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OBJECTIVE To study the mechanism of curcumol inhibiting the pro liferation of breast cancer cells T 47D. METHODS MTT assay was used to detect the inhibitory effects of different doses of curcumol （0，6.25，12.5，25，50，100 μg/mL）on the proliferation of T 47D cells. After treated with curcumol （12.5，25，50，100 μg/mL），the morphology of T 47D cells was observed by inverted phase contrast microscope. The cell cycle and the levels of reactive oxygen species （ROS）were detected by flow cytometry. Quantitative real-time PCR （qRT-PCR）was used to detect the expressions of proliferating cell nuclear antigen （PCNA），cell cycle regular p 21 and cyclin-dependent kinase 2（CDK2）mRNA. Western blot assay was used to detect the protein expression of CDK 2，CDK6，Cyclin D ，PCNA，nucler transcription factor E 2-related factor （Nrf2）and Kelch-like ECH associated protein 1（Keap1）. Breast cancer cells T 47D were divided into 2 groups，one group was given different doses of curcumol ，and another group was given curcumol 33 μg/mL for 6，12，24，48 h. After the optimal oxidation time and administration concentration were determined according to the results of the above two groups ，the blank control group ，N-acetylcysteine（NAC）group（ROS antioxidant NAC alone ），curcumol group （curcumol alone ），curcumol combined with NAC group （pretreatment with ROS antioxidant NAC ，and then adding into curcumol ）. Cell cycle and fluorescence intensity of ROS were detected. RESULTS Curcumol could significantly increase the inhibitory rate of the proliferation of T 47D cells （P＜0.05 or P＜0.01），and showed a certain dose and time dependent trend. Curcumol blocked the ， cycle in the G 1 phase and significantly increased the level of ROS （P＜0.05 or P＜0.01）；ROS antioxidant NAC could significantly reverse above inductive effect of curcumol （P＜ 0.01）. qRT-PCR showed that curcumol down-regulated the com expression of PCNA and CDK 2 mRNA and up-regulated the expression of p 21 mRNA（P＜0.05 or P＜0.01）. Western blot assay showed that curcumol significantly down-regulated the edu.cn protein expression of Keap 1，Nrf2，CDK2，CDK6 and Cyclin D（P＜0.05，P＜0.01）；ROS antioxidant NAC could reverse the down-regulation effects of curcumol on the expression of these proteins（P＜0.05 or P＜0.01）. CONCLUSIONS Curcumol may induce oxidative stress and cell arrest in G 1 phase to inhibit the proliferation of T 47D cells.
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OBJECTIVE:To establish the method for contents determination of catechins active components in lipid-lowering slimming health products. METHODS:HPLC method was adopted. Using epigallocatechin gallate(EGCG)as a reference,relative correction factor(RCF)of EGCG to gallocatechin(EGC),catechin(C),epicatechin(EC),gallocatechin gallate(GCG)and gal-loylepicatechin(ECG)were calculated. The contents of EGC,C,EC,GCG and ECG in 5 batches of samples were calculated through RCF. The contents of EGC,C,EC,GCG and ECG determined by external standard method were used as measured value. The similarity of the value determined by external standard method with the value calculated by quantitative analysis of multi-com-ponents via single marker method(QAMS)was evaluated with vector included angle cosine method. RESULTS:The linear ranges of EGCG,EGC,C,EC,GCG and ECG were 0.0065-0.1305 mg/mL(r=0.9998)、0.0005-0.0107 mg/mL(r=0.9997)、0.0020-0.0400 mg/mL(r=0.9999)、0.0153-0.3053 mg/mL(r=0.9998)、0.0008-0.0155 mg/mL(r=0.9998)、0.0040-0.0792 mg/mL (r=0.9999);RSDs of precision,stability and reproducibility tests were all lower than 2.0%.The recoveries were 95.07%-100.35%(RSD=1.94%,n=6)、95.24%-101.87%(RSD=2.79%,n=6)、96.08%-103.86%(RSD=3.01%,n=6)、97.51%-101.06%(RSD=1.45%,n=6)、96.01%-101.66%(RSD=2.27%,n=6)、96.20%-102.89%(RSD=2.71%,n=6),respectively. There was no signifi-cant difference between measured value and calculated value. CONCLUSIONS:The method is simple,precise,stable and repro-ducible,and can be used for contents determination of catechins active components in lipid-lowering slimming health products.
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AIM To establish a quantitative analysis of multi-components by single marker(QAMS) method for the content determination of six catechins in Xinnaojian Capsules (Tablets) (tea extract).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Shimadzu Wonda Cract ODS-2 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of 0.5％ acetic acid (A)-acetonitrile (B) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.With epigallocatechin gallate as an internal standard,the relative correction factors of epigallocatechin,catechin,epicatechin,gallocatechin gallate and epicatechin gallate were calculated,from which the content determination was made.RESULTS Six constituents showed good linear relationships within their own ranges (r ≥ 0.999 8),whose average recoveries were 96.00％-98.47％ with the RSDs of 2.09％-2.91％.The results obtained by QAMS method approximated those obtained by external standard method.CONCLUSION This simple and reliable method can be used for the quality control of Xinnaojian Capsules (Tablets).
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Near infrared chemical imaging (NIR-CI) is an emerging technology for rapidly analyzing the critical quality attribute of Chinese materia medica (CMM). It integrates NIR spectroscopy with chemical imaging. In this paper, it provided a systematic introduction to NIR-CI, such as the core part of instrument, the reliability, transformation, analysis and application of high-dimensional data acquisition. In addition, current studies of NIR-CI application in pharmaceutical field were analyzed. Finally, future opportunities and challenges of NIR -CI applications in the quality control of CMM preparation were prospected.
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This study was aimed to optimize the near infrared (NIR) variable selection method based on multivariate detection limit (MDL). Using Qing-Kai-Ling (QKL) injection as object, three variable selection methods (interval par-tial least-squares, iPLS; backward interval partial least squares, BiPLS; moving window interval partial least squares, mwPLS) were used to establish the PLS models of baicalin in QKL injection, respectively. The prediction ability of different variable selection method was compared. MDL of all models were calculated in contrast to the MDL value of full spectra PLS model, to select optimal variable selection method. The results showed that different variable selec-tion methods had different prediction ability. Among them, iPLS had the best performance which determination coef-ficient of prediction (Rpre2) and the root mean square errors of prediction (SEP) were 0.996 5 and 602.3 μg·mL-1, re-spectively. All MDLs of different variable selection methods were reduced compared with the full spectra PLS model. The value of iPLS was the lowest comes to be 1.19 μg·mL-1. The results above indicated that the best variable se-lection method for baicalin in QKL injection was iPLS. MDL theory took the error of calibration and validation set and the leverage of external sample into account, which can comprehensively evaluate model detection performance compared to the classic chemical indicator parameters. This method was particularly suitable for the variable selec-tion method optimization of NIR quantitative model of low concentration sample such as Chinese herbal medicine.
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The system of meridians and acupoints in the traditional Chinese medication (TCM) builds up a complex network. We analyze the features and properties of the system of meridians and acupoints as a complex network based on knowledge of TCM and mathematics. The system shows some important features of complex networks such as the small-world effect and the scale-free property. A preliminary discussion on the robustness of the meridian system as a complex network, and the biomedical functions of such robustness is also given.