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Objective To investigate the effect of ALA-PDT on the proliferation and apoptosis proteins of KFB.Methods Tissue adherent method was used for the culture of KFB in vitro.The cultured KFB was divided into four groups:the experimental group treated with ALA at 4.0 mmol/L concentration for 5 hours were irradiated by red laser with energy density at 64 J/cm2,and KFB survival was observed after ALA-PDT by CCK-8 method,and then the levels of caspase 3,caspase 8 and caspase 9 proteins were detected after irradiated by PDT.Results When the ALA concentration was 4 mmol/L and the laser energy was 64 J/cm2,the inhibitory effect on the cells was strongest in group A.ALA-PDT could obviously increase the activation of caspase 3,caspase 8 and caspase 9 protein levels and increase the levels of expression [(0.17±0.03) vs (0.58±0.01),(0.21 ±0.02) vs (0.69± 0.15),and (0.13±0.15) vs (0.15±0.02),P＜0.05],respectively.Conclusions ALA-PDT can obviously inhibit the proliferation of KFB.Concentration and laser energy are important factors affecting cell survival.ALA-PDT can significantly increase the activity of caspases 3,8 and 9 proteins in KFB.
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Aim To investigate the anti-angiogenesis and anti-xenograftes of UA in zebrafish larvaes. Meth-ods 24 hour post-fertilization ( hpf ) TG zebrafish was treated with different concentration of UA for 24h, and the number of intersegmental vessels( IVS) were detec-ted under fluorescent microscope respectively;then the models of AB/TG zebrafish xenografts were established by be micro-injected with SMMC-7721 or HT-29 cell at 48hpf respectively, and after cocultured with UA for 48h, optical density (OD) of the SMMC-7721 cell and subintestinal veins ( SIVs) induced by HT-29 were e-valuated under confocal microscope. Results ISV as-say showed that UA could cause IVS missing or disap-perance, inhibition ratio reaching 20. 25% and 26. 65%. UA blocked the spread of SMMC-7721 cells in zebrafish and OD value,and inhibition ratios at three concentrations were 38. 01%, 54. 69%, 61. 88%, re-spectively; in another SIVs assay induced by xeno-grafts, UA at concentration 10 and 15mg·L-1 showed that SIVs were inhibited (P<0. 01) obviously. Con-clusion UA could inhibit the angiogenesis and the growth of SMMC-7721/HT-29 xenografts,and the anti-tumor mechanism may be related with VEGFR2 expres-sion.
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Objective The aim of the study was to detect the mutation of Fms-like tyrosine kinase 3 internal tandem duplication (FLT3-1TD) rate in older de novo acute myeloid leukemia (AML) patients, to evaluate the role of FLT3-ITD in AML and its clinical significance. Methods The mutations of FLT3-1TD in bone marrow mononuelear cells (MNCS) from 30 cases of older AML were screened by polymerase chain reaction denaturing-high performance liquid chromatography (PCR-DHPLC). Results FLT3-1TD mutations were identified in 26.67 %(8/30) patients, while there were no mutations identified in control cases. And these kinds of mutations were likely to attend in M3 types. All mutations of FLT3-ITD were heterozygous and rearrangement fragment located in reading frame. Different karyomite groups had different FLT3-ITDmutations rate. We could see that FLT3-ITD positive patients were more prevalent in patients with normal karyotype. Clinical researches indicated that FLT3-ITD mutations had the characteristics of a higher peripheral white cell count, higher blast cells and lower complete remission rate in older AMKA Conclusion FLT3-ITD positive older AML patients conferred a poor prognosis and were likely to attend in normal karyomite group. The detection of FLT3-ITD mutations could make up for the deficiency of cytogenetics to some extent, and may become a routine examination of AML in older, which can direct their treatment and predict their prognosis.
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Objective To examine the expression levels of cyclin Dl in the patients with chronic myelogenous leukemia (CML), and evaluate the pathogenesis and clinical significance of cyclin Dl in CML Methods The real-time quantitative polymerase chain reaction (RQ-PCR) was performed to detect the expression levels of cyclin Dl in the bone marrow samples of 18 patients with CML, and 16 samples of benign hemopoietic patients. The relationship between the expression levels of cyclin Dl and the progression and prognosis of patients with CML were analyzed. Results The level of cyclin Dl was higher expressed in 18 patients with CML than the control group (P <0.001). The levels of cyclin Dl was apparently higher expressed in accelerated phase /blast crisis phase than in chronic phase (P <0.05). And the RQ-PCR method showed the tendency that a significant increase was observed in the levels of cyclin Dl from 0.1980 in control group to 1.4002 in chronic phase and 5.4540 in accelerated phase /blast crisis phase. Conclusion The cyclin Dl overexpressed in CML, the roles of cyclin Dl in CML might be an oncogene expressed. The expression level is correlated with the progression and prognosis of patients with CML.
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Objective To study the quality control method for Chanshukang granule. Methods TLC and HPLC method were used to identify the Chinese herbal medicines. The determination was carried on HPLC, using Kromasil C18 column (200 mm?4.6 mm, 5 ?m), ethanol-2% glacial acetic acid (40∶60) as mobile phase and detected at the wavelength of 324 nm. Results The Chinese herbal medicines, Radix Angelicae Sinensis, Rhizoma Chuanxiong, Placenta Hominis and Radix Glycyrrhizae Preparata, in granules were identified by TLC. The calibration curve was linear in the range of 0.625～3.750 ?g for ferulic acid. The contents in three batches were 0.054%, 0.037%, 0.042% respectively. The average recovery was 96.2% and RSD=2.37%. Conclusion The TLC method can be used for the quality control of four herbal medicines in the granule. The HPLC method is simple, sensitive, accurate and provides a certain evidence for controlling the quality of Chanshukang Granule.
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Objective To establish an HPLC method for the determination of isopimaric acid in the leaves of Platycladus orientalis and its preparations Cebai No.V Capsula and Isopimaric Acid Capsula to evaluate the quality of them. Methods HPLC Method was developed. Kromasil C18(200 mm?4.6 mm, 5 ?m) column was used and the mobile phase: methyl alcohol-water-acetic acid (60∶37∶3), float rate: 1 mL/min, temperature: 25 ℃, wavelength: 231 nm, injected volume: 10 ?L. Content of isopimaric acid in samples was calculated by external standard. Results The linearity relationship of isopimaric acid was good in the range of 1.25-10.00 ?g. The equation of standard curve: Y=66 214 X+2 487.9, r=1.000. The average recovery was 95.8%, RSD=2.7% (n=5). The determination results of isopimaric acid in three batches of P. orientalis were 3.09, 3.20, and 3.56 mg/g; three batches of Cebai No.V Capsula were 10.39, 9.67, and 10.99 mg/g; three batches of Isopimaric Acid Capsula were 553.89, 541.50, and 528.83 mg/g. Conclusion The method is reliable, simple, and precise, which could be used for the quality control of P. orientalis and its capsules.