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Objective:To make statistics on the annual frequency of patients with eczema by traditional Chinese medicine (TCM) physique, syndrome differentiation, and western medicine staging based on questionnaire survey, in order to infer the distribution and characteristics of the annual frequency by different TCM physique, syndrome differentiation and western medicine stage, and provide new ideas and new methods for the prevention and treatment of Eczema. Method:According to the Dermatovenerology of Traditional Chinese Medicine edited by QU Xing, and Clinical Diagnosis and Treatment of Chinese Medicine for Dermatovenerology edited by CHEN Da-can, and Traditional Chinese Medicine for Dermology edited by YU Wen-qiu, and Physique Classification and Patient Self-Testing Table formulated by China Association of Chinese Medicine and Professor WANG Qi's nine categories of physique, the TCM Physique Classification and Patient Self-Testing Table for eczema patients and the Syndrome Differentiation and Classification Table for Eczema Patients were formulated. General conditions of 482 cases of eczema patients treated at Tianjin Academy of Traditional TCM Affiliated Hospital and their types of TCM physique, TCM syndrome differentiation, western medicine staging and annual frequency were surveyed. Result:There were significant differences in the annual frequency of patients with different physical constitutions. By single physique, Tanshi, Shire and Qiyu had more frequent occurrences every year, and Pinghe had the lowest annual frequency. There were differences in the annual occurrences among cases with different TCM syndrome types. Shire syndrome patients have more frequent annual occurrences than other types of eczema patients. There were significant differences in the annual occurrences among cases of different western medicine staging, and the annual occurrences of acute eczema were more than those of subacute and chronic eczema. Conclusion:The annual occurrences of patients with Tanshi, Shire and Qiyu physique were higher than those of other physiques. There are fewer outpatients with Pinghe physique than other physiques. The annual occurrences of Shire syndrome patients are higher than that of other types of eczema patients. The annual occurrences of acute eczema are more than those of subacute and chronic eczema. The annual occurrences of chronic eczema are less than acute and subacute eczema.
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AIM: To evaluate the therapeutic efficacy and safety of heavy silicone oil (HSO), Densiron 68, was used as internal tamponade to treat complex vitreoretinopathy.METHODS: A retrospective study of 30 patients (30 eyes) who underwent vitrectomy and HSO tamponade for complex retinal detachment between January 2015 and January 2016.The best corrected visual acuity (BCVA), intraocular pressure (IOP), retinal reattachment and complications after surgery were observed.RESULTS: There were statistical significances in both the BCVA difference between pre-operation and HSO tamponade, and the BCVA difference between pre-operation and the removal of HSO for 3mo (z=-2.198, P=0.028;z=-2.682, P=0.007).The average intraocular pressure of HSO tamponade group was 20.233±8.007mmHg, and the average intraocular pressure of pre-operation group was 16.067±4.025mmHg, showing significant difference(t=-2.913, P=0.005).Between the pre-operation group and the HSO removed group 14.933±3.423mmHg, there was no significant statistical difference in the analysis of IOP (t=2.635, P=0.430).Anatomical success was achieved in 90% of cases after the removal of HSO.Most common complications were cataract formation and oil emulsification.CONCLUSION: Densiron-68 is a safe and effective tamponade material for the treatment of complex vitreoretinopathy.However, most common complications are cataract formation and oil emulsification.So clinicians should strictly handle indications and usage during the clinical applications.
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Acute myeloid leukemia (AML) is a malignant clonal hematologic disease from hematopoietic stem and progenitor cells. The isocitrate dehychogenase 2 (IDH2) gene mutation has been recently found, which may be associated with the course of AML. The incidence of IDH2 gene mutation in the patients with acute myeloid leukemia is high, especially in the AML patients with normal karyotype. Different subtypes of IDH2 mutation, or companing other molecular biology, will make different influence on clinical features and progress of patients with AML. IDH2 mutation is stable, which can be used as the test sign of AML and minimal residual disease (MRD), and for guiding the clinical treatment and predicting the progress. In this article, the research progress of IDH2 mutation in acute myeloid leukemia is reviewed.
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Humans , Isocitrate Dehydrogenase , Genetics , Leukemia, Myeloid, Acute , Genetics , Mutation , Neoplasm, Residual , PrognosisABSTRACT
OBJECTIVE@#To discuss the effect of insulin and metformin on a methylation and glycolipid metabolism of peroxisome proliferator-activated receptor γ coactivator-1A (PPARGC1A) of rat offspring with gestational diabetes mellitus (GDM).@*METHODS@#A total of 45 pregnant rats received the intraperitoneal injection of streptozotocin to establish the pregnant rat model of GDM. A total of 21 pregnant rats with GDM were randomly divided into three groups, with 7 rats in each group, namely the insulin group, metformin group and control group. Rats in the insulin group received the abdominal subcutaneous injection of 1 mL/kg recombinant insulin glargine at 18:00 every day. Rats in the metformin group received the intragastric infusion of metformin hydrochloride at 18:00 every day, with the first dose of 300 mg/kg. The doses of two groups were adjusted every 3 d to maintain the blood glucose level at 2.65-7.62 mmol/L. Rats in the control group received the intragastric infusion of 1 mL normal saline at 18:00 every day. After the natural delivery of pregnant rats, 10 offspring rats were randomly selected from each group. At birth, 4 wk and 8 wk after the birth of offspring rats, the weight of offspring rats was measured. The blood glucose level of offspring rats was measured at 4 wk and 8 wk, while the level of serum insulin, triglyceride and leptin was measured at 8 wk.@*RESULTS@#The weight of offspring rats at birth in the insulin group and metformin group was significantly lower than the one in the control group (P 0.05). The fasting blood glucose and random blood glucose in the insulin group and metformin group at 4 wk and 8 wk were all significantly lower than ones in the control group (P 0.05). The expression of PPARGC1A mRNA in the insulin group and metformin group was significantly higher and the methylation level of PPARGC1A was significantly lower than the one in the control group (P 0.05). Insulin and leptin at 8 wk in the insulin group and metformin group were significantly higher, while triglyceride was significantly lower than the one in the control group (P 0.05).@*CONCLUSIONS@#GDM can induce the methylation of PPARGC1A of offspring rats to reduce the expression of PPARGC1A mRNA and then cause the disorder of glycolipid metabolism when the offspring rats grow up; the insulin or metformin in the treatment of pregnant rats with GDM can reduce the methylation level of PPARGC1A and thus improve the abnormal glycolipid metabolism of offspring rats.
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Objective: To discuss the effect of insulin and metformin on a methylation and glycolipid metabolism of peroxisome proliferator-activated receptor γ coactivator-1A (PPARGC1A) of rat offspring with gestational diabetes mellitus (GDM). Methods: A total of 45 pregnant rats received the intraperitoneal injection of streptozotocin to establish the pregnant rat model of GDM. A total of 21 pregnant rats with GDM were randomly divided into three groups, with 7 rats in each group, namely the insulin group, metformin group and control group. Rats in the insulin group received the abdominal subcutaneous injection of 1 mL/kg recombinant insulin glargine at 18:00 every day. Rats in the metformin group received the intragastric infusion of metformin hydrochloride at 18:00 every day, with the first dose of 300 mg/kg. The doses of two groups were adjusted every 3 d to maintain the blood glucose level at 2.65-7.62 mmol/L. Rats in the control group received the intragastric infusion of 1 mL normal saline at 18:00 every day. After the natural delivery of pregnant rats, 10 offspring rats were randomly selected from each group. At birth, 4 wk and 8 wk after the birth of offspring rats, the weight of offspring rats was measured. The blood glucose level of offspring rats was measured at 4 wk and 8 wk, while the level of serum insulin, triglyceride and leptin was measured at 8 wk. Results: The weight of offspring rats at birth in the insulin group and metformin group was significantly lower than the one in the control group (P 0.05). The fasting blood glucose and random blood glucose in the insulin group and metformin group at 4 wk and 8 wk were all significantly lower than ones in the control group (P 0.05). The expression of PPARGC1A mRNA in the insulin group and metformin group was significantly higher and the methylation level of PPARGC1A was significantly lower than the one in the control group (P 0.05). Insulin and leptin at 8 wk in the insulin group and metformin group were significantly higher, while triglyceride was significantly lower than the one in the control group (P 0.05). Conclusions: GDM can induce the methylation of PPARGC1A of offspring rats to reduce the expression of PPARGC1A mRNA and then cause the disorder of glycolipid metabolism when the offspring rats grow up; the insulin or metformin in the treatment of pregnant rats with GDM can reduce the methylation level of PPARGC1A and thus improve the abnormal glycolipid metabolism of offspring rats.
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Abstract?AlM: To study the effect of latanoprost combined with timolol treatment on visual function, intraocular pressure and ocular blood flow of open angle glaucoma.?METHODS:A total of 50 cases ( 59 eyes ) with open angle glaucoma were enrolled from January 2012 to May 2014 in our hospital department of ophthalmology, and divided into observation group and control group by adopting the random number table method. Patients in observation group were treated with latanoprost combined with timolol, patients in control group were treated only with timolol. Visual function, intraocular pressure and ocular blood flow were compared.?RESULTS:1, 2, 3 and 4wk after treatment, observation group during the day intraocular pressure, night intraocular pressure were significantly lower than that in control group, vision levels (0. 27±0. 03, 0. 36±0. 06, 0. 44± 0. 06, 0. 63 ± 0. 13 ) were significantly higher than that in control group; observation group peak systolic velocity (14. 41± 1. 73) cm/s, end diastolic velocity (4. 18 ± 0. 67) cm/s were significantly higher than that in control group;vascular resistance index ( 0. 58 ± 0. 07 ) was significantly lower than that in control group.? CONCLUSlON: Latanoprost combined with timolol treatment can reduce intraocular pressure, increase blood flow of central retinal artery, reduce vascular resistance, improve visual acuity.
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Objective: To optimize the extracting technology of assessing the maximum yield of phenolic compounds (PC) from Inonotus obliquus by single factor experiments and orthogonal array design methods through aqueous two-phase systems combined with ultrasonic extraction. Methods: The range of the independent variables, namely levels of acetone and ammonium sulfate, and ultrasonic time were identified by a first set of single factor experiments. The actual values of the independent variables coded at four levels and three factors were selected based on the results of the single factor experiments. Subsequently, the levels of acetone and ammonium sulfate, and ultrasonic time were optimized using the orthogonal array method. Results: The optimum conditions for the extraction of PC were found to use 7.0 mL acetone, 5.5 mg ammonium sulfate, with ultrasonic time for 5 min. Under these optimized conditions, the experimental maximum yield of PC was 37.8 mg/g, much higher than that of the traditional ultrasonic extraction (UE, 29.0 mg/g). And the PC obtained by this method had stronger anti-oxidative activities than those by traditional UE method. Conclusion: These results indicate the suitability of the models developed and the success in optimizing the extraction conditions. This is an economical and efficient method for extracting polyphenols from I. obliquus. © 2013 Tianjin Press of Chinese Herbal Medicines.
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This study explored the role of radiation-induced autophagy in low-dose hyperradiosensitivity (HRS) in the human lung cancer cell line A549. A549 cells, either treated with an autophagic inhibitor 3-methyladenine (3-MA), or with a vehicle control, were irradiated at different low doses (≤0.5 Gy). The generation of autophagy was examined by laser scanning confocal microscopy. Western blotting was used to detect the expression of microtubule-associated protein l light chain 3B II (LC3B-II). Flow cytometry (FCM) and clonogenic assays were used to measure the fraction of surviving cells at the low irradiation doses. Our results showed that there was a greater inhibition of autophagic activity, but a higher degree of low-dose HRS in A549 cells treated with 3-MA than in control group. Our data demonstrated that radiation-induced autophagy is correlated with HRS in A549 cells, and is probably one of the mechanisms underlying HRS.
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The present study was aimed to investigate the effect of different altitudes on telomere length of rat peripheral blood leukocyte and possible mechanism. Sixty male rats were randomly divided into three groups, lower altitude control group (10 m), moderate altitude group (2 260 m) and very high altitude group (simulated 5 000 m). The moderate altitude group and very high altitude group rats were transported to Xining and hypobaric chamber in Qinghai University, respectively. The peripheral blood specimens were extracted 30 d after the transportation. By means of real-time PCR, automatic blood cell analyzer, ELISA, TBA and WST-1 methods, the telomere lengths of blood leukocyte, the hemoglobin (Hb) contents, the plasma levels of telomerase reverse transcriptase (TERT) and hypoxia-inducible factor 1α (HIF-1α), the plasma content of malondialdehyde (MDA) and superoxide dismutase (SOD) activity were measured, respectively. The results showed that the telomere lengths of peripheral blood leukocyte in moderate altitude group were longer than those in control group and very high altitude group. The changes of TERT were compatible with the telomere length of peripheral blood leukocyte under different altitudes. The levels of HIF-1α in moderate altitude group and very high altitude group were higher than that of control group. The very high altitude group showed decreased SOD activities and increased level of MDA, compared with the other two groups. These results suggest that the telomere lengths of rat peripheral blood leukocyte in moderate altitude are elongated, and that the telomere-elongating effect is lost under very high altitude. The changes of HIF-1α, TERT and oxidative stress damage are the main mechanisms of telomere length changes. Moderate altitude living might be beneficial to increasing the life span in mammals.
Subject(s)
Animals , Male , Rats , Altitude , Hemoglobins , Metabolism , Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit , Blood , Leukocytes , Physiology , Malondialdehyde , Blood , Oxidative Stress , Superoxide Dismutase , Metabolism , Telomerase , Blood , Telomere , PhysiologyABSTRACT
This study explored the role of radiation-induced autophagy in low-dose hyperradiosensitivity (HRS) in the human lung cancer cell line A549. A549 cells, either treated with an autophagic inhibitor 3-methyladenine (3-MA), or with a vehicle control, were irradiated at different low doses (≤0.5 Gy). The generation of autophagy was examined by laser scanning confocal microscopy. Western blotting was used to detect the expression of microtubule-associated protein l light chain 3B II (LC3B-II). Flow cytometry (FCM) and clonogenic assays were used to measure the fraction of surviving cells at the low irradiation doses. Our results showed that there was a greater inhibition of autophagic activity, but a higher degree of low-dose HRS in A549 cells treated with 3-MA than in control group. Our data demonstrated that radiation-induced autophagy is correlated with HRS in A549 cells, and is probably one of the mechanisms underlying HRS.
Subject(s)
Humans , Adenine , Pharmacology , Autophagy , Radiation Effects , Blotting, Western , Cell Line, Tumor , Cell Survival , Radiation Effects , Dose-Response Relationship, Radiation , Flow Cytometry , Green Fluorescent Proteins , Genetics , Metabolism , Lung Neoplasms , Genetics , Metabolism , Pathology , Microscopy, Confocal , Microscopy, Electron, Transmission , Microtubule-Associated Proteins , Genetics , Metabolism , Phagosomes , Radiation Effects , Radiation Tolerance , Radiation EffectsABSTRACT
This study was purposed to investigate the effect of crocin on the proliferation in vitro and immune function of dendritic cells (DC) derived from the bone marrow of children with acute leukemia. The mononuclear cells were isolated from bone marrow of leukemia children by Ficoll-Hypaque. The experiment was divided into six groups: blank control group (A), crocin 1.25 mg/ml group (B), cytokines (rhGM-CSF 75 ng/ml+rhIL-4 75 ng/ml+rhTNF-α 50 ng/ml) group (C), cytokines+crocin 0.3125, 1.25 or 5.0 mg/ml groups (D, E, F). The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture. The DC of different groups were mixed with T cells just separated from peripheral blood of another children with acute lymphoblastic leukemia, and cultured with rhIL-2 200 U/ml for 5 d. The function of DC was detected by mixed lymphocyte reaction (MLR). The results indicated that the test groups and control group all obtained a certain amount of typical DC, but the DC numbers in test groups were all higher than those in control group (P < 0.01). Cultured for 9 days, the rates of CD1a(+), CD83(+), and HLA-DR(+) in group C, D, E, F were higher than group A (P < 0.01). There was no statistically significant difference between A and B groups (P > 0.05). MLR showed that with the increasing of DC, the stimulation index of T cells in group A and B was not rising (P > 0.05); the stimulated index of T cells in group C and E was significantly rising, there was statistically significant difference between them (P < 0.01). When the number of stimulated cells was the same, the stimulation index of T cell in group E was the highest (P < 0.01). It is concluded that the capability of DC proliferation promoted by crocin alone is lower than that of its combination with rhGM-CSF, rhIL-4 and rhTNF-α, but the crocin can synergically promote the maturity of DC cooperating with rhGM-CSF, rhIL-4 and rhTNF-α. The DC induced by crocin can particularly enhance the proliferation of T cells.
Subject(s)
Child , Humans , Bone Marrow Cells , Cell Biology , Carotenoids , Pharmacology , Cell Proliferation , Dendritic Cells , Cell Biology , Leukemia , Pathology , Lymphocyte Culture Test, Mixed , T-Lymphocytes , Cell Biology , Tumor Cells, CulturedABSTRACT
This study was to investigate the proliferative inhibition and apoptosis of human leukemia HL-60 cells induced by crocin and their possible mechanisms. The cell viability was tested by cell counting. The morphology of HL-60 cells was observed by fluorescence microscopy. The MTT assay was used to evaluate the inhibitory effect of crocin on the growth of HL-60 cells. Flow cytometry was used to measure the cell cycle. RT-PCR was used to detect bcl-2 and bax expression. The results indicated that the growth of HL-60 cells was inhibited remarkably in the dose and time dependent way. When the crocin concentration was higher than 5 mg/ml, the percentage of apoptotic HL-60 cells was not increased, on the contrary this percentage decreased, the cells manifested necrosis. Flow cytometry profiles revealed that cells were blocked in G₀/G₁ phase, the cell proliferation was inhibited obviously at 5 mg/ml. RT-PCR detection revealed that the expression of bcl-2 was down-regulated strikingly and bax was up-regulated. It is concluded that the crocin can inhibit the proliferation of HL-60 cells effectively, and block cells in G₀/G₁ phase. The mechanisms by which crocin induced apoptosis in HL-60 cells may be related to the inhibition of bcl-2 and activation of bax.
Subject(s)
Humans , Apoptosis , Carotenoids , Pharmacology , Therapeutic Uses , Cell Cycle , Cell Proliferation , Gene Expression Regulation, Leukemic , HL-60 Cells , Leukemia , Drug Therapy , Metabolism , Phytotherapy , Proto-Oncogene Proteins c-bcl-2 , Metabolism , bcl-2-Associated X Protein , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of Rho-kinase signaling pathway in human airway smooth muscle cell (ASMCs) migration and cytoskeletal reorganization induced by endothelin-1 (ET-1).</p><p><b>METHODS</b>Primary cultured human ASMCs obtained by tracheal explant culture method were examined for cell migration in response to ET-1 treatment using modified Boyden chambers. The changes in actin cytoskeletal reorganization were observed under confocal laser scanning microscope, and the phosphorylation of myosin-phosphatase target 1 (p-MYPT1) was examined using Western blot analysis.</p><p><b>RESULTS</b>At the concentration of 0.1, 1, 10, and 100 nmol/L, ET-1 induced migration of the ASMCs, and 10 nmol/L ET-1 produced the most obvious effect (P<0.01). Rho-kinase inhibitor Y-27632 showed a dose-dependent inhibitory effect on ET-1-induced ASMC migration, and in cells exposed to 10 nmol/L ET-1, Y-27632 at 10 micromol/L significantly blocked ASMC migration (P<0.01). ET-1 (10 nmol/L) exposure resulted in reorganization of actin cytoskeleton and formation of stress fibers in the ASMCs, which were obviously inhibited by Y-27632. Compared with the control group, the AMSCs showed significant enhancement of p-MYPT1 protein expression after ET-1 exposure for 15 and 30 min (P<0.01), but prolonged exposure failed to result in the expression enhancement (P>0.05).</p><p><b>CONCLUSION</b>Rho-kinase signaling pathway may play an important role in ET-1-induced ASMC migration and reorganization of actin cytoskeleton.</p>
Subject(s)
Humans , Amides , Pharmacology , Bronchi , Cell Biology , Cell Movement , Cells, Cultured , Cytoskeleton , Metabolism , Endothelin-1 , Pharmacology , Enzyme Inhibitors , Pharmacology , Microscopy, Confocal , Muscle, Smooth , Cell Biology , Pyridines , Pharmacology , Signal Transduction , rho-Associated Kinases , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To understand the nutrition status of children under three years in rural area of western China and to explore the influencing factors so as to provide reasonable suggestions for policy making.</p><p><b>METHODS</b>Use PPS sampling method to investigate the 13,532 children under three years old. Height and weight were used as nutritional indexes.</p><p><b>RESULTS</b>The prevalence of stunting (height for age Z-score < -2), underweight (weight for age Z-score < -2) and wasting (weight for height Z-score < -2) were 12.4%,11.8% and 5.7% respectively. Boys, minority and the children from western China had higher prevalence rate. The prevalence rates of the Han nationality children's underweight and stunting were 9.5% and 9.8%, but these rates of the minority children were 15.6% and 16.5% respectively, which were obviously higher than the Han ethnicity children with significant differences between them (P <0.01).The prevalence of malnutrition was rising with age and the peak age of stunting, underweight and wasting appeared at 21 months, 12 months and 15 months, respectively. Compared with growth reference of NCHS/WHO, the HAZ, WAZ and WHZ left moved 0.59,0.60 and 0.26 units which indicated the whole nutritional status of children from program area impaired to some extent. Underweight inclined to have higher two-week prevalence rates of diarrhea and flu than in the normal children,achieving 15.9% and 13.5%, but with significant differences between them (P<0.01). We administered non-conditional logistic regression analysis to identify the influencing factors of malnutrition. Under-6-month children who were not taken care by their mothers showed higher risk of stunting. Over-6-month children stunting had significant relationship with age, gender, sibling order, nationality,maternal educational level,special cooking for children and residential region. Underweight of over-6-month children significantly related to age, nationality, and maternal educational level, yolk supply during 6-8 month old and living region.</p><p><b>CONCLUSION</b>Malnutrition was really prevalent among children in China,suggesting that intervention should be done according to the influencing factors.</p>
Subject(s)
Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Child Nutrition Disorders , Epidemiology , China , Epidemiology , Nutrition Assessment , Nutritional Status , PrevalenceABSTRACT
<p><b>OBJECTIVE</b>To study the inhibitory effect of shikonin on the proliferation of human airway smooth muscle cells (HASMCs) in vitro.</p><p><b>METHODS</b>HASMCs from the trachea were obtained by primary culture of the tissue explants and adherent culture. The HASMCs were exposed to shikonin at 0 (control group), 0.5, 1, 2, 5, 10, 20, and 40 micromol/L for 12, 24, and 48 h, after which the cell proliferation was assessed by 3-(4,5-carboxymethoxypheny1)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetric assay. Flow cytometry was used for cell cycle analysis of the HASMCs exposed to shikonin at 40, 20, 10, 5 micromol/L and 0 micromol/L (control group) for 24 h. Immunocytochemistry with SP method was performed to detect the expression of proliferating cell nuclear antigen (PCNA) in the HASMCs treated with shikonin at 20 micromol/L and 0 micromol/L (control group) for 24 h.</p><p><b>RESULTS</b>Shikonin at the concentrations of 20 and 40 micromol/L significantly inhibited the proliferation of the cells (P<0.05), and the effect was especially obvious after 48 h of cell exposure, with inhibition rates of 30.1% and 42.9%, respectively. No significant difference was found between the two concentrations for their cell growth inhibition effects (P>0.05). Shikonin at the concentrations of 20 and 40 micromol/L caused significant cell cycle arrest in G(0)/G(1) phase (P<0.05), the effect of which, however, was not concentration-dependent (P>0.05). Shikohin at 20 micromol/L significantly down-regulated the expression of PCNA in the cells (P<0.05).</p><p><b>CONCLUSION</b>Shikonin can inhibit the proliferation of HASMCs in vitro.</p>
Subject(s)
Humans , Cell Cycle , Cell Proliferation , Cells, Cultured , Dose-Response Relationship, Drug , Flow Cytometry , Immunohistochemistry , Muscle, Smooth , Cell Biology , Metabolism , Naphthoquinones , Pharmacology , Proliferating Cell Nuclear Antigen , Metabolism , Trachea , Cell BiologyABSTRACT
To explore the effects of tetra-arsenic tetra-sulfide (As(4)S(4)) in treatment of human chronic myelogenous leukemia K562 cells and its mechanism, trypan blue staining and microculture MTS assay were used to measure the effects of As(4)S(4) on growth inhibition of K562 cells; the morphologic change was determined by Wright's staining assay. The apoptosis rate and cell cycle were detected by flow cytometry; the changes of transcript and protein level were determined by real-time quantitative RT-PCR and Western blot analysis, respectively. The results indicated that As(4)S(4) had significant cytotoxicity on K562 cells. At the concentration of 0.5 micromol/L, the cell viability decreased significantly after being cultured with As(4)S(4) for 24 hours. When the concentration was lower than 0.1 micromol/L, As(4)S(4) had a little effect on K562 cells. The effect of As(4)S(4) on K562 was time- and concentration- dependent. After being cultured with As(4)S(4) at the concentration of 1.0 micromol/L for 24 to 48 hours, K562 cells displayed typical morphological changes of apoptosis. At a concentration greater than or equal to 1.0 micromol/L, As(4)S(4) could induce apoptosis significantly. After 12 hours of incubation with 1.0 micromol/L As(4)S(4), the apoptosis rate increased from (3.47 +/- 0.42)% to (6.16 +/- 0.98%). At the same time, the percentage of cells in G(1) phase decreased from (69.65 +/- 3.24)% to (50.53 +/- 2.86)%, whereas the percentage of cells in G(2)/M phase increased from (9.56 +/- 2.51)% to (12.91 +/- 2.13)%. The mRNA level of Bcl-X(L) and the protein level of pAkt were down-regulated after the inhibition of As(4)S(4), while the mRNA expression of Bcl-2, Bad and Bax had no change. Both of the transcript and protein level of bcr-abl had no change after incubation with As(4)S(4). It is concluded that As(4)S(4) can inhibit the growth of K562 cells efficiently through inducing apoptosis and cell cycle arrest. It seems that As(4)S(4) interferes with pAkt pathway and down-regulates Bcl-X(L), which may be involved in the response of K562 to this agent.
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Apoptosis , Arsenicals , Chemistry , Pharmacology , Blotting, Western , Cell Cycle , Cell Survival , Dose-Response Relationship, Drug , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Genetics , Metabolism , Pathology , Proto-Oncogene Proteins c-akt , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sulfides , Chemistry , Pharmacology , Time Factors , bcl-X Protein , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the expressions of costimulators on peripheral T and B lymphocytes in patients with idiopathic thrombocytopenic purpura (ITP).</p><p><b>METHODS</b>The expression of B7-CD(28) and CD(40) of peripheral lymphocytes was measured by flow cytometry in 21 ITP patients and 9 normal subjects. The expression of PAIgG was measured by ELISA method.</p><p><b>RESULTS</b>The expression of CD(4)(+)CD(28)(+) was lower in ITP patients than in normal controls, but the expression of CD(86)(+) and CD(86)(+)CD(19)(+) was higher in ITP patients than in normal controls, while the expression of CD(80)(+), CD(40)(+), CD(28)(+), CD(19)(+)CD(86)(+), CD(19)(+)CD(40)(+), CD(4)(+)CD(28)(+)/CD(4)(+), CD(19)(+)CD(80)(+)/CD(19)(+) and CD(19)(+)CD(40)(+)/CD(19)(+) in ITP patients was normal. The PAIgG level was higher in 16 patients with a mean of (184.62 +/- 38.00) ng/10(7) plt. A positive correlation was found between PAIgG and CD(19)(+)CD(86)(+)/CD(19)(+) expression.</p><p><b>CONCLUSION</b>There is no deficiency in expression of CD(28) on CD(4)(+) T lymphocytes in ITP patients. The change of CD(86) expression on B lymphocytes is possibly involved in pathophysiology of ITP, which may provide a theoretical instruction for ITP patients immunological therapy.</p>