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Objective:To systematically evaluate the efficacy of oral Chinese herbal prescriptions combined with transcatheter arterial chemoembolization （TACE） against primary hepatic carcinoma （PHC） and screen the basic Chinese herbs，in order to provide certain reference for clinical medication. Method:The randomized controlled trials concerning the treatment of PHC with oral Chinese herbal prescriptions plus TACE were retrieved from CBM，China National Knowledge Infrastructure （CNKI），Chongqing Weipu Database for Chinese Technical Periodicals （VIP），and Wanfang Data Knowledge Service Platform.The quality of the included trials was evaluated by Cochrane handbook，and the Meta-analysis was performed using RevMan 5.3.The enumeration data were expressed by odds ratio （OR），the measurement data by mean difference （MD） or standardized mean difference （SMD），and the effect size by 95% confidence interval （CI）.The data of oral Chinese herbal prescriptions involved in trials were sorted out and subjected to association rule analysis and frequency analysis based on the Traditional Chinese Medicine Inheritance Support System （TCMISS），for exploring the basic Chinese herbs and their dosages against PHC. Result:A total of 75 randomized controlled trials were included，involving 7 406 cases. As revealed by the Meta-analysis，oral Chinese herbal prescriptions combined with TACE was significantly better than TACE alone in improving the short-term curative effect ［OR=2.05，95%CI（1.83，2.29）］，decreasing alpha fetoprotein （AFP） ［MD=-59.02，95%CI（-79.03，-39.01）］，ameliorating liver function ［SMD=-1.23，95%CI（-1.58，-0.88）］，boosting immunity ［SMD=1.08，95%CI（0.84，1.32）］，adjusting Karnofsky Performance Status （KPS） scale score ［OR=2.7，95%CI（1.11，11.02）］，elevating survival rate ［OR=2.31，95%CI（1.96，2.71）］，and reducing adverse reactions ［OR=0.38，95%CI（0.34，0.43）］.Data mining results showed that the basic Chinese herbs against PHC were Bupleuri Radix，Paeoniae Alba Radix，Atractylodis Macrocephalae Rhizoma，Poria，and Glycyrrhizae Radix et Rhizoma，with their clinical dosages listed as follows：6-15 g for Bupleuri Radix，10-15 g for Paeoniae Alba Radix，9-15 g for Atractylodis Macrocephalae Rhizoma，10-15 g for Poria，and 3-10 g for Glycyrrhizae Radix et Rhizoma. Conclusion:The oral Chinese herbal prescriptions combined with TACE produce better effects in treatment of PHC as compared with TACE alone.These five basic Chinese herbs have anti-cancer effect，and their dosages are within the ranges stipulated in 2020 edition of <italic>Chinese Pharmacopoeia.</italic>This Meta-analysis has provided certain reference for clinical medication.
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This study was designed to evaluate the effects of drilling through the growth plate and using adipose-derived stem cells (ADSCs) and bone morphogenetic protein-2 (BMP-2) to treat femoral head epiphyseal ischemic necrosis,which can be done in juvenile rabbits.Passage-four bromodeoxyuridine (BrdU)-labeled ADSCs were cultured,assayed with MTT to determine their viability and stained with alizarin red dye to determine their osteogenic ability.Two-month-old,healthy male rabbits (1.2 to 1.4 kg,n=45) underwent ischemic induction and were randomly divided into five groups (group A:animal model control;group B:drilling;group C:drilling & ADSCs;group D:drilling & BMP-2;and group E:drilling & ADSCs & BMP-2).Magnetic resonance imaging (MRI),X-ray imaging,hematoxylin and eosin staining and BrdU immunofluorescence detection were applied 4,6 and 10 weeks after treatment.Approximately 90％ of the ADSCs were labeled with BrdU and showed good viability and osteogenic ability.Similar results were observed in the rabbits in groups C and E at weeks 6 and 10.The animals of groups C and E demonstrated normal hip structure and improved femoral epiphyseal quotients and trabecular areas compared with those of the groups A and B (P＜0.01).Group D demonstrated improved femoral epiphyseal quotients and trabecular areas compared with those of groups A and B (P＜0.05).In summary,drilling through the growth plate combined with ADSC and BMP-2 treatments induced new bone formation and protected the femoral head epiphysis from collapsing in a juvenile rabbit model of femoral head epiphyseal ischemic necrosis.
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This study was designed to evaluate the effects of drilling through the growth plate and using adipose-derived stem cells (ADSCs) and bone morphogenetic protein-2 (BMP-2) to treat femoral head epiphyseal ischemic necrosis,which can be done in juvenile rabbits.Passage-four bromodeoxyuridine (BrdU)-labeled ADSCs were cultured,assayed with MTT to determine their viability and stained with alizarin red dye to determine their osteogenic ability.Two-month-old,healthy male rabbits (1.2 to 1.4 kg,n=45) underwent ischemic induction and were randomly divided into five groups (group A:animal model control;group B:drilling;group C:drilling & ADSCs;group D:drilling & BMP-2;and group E:drilling & ADSCs & BMP-2).Magnetic resonance imaging (MRI),X-ray imaging,hematoxylin and eosin staining and BrdU immunofluorescence detection were applied 4,6 and 10 weeks after treatment.Approximately 90％ of the ADSCs were labeled with BrdU and showed good viability and osteogenic ability.Similar results were observed in the rabbits in groups C and E at weeks 6 and 10.The animals of groups C and E demonstrated normal hip structure and improved femoral epiphyseal quotients and trabecular areas compared with those of the groups A and B (P＜0.01).Group D demonstrated improved femoral epiphyseal quotients and trabecular areas compared with those of groups A and B (P＜0.05).In summary,drilling through the growth plate combined with ADSC and BMP-2 treatments induced new bone formation and protected the femoral head epiphysis from collapsing in a juvenile rabbit model of femoral head epiphyseal ischemic necrosis.
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This study aimed to examine the biocompatibility of calcium titanate (CaTiO3) coating prepared by a simplified technique in an attempt to assess the potential of CaTiO3 coating as an alternative to current implant coating materials.CaTiO3-coated titanium screws were implanted with hydroxyapatite (HA)-coated or uncoated titanium screws into medial and lateral femoral condyles of 48 New Zealand white rabbits.Imaging,histomorphometric and biomechanical analyses were employed to evaluate the osseointegration and biocompatibility 12 weeks after the implantation.Histology and scanning electron microscopy revealed that bone tissues surrounding the screws coated with CaTiO3 were fully regenerated and they were also.well integrated with the screws.An interfacial fibrous membrane layer,which was found in the HA coating group,was not noticeable between the bone tissues and CaTiO3-coated screws.X-ray imaging analysis showed in the CaTiO3 coating group,there was a dense and tight binding between implants and the bone tissues;no radiation translucent zone was found surrounding the implants as well as no detachment of the coating and femoral condyle fracture.In contrast,uncoated screws exhibited a fibrous membrane layer,as evidenced by the detection of a radiation translucent zone between the implants and the bone tissues.Additionally,biomechanical testing revealed that the binding strength of CaTiO3 coating with bone tissues was significantly higher than that of uncoated titanium screws,and was comparable to that of HA coating.The study demonstrated that CaTiO3 coating in situ to titanium screws possesses great biocompatibility and osseointegration comparable to HA coating.
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<p><b>OBJECTIVE</b>To explore the effect of arsenic trioxide combined with itraconazole on proliferation and apoptosis of KG1a cells and its potential mechanism.</p><p><b>METHODS</b>The cell morphology was observed with Wrighe-Giemsa staining; cell survival rate was examined by CCK-8; and colony formation capacity was measured by methylcellulose colony formation test; the flow cytometry was used to analyse the cell apoptosis rate and cell cycle; the protein expressions of BCL-2,caspase-3,BAX,SMO,Gli1 and Gli2 were detected by Western-blot.</p><p><b>RESULTS</b>The arsenic trioxide and itraconazole alone both could inhibit the KG1a cell proliferation in dose-and time-dependent manner. In comparison between single and combined drug-treatment group, both the cell survival rate and the colony number of the single drug-treatment group were significantly lower(P<0.05), and the apoptosis rate was higher in the combined drug-treatment group. In the combined-treatment group, the protein expression of Caspase-3 and BAX was upregulated, while the protein expression of BCL-2,SMO,Gli1 and Gli2 was downregulated.</p><p><b>CONCLUSION</b>Arsenic trioxide combined with itraconazole can inhibit the KG1a cell proliferation and induce apoptosis, which may be related with the inhibition of Hh signaling pathway and upregulation of both Caspase-3 and BAX protein expression, and provided experimental data of arsenic trioxide combined with itraconazole for the treatment of refractory AML.</p>
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OBJECTIVE@#To explore the regulation effect of Rheum palmatum (R. palmatum) L. on NF-κB signaling pathway of ALF mice.@*METHODS@#The intraperitoneal injection of d-GalN/LPS was employed for the model building. Mice in the treatment group and positive control group were given the R. palmatum L. and bifendate before the model building. Mice in the normal group were given the intraperitoneal injection of equivalent normal saline for continuously 3 d. After 16 h of model building, the blood was collected from eyeballs of mice and then mice were executed. The measurement was performed on the content of ALT, AST, NO and Il-1β in the serum of mice in each group, as well as the activity of Caspase 3 and Caspase 8 in the liver tissue. HE staining was employed to detect the pathological morphology of liver; and the western blot was used to detect the expression of iNOS, COX-2, Bax, Bcl-2, PCNA, NF-κB p65 and IκBα.@*RESULTS@#The content of ALT, AST, NO and Il-1β in the serum and the activity of Caspase 3 and Caspase 8 in the liver tissue were increased in the mice of ALF model group. Besides, the expression of iNOS, COX-2 and Bax was increased, the expression of Bcl-2 and PCNA was decreased, the phosphorylation of NF-κB p65 and IκBα was significant and the treatment group of R. palmatum L. could inhibit such change.@*CONCLUSIONS@#Through NF-κB signaling pathway, the R. palmatum L. could reduce the content of enzyme of liver function and inflammation factor in the serum of ALF mice, regulate the expression of cell apoptosis-related protein and improve the symptoms of ALF mice.
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Objective: To explore the regulation effect of Rheum palmatum (R. palmatum) L. on NF-κB signaling pathway of ALF mice. Methods: The intraperitoneal injection of d-GalN/LPS was employed for the model building. Mice in the treatment group and positive control group were given the R. palmatum L. and bifendate before the model building. Mice in the normal group were given the intraperitoneal injection of equivalent normal saline for continuously 3 d. After 16 h of model building, the blood was collected from eyeballs of mice and then mice were executed. The measurement was performed on the content of ALT, AST, NO and Il-1β in the serum of mice in each group, as well as the activity of Caspase 3 and Caspase 8 in the liver tissue. HE staining was employed to detect the pathological morphology of liver; and the western blot was used to detect the expression of iNOS, COX-2, Bax, Bcl-2, PCNA, NF-κB p65 and IκBα. Results: The content of ALT, AST, NO and Il-1β in the serum and the activity of Caspase 3 and Caspase 8 in the liver tissue were increased in the mice of ALF model group. Besides, the expression of iNOS, COX-2 and Bax was increased, the expression of Bcl-2 and PCNA was decreased, the phosphorylation of NF-κB p65 and IκBα was significant and the treatment group of R. palmatum L. could inhibit such change. Conclusions: Through NF-κB signaling pathway, the R. palmatum L. could reduce the content of enzyme of liver function and inflammation factor in the serum of ALF mice, regulate the expression of cell apoptosis-related protein and improve the symptoms of ALF mice.
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Objective To reversing methicillin-resistant Staphylococcus(MRS) to methicillin-susceptible Staphylococcus(MSS) by changing nutritional conditions and continuous transfer of culture .Methods MRS trains separating from clinical specimens were cultured in different conditions ,continuous cultural transfer ,and drug sensitive test were proceeded periodically to observe the phe-notypic and chemical reaction change of MRS .The mecA gene were detected of the original and mutant strains by polymerase chain reaction(PCR) ,then the gene sequenced and compared .Results 53 MRS strains were studied .6 strains were phenotype successful-ly converted to MSS in different cultural conditions ,among them mecA gene was undetected in 2 strains ,and down expressed in 4 strains .Conclusion The MRS strains separated from clinical specimens may revert to MSS by culture under different nutritional conditions .The mecA gene of MRS may be lost or lower expressed and the MRS and mutant strains may be different in genomics .
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Melanocyte loss in vitiligo vulgaris is believed to be an autoimmune process. Macrophage migration inhibitory factor (MIF) is involved in many autoimmune skin diseases. We determined the possible role of MIF in the pathogenesis of vitiligo vulgaris, and describe the relationship between MIF expressions and disease severity and activity. Serum MIF concentrations and mRNA levels in PBMCs were measured in 44 vitiligo vulgaris patients and 32 normal controls, using ELISA and real-time RT-PCR. Skin biopsies from 15 patients and 6 controls were analyzed by real-time RT-PCR. Values are reported as median (25th-75th percentile). Serum MIF concentrations were significantly increased in patients [35.81 (10.98-43.66) ng/mL] compared to controls [7.69 (6.01-9.03) ng/mL]. MIF mRNA levels were significantly higher in PBMCs from patients [7.17 (3.59-8.87)] than controls [1.67 (1.23-2.42)]. There was also a significant difference in MIF mRNA levels in PBMCs between progressive and stable patients [7.86 (5.85-9.13) vs 4.33 (2.23-8.39)] and in serum MIF concentrations [40.47 (27.71-46.79) vs 26.80 (10.55-36.07) ng/mL]. In addition, the vitiligo area severity index scores of patients correlated positively with changes of both serum MIF concentrations (r = 0.488) and MIF mRNA levels in PBMCs (r = 0.426). MIF mRNA levels were significantly higher in lesional than in normal skin [2.43 (2.13-7.59) vs 1.18 (0.94-1.83)] and in patients in the progressive stage than in the stable stage [7.52 (2.43-8.84) vs 2.13 (1.98-2.64)]. These correlations suggest that MIF participates in the pathogenesis of vitiligo vulgaris and may be useful as an index of disease severity and activity.
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Leukocytes, Mononuclear/chemistry , Macrophage Migration-Inhibitory Factors/metabolism , RNA, Messenger/metabolism , Vitiligo/metabolism , Case-Control Studies , Enzyme-Linked Immunospot Assay , Macrophage Migration-Inhibitory Factors/analysis , Reverse Transcriptase Polymerase Chain Reaction , Severity of Illness Index , Vitiligo/etiology , Vitiligo/pathologyABSTRACT
<p><b>OBJECTIVE</b>To explore the associated biomarkers influencing recurrence, metastasis and prognosis in patients with gastrointestinal stromal tumors (GIST) after complete resection.</p><p><b>METHODS</b>Tumor tissue samples of 148 patients with GIST undergoing complete resection from January 1990 to December 2008 in Sun Yat-sen University Cancer Center were collected. The expressions of Ki-67, E-cadherin, MMP7, CD44, nm23, P53, survivin, Cyclin D1, COX-2, and VEGF in tumor tissue samples were detected by tissue microarray and immunohistochemistry (IHC). The association of above factors expressions with recurrence, metastasis and prognosis was examined.</p><p><b>RESULTS</b>Log-rank test showed that Ki-67, E-cadherin, MMP7, CD44, P53 and survivin were associated to disease-free duration after complete GIST resection (all P<0.05), and the Ki-67, E-cadherin, P53 and survivin were associated to overall survival (all P<0.05). Cox multivariate analysis revealed that disease-free survival was associated with Ki-67, CD44 and P53 (all P<0.05), and the overall survival was only associated with Ki-67 (P<0.05).</p><p><b>CONCLUSION</b>Ki-67, CD44 and P53 are closely associated with recurrence and metastasis after complete GIST resection, and Ki-67 can predict the prognosis of GIST.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Metabolism , Gastrointestinal Neoplasms , Metabolism , General Surgery , Gastrointestinal Stromal Tumors , Metabolism , General Surgery , Hyaluronan Receptors , Metabolism , Ki-67 Antigen , Metabolism , Neoplasm Metastasis , Neoplasm Recurrence, Local , Prognosis , Tumor Suppressor Protein p53 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To observe the inhibitory effect of 4-chlorobenzoyl berbamine (BBD9) on imatinib-resistant cell line K562 (K562/IR) in vitro and in vivo and explore the mechanisms.</p><p><b>METHODS</b>The IC50 of BBD9 and berbamine (BBM) was determined by MTT assay. The expressions of p210(Bcr-Abl), IKKa, cytoplasmic and nuclear NF-κBp65 were determined using Western blotting in K562/IR cells following a 48-h exposure to 0.5 µg/ml BBD9 or 8 µg/ml BBM. Flow cytometry was used to analyze the cell viability, apoptosis and necrosis; Western blotting was employed to determine the expressions of PARP, caspase-3, caspase-9 and LC3II in K562/IR cells exposed to different concentrations of BBD9 for 48 h. In nude mouse models bearing K562/IR cell xenograft, the tumor weight, tumor regression, and body weight changes of the mice were measured after treatments with 15 mg/kg and 30 mg/kg BBD9 and 100 mg/kg imatinib.</p><p><b>RESULTS</b>The IC50 of BBD9 and BBM was 0.73 µg/ml and 5.43 µg/ml, respectively. In K562/IR cell cultures, the expressions of p210(Bcr-Abl), IKKa and nuclear NF-κB p65 were all decreased following BBD9 and BBM treatments, but BBD9 produced more potent effect; cytoplasmic NF-κB p65 showed no obvious changes after the treatments. The cell apoptosis and necrosis increased with the concentrations of BBD9, which also dose-dependently increased the levels of cleaved caspase-3, csapase-9, PARP, and LC3II expression. In the tumor-bearing mouse model, BBD9 showed stronger effects than imatinib in reducing the tumor weight, promoting tumor regression, and increasing the body weight.</p><p><b>CONCLUSION</b>BBD9 can effectively inhibit the growth of K562/IR cells in vitro and in vivo by activating cell apoptosis, necrosis and autophage pathways, down-regulating expressions of p210(Bcr-Abl) and IKKa and suppressing the cytoplasm-to- nucleus translocation of NF-κBp65.</p>
Subject(s)
Animals , Female , Humans , Mice , Antineoplastic Agents , Pharmacology , Therapeutic Uses , Benzamides , Benzylisoquinolines , Pharmacology , Therapeutic Uses , Drug Resistance, Neoplasm , Fusion Proteins, bcr-abl , Metabolism , Gene Expression Regulation, Neoplastic , I-kappa B Kinase , Metabolism , Imatinib Mesylate , K562 Cells , Liver Neoplasms, Experimental , Drug Therapy , Metabolism , Mice, Nude , Piperazines , Pharmacology , Protein-Tyrosine Kinases , Pyrimidines , Pharmacology , Transcription Factor RelA , Metabolism , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To study the effects of pulsed magnetic field on insulin-like growth factor-1 (IGF-1) level in the cerebrospinal fluid (CSF) and the association of IGF-1 alterations with the activities of daily living (ADL) of patients with brain injury.</p><p><b>METHODS</b>Sixty-five patients with brain injury were divided randomly into the control group (n=30) and magnetic therapy group (n=35), both receiving conventional therapy and in the latter group, daily pulsed magnetic field treatment (20-40 mT, 50 Hz, 20 min per time, 1 time per day) for 14 consecutive days were administered. On the first and 14th days of the treatment, 2 ml CSF was collected from the cases patients for IGF-1 measurement by radioimmunoassay, and Barthel index (BI) was used to assess the ADL of the patients.</p><p><b>RESULTS</b>After a 14-day treatment, IGF-1 level in the CSF were significantly increased in the magnetic group in comparison with the level before the treatment and with those in the control group (P<0.05). IGF-1 in the CSF underwent no significant changes in the control group (P>0.05). The scores of BI increased significantly in both groups after the treatment (P<0.01), but the increment was more obvious in the magnetic therapy group (P<0.05). A significant positive correlation was found between IGF-1 level in the CSF and BI in these patients (r=0.283, P=0.022).</p><p><b>CONCLUSION</b>Pulsed magnetic field might increase IGF-1 level in the CSF of patients with brain injury to promote the recovery of the patients ADL, suggesting its potential clinical value in the treatment of brain injury.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Brain Injuries , Cerebrospinal Fluid , Rehabilitation , Therapeutics , Insulin-Like Growth Factor I , Cerebrospinal Fluid , Magnetic Fields , Recovery of FunctionABSTRACT
Objective To observe the effects of T-lymphocyte function subsets,IL-4 and IFN-γ cell factor in different dose and stage after irradiation.Methoda The C57BL/6j mice were divided into shammed irradiation group and model groups.The radiation hurt model was induced by 60Co gamma rays(0.7,1.4,2.8 and 5.6 Gy).The changes of T-lymphocyte subsets CD3,CD4,CD8,IL-4 and IFN-γ in spleen cells were analyzed by flow cytometry in acute injury stage and recovery stage after irradiation.Results The lymphocyte subsets CD3 +,CD4+ and CD8 + decreased after irradiation,which were related to the irradiation dose.At 1 day after irradiation,the decreasing level of IFN-γ was higher than that of IL-4.When irradiation dose was over 2.8 Gy,IL-4 / IFN-γ showed a markedly increased compared with control group.At 25 days after irradiation,CD3 +,CD4+,CD8 +,CD4 +/CD8 +,IL-4 and IFN-γ recovered obviously,but they did not recover to the normal level of shammed irradiation group.Conclusions The depression of mouse immune function induced by γ-irradiation might be caused by changes of CD3,CD4,CD8 and CD4/CD8 ratio,especially the imbalance of IL4 and IFN-γ.
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<p><b>OBJECTIVE</b>To investigate the expressions of epidermal growth factor receptor (EGFR) in primary nasopharygeal carcinoma (NPC) and lymph node metastases.</p><p><b>METHODS</b>Archived samples of primary NPC and paired lymph node metastases from 86 patients were examined immunohistochemically for the protein expression of EGFR.</p><p><b>RESULTS</b>EGFR expression positivity was detected in the primary NPC and lymph node metastases at the rate of 73.3% and 60.5%, respectively, and primary and metastatic foci showed significant difference in the expression levels (P=0.001). A discrepancy of EGFR expression between the primary and metastatic foci was found in 25 patients, with a discrepancy rate of 29.1% (25/86).</p><p><b>CONCLUSION</b>The difference in EGFR expression between the primary and lymph node metastastic foci of NPC needs to be evaluated when performing EGFR-targeted therapies especially in advanced NPC cases.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Lymph Nodes , Metabolism , Lymphatic Metastasis , Nasopharyngeal Neoplasms , Metabolism , Pathology , ErbB Receptors , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of berbamine (BBM) on multiple myeloma (MM) cell line RPMI 8226 and its mechanism.</p><p><b>METHODS</b>MTT bioassay was used to examine the effect of berbamine on cell growth and IC(50) was calculated. Apoptosis was observed by flow cytometry (FCM) and DNA gelose electrophoresis. p53, p21, GADD45 mRNA were measured by RT-PCR. The alterations in p53, J NK, p-JNK and c-Jun proteins were detected by Western blot method.</p><p><b>RESULT</b>The growth of RPMI 8226 cells was suppressed in a dose-dependent manner after treatment with BBM(P<0.05), and its IC(50) value was 3.83 microg/ml at 48 h. Both DNA ladder and FCM results showed that BBM induced apoptosis of RPMI 8226 cells with concomitant increase of activated p53, p21 and GADD45gamma mRNA. After treatment with BBM at 8 microg/ml for 24 h, the percentage of apoptotic cells increased from 1.07% to 24.84%. p-JNK and c-Jun proteins were activated.</p><p><b>CONCLUSION</b>BBM can inhibit the growth of RPMI 8226 cells, which is associated with activation of GADD45/JNK signaling pathway and induction of cell apoptosis.</p>
Subject(s)
Humans , Apoptosis , Benzylisoquinolines , Pharmacology , Cell Cycle Proteins , Metabolism , Cell Line, Tumor , Dose-Response Relationship, Drug , JNK Mitogen-Activated Protein Kinases , Metabolism , Multiple Myeloma , Pathology , Nuclear Proteins , Metabolism , Signal TransductionABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of berbamine on human hepatoma cell line SMMC7721.</p><p><b>METHODS</b>The effects of 24 h and 48 h incubation with different concentrations (0 to approximately 64 microg/ml) of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI (propidium iodide) staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential (psi(m)); the expression of activated caspase3 and caspase9 was analyzed by Western-blot.</p><p><b>RESULTS</b>The proliferation of SMMC7721 was decreased after treatment with berbamine in a dose- and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential (psi(m)) and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk.</p><p><b>CONCLUSION</b>Berbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.</p>
Subject(s)
Humans , Alkaloids , Pharmacology , Apoptosis , Benzylisoquinolines , Pharmacology , Carcinoma, Hepatocellular , Metabolism , Pathology , Caspase 3 , Metabolism , Caspase 9 , Metabolism , Cell Line, Tumor , Enzyme Activation , Liver Neoplasms , Metabolism , Pathology , Medicine, Chinese Traditional , Membrane Potentials , Mitochondria, Liver , MetabolismABSTRACT
<p><b>BACKGROUND</b>Currently, resistance and relapse are still major problems in acute promyelocytic leukemia (APL) cases. Thus, new agents that override the resistance are crucial to the development of curative therapies for APL. In this study, we investigated the effects of berbamine on the proliferation of APL cell line NB4 and its possible mechanisms.</p><p><b>METHODS</b>NB4 cells were treated with berbamine at different concentrations (0-64 microg/ml) for 72 hours. MTT assay was used to determine proliferation inhibition of NB4 cells. Cell apoptosis was evaluated by both flow cytometry (FCM) and morphological examination. PML/RAR-alpha and survivin mRNAs were measured by nested-RT-PCR and RT-PCR, respectively. Activated-caspase 3 was determined by FCM.</p><p><b>RESULTS</b>Berbamine greatly inhibited the proliferation of NB4 cells in dose- and time-dependent manners, and its IC50 value was 3.86 microg/ml at 48 hours. Both morphological observations and FCM results showed that berbamine induced apoptosis of NB4 cells with concomitant increase of activated caspase-3 and decrease of survivin mRNA. After treatment with berbamine at 8 microg/ml for 48 hours, the percentage of apoptotic cells increased from 2.83% to 58.44% (P<0.01), and the percentage of cells with activated-caspase 3 elevated from 2.06% to 70.89% (P<0.01), whereas, level of survivin mRNA was reduced to 38.24% of control (P<0.01). However, no significant change was observed in PML/RAR-alpha mRNA.</p><p><b>CONCLUSIONS</b>Berbamine induces caspase-3-dependent apoptosis of leukemia NB4 cells via survivin-mediated pathway, suggesting that berbamine may be a novel potential agent against APL with a mechanism distinct from that of all-trans retinoic acid (ATRA) and arsenic trioxide (ATO).</p>
Subject(s)
Humans , Alkaloids , Pharmacology , Antineoplastic Agents , Pharmacology , Apoptosis , Benzylisoquinolines , Pharmacology , Caspase 3 , Physiology , Cell Line, Tumor , Cell Proliferation , Dose-Response Relationship, Drug , Inhibitor of Apoptosis Proteins , Leukemia, Promyelocytic, Acute , Drug Therapy , Pathology , Microtubule-Associated Proteins , Genetics , Physiology , Neoplasm Proteins , Genetics , Physiology , Oncogene Proteins, Fusion , Genetics , Transcription, GeneticABSTRACT
<p><b>OBJECTIVE</b>To analyze the effects of surgical treatment for gastrointestinal stromal tumors (GISTs) and influential factors of survival.</p><p><b>METHODS</b>The clinical data and the tissue slices including immunohistochemical staining of 153 cases of GISTs from January 1990 to March 2006 were rechecked retrospectively. All patients were followed up carefully. More attention was paid to the surgical effects and the influential factors of survival.</p><p><b>RESULTS</b>The overall survival rates at 1-, 2-, 3-, 4- and 5-year were 94.9%, 83.3%, 73.3%, 70.5% and 64.3%, respectively. The median survival time for patients with tumor resected completely was 66.0 months, and the 2- and 5-year survival rate were 89.4% and 70.9% respectively. The median survival time was 23.8 months for the patients with tumor resected partly, and only two of these patients survived over 2 years. Gender, tumor sites, preoperative metastasis, tumor size, pathological type, karyokinesis and recurrence and metastasis were related with survival rates for the patients with tumor resected completely on univariate analysis, but tumor size, pathology type, recurrence and metastasis were related with survival rates on Cox regression multivariate analysis (P < 0.05).</p><p><b>CONCLUSIONS</b>Surgery should still be the main therapy for GISTs. Local complete resection is the principal treatment. The survival cannot be improved by extensive resection and lymph nodes clearance.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Antigens, CD34 , Follow-Up Studies , Gastrointestinal Stromal Tumors , Metabolism , Mortality , General Surgery , Immunohistochemistry , Kaplan-Meier Estimate , Proto-Oncogene Proteins c-kit , Retrospective Studies , Survival Rate , Treatment OutcomeABSTRACT
<p><b>BACKGROUND</b>It has been identified that in patients with pre-eclampsia, several factors were released into the serum, which can regulate the proliferation of vascular smooth muscle cells and stimulate the synthesis of extracellular matrix (ECM). However, the signal transduction pathway of the growth factors and cytokines synthesized by endothelial cells leading to the proliferation and ECM synthesis of human umbilical arterial smooth muscle cell (HUASMC) is unknown. The aim of this study was to research the effects of protein kinase C (PKC) inhibitor, polymyxin B (PMB), on the proliferation, nuclear factor-kappa B (NF-kappaB) activation, and expression of procollagen I and III mRNA in HUASMC cultured with pre-eclamptic umbilical sera.</p><p><b>METHODS</b>Normal HUASMCs were treated with pre-eclamptic umbilical serum (pre-eclamptic group), pre- eclamptic umbilical serum plus PMB (PMB group), or normal umbilical serum (normal group). The expression of I-kappaB, NF-kappaB was detected by Western blotting after the HUASMC was incubated for 2 hours. The proliferation of HUASMC was evaluated by MTT, the cell cycle was analyzed by flow cytomerty, and the expression of procollagen I, III mRNA was measured by RT-PCR assay after HUASMC was incubated for 48 hours. ANOVA was used for statistical analysis.</p><p><b>RESULTS</b>Umbilical sera in pre-eclampsia could stimulate the proliferation, the DNA synthesis, the transition of G(0) + G(1) phase or G(2)/M phase to S and phase, the activation of NF-kappaB, and the expression of procollagen I mRNA of HUASMC as compared with normal umbilical sera (P < 0.01). PMB could inhibit the proliferation, the DNA synthesis, the transition of G(0) + G(1) or G(2)/M phase phase to S phase, the activation of NF-kappaB, and the expressions of procollagen I mRNA of HUASMC stimulated by umbilical sera in pre-eclampsia (P < 0.01).</p><p><b>CONCLUSION</b>PKC-NF-kappaB signal transduction pathway may play a key role in SMC phenotype modulation, which is more important in the pathogenesis of placental blood vessel in pre-eclampsia.</p>
Subject(s)
Female , Humans , Pregnancy , Cell Cycle , Cell Proliferation , Cells, Cultured , Collagen Type I , Genetics , Collagen Type III , Genetics , Muscle, Smooth, Vascular , Cell Biology , Metabolism , Myocytes, Smooth Muscle , Metabolism , NF-kappa B , Physiology , Polymyxin B , Pharmacology , Pre-Eclampsia , Metabolism , Pathology , Protein Kinase C , Physiology , RNA, Messenger , Signal Transduction , Umbilical Arteries , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To determine effects of berbamine on the growth of leukemia cell line NB4 and explore its possible mechanisms.</p><p><b>METHODS</b>The growth of NB4 cells was examined with MTT assay. Morphological analysis and DNA agarose electrophoresis were used to detect apoptosis in NB4 cells, and the apoptosis rate was measured by flow cytometry. The PML/RAR alpha mRNA was determined by nested-PCR, and the Survivin mRNA was tested by RT-PCR. The expression of caspase 3 protein in NB4 cells was evaluated by flow cytometry.</p><p><b>RESULT</b>The growth of NB4 cells was inhibited significantly after treated with berbamine at different concentrations for different time points, the IC(50)value was 3.860 microg/ml at 48 hours. Morphology analysis showed the characteristics of apoptosis, and the DNA agarose electrophoresis showed the typical DNA ladder. The apoptosis rate increased from 2.83% to 58.44% after treated with berbamine at 12 microg/ml for 48 hours. The expression of PML/RAR alpha mRNA presented no significant changes, however, Survivin mRNA was decreased dramatically. The protein expression of Caspase 3 increased significantly from 2.06% to 70.89% after treated with berberine at a concentration of 12 mug/ml for 48 hours.</p><p><b>CONCLUSION</b>Berbamine could inhibit the growth of leukemia cell line NB4. The induction of cell apoptosis may be one of the mechanisms for suppressing the growth of leukemia cell line NB4. Inhibition of Survivin mRNA and upregulation of Caspase 3 protein might be also involved in cell apoptosis.</p>