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OBJECTIVE@#To analyze the expression and clinical characteristics of CD68 in bone marrow and peripheral blood of patients with acute myeloid leukemia (AML).@*METHODS@#The expression of CD68 in bone marrow blast cells was detected by four-color flow cytometry in 50 newly diagnosed AML patients and 23 controls. The expression of CD68 in peripheral blood of 85 newly diagnosed AML patients, 29 remission AML patients and 24 controls was detected by ELISA. The correlation between the expression rate of non-M3 AML bone marrow CD68, peripheral blood CD68 concentration and white blood cell count and other clinical data was compared respectively.@*RESULTS@#The median CD68 expression rate in myeloid leukemia cells of non-M3 AML patients was 19.7%, significantly higher than control (0.2%) (P<0.001). The median concentration of non-M3 CD68 in peripheral blood was 67.97 pg/ml, significantly higher than in control (29.94 pg/ml)（P<0.01）. There was no statistically significant difference in the plasma CD68 concentration of the peripheral blood between the newly diagnosed (45.72 pg/ml) and the remission stage (55.12 pg/ml) of non-M3 AML patients by paired analysis (P>0.05). The results showed that the higher the expression rate of CD68 in bone marrow, the higher the count of white blood cells in peripheral blood, and the lower the count of hemoglobin and platelet in peripheral blood. The higher the plasma concentration of CD68 in peripheral blood, the higher the white blood cell count and the lower the complete remission rate.@*CONCLUSION@#The expression of CD68 both in bone marrow and peripheral blood of patients with non-M3 AML is higher than that of control group. Patients with high expression of CD68 show a low rate of complete remission, suggesting that the expression level of CD68 is correlated with treatment response.
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Humans , Bone Marrow , Flow Cytometry , Leukemia, Myeloid, Acute , Leukocytes , Prognosis , Remission InductionABSTRACT
OBJECTIVE@#To investigate the mechanism of Radix Kansui (RK) stir-fried with vinegar (VRK) decreased hepatotoxicity in mice.@*METHODS@#According to a random number table, 40 mice were randomly divided into negative control group (0.5% carboxymethylcellulose sodium, 20 mL/kg), positive control group (0.1% mixture of carbon tetrachloride in soybean oil, 20 mL/kg), RK group (the ethyl acetate extracts of RK, 250 g crude drug/kg) and VRK group (the ethyl acetate extracts of VRK, 250 g crude drug/kg) with 10 mice per group. All mice were administered orally by gavage daily for 7 continuous days. The morphology of liver tissues was examined to assess the liver injury by a transmission electron microscope. Hepatocyte apoptosis in vivo was determined by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nickend labeling (TUNEL) assay. Immunohistochemical technique was adopted to detect the expression of particular antiapoptotic and proapoptotic proteins in the mitochondrial pathways, including B-cell lymphoma (Bcl-2) and caspase-3, as well as the expression of inflammatory mediators, including nuclear factor kappa B (NF- κ B) and intercellular adhesion molecule-1 (ICAM-1).@*RESULTS@#Liver injury and hepatocyte apoptosis were observed in RK mice, and the liver injury were significantly reduced in VRK-treated mice. In immunohistochemistry study, compared with the negative control group, RK inhibited dramatically the Bcl-2 protein expression and significantly increased the expression of caspase-3, NF- κ B and ICAM-1 (all P<0.01). Compared with the RK group, VRK group induced significant increase on Bcl-2 protein expression, and decreased the caspase-3, NF- κ B and ICAM-1 protein expression (P<0.05 or P<0.01).@*CONCLUSION@#The mechanism of reduced hepatotoxicity of VRK may be associated with the reduced inflammation, regulation of antiapoptotic and proapoptotic mediators in the mitochondrial pathway.
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Objective: To compare the chemical components in raw products and characteristic processed products with porcine cardiac blood of Salviae Miltiorrhizae Radix et Rhizoma from Menghe medical school.Method: The ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF/MS) was performed on an ACQUITY UPLC® C18 column (2.1 mm×100 mm,1.7 μm),mobile phase was 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) for gradient elution (0-1 min,8% B,1-1.5 min,8%-20% B,1.5-4 min,20% B,4-5 min,20%-60% B,5-9 min,60%-70% B,9-10 min,70%-95% B,10-13 min,95% B).The column temperature was 40℃ and the flow rate was 0.3 mL·min-1.Q-TOF/MS with electrospray ionization (ESI) and scanning range of m/z 50-1 200 were applied for analysis under positive and negative ion mode,respectively.All ionic peaks were assigned by comparison of reference substances,mass spectra data,database matching and literature reference,changes of chemical components in Salviae Miltiorrhizae Radix et Rhizoma were investigated by comparing number and area of ionic peaks before and after processing.Result: A total of 59 components were identified from raw products and characteristic processed products,and peak areas of 25 components showed obvious change.However,there were no new compound was found in characteristic processed products.After being processed with porcine cardiac blood,the contents of water-soluble ingredients (protocatechuic aldehyde,salvianolic acid C,F,G),fat-soluble ingredients (tanshinaldehyde,tanshindiol A,tanshinone Ⅰ) and amino acid (L-phenylalanine) in Salviae Miltiorrhizae Radix et Rhizoma were significantly increased.Conclusion: Changes of contents of chemical components in raw products and processed products of Salviae Miltiorrhizae Radix et Rhizoma are remarkable,part of water-soluble ingredients,fat-soluble ingredients and amino acids have quantitative change,which may be related to promoting treatment of Salviae Miltiorrhizae Radix et Rhizoma on cerebral ischemia after being processed with porcine cardiac blood.
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The porcine cardiac blood processed Salvia miltiorrhiza (PCB Danshen) is the characteristic processing of Menghe medical school and has been inherited for hundreds of years, commonly used in the treatment of brain ischemia-induced agitation, palpitation and phlegm confusing heart. Ancient and modern physicians believe that porcine cardiac blood is a guiding for heart nourishing drugs, which could enhance the effects of Salvia miltiorrhiza by nourishing and soothing the nerves. However, the material basis and processing mechanism of PCB Danshen are still unclear. This paper investigated the historical evolution and modern research of PCB Danshen, including the clinical application, the intention of clinic processing, the processing technology and recent research of PCB Danshen. In addition, the major problems and significance in research and development of PCB Danshen were further thought and prospected, hoping to provide basic data for material basis and processing mechanism of PCB Danshen, and provide effective support for inheriting and carrying forward the characteristic processing technology of Menghe medical school.
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<p><b>OBJECTIVE</b>To explore the expression of insulin-like growth factor binding protein 3 (IGFBP3) in acute myeloid leukemia and its clinical significance.</p><p><b>METHODS</b>Using GAPDH as internal reference, IGFBP3 gene expression was detected in the bone marrow mononuclear cells of 43 de novo AML patients, 36 patients with complete remission (CR) and 28 cases of controls by using SYBR-Green I real-time quantitative PCR, the differences of gene expression between the three groups were compared.</p><p><b>RESULTS</b>IGFBP3 gene expression level in the de novo group were lower than that in CR group and control group (P < 0.05), but there was no statistically significant difference of IGFBP3 expression level in CR group and control group (P > 0.05).</p><p><b>CONCLUSIONS</b>The IGFBP3 as a tumor suppressor gene may play a role in the pathogenesis of acute myeloid leukemia, its expression level is recovered after complete remission, therefore, IGFBP3 can be used as an indicator of evaluating clinical curative effect.</p>
Subject(s)
Humans , Bone Marrow Cells , Gene Expression , Insulin-Like Growth Factor Binding Protein 3 , Leukemia, Myeloid, Acute , Real-Time Polymerase Chain ReactionABSTRACT
This study aims to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 using the orthogonal design method, and to investigate its effects of the component formula on cell proliferation, apoptosis and cytoskeleton in lung cancer A549 cells. The orthogonal design method was introduced to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 cells. CCK-8 assay and Real-time cell analysis were adapted to analyze the effect of component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on A549 cells viability at different time and dose. Cell apoptosis was measured by Annexin V- FITC/PI double staining and flow cytometry. Cell skeleton protein F-actin was detected by high content screening (HCS). The optimizing component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma for total salvianolic acid, total saponins of panax ginseng and ginseng polysaccharide doses were 5, 10, 5 mg L(-1). CCK-8 assay and real-time cell analysis demonstrated that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma treatment could significantly decrease the A549 cell viability in both dose- and time-dependent manner compared with control group (P < 0.01). Moreover, the increase of cell apoptosis was detected by Annexin V-FITC/PI double staining and flow cytometry when cells treated with the component formula, which indicating that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma could induce A549 cell apoptosis in a time-dependent manner compared with control group (P < 0.01). Furthermore, compared with control group, a significant decrease in A549 cell skeleton area was found in the component formula-exposed cells in the dose-dependent manner (P < 0.01). In summary, the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma inhibits A549 cell proliferation by inducing cell apoptosis and decreasing cell microfilament formation. All of these results will be helpful to reveal antitumor mechanism of the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma, which provides a basis for the exploration of antitumor mechanism of the component formula on lung cancer.
Subject(s)
Humans , Apoptosis , Cell Line, Tumor , Cell Proliferation , Drugs, Chinese Herbal , Chemistry , Pharmacology , Lung Neoplasms , Drug Therapy , Panax , Chemistry , Plant Extracts , Chemistry , Pharmacology , Plant Roots , Chemistry , Rhizome , Chemistry , Salvia miltiorrhiza , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To compare the difference of Euphorbia Pekinensis Radix before and after being processed with vinegar in the toxicity on rat small intestinal crypt epithelial cells IEC-6, and make a preliminary study on the mechanism of detoxication of Euphorbia Pekinensis Radix processed with vinegar.</p><p><b>METHOD</b>With rat small intestinal crypt epithelial cells IEC-6 as the study object, the MTT method was adopted to detect the effect of Euphorbia Pekinensis Radix before and after being processed with vinegar on IEC-6 cell activity. The morphology of cells were observed by the inverted microscope. The down-regulated mitochondrial apoptosis pathway of enterocytes caused by the vinegar processing was analyzed by using the high content screening.</p><p><b>RESULT</b>Compared with the negative control group, the proliferation inhibition experiment showed that Euphorbia Pekinensis Radix showed a relatively high intestinal cell toxicity (P < 0.01). The results of HCS analysis showed that Euphorbia Pekinensis Radix could significantly reduce the cell nucleus Hoechst fluorescence intensity and mitochondria membrane (P < 0.05, P < 0.01), and increase Annexin V-FITC and PI fluorescence intensity and membrane permeability (P < 0.01, P < 0.01, P < 0.01). After being processed with vinegar, compared with Euphorbia Pekinensis Radix groups with different doses, Euphorbia Pekinensis Radix processed with vinegar could significantly decrease the cell proliferation inhibition effect on enterocytes, increase the cell nuclear Hoechst fluorescence intensity and mitochondria membrane (P < 0.05, P < 0.05), and decrease Annexin V-FITC and PI fluorescence intensity and membrane permeability (P < 0.01, P < 0.01, P < 0.05), and showed a certain dose-effect relationship.</p><p><b>CONCLUSION</b>The vinegar processing can further reduce the toxicity of Euphorbia Pekinensis Radix on enterocytes. Its possible mechanism can decrease the effect of Euphorbia Pekinensis Radix on the permeability of IEC-6 cell membrane, so as to provide a basis for further explanation of the detoxication mechanism of Euphorbia Pekinensis Radix processed with vinegar.</p>
Subject(s)
Animals , Rats , Acetic Acid , Chemistry , Apoptosis , Cell Line , Cell Proliferation , Cell Survival , Chemistry, Pharmaceutical , Methods , Drugs, Chinese Herbal , Chemistry , Toxicity , Epithelial Cells , Cell Biology , Euphorbia , Chemistry , Intestine, Small , Cell BiologyABSTRACT
Endoplasmic reticulum stress (ERS) is a new pathway inducing cell apoptosis that has been discovered in recent years. This study focused on the protective effect of Liangxue Huayu recipe (LHR) on tumor necrosis factor-alpha (TNF-alpha) and D-GalN-induced hepatocyte apoptosis. It found that TNF-alpha and D-GalN could obviously inhibit hepatocyte proliferation, induce cell apoptosis, and significantly increase free calcium ions in cytoplasms, as well as protein expressions of ERS apoptosis-related signal molecules phosphorylated PERK, phosphorylated elF2alpha, cleaved Caspase-12, GRP78 and CHOP. After the administration of LHR of different concentrations, compared with the TNF-alpha/GalN injury group, LHR could significantly alleviated L02 hepatocyte proliferation, decreased cell apoptosis, inhibited growth of intracytoplasmic free calcium content, and gradually reduced the protein expressions of phosphorylated PERK, phosphorylated elF2alpha, cleaved Caspase-12, GRP78 and CHOP. These findings indicated that LHR has the inhibitory effect on TNF-alpha and D-GalN-induced hepatocyte apoptosis. Its mechanism may be related to down-regulation of ERS apoptosis-related signal molecules phosphorylated PERK, phosphorylated elF2alpha, cleaved Caspase-12, GRP78 and CHOP that maintain calcium homeostasis in endoplasmic reticulum.
Subject(s)
Humans , Apoptosis , Cell Line , Cell Proliferation , Drugs, Chinese Herbal , Pharmacology , Endoplasmic Reticulum Stress , Heat-Shock Proteins , Genetics , Metabolism , Hepatocytes , Cell Biology , Nitric Oxide Synthase Type II , Genetics , Metabolism , Tumor Necrosis Factor-alpha , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To discuss the effect of Kansui Radix prepared by different processes on cell cycle and apoptosis of normal human liver cell lines LO2.</p><p><b>METHOD</b>With normal human liver cell lines LO2 as the study object, the MTT method was adopted to study the effect of Kansui Radix prepared by different processes, including Kansui Radix, stir-baking Kansui Radix, Kansui Radix moistening with vinegar and Kansui prepared by different processes, on LO2 cell activity. The cellular morphological changes were observed by inverted microscope. The effect of Kansui Radix stir-baked with vinegar on LO2 cell cycle and apoptosis was observed by flow cytometry.</p><p><b>RESULT</b>Compared with the negative control group, Kansui could obviously inhibit the activity of human normal liver cell lines LO2 (P <0.01) , and significantly increase the percentage of LO2 cells in S phase (P <0.05) , notably decrease the percentage of LO2 cells in G2/M phase (P <0.01) , significantly increase the early apoptosis rate, late apoptosis rate and necrosis rate and total apoptosis rate of human normal liver cell lines LO2 (P <0.01). Compared with the Kansui group, all of the other processed Kansui samples could significantly decrease the cell proliferation inhibition (P <0.01) , and the trend of morphological degradation. Besides, they could significantly increase the percentage of LO2 cells in G2/M phase (P <0.05, P <0.05, P <0. 01) , significantly decrease the early apoptosis rate, late apoptosis rate and necrosis rate, and total apoptosis rate of human normal liver cell lines LO2 (P < 0.01). The order of the increase in the percentage of cells in G2/M phase and the decrease in apoptosis rate was Kansui Radix stirbaked with vinegar > Kansui Radix moistening with vinegar > stir-baking Kansui Radix.</p><p><b>CONCLUSION</b>The toxicity of processed Kansui could be reduced by affecting LO2 cell cycle and apoptosis. The processes of stir-baking and moistening with vinegar can play a synergistic effect in the detoxication of human normal liver cell lines LO2, which provides a basis for unveiling the rationality of stirbaking with vinegar of Kansui in the detoxication, as well as the optimizing the process.</p>
Subject(s)
Humans , Apoptosis , Cell Cycle , Cell Line , Cell Proliferation , Chemistry, Pharmaceutical , Methods , Drugs, Chinese Herbal , Chemistry , Euphorbia , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To compare the toxicity of Euphorbia pekinensis before and after being processed by vinegar on normal liver cells LO2, and discuss its possible mechanism.</p><p><b>METHOD</b>LO2 cells were cultured in vitro, and processed with different concentrations of crude and vinegar-processed E. pekinensis. MTT assay was used to measure the inhibitory effect of LO2 cell; Hoechst 33258 staining was used to observe the morphological changes in apoptosis cell; Annexin V-FITC flow cytometry was used to analyze the apoptotic rate of LO2 cell; PI staining flow cytometry was used to analyze its impact on cell cycle. The level or content of ALT, AST, LDH, SOD, MDA and GSH were observed as well.</p><p><b>RESULT</b>Compared with the negative control group, crude E. pekinensis at all concentrations could obviously inhibit LO2 cell proliferation, induce LO2 cell apoptosis and cause cell arrest in S phase, with significant differences (P <0.05). E. pekinensis could significantly increase the levels of ALT, AST and LDH (P <0.05) in the supernatant of cell culture fluid, significantly decrease the level of SOD and the content of GSH (P <0.05) , and significantly increase the content of MDA (P <0.05). Compared with the crude E. pekinensis group, E. pekinensis after being vinegar-processed can significantly reduce cell apoptotic rate, cell cycle arrest, activities of ALT, AST, LDH in the supernatant of cell culture fluid (P <0.05) , and remarkably increase the level of SOD and the content of GSH, but reduce the content of MDA in the supernatant of cell culture fluid.</p><p><b>CONCLUSION</b>Vinegar-processed E. pekinensis can release the cytotoxicity of LO2 cell. Its mechanism may be related to the decrease in the oxidative damage of LO2 cells, thereby reducing the cell cycle arrest and apoptosis.</p>
Subject(s)
Humans , Acetic Acid , Chemistry , Apoptosis , Cell Cycle Checkpoints , Cell Line , Cell Membrane , Metabolism , Cell Proliferation , Chemistry, Pharmaceutical , Drugs, Chinese Herbal , Chemistry , Toxicity , Euphorbia , Chemistry , Liver , Cell Biology , Oxidative StressABSTRACT
<p><b>OBJECTIVE</b>To study the mechanism of the reducing mechanism of hepatotoxicity induced by ethyl acetate fractions of Kansui Radix stir-baked with vinegar in mice.</p><p><b>METHOD</b>Mice with normal ICR were orally administered with ethyl acetate fractions of Kansui Radix and Kansui Radix stir-baked with vinegar. Their blood and liver homogenate were collected to detect the level of AST, ALT, LDH, SOD, activities of Na(+) -K(+) -ATPase and Ca(2+) -Mg(2+) -ATPase, GSH and MDA. Liver tissues were collected for HE staining and morphological observation under light microscope.</p><p><b>RESULT</b>According to the results of pathological sections, compared with the control group, all of Kansui groups showed a significant increase in the hepatic tissues injury (P < 0.01). Compared with Kansui groups, all of vinegar-baked groups showed a significant decrease in the hepatic tissues injury (P < 0.01). Compared with the control group, all of Kansui groups showed a significant increase in ALT, AST and LDH (P < 0.05, P < 0.001) in serum and hepatic tissues, and significantly decrease in the activity of SOD (P < 0.001) and the content of GSH. They also showed a significant increase in MDA (P < 0.001) and a significant decrease in the level of Na(+) -K(+) -ATPase and Ca(2+) -Mg(2+) -ATPase (P < 0.01) in hepatic tissues, with a certain dose-effect relationship. Compared with all of Kansui groups, all of vinegar-baked groups showed a significant decrease in ALT, AST and LDH (P < 0.05, P < 0.001), and a notable increase in SOD (P < 0.001) and GSH in serum and hepatic tissues. They also showed a remarkable decrease in MDA (P < 0.001), and a significant increase in the level of Na(+) -K(+) -ATPase and Ca(2+) - Mg(2+) -ATPase (P < 0.01) in hepatic tissues, with a certain dose-effect relationship.</p><p><b>CONCLUSION</b>Being stir-baked with vinegar can significantly reduce the hepatotoxicity of Kansui Radix. Its mechanism may be related to the reduction of the effect of Kansui Radix on the permeability of hepatic tissues cell membranes and the oxidative injury.</p>