Integrative Medicine ; education ; Internal Medicine ; education

To evaluate the expression of cyclin dependent kinase inhibitor P27(Kip1) in leukemia and to investigate its clinical significance, the P27(Kip1) protein in bone marrow or peripheral blood samples from 82 cases of leukemia was measured by Western blot and enhanced chemoluminescence (ECL). The results showed that the expression of P27(Kip1) protein in ALL was higher than that in ANLL (P = 0.033) and also that in CML (P = 0.008). P27(Kip1) expression in CLL was higher than that in CML too (P = 0.017). In acute leukemia, the effective rate (CR and PR) of initial chemical therapy in the group of P27(Kip1) > 0.655 was higher than that in the group of P27(Kip1) < or = 0.655, P = 0.041. For ANLL and ALL patients, the survival time in the group of P27(Kip1) > 0.655 was longer than that in the group of P27(Kip1) < or = 0.655, P = 0.0065. There were similar statistical significance for ANLL and ALL patients, P = 0.0271 and P = 0.0266 respectively. There was a negative correlation between chromosomal abnormalities and P27(Kip1) expression in ALL patients (r = -0.775, P = 0.04). The expression of P27(Kip1) protein appeared nothing to do with sex, age, white blood cell number, blast cell number in peripheral blood, serum LDH or uric acid. In conclusion, the expression level of P27(Kip1) protein is in relation to the effect of initial chemical therapy and survival time, so that the lower P27(Kip1) expression may associated with poor prognosis in acute leukemia.
Adolescent ; Adult ; Aged ; Blotting, Western ; Cell Cycle Proteins ; analysis ; Child ; Child, Preschool ; Chromosome Aberrations ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; Leukemia, Lymphocytic, Chronic, B-Cell ; drug therapy ; genetics ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; metabolism ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; metabolism ; Survival Rate ; Tumor Suppressor Proteins ; analysis

In order to establish a new more rapid, safe and sensitive colorimetric assay for the proliferation of leukemic cells, MTS/pms has been developed. This automated colorimetric assay is based on the characteristic of viable and metabolically active leukemic cells to cleave MTS/pms into a water-soluble product whose optical density is determined at 492 nm by an automated microtiter-plate reader photometer. The results indicated that only active leukemic cells cleaved MTS/pms into product measured, and dead cells did not reduce MTS/pms. A linear relations hip were found between the viable cell number and optical density of MTS/pms cleaved by HL-60 and K562 cell (r = 0.963). Compared with MTT and INT assays, the reduced product of MTS/pms is water-soluble. It is concluded that MTS/pms colorimetric assay is more rapid, accurate and sensitive for the bioassay of proliferation of leukemic cells.
Cell Division ; Colorimetry ; methods ; Formazans ; metabolism ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; pathology ; Methylphenazonium Methosulfate ; metabolism ; Tetrazolium Salts ; metabolism ; Thiazoles ; metabolism

The study was aimed to explore the role of gene JWA, a novel retinoic acid responsible and cytoskeleton associate gene, in regulating committed differentiation of HL-60 cell and the molecular mechanism in the course of differentiation and apoptosis of leukemic cells. By using FCM, the changes of CD13, CD14, CD15, CD11b and cell cycles were detected in HL-60 cells treated with ATRA (10(-6) mol/L), Ara-C (10 ng/ml) and TPA (10(-8) mol/L) respectively. The samples were determined by semi-quantitative reverse transcript-polymerase chain reaction (RT-PCR) and Western blot for the expression of JWA, Bcl-2, HSP27 and HSP70 at day 0, 2, 4, 6, 8. The results showed that HL-60 cells committedly differentiated into granulocyte-, monocyte-, macrophage-like cells. As a result, JWA was up-regulated in a time-dependent manner, while Bcl-2 was down- regulated at the same time. In ATRA and TPA group, the change of HSP70 had positive correlation with JWA, and negative correlation with Bcl-2. The expression of HSP27 was not detected. Contrast to the cells from APL patient, the expression of JWA need not be activated by ATRA in advance. In this study, we also exposed HL-60 cells in higher dose of Ara-C (20 ng/ml), and JWA expression underwent opposite trend comparing with in lower dose of Ara-C (10 ng/ml). It is concluded that JWA may play double important roles in regulating ATRA and TPA-induced differentiation and apoptosis in leukemic cells. The JWA expression had a negative correlation between induction and cytotoxic response. The difference of JWA expressions between HL-60 cell and ANLL patient cells would be involved in different leukemia pathogenesis.
Antineoplastic Agents ; pharmacology ; Blotting, Western ; Cell Differentiation ; drug effects ; Cytarabine ; pharmacology ; HL-60 Cells ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tetradecanoylphorbol Acetate ; pharmacology ; Time Factors ; Tretinoin ; pharmacology

OBJECTIVETo explore the acting mechanism of viscera purging method (VP) in purging Fu-organs and benefiting Fei from integral, cellular and molecular levels.

METHODSForty SD rats were equally divided into four groups randomly: the normal group, the model group, the unhitch group and the VP group. Except those in the normal group were untreated, rats were established to intestinal obstruction model by incomplete ligation of the rectum in vitro. The ligation was relieved 48 h after operation in the unhitch group and the VP group, and the animals were fed continuously on routine. Meanwhile, Dachengqi Decoction (DD) 2 ml was given twice a day to the VP group for 2 days. Finally, the serum interleukin 8 (IL-8) and mRNA expression of tumor necrosis factor-alpha (TNF-alpha) in the lung tissue were detected by radioimmunoassay and RT-PCR respectively. Besides, the number of pulmonary alveolar macrophages (PAM) in the bronchial alveolus lavage fluid (BALF) was counted and their death rate calculated.

RESULTSCompared with the normal control, the serum IL-8 content in lung tissue in the model rats were remarkably higher (P < 0.01); however, the VP group showed the lowest level of IL-8 content and the highest was shown in the model group. Number of PAM in BALF was higher and its death rate was lower in the VP group than that in the unhitch groups (both P< 0.05). The expression of TNF-alpha mRNA was sinificantly higher as compared with that in the normal group, and lowered after administration of DD.

CONCLUSIONViscera purging method could protect the injured lung tissue to some extent.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Interleukin-8 ; blood ; Intestinal Obstruction ; complications ; drug therapy ; Lung ; metabolism ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Plant Extracts ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; etiology ; prevention & control ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

To observe the therapeutic effect and safety of rituximab (anti-CD20 monoclonal antibody) in the treatment of refractory autoimmune hemolytic anemia (AIHA). One AIHA patient refractory to corticosteroid and splenectomy was treated with rituximab, 375 mg/m(2) weekly for four times. Her hemolytic symptoms, adverse effects, hemoglobin (Hb) concentration and other laboratory data were monitored. The results showed that concentration of lactic dehydrogenase (LDH), total bilirubin (TBIL) and indirect bilirubin (IBIL) began to decrease at 11 days after the first dose of rituximab, and decreased to normal range after 45 days. Concentration of hemoglobin increased up to 95 - 100 g/L. The patient remained disease-free 4 months after treatment. No adverse effect was found during the process of treatment. It is concluded that anti-CD20 monoclonal antibody (rituximab) is both effective and safe for the treatment of refractory autoimmune hemolytic anemia.
Anemia, Hemolytic, Autoimmune ; drug therapy ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Murine-Derived ; Antigens, CD20 ; immunology ; Female ; Humans ; Middle Aged ; Rituximab

To investigate the expressions of proliferative antigen Ki-67 and apoptosis-antagonizing protein Bcl-2 in patients with chronic lymphocytic leukemia (CLL) and their clinical significance, immunohistochemistry method was used for detection of Ki-67 and Bcl-2 in bone marrow or peripheral blood of 29 patients with CLL. The results showed that the level of Ki-67 expression in advanced stage of CLL was higher than that in early stage of CLL and there was significant difference between these stages of CLL, but there was no significant difference between expression levels of Bcl-2 in two stages. The survival time in the group with Ki-67 expression < or = 8% was longer than that in the group of Ki-67 > 8%, and there was no significant difference of survival time between high and low groups in terms of Bcl-2 expression. It is concluded that detection of Ki-67 antigen and Bcl-2 protein for CLL patients can reflect the status of proliferation activity and apoptosis suppression of leukemia cells in patients; the level of Ki-67 expression closely correlate with the Binet stage and prognosis of CLL.
Aged ; Aged, 80 and over ; Apoptosis ; physiology ; Bone Marrow Cells ; metabolism ; Cell Proliferation ; Female ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; analysis ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; analysis

Adult ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Hemoglobinuria, Paroxysmal ; therapy ; Humans ; Male

OBJECTIVETo observe the effect of salvianolic acid B (SAB), an extract from Radix Salviae miltiorrhizae, on expression of leucocyte differentiation antigen 14 (CD14) in the liver tissue of experimental rats with carbon tetrachloride (CCl4)-induced liver fibrosis.

METHODSThirty SD rats were randomly divided into three groups, the model group, the treated group, and the control group. The pathological fibrosis changes in liver of rats were observed. Meantime, their liver function was detected by automatic biochemical analyzer. Serum content of endotoxin was assayed by matrix staining, and plasma content of tumor necrosis factor-alpha (TNF-alpha) was detected by radioimmunoassay. mRNA and protein expressions of CD14 in the liver tissue were measured using reverse transcriptional-polymerase chain reaction and immunohistochemistry respectively.

RESULTSAll the laboratory parameters, including liver function, degree of liver fibrosis, serum endotoxin levels, plasma TNF-alpha contents, and CD14 mRNA and protein expressions in the model group were higher than those in the control group (all P<0.01). All the aforesaid indices were lowered more in the treated group than in the model group (all P<0.01).

CONCLUSIONSSAB could antagonize the CCl4, induced liver fibrosis in rats. Its mechanism of action was possibly correlated with its effects on down-regulating hepatic CD14 expression and blocking the endotoxin signal transduction pathway.

Animals ; Benzofurans ; pharmacology ; Lipopolysaccharide Receptors ; metabolism ; Liver ; drug effects ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

In order to study the relationship between the expression of glutathione S-transferase (GST) in leukemic cells and the chemoresistance in patients with acute leukemia, the expressions of GST activity and GST mRNA were measured according to spectrophotometric assay based on the use of 1-choloro-2, 4-dinitro benzene and in situ hybridization. The results were studied in correlation with some clinical and pathological data. Results showed that: 1. There is no significant differences between activities of the enzyme with the different leukemia types according to the FAB classification. 2. GST activity and GST mRNA expression in the patients, both untreated and relapse, were (4.5 +/- 1.0) U, 33.3% and (7.9 +/- 15) U, 66.3% respectively. 3. In 56 patients, GST activity was 1.7 +/- 0.7, 5.9 +/- 2.0 and 9.3 +/- 1.7 U and GST mRNA expression was 13.3%, 29.7% and 76.6%, respectively, in CR, PR and NR groups. The lowest values of GST activity and GST mRNA expression were observed in those patients who achieved complete remission. The highest values of GST activity and GST mRNA expression were observed in those patients with no response to treatment. It was concluded that the expression of GST in patients with acute leukemia is closely related to the chemosensitivities clinically. Determinations of GST activity and GST mRNA are useful for predicting the chemosensitivities and the prognosis of the disease.
Adolescent ; Adult ; Aged ; Drug Resistance, Neoplasm ; Female ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Glutathione Transferase ; genetics ; metabolism ; Humans ; Isoenzymes ; genetics ; metabolism ; K562 Cells ; Leukemia ; drug therapy ; enzymology ; genetics ; Leukemia, Lymphoid ; drug therapy ; enzymology ; genetics ; Leukemia, Monocytic, Acute ; drug therapy ; enzymology ; genetics ; Leukemia, Myeloid, Acute ; drug therapy ; enzymology ; genetics ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism