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The quality difference of pharmaceutical excipients from different sources affects the molding properties of the powder, resulting in changes in the properties of the final product. In this study, the critical quality attributes of hydroxypropyl methylcellulose (HPMC) with different specifications from two manufacturers (manufacturer A and manufacturer B) were characterized including particle size, physical morphology, viscosity and powder physical quality attributes. Aminophylline, diclofenac sodium, and metformin hydrochloride were utilized as model drugs with different solubility to prepare sustained-release tablets, and the effect of HPMC from different sources on drug release of sustained-release tablets in vitro was investigated. The results showed that HPMC with the same viscosity specification from different sources had outstanding differences in the physicochemical properties (including particle size, physical morphology, viscosity, dimension, compressibility and powder flow), which could change the hardness and friability of the sustained-release tablets. The differences in the physicochemical properties of HPMC had different effects on the dissolution of different sustained-release tablets in vitro. It had no significant effect on the release of easily soluble aminophylline and metformin hydrochloride, but had a greater impact on the release of poorly soluble diclofenac sodium. Compared with manufacturer A, the sustained-release effect of matrix tablets prepared by HPMC from manufacturer B was more excellent. The results of this study will provide a theoretical reference on selecting the appropriate excipients for formulation design.
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Objective:To investigate the differences in matrix metalloproteinases (MMPs) and matrix metalloproteinase inhibitors (TIMPs) from diseased splenic vein (DSVs) and varicose great saphenous vein (VGSVs) under high hemodynamics.Methods:Seventy-two specimens of DSVs,normal splenic veins (SVs),VGSVs,and normal great saphenous vein (GSVs) were collected.Venous wall in the four groups,MMP-2,MMP-9,TIMP-1,and TIMP-2 protein expression were observed and MMP-2,MMP-9,TIMP-1,TIMP-2 proteins positive expression ratio and mRNA expression were determined.Results:DSVs and VGSVs in the two groups,MMP-2,MMP-9,TIMP-1,TIMP-2 proteins with clustered strong expression were observed;In DSVs group,MMP-2,MMP-9,TIMP-1,TIMP-2 protein positive expression ratio and mRNA expression were significantly increased compared with SVs group,while in VGSVs group,MMP-2,MMP-9,TIMP-1,TIMP-2 protein positive expression ratio and mRNA expression were significantly increased compared with GSVs group (P＜0.05).VGSVs/GSVs ratio was significantly increased compared with DSVs/SVs ratio (P＜0.05).Conclusion:Under high hemodynamics,the dysequilibrium of MMPs and TIMPs from splenic vein and great saphenous vein,These results may be one of the molecular mechanism in vascular remodeling.
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A strategy based on immunomagnetic nanospheres ( IMNs ) for rapid, efficient and accurate detection of lymphnode metastasis carcinoma cells ( LNMCCs ) was developed in this study. First, IMNs processing magnetism and biological targeting were fabricated by the approach developed by our group previously. Then, LNMCCs in lymph node fine needle aspiration ( LNFNA) specimens were separated and enriched by the immunomagnetic isolation using IMNs. At last, the captured cells were identified with Wright's staining and immunocytochemistry ( ICC) . The separation and enrichment of LNMCCs with immunomagnetic isolation could reduce the background interference of LNFNA specimens effectively; the identification with Wright ' s staining and ICC offered more reliable information for accurate diagnosis, so the sensitivity, specificity and overall diagnostic accuracy had an obvious improvement compared with the conventional cytologic diagnosis. Besides, the simple and rapid incubation of LNFNA specimens with IMNs needed just 5 min, so the cytomorphology of captured LNMCCs could be intactly retained, which enabled to provide important basis for classifying lymphnode metastasis carcinoma ( LNMC ) and the subsequent pathological study. Moreover, the specific capture of epithelial carcinoma cells in LNFNA specimens with IMNs could make a definite diagnosis of the captured cells as LNMCCs, thus realizing the differentiated diagnosis of LNMC and malignant lymphoma. Additionally, this strategy exhibited successful LNMCCs detection in LNFNA specimens from 110 patients and had higher sensitivity ( 98 . 0%) , specificity ( 100 . 0%) , and overall diagnostic accuracy (98. 2%) than the conventional cytologic diagnosis. Therefore, it was a new attempt to use IMNs for detection of LNMCCs in LNFNA specimens from LNMC patients, and offered new ideas for LNMC diagnosis and study.
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To synthesize and characterize a novel metal complex of Mn (II) with emodin, and evaluate its anti-cancer activity. The elemental analyses, IR, UV-vis, atomic absorption spectroscopy, TG-DSC, (1)H NMR, and (13)C NMR data were used to characterize the structure of the complex. The cytotoxicity of the complex against the human cancer cell lines HepG2, HeLa, MCF-7, B16, and MDA-MB-231 was tested by the MTT assay and flow cytometry. Emodin was coordinated with Mn(II) through the 9-C=O and 1-OH, and the general formula of the complex was Mn(II) (emodin)2·2H2O. In studies of the cytotoxicity, the complex exhibited significant activity, and the IC50 values of the complex against five cancer cell lines improved approximately three-fold compared with those of emodin. The complex could induce cell morphological changes, decrease the percentage of viability, and induce G0/G1 phase arrest and apoptosis in cancer cells. The coordination of emodin with Mn(II) can improve its anticancer activity, and the complex Mn(II) (emodin)2·2H2O could be studied further as a promising anticancer drug.
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Therapeutic Uses , Antineoplastic Agents, Phytogenic , Pharmacology , Therapeutic Uses , Emodin , Pharmacology , Therapeutic Uses , HeLa Cells , Hep G2 Cells , MCF-7 Cells , Manganese , Pharmacology , Therapeutic Uses , Melanoma, Experimental , Drug Therapy , Molecular Structure , Neoplasms , Drug Therapy , Phytotherapy , Plant Extracts , Pharmacology , Therapeutic Uses , Polygonaceae , ChemistryABSTRACT
The aim of this paper is to develop a rapid and highly sensitive quantitative HPLC fingerprint method with multiple indicators by using the Compound Chinese Medicine Wuwei Changyanning granule and 5 herbs in the prescription. The quantitative fingerprint chromatogram with multiple indicators was investigated. i]6 compositions included rutin, gallic acid, chlorogenic acid, atractylenolide I, pachymic acid and apigenin, which originated from 5 herbs respectively, were selected as quantitative compositions, and their contents were determined using HPLC from 11 batches granules and the corresponding 5 medicinal materials. ii] The precision, stability and repeatability of fingerprinting were investigated. In addition, common peaks number, the percentage of non-common peaks and similarity were also studied. Among them, 21 common peaks in the granule could find the source of peaks from the 5 herbs, among of 10 peaks from Niuerfeng, 9 peaks from Laliao, 3 peaks from Baishu, 3 peaks from Fuling and 5 peaks from Guanghuoxiang. The results showed that the identification method of fingerprinting was reliable
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This study examined the expression of brain-derived neurotrophic factor (BDNF) in multiple myeloma (MM) and its role in bone marrow angiogenesis. The peripheral blood plasma was harvested from 71 MM patients and 63 patients without hematological malignancy. The BDNF level in the blood plasma was determined by ELISA. Human bone marrow endothelial cells (HBMECs) were cultured. The mRNA and protein expression levels of the BDNF receptor TrkB in HBMECs were detected by using RT-PCR and flow cytometry, respectively. The viability of HBMECs treated with recombinant human (rh) BDNF or not was measured by using MTT assay. The migration of HBMECs in the presence of rhBDNF or not was determined by modified Boyden chamber assay. In vitro tube formation assay was used to assess the effect of rhBDNF on HBMECs differentiation. The results of ELISA revealed that the BDNF level was significantly higher in peripheral blood plasma of MM patients than in that of control patients (4.39±0.67 vs. 1.96±0.39 ng/mL, P<0.05). The BDNF receptor TrkB was expressed in HBMECs at mRNA and protein level. MTT assay manifested that rhBDNF could significantly concentration-dependently promote the HBMECs proliferation. The number of HBMECs treated with 160 ng/mL rhBDNF for 48 h was 1.57±0.10 folds higher than that in control group (P<0.05). Moreover, rhBDNF could enhance HBMECs migration in a concentration-dependent manner and the maximal migration was reached in the presence of 100 ng/mL rhBDNF. The migration indexes were 1.40±0.11, 1.64±0.16, 2.06±0.25 and 2.18±0.21 in 25, 50, 100 ng/mL rhBDNF groups and 25 ng/mL rhVEGF group, respectively. In vitro tube formation assay demonstrated that the area of the formed tubular structure was increased with the rhBDNF concentration. In control group, there was no formation of intact tubular structure and the HBMECs on the matrigel were irregularly dispersed. HBMECs treated with 100 ng/mL rhBDNF could form intact tubular structure and the area and the diameter of tubes were significantly greater than those in control group (P<0.05). There was no significant difference in the formed tubular area between 25 ng/mL VEGF group and 100 ng/mL rhBDNF group. It was concluded that BDNF plays an important role in myeloma cell-induced angiogenesis, and it may become a new target of anti-angiogenesis treatment for MM.
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Objectives To explore the clinical features, diagnosis and treatment of Crohn’s disease complicated by erythromelalgia (EM) in a pediatric case. Methods The clinical manifestation, results of laboratory testing and endoscopy, mutational analysis of the SCN9A gene, and the follow-up record were collected and analyzed based on review of literature to a thirteen-year-old girl with Crohn’s disease complicated by erythromelalgia. Results The patient experienced symptoms of anorexia, fatigue, diarrhea, dark red and swelling skin, increased skin temperature and burning pain in her both lower extremities during the course of disease. The endoscopic ifndings included multiple ulcerations and polypoid protrusion lesion in colon, and the pathological examination showed the local abscess formation in colonic mucosa. The mutation in SCN9A gene of the child was excluded by gene analysis. The symptoms were gradually ameliorated after treatment using prednisone and mesalazine combined with dipyridamole and low-molecular-weight heparin calcium. Conclusions Crohn’s disease complicated by erythromelalgia is rare. The pathogenesis may relate to immune factors, thrombocytosis, and hyper-coagulable states, etc. The combination of glucocorticoids, 5-aminosalicylic acid and anticoagulants may lead to a better therapeutic effect.
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This study examined the expression of brain-derived neurotrophic factor (BDNF) in multiple myeloma (MM) and its role in bone marrow angiogenesis. The peripheral blood plasma was harvested from 71 MM patients and 63 patients without hematological malignancy. The BDNF level in the blood plasma was determined by ELISA. Human bone marrow endothelial cells (HBMECs) were cultured. The mRNA and protein expression levels of the BDNF receptor TrkB in HBMECs were detected by using RT-PCR and flow cytometry, respectively. The viability of HBMECs treated with recombinant human (rh) BDNF or not was measured by using MTT assay. The migration of HBMECs in the presence of rhBDNF or not was determined by modified Boyden chamber assay. In vitro tube formation assay was used to assess the effect of rhBDNF on HBMECs differentiation. The results of ELISA revealed that the BDNF level was significantly higher in peripheral blood plasma of MM patients than in that of control patients (4.39±0.67 vs. 1.96±0.39 ng/mL, P<0.05). The BDNF receptor TrkB was expressed in HBMECs at mRNA and protein level. MTT assay manifested that rhBDNF could significantly concentration-dependently promote the HBMECs proliferation. The number of HBMECs treated with 160 ng/mL rhBDNF for 48 h was 1.57±0.10 folds higher than that in control group (P<0.05). Moreover, rhBDNF could enhance HBMECs migration in a concentration-dependent manner and the maximal migration was reached in the presence of 100 ng/mL rhBDNF. The migration indexes were 1.40±0.11, 1.64±0.16, 2.06±0.25 and 2.18±0.21 in 25, 50, 100 ng/mL rhBDNF groups and 25 ng/mL rhVEGF group, respectively. In vitro tube formation assay demonstrated that the area of the formed tubular structure was increased with the rhBDNF concentration. In control group, there was no formation of intact tubular structure and the HBMECs on the matrigel were irregularly dispersed. HBMECs treated with 100 ng/mL rhBDNF could form intact tubular structure and the area and the diameter of tubes were significantly greater than those in control group (P<0.05). There was no significant difference in the formed tubular area between 25 ng/mL VEGF group and 100 ng/mL rhBDNF group. It was concluded that BDNF plays an important role in myeloma cell-induced angiogenesis, and it may become a new target of anti-angiogenesis treatment for MM.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Bone Marrow , Metabolism , Pathology , Brain-Derived Neurotrophic Factor , Metabolism , Multiple Myeloma , Metabolism , Pathology , Neovascularization, Pathologic , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To explore an innovative technique that is aided by multi-disciplinary hybrid approach in identification and treatment of tracheoesophageal fistula (TEF) in children intraoperatively.</p><p><b>METHOD</b>From April 2008 to October 2011, 4 patients with isolated TEF were presented with 2 H-type fistulas and 2 recurrent TEF. For all the four cases, with the cooperation of the gastroenterologists, respiratory physician and surgeon, methylene blue was first injected into the trachea for detecting the dye in the esophagus by the gastroscopy. Bronchoscopy was performed where the fistula tract was shown by the methylene blue and a guide wire was passed through the fistula. The patients underwent rigid gastroscopy and the guide wire was identified and brought out through the mouth by biopsy pliers. This created a wire loop through the fistula. X-ray was then used to identify the level of the fistula. According to the level of the fistula it was determined whether surgical incision and approach should be used. The fistula was then repaired successfully by surgery.</p><p><b>RESULT</b>In the 4 patients, with the aid of gastroscopy and bronchoscopy, identification of the fistula intraoperatively was then facilitated by traction on the loop. The fistula was identified and repaired. There were no fistula recurrences.</p><p><b>CONCLUSION</b>Multi-disciplinary hybrid therapy for tracheoesophageal fistula in children is beneficial for the precise localization of the fistula. This new technique is an effective and definitive method in identification and treatment of TEF in children.</p>
Subject(s)
Child, Preschool , Female , Humans , Bronchoscopy , Methods , Gastroscopy , Methods , Minimally Invasive Surgical Procedures , Methods , Patient Care Team , Retrospective Studies , Suture Techniques , Tracheoesophageal Fistula , Diagnosis , General Surgery , Treatment OutcomeABSTRACT
Objective To investigate the mechanical mechanism of bypass graft for the treatment of DeBakey III aortic dissection and explore the valid surgical planning. Methods Patient-specific models of DeBakey III aortic dissection, including the models of through lumen and blind lumen, before and after bypassing between ascending aorta and abdominal aorta, between left subclavian artery and abdominal aorta, were constructed, and then numerical simulations were performed using computational fluid dynamics (CFD) method under physiological flow conditions based on fluid-structure interaction (FSI). Results Blood flow velocity, pressure, vessel wall displacement of the false lumen after bypass graft were reduced by 38.86%, 15.347 kPa and 39.46% on average, respectively. Conclusions Bypass graft is an effective surgical method for the treatment of DeBakey III aortic dissection under specific conditions with good prospects in clinical application.
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<p><b>OBJECTIVE</b>To study the effects of vitamin A on the development of T lymphocytes in peripheral blood and small intestine and on the cytokine response of intestinal mucosa in mice.</p><p><b>METHODS</b>Twenty young mice were randomly fed with forage containing vitamin A 250 or 4 IU/g (n=10 each). Three weeks later, the levels of CD4+ CD25+ T subsets in peripheral blood and intestinal mucosa were measured by flow cytometry. The levels of cytokines IFN-γ, IL-4, IL-17 and IL-23 in stool were measured using ELISA.</p><p><b>RESULTS</b>The levels of CD4+ CD25+ T subsets in peripheral blood and intestinal mucosa in the 250 IU/g vitamin A group were significantly higher than those in the 4 IU/g vitamin A group (P<0.05). The IL-4 level in stool increased, in contrast, the IL-23 level in stool decreased significantly in the 250 IU/g vitamin A group when compared with the 4 IU/g vitamin A group (P<0.05).</p><p><b>CONCLUSIONS</b>vitamin A may promote the development of CD4+ CD25+ T lymphocytes in peripheral blood and small intestine. Moreover, it may be involved in intestinal mucosa-associated immune response by regulating cytokines IL-4 and IL-23.</p>
Subject(s)
Animals , Mice , CD4-Positive T-Lymphocytes , Cytokines , Flow Cytometry , Interleukin-4 , Intestinal Mucosa , T-Lymphocytes, Regulatory , Vitamin AABSTRACT
Aortic dissection is a common disease which is very dangerous,with high mortality rate.Bypass graft for the treatment of DeBakey Ⅲ dissection has outstanding advantages than the ordinary replacement of thoracic aorta,and some patients will inevitably require the use of the procedure.The purpose of this study was to explore the impact of the subclavian artery-abdominal aorta bypass graft on hemodynamic parameters in the false lumen and the effectiveness of surgical treatment.First of all,the idealized three-dimensional geometric models of DeBakey Ⅲ aortic dissection and its subclavian artery-abdominal aorta bypass graft operation were constructed,respectively.Then the models were imported into ANSYS 11.0 for finite element analysis.Results of numerical simulation showed that both velocity and pressure of the blood flow were reduced after bypass graft at the entrance and in the internal sac of false lumen,which is very favorable for reducing the impact of blood flow on false lumen,slowing down the further expansion of entrance,preventing the breakdown of false lumen,and promoting the healing of dissection.Therefore,the subclavian artery-abdominal aorta bypass graft operation is an effective surgical method for the treatment of DeBakey Ⅲ aortic dissection in some particular cases.This operation is with great prospects for clinical application.
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<p><b>OBJECTIVE</b>This study aimed to investigate the value of the liver function test in the differential diagnosis of infantile hepatitis syndrome (IHS) and biliary atresia (BA) by analyzing seven conventional serological markers in this test using receiver operating characteristic (ROC) curves.</p><p><b>METHODS</b>Serum levels of seven conventional serological markers: alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (gamma-GT), alkaline phosphatase (ALP), total bilirubin (TB), conjugated bilirubin (CB) and serum albumin (ALB) were measured in 103 children with IHS and 60 children with BA. ROC curves were used to evaluate the sensitivity, specificity, positive and predictive values and optimal cut-off. The united tests (parallel test and serial test) of gamma-GT, TB and CB were performed to elevate diagnostic efficiency.</p><p><b>RESULTS</b>Compared with the IHS group, the BA group had significantly increased serum ALT, AST, gamma-GT, TB and CB levels (p<0.01). The area under ROC (AUCROC) of AST, gamma-GT, CB and TB was 0.77, 0.881, 0.841 and 0.87, respectively. gamma-GT showed the highest AUCROC, specificity, positive predictive value and positive likelihood ratio in the diagnosis of BA, followed by CB, TB and AST in turn. The negative predictive value of CB was the highest, followed by TB. The negative likelihood ratio of CB was the lowest but its Youden index was the highest. The Youden index of gamma-GT and TB was lower than that of CB. After the parallel tests, the sensitivity and negative predictive value of gamma-GT, CB and TB increased to 100%. After the serial tests, the specificity of gamma-GT, CB and TB increased to 90.4% and the positive predictive value increased to 87.5%.</p><p><b>CONCLUSIONS</b>The measurement of gamma-GT, TB and CB levels are valuable in the differential diagnosis of BA and IHS. An imaging examination is required in the parallel test positive patients.</p>
Subject(s)
Female , Humans , Infant , Infant, Newborn , Male , Alanine Transaminase , Blood , Aspartate Aminotransferases , Blood , Biliary Atresia , Blood , Diagnosis , Bilirubin , Blood , Diagnosis, Differential , Hepatitis , Blood , Diagnosis , Liver Function Tests , ROC Curve , gamma-Glutamyltransferase , BloodABSTRACT
The aircraft industry is crucial to the economy and security of a nation. In this paper, the spatial characteristics and patterns of the aircraft industry are analyzed on different spatial scales. It is found that there is a 'Matthew effect' in the global aircraft industry and the spatial evolution of the industry is consistent with the industrialization process of the whole country. It is also revealed that the spatial evolution of the country is driven by both the centripetal forces including capital, talents, technology and agglomeration economies and the centrifugal forces including the comparative advantage, cost &risk sharing, emerging markets, development policy for less-developed regions and the military imperative. These forces have both market-stabilizing and market-disrupting effects on the spatial evolution of the aircraft industry. The study suggests that lessons drawn from the experiences in the United States and France are expected to be conducive to the rise of China's aircraft industry in the future.
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<p><b>OBJECTIVE</b>To screen effective sequences of short hairpin RNA on brain-derived neurotrophic factor (BDNF) gene and the effect of RNA interference on the proliferation and apoptosis of HeLa cells, a cervix carcinoma cell line with high expression of BDNF.</p><p><b>METHODS</b>Two recombinant eukaryotic human-BDNF siRNA expression vectors were designed and constructed. Sequences were confirmed by restrictive endonuclease digestion and DNA sequencing. The empty vector pGenesil-1 and two recombinant plasmids, pGenesil-shRNA-BDNF1 and pGenesil-shRNA-BDNF2, were transfected into HeLa cells using Lipofectamine 2000 (groups: P(0), P(1) and P(2), respectively). The mRNA and protein levels of BDNF in HeLa cells were detected by RT-PCR and Western blot, respectively. The cellular proliferation rates were determined by MTT assay and the apoptotic rates were measured by flow cytometry and Hoechest 33258.</p><p><b>RESULTS</b>The recombinant eukaryotic BDNF siRNA expression vectors were successfully constructed. The expression of mRNA and protein of BDNF in P(1) group were significantly decreased, comparing with non-transfected group, P(0) and P(2) groups (F = 48.19, P < 0.01). P(2) group failed to meet the expected results (P > 0.05). In addition, the proliferation activity was reduced in P(1) group and the peak point of proliferation curve was prolonged. Moreover, the early cell apoptotic rates were statistically increased in P(1)[(53.4 +/- 4.2)%] VS. non-transfected [(0.8 +/- 0.4)%], P(0) [(5.1 +/- 1.8)%] and P(2)[(7.9 +/- 2.4)%] groups (F = 269.77, P < 0.01).</p><p><b>CONCLUSION</b>HeLa cells express a high level of BDNF. BDNF gene silencing by RNA interference increases the apoptosis of HeLa cells and inhibits cell proliferation, offering a possible target for efficient tumor therapy.</p>
Subject(s)
Humans , Apoptosis , Brain-Derived Neurotrophic Factor , Genetics , Cell Proliferation , Gene Expression Regulation, Neoplastic , Genetic Vectors , HeLa Cells , RNA Interference , RNA, Messenger , Metabolism , RNA, Small Interfering , Genetics , Recombinant Proteins , Genetics , Metabolism , TransfectionABSTRACT
Lentiviral vectors (Lv), known as holding lots of virtues (e.g. transfection to the dividing or non-dividing cells;large capacity of transfer gene fragments; long-term expression transfer genes; and low rate of immunological response) has come to be one of the hot-spots in gene therapy research. In this paper, taking the Lv derived from HIV-1 for example, we review the characteristics of frame, the developments and the advances in application of Lv.
Subject(s)
Animals , Humans , Animals, Genetically Modified , Genetic Therapy , Methods , Genetic Vectors , Genetics , HIV-1 , Genetics , RNA, Small Interfering , Genetics , Transduction, Genetic , MethodsABSTRACT
An automatic inspection system for biochip's print quality is presented in this paper. It consists of an automatic mechanical control, a CCD sensor for getting the image of PET boart, and the special computer software for image processing and recognition. Experimental results indicate that this system is capable of providing a precise and effective realtime inspection for biochips' print quality.
Subject(s)
Biosensing Techniques , Methods , Equipment Design , Image Processing, Computer-Assisted , Methods , Microchip Analytical Procedures , Methods , Pattern Recognition, Automated , Methods , Quality Control , SoftwareABSTRACT
<p><b>OBJECTIVE</b>To explore the significance of abnormal expression of brain-derived neurotrophic factor (BDNF)/TrkB in the development and evolution of multiple myeloma (MM) and the involved signaling pathways.</p><p><b>METHODS</b>The effect of BDNF on the cell viability of human myeloma cell line (HMCL) (RPMI8226, U266, KM3) was determined by trypan blue dye-exclusion. MTT assay was used to evaluate the cytotoxicity of tested chemotherapeutic agents. The effect of BDNF on the phosphorylation of TrkB was determined by Western blot. A human myeloma xenograft animal model was used to evaluate the effects of BDNF on tumor growth and survival time.</p><p><b>RESULTS</b>BDNF at 50 microg/L triggered significant increase in cell viability of HMCL. BDNF protected KM3 cells from melphalan and vincristine. The viability of KM3 cells exposed to varying concentrations of melphalan with and without 50 microg/L BDNF showed that BDNF induced almost a 2-fold and a 3-fold increase in melphalan and vincristine toxicity respectively. BDNF treatment increased MM cell growth in xenografted MM model (3240.9 mm3 vs 1032.7 mm3 ) (P <0.05). Intratumoral injection of BDNF also significantly reduced survival time (13 d vs 21 d) (P <0.05). The phosphorylated TrkB level was increased significantly after treated by exogenous BDNF. BDNF-triggered migration in RPMI8226 cells was completely abrogated by a Trk tyrosine kinase inhibitor K252a.</p><p><b>CONCLUSION</b>BDNF can activate TrkB signaling cascades resulting in MM cells growth, migration, and chemoprotection and appears to have a major contribution to the pathogenesis of MM.</p>
Subject(s)
Animals , Humans , Mice , Brain-Derived Neurotrophic Factor , Pharmacology , Cell Line, Tumor , Cell Proliferation , Cell Survival , Mice, Inbred BALB C , Multiple Myeloma , Metabolism , Pathology , Phosphorylation , Receptor, trkB , Metabolism , Signal Transduction , Xenograft Model Antitumor AssaysABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of brain-derived neurotrophic factor (BDNF) promoting human multiple myeloma (MM) cells secreting matrix metalloproteinase-9 ( MMP-9).</p><p><b>METHODS</b>Gelatin zymography of culture supernatants was performed to visualize the content of MMPs in myeloma RPMI 8226 cells stimulated by BDNF. NF-kappaB activity was determined by chemiluminescent electrophoretic mobility shift assay (EMSA).</p><p><b>RESULTS</b>Treatment with 25, 50, 100 and 200 microg/L BDNF for 24 h significantly (P < 0.01) enhanced the level of MMP-9 (2.03+/-0.48, 2.99+/-0.046, 4.63+/-0.62 and 5.62+/-1.29 microg/L, respectively, vs 1.00 microg/L of the control) secreted by RPMI8226 cells in a dose-dependent manner, while that of MMP-2 was not changed significantly (P > 0.05). The BDNF-induced activation of MMP-9 was inhibited by pretreatment with pyrrolidine dithiocarbamate (PDTC), a NF-kappaB inhibitor, or K252 alpha, a specific tyrosine inhibitor of TrkB which is the receptor for BDNF. Pretreated with 1 mmol/L PDTC or 500 nmol/L K252 alpha significantly downregulated MMP-9 secreted by the 100 microg/L of BDNF stimulated RPMI 8226 cells (the optical density values were 867.52+/-101.81 and 727.98 +/-92.05, respectively, vs 1,159.01+/-233.15 of the control). The activity of NF-kappaB was enhanced by BDNF in a dose-dependent manner, and pretreatment with K252 alpha could significantly inhibit this activation at 1, 6, 12 and 24 h (P < 0.05) in a time-dependent manner.</p><p><b>CONCLUSION</b>BDNF plays an important role in the angiogenesis of MM to promote the up-regulation of MMP-9, which may be induced by enhanced NF-kappaB activity in MM cells.</p>
Subject(s)
Humans , Brain-Derived Neurotrophic Factor , Pharmacology , Cell Line, Tumor , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Multiple Myeloma , Metabolism , NF-kappa B , MetabolismABSTRACT
<p><b>BACKGROUND</b>In multiple myeloma (MM), bone marrow angiogenesis parallels tumour progression and correlates with disease activity. Recent studies have proved resveratrol possesses antiangiogenic activity in vitro and in vivo. In this study, we examined the effects of resveratrol on myeloma cell dependent angiogenesis and the effects of resveratrol on some important angiogenic factors of RPMI 8226 cells.</p><p><b>METHODS</b>RPMI 8226 cells were cocultured with human umbilical vein endothelial cells (HUVECs) to evaluate the effects of myeloma cells on angiogenesis. The RPMI 8226 cells were treated with various concentrations of resveratrol (6.25 - 50.00 micromol/L) for different times (12 - 72 hours). Reverse transcriptase polymerase chain reaction (RT-PCR) was used to assay vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), metalloproteinases (MMP)-2 and MMP-9 mRNA. Gelatin zymography was used to analyze MMP-2 and MMP-9 activity. VEGF and bFGF proteins secreted by the cells in the medium were quantified by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Cell proliferation, migration and differentiation of HUVECs markedly increased by coculture with RPMI 8226 cells. Resveratrol inhibited proliferation, migration and tube formation of HUVECs cocultured with myeloma cells in a dose dependent manner. Treatment of RPMI 8226 cells with resveratrol caused a decrease in MMP-2 and MMP-9 activity. Resveratrol inhibited VEGF and bFGF protein expression in a dose and time dependent manner. Furthermore, decreased levels of VEGF, bFGF, MMP-2 and MMP-9 mRNA from cells treated with various concentrations of resveratrol confirmed its antiangiogenic action at the level of gene expression.</p><p><b>CONCLUSIONS</b>Resveratrol inhibits multiple myeloma angiogenesis by regulating expression and secretion of VEGF, bFGF, MMP-2 and MMP-9. Resveratrol may be a potential candidate for the treatment of multiple myeloma.</p>