patients of inter-accumulation of blood stasis and phlegm syndrome and toxic heat inflaming syndrome were more likely to have ID4 gene methylation. The ID4 methylation positive expression has verified the essence of evil domination in the early AML at the molecular level. It can also reflect the degree of malignancy of AML to some extent.</p>


Subject(s)
Adolescent , Adult , Aged , Case-Control Studies , DNA Methylation , Female , Humans , Inhibitor of Differentiation Proteins , Genetics , Metabolism , Leukemia, Myeloid, Acute , Diagnosis , Genetics , Metabolism , Male , Medicine, Chinese Traditional , Middle Aged , Promoter Regions, Genetic , Young Adult
8.
Article in Chinese | WPRIM | ID: wpr-313939

ABSTRACT

Objective of this study was to investigate the ID4 gene methylation status in patients with acute myeloid leukemia (AML). Methylation-specific PCR (MS-PCR) was used to detect the promoter methylation status of ID4 gene in bone marrow samples from 46 AML patients with different subtypes and stage of disease and from 10 patients with iron deficiency anemia (IDA) as a control. The results showed that ID4 gene in bone marrow of IDA patients was completely non-methylated, while ID4 gene methylation was found in 39 out of 46 AML patients (positive rate 84.8%). The positive rates of ID4 gene methylation in different FAB types M₁, M₂, M₃, M₄, M₅, M₆ were 100% (4/4), 75% (9/12), 100% (8/8), 77.8% (7/9), 81.8% (9/11), 100% (2/2) respectively. The positive rates of ID4 gene methylation in newly diagnosed and complete remitted of AML patients were 90% (27/30) and 63.3% (7/11) respectively; ID4 methylation was detected in 5 relapsed and refractory AML patients. There were statistically significant differences in ID4 gene methylation between AML and IDA patients (p < 0.01). It is concluded that compared with IDA patients, ID4 gene methylation changes of different degrees occur in AML patients with different subtypes and stages, which suggests that ID4 gene methylation may be an early molecular event in the process of AML.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , DNA Methylation , Female , Humans , Inhibitor of Differentiation Proteins , Genetics , Leukemia, Myeloid, Acute , Genetics , Male , Middle Aged , Promoter Regions, Genetic , Young Adult
9.
Article in Chinese | WPRIM | ID: wpr-313245

ABSTRACT

<p><b>OBJECTIVE</b>To study the action mechanism of Yiqi Yangyin Recipe (YYR) in treating leukemia by observing the effects of YYR and its different assembling, energy supporting part (P1) and evil dispelling part (P2) on expressions of Flt3 and N-ras gene in acute myeloid leukemic (AML) cells.</p><p><b>METHODS</b>The mononuclear cells collected from bone marrow of 60 AML patients were assigned to four groups: the blank was untreated for control and the three tested groups were treated with YYR, P1 and P2, respectively. The effects on Flt3 and N-ras gene expressions and FLT3 protein expression were observed by RT-PCR and Western bloting.</p><p><b>RESULTS</b>RT-PCR test showed the expression of Flt3 in the control, YYR, P1 and P2, group was 90.78% +/- 6.92%, 38.18% +/- 4.50%, 65.57% +/- 5.55% and 61.35% +/- 6.39%, respectively; and that of N-ras in them 93.28% +/- 5.54%, 34.38% +/- 6.69%, 59.42% +/- 7.35% and 65.28% +/- 7.64%, respectively, both showed significant difference as compared the data in the tested groups with those in the control group (P < 0.05). Western bloting test showed the FLT3 protein gray value in the four groups was 0.8127 +/- 0.0284, 0.4265 +/- 0.0353, 0.5396 +/- 0.0274 and 0.5473 +/- 0.0282, respectively, also showed significant difference between the control and the tested groups (P < 0.01).</p><p><b>CONCLUSION</b>YYR can inhibit the colonic proliferation of AML cells, decrease the expressions of FLT3 and N-ras in cells, therefore shows a therapeutic effect on AML.</p>


Subject(s)
Adolescent , Adult , Bone Marrow Cells , Pathology , Child , Drugs, Chinese Herbal , Pharmacology , Female , Genes, ras , Humans , Leukemia, Myeloid, Acute , Genetics , Pathology , Leukocytes, Mononuclear , Metabolism , Pathology , Male , Middle Aged , Tumor Cells, Cultured , Young Adult , fms-Like Tyrosine Kinase 3 , Genetics , Metabolism , ras Proteins , Genetics , Metabolism
10.
Journal of Experimental Hematology ; (6): 1100-1102, 2008.
Article in Chinese | WPRIM | ID: wpr-234291

ABSTRACT

The objective of this study was to investigate the expressions of hematopoietic cytokines EPO, SCF and GM-CSF in bone marrow cells of patients with chronic aplastic anemia (CAA) and their significance. The mRNA expressions of EPO, SCF and GM-CSF from 35 CAA patients and 10 healthy individuals were detected by semi-quantitative RT-PCR. The results showed that the levels of EPO, SCF and GM-CSF in CAA patients were obviously lower than those in the healthy individuals (p < 0.01). It is concluded that the immunity disorder mediated by hematopoietic cytokines is one of the pathogenesis in the CAA patients, it provided theoretical basis for guiding clinical treatment.


Subject(s)
Adolescent , Adult , Aged , Anemia, Aplastic , Genetics , Metabolism , Pathology , Bone Marrow , Metabolism , Pathology , Child , Chronic Disease , Erythropoietin , Metabolism , Female , Granulocyte-Macrophage Colony-Stimulating Factor , Metabolism , Humans , Male , Middle Aged , RNA, Messenger , Genetics , Stem Cell Factor , Metabolism , Young Adult
11.
Article in Chinese | WPRIM | ID: wpr-267881

ABSTRACT

The study was purposed to explore the effect and mechanisms of decitabine and/or Trichostatin A (TSA) on SKM-1 cells in vitro. The effect of decitabine and/or TSA on proliferation of SKM-1cells was analyzed with trypan blue exclusion; the differentiation of SKM-1 cells was detected by nitro-blue tetrazolium (NBT) reduction and flow cytometry; the apoptosis of cells was measured by Annexin V-FITC; the mRNA expression of Fas, survivin and P15(INK4B) in cells treated with decitabine and/or TSA was evaluated by RT-PCR. The results showed that decitabine and/or TSA were capable of inhibiting SKM-1 cell growth and promoting cell differentiation; they stimulated the expression of CD14 and CD11b and inhibited HLA-DR expression; meanwhile and decitabine or/and TSA could induce cell apoptosis, up-regulate mRNA expression of Fas and P15(INK4B), and down-regulate survivin mRNA expression. It is concluded that decitabine can induce apoptosis/differentiation of SKM-1 cells, whose mechanisms may related to the expression of Fas, survivin and P15(INK4B). Decitabine has the synergistic effect with TSA.


Subject(s)
Antimetabolites, Antineoplastic , Pharmacology , Apoptosis , Azacitidine , Pharmacology , Cell Differentiation , Cell Line, Tumor , Cell Proliferation , Drug Synergism , Humans , Hydroxamic Acids , Pharmacology , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Metabolism , Myelodysplastic Syndromes , Pathology , fas Receptor , Genetics , Metabolism
12.
Article in Chinese | WPRIM | ID: wpr-269106

ABSTRACT

In this article, the mechanism of hand-foot syndrome (HFS) and its prevention and treatment measures with Chinese and modern medicine in recent years were reviewed. Although standard preventive and therapeutic program on HFS is still lack so far, both TCM and modern medicine have achieved certain effects in this aspect, which is of important significance for improving the quality of life and the effect of chemotherapy in patients with HFS.


Subject(s)
Antimetabolites, Antineoplastic , Capecitabine , Deoxycytidine , Drug Therapy , Methods , Drugs, Chinese Herbal , Therapeutic Uses , Fluorouracil , Foot , Hand , Humans , Paresthesia , Pathology , Phytotherapy , Methods , Syndrome
13.
Article in Chinese | WPRIM | ID: wpr-280685

ABSTRACT

The aim of this study was to investigate the inhibition effect of arsenic sulfide (As2S2) on the growth of in vitro cultured BMMNC from MDS patients and to explore its possible cellular and molecular mechanisms. The apoptosis of MDS cells induced by As2S2 solution of different concentrations were studied with MTT, flow cytometry, and RT-PCR. The results showed that (1) low concentration of As2S2 (0-0.6 mg/L) had no marked inhibition effect on proliferation of MDS cells; (2) after treatment with 1.5-50 mg/L of As2S2, both low risk MDS cells and high risk MDS cells presented typical features of apoptosis with a dose-dependent manner, the expression of bcl-2 mRNA and the ratio of bcl-2/bax obviously decreased after As2S2 treatment (P < 0.05); (3) BMMNC from MDS patients had higher apoptosis ratio than that of BMMNC from control. It is concluded that BMMNC excessive apoptosis exists in MDS patients; low concentration of As2S2 (0-0.6 mg/L) shows no inhibition effect on proliferation of MDS cells; high concentration of As2S2 (1.5-50 mg/L) induces apoptosis of MDS cells.


Subject(s)
Adult , Aged , Aged, 80 and over , Antineoplastic Agents , Pharmacology , Apoptosis , Arsenicals , Pharmacology , Bone Marrow Cells , Pathology , Cyclin D1 , Genetics , Female , Humans , Leukocytes, Mononuclear , Pathology , Male , Middle Aged , Myelodysplastic Syndromes , Pathology , RNA, Messenger , Genetics , Sulfides , Pharmacology , bcl-2-Associated X Protein , Genetics
14.
Article in Chinese | WPRIM | ID: wpr-352008

ABSTRACT

To explore the difference of negative regulatory factors among T lymphocyte subsets in bone marrow (BM) of myelodysplastic syndromes (MDS) and their relations to apoptotic gene Fas, different lymphocyte subsets in BM were categorized by monoclonal antibodies with 3 color fluorescence using flow cytometry, and the intracellular cytokines such as tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) were determined following marrow cells culture. Then Fas mRNA of bone marrow mononuclear cells (BMMNC) were examined by RT-PCR. The results showed that TNF-alpha, IFN-gamma levels in BM of MDS both increased, the former produced by cells CD4+CD45RO+, CD8+CD45RO+, the latter by cells CD4+CD45RO+, CD8+CD45RO+, CD8+CD45RA+, in which the cells CD8+CD45RO+ were dominant. Fas mRNA expression had relationship with IFN-gamma produced by T cells but not with TNF-alpha. It is concluded that in hematopoietic microenvironment of MDS, not only the T lymphocyte subsets are in disorder, but also negative regulatory factors secreted by T lymphocyte increase. T lymphocytes play an important role in producing IFN-gamma in patients with MDS.


Subject(s)
Adult , Aged , Bone Marrow Cells , Metabolism , Female , Humans , Interferon-gamma , Male , Middle Aged , Myelodysplastic Syndromes , Metabolism , Pathology , RNA, Messenger , T-Lymphocyte Subsets , Metabolism , Tumor Necrosis Factor-alpha , fas Receptor , Genetics
15.
Article in Chinese | WPRIM | ID: wpr-326735

ABSTRACT

<p><b>OBJECTIVE</b>To observe the clinical efficacy of TCM with supplementing Qi, nourishing Yin and clearing heat principle (SQNYCH) combined with chemotherapy in treating myelocytic leukemia.</p><p><b>METHODS</b>One hundred and fourteen patients were randomly divided into the treated group (n = 68) and the control group (n = 46). To the treated group, SQNYCH was applied as the basic treatment, with combined chemotherapeutic protocol, using DA, HA and IA, to induce remission, and to the M3 patients, all-trans retinoic acid and arsenic trioxide were given. As for patients in the control group, only western medicine was administered.</p><p><b>RESULTS</b>In the treated group 49 patients (72.1%) were completely remitted, 9 (13.2%) partially remitted and the total remission rate being 85.3%, which was significantly different from that in the control group. After treatment, the blood and bone marrow picture were obviously improved in both groups, but the increase of hemoglobin and platelet were better in the treated group than in the control group (P < 0.05 or P < 0.01). Immune functions were enhanced in both groups, but the elevation of CD4, CD4/CD8 ratio and NK cells were higher in the treated group than in the control group (P < 0.05 and P < 0.01).</p><p><b>CONCLUSION</b>Application of SQNYCH principle in treating acute myelocytic leukemia could elevate the clinical efficacy, which is of great value in clinical practice.</p>


Subject(s)
Adolescent , Adult , Aged , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Child , Diagnosis, Differential , Drug Therapy, Combination , Drugs, Chinese Herbal , Therapeutic Uses , Female , Humans , Leukemia, Myeloid, Acute , Drug Therapy , Male , Medicine, Chinese Traditional , Middle Aged , Phytotherapy , Yin Deficiency , Drug Therapy
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