1.Studies on Ultrasound-assisted Extraction Process of Total Flavonoids in Vaccinium Bracteatum Thunb. Leaves
Xiaoxian WANG ; Fei TAN ; Shun YAO ; Yifeng ZHOU
China Pharmacist 2015;(1):66-68
Objective:To optimize the ultrasound-assisted extraction process of total flavonoids in Vaccinium bracteatum Thunb. leaves. Methods:The single factor test and orthogonal test were adopted with the content of total flavonoids as the index, and the ultra-sound-assisted extraction was optimized with the concentration of solvent, extracting time, ratio of raw material to liquid and extraction times as the factors. Results:The optimal extraction conditions were 70% ethanol with 40-fold volume, extracting 2 times with 30 mi-nutes per time. Conclusion:The extraction process is stable and feasible, and all the results are useful references for the extraction process improvement.
2. Preparation of sialic acid-modified chlorogenic acid liposomes and its anti-tumor activity in vitro
Chinese Traditional and Herbal Drugs 2020;51(24):6178-6187
Objective: Sialic acid (SA)-modified chlorogenic acid (CA) liposomes (CA-SAL) was prepared by response surface design to investigate its in vitro cytotoxicity and uptake. Methods: CA-SAL was prepared by a modified reverse-phase ethanol injection method. Sephadex G50 column was used to separate the CA-loaded liposomes and the free CA. The drug concentration was determined by HPLC method and the encapsulation efficiency was calculated. With encapsulation efficiency and drug loading as indicators, Box-Behnken response surface design experiments were used to optimize the prescription process of CA-SAL. The MTT method was used to evaluate the cytotoxicity of CA-SAL on human lung cancer cells A549. Inverted fluorescence microscope was used to investigate the uptake of CA-SAL by A549 cells. Results: The optimized preparation conditions: hydrogenated soybean lecithin-CA ratio at 15:1, hydration temperature 60 ℃, ultrasonic power 400 W. The average particle size of CA-SAL was (90.13 ± 0.51) nm, the polydispersity index (PDI) was 0.16 ± 0.01, the zeta potential was (-25.3 ± 0.5) mV, the encapsulation efficiency was 57.8%, RSD was 0.1%. MTT results showed that the inhibitory effect of CA-SAL on A549 cells was significantly greater than CA-CL. Greater cellular uptake of CA-SAL was observed compared with CA-CL. Conclusion: CA-SAL prepared by response surface optimization has a uniform particle size and good stability. SA-modified CA-loaded liposomes could enhance cellular uptake and cytotoxicity of human lung cancer cell A549 in vitro.
3. Mogroside v stimulates osteoblast proliferation and differentiation by promoting lncRNA TUG1 expression
Chinese Journal of Tissue Engineering Research 2020;24(26):4129-4134
BACKGROUND: Osteoporosis is a balance disorder between bone formation of osteoblasts and bone resorption of osteoclasts during bone remodeling. Strict control of bone remodeling at the cellular level is important to maintain bone homeostasis and avoid osteoporosis. Previous studies have shown that 1.25×10-2 g/L mogroside V can promote osteoblast proliferation and differentiation, and its mechanism may be related to LncRNA TUG1. OBJECTIVE: To investigate the role of LncRNA TUG1 in the promotion of osteoblast proliferation and differentiation by mogroside V. METHODS: Osteoblasts from neonatal Sprague-Dawley rats were extracted by two-step enzymatic digestion. The cells were divided into two groups and treated with 0 and 1.25×10-2 g/L Mogroside V. The LncRNA was detected after 2 days of culture. LncRNA TUG1 silencing virus was designed and synthetized. The newly extracted osteoblasts were divided into normal cell control group, mogroside V intervention group, mogroside V+negative virus group, TUG1 silent group, and mogroside V+TUG1 silent group. The proliferation of osteoblasts was observed by FDA fluorescence staining at 2, 4, and 6 days after processing according to the above grouping conditions. After 6 days of treatment on osteoblasts, the effect of TUG1 on osteoblast proliferation and differentiation was studied by alkaline phosphatase staining, alizarin red staining and qRT-PCR. RESULTS AND CONCLUSION: LncRNA detection showed that 1.25×10-2 g/L Mogroside V significantly promoted the expression of LncRNA TUG1 in osteoblasts. FDA fluorescent staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on osteoblast proliferation. After 6 days of treatment, alkaline phosphatase staining and alizarin red staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on mineralization of osteoblasts. The results of qRT-PCR showed that Runx2, BSP, OCN and COL1A1 genes were highly expressed in the mogroside V intervention group, but their expression was inhibited in the mogroside V+TUG1 silent group. Overall findings indicate that mogroside V stimulates the proliferation and differentiation of osteoblasts by promoting the expression of LncRNA TUG1.
4.A study for the time course of cerebral infarction with diffusion kurtosis imaging
Shun ZHANG ; Yihao YAO ; Shuixia ZHANG ; Jingjing SHI ; Yan ZHANG ; Suiqiang ZHU ; Wenzhen ZHU
Chinese Journal of Radiology 2014;(6):443-447
Objective To investigate the time course of cerebral infarction with diffusion kurtosis imaging-related parameters.Methods According to the time interval from symptom onset to MRI examination, 114 cases of cerebral infarction were divided into five groups:8 cases of hyperacute phase(less than 6 hours), 14 cases of acute(>6-24 h), 60 cases of early subacute(>24 h-7 d), 20 cases of late subacute(>7-14 d), and 12 cases of chronic phase ( >14 d-2 months).They underwent routine diffusion weighted imaging (DWI) and diffusion kurtosis imaging (DKI) scanning, and apparent diffusion coefficient ( ADC) and DKI-derived parameters were obtained from them.The derived diffusion parameters were compared among different phases in the patients , and the percent of changes in the infarcted regions were calculated.Paired t-test was used to compare the difference of each parameter between the infarcted region and contralateral normal region , and their correlation with time interval was tested using Pearson correlation analysis.Results Except for chronic phase , mean kurtosis ( MK) , axial kurtosis ( K∥) , radial kurtosis (K⊥)map showed uneven high signal in the infarcted regions , while mean diffusion(MD), axial diffusion(D∥), radial diffusion(D⊥) showed uniform low signal.MK values in the infarcted regions of hyperacute, acute, early subacute and late subacute phase (1.331 ±0.357,1.578 ±0.453,1.519 ±0.455, 1.403 ±0.275 ) increased significantly , compared with the contralateral normal mirror regions ( 0.850 ± 0.236,0.827 ±0.194,0.865 ±0.144,0.939 ±0.212) (t values were 5.242,6.907,12.416,5.629, respectively.P values were all less than 0.01 ).MK, K∥, K⊥ achieved a peak in the acute and early subacute phase , and showed more amplitude than the decrease of MD , D∥, D⊥.Then they gradually reduced, and tended to normalize.MK, MD, ADC had a significant correlation with the time onset of cerebral infarction ( r was 0.354, 0.747, 0.723, respectively, P values were all less than 0.05 ).Conclusion Diffusion kurtosis imaging can provide more diffusion information than conventional DWI , which can better reflect the microstructure changes in tissue.
5.Meta-analysis of effect comparison between video endoscopic inguinal lymphadenectomy and open inguinal lymphadenectomy
Zixiang YAO ; Delin WANG ; Fan YANG ; Shun LIU ; Wencong LIU ; Xin ZHANG
Chongqing Medicine 2014;(12):1422-1424,1427
Objective To evaluate the effects between video endoscopic inguinal lymphadenectomy (VEIL ) and open inguinal lymphadenectomy(OIL) to provide the evidence-based basis for the selection of the clinical therapy schemes .Methods The related clinical controlled trial literature on the effective comparison of VEIL and OIL were retrieved from the databases of PubMed ,Co-chrane library ,Elsevier ,CNKI and Wanfang database .The screening was independently performed by 2 reviewers according to the including and excluding criteria .The related data were extracted and performed the meta analysis by the RevMan 5 .2 software .Re-sults A total of 4 trials were included .There were 146 cases of inguinal lymphadenectomies ,in which 61 cases were VEIL and 85 cases were OIL .The meta-analysis results showed that there were no statistical differences between the two operation modes in terms of the operative time(WMD=32 .33 ,95% CI -25 .70-90 .36 ,P=0 .27) ,intraoperative blood loss(WMD=9 .10 ,95% CI -76 .03-94 .23 ,P=0 .83) ,number of removed lymph nodes(WMD=0 .77 ,95% CI -1 .66-3 .20 ,P=0 .53) ,number of positive re-moved lymph nodes(WMD=0 .08 ,95% CI -0 .23-0 .40 ,P=0 .61) ,postoperative drainage time(WMD= -1 .30 ,95% CI -6 .40 -3 .80 ,P=0 .62) ,postoperative hospital stay (WMD= -4 .02 ,95% CI -10 .19-2 .15 ,P=0 .20) ,but the difference between VEIL and OIL in term of surgical complications had statistical significance (OR=0 .08 ,95% CI 0 .03-0 .26 ,P<0 .01) .Conclusion VEIL has equivalent efficacy to OIL ,but has less surgical complications .
6.Study of the lateral ventricular in patients with Alzheimer's disease at 3.0T MR and MIMICS reconstruction
Danqing WANG ; Yuefeng LI ; Shun YAO ; Yuhao XU ; Tian ZHAO ; Qiong JI ; Yu CHEN
Journal of Practical Radiology 2017;33(3):349-352,364
Objective To evaluate cerebral parenchymal atrophy of patients with Alzheimer's disease(AD)through the compara-tive analysis of the volume and morphology of the brain ventricle between patients with AD and normal elderly.Methods 20 patients with AD and 20 normal elderly people were scanned at 3.0T MR,and lateral ventricle section images were achieved,and the lateral ventricle volume and the anterior horn,posterior horn and temporal horn of the lateral ventricle were calculated by analyzing the re-construction of section images with MIMICS software from Belgian.Results As compared with normal elderly group,the patients with AD exhibited significantly increased the volume of left ventricular volume(LV),right ventricular volume (RV)and total vol-ume (TV)(P<0.05).Angle of bilateral anterior horn and temporal horn but not posterior horn of the lateral ventricle in patients with AD were significantly higher than that in normal elderly (P<0.05).The volume of the left,right and total cerebral ventricle, the angle of the anterior horn of the left and right lateral ventricle and the angle of the temporal horn of the left and right lateral ven-tricle were negatively correlated with MMSE (P<0.05).Conclusion Patients with AD exhibites significantly greater volume and an-gle of the lateral ventricular than normal elderly people.These related data measured can predict brain parenchymal atrophy of pa-tients with AD more conveniently and accurately.
7.Some leukocyte differentiation antigens express in side population cells in leukemia K562 cell line
Ruihua FAN ; Dianxuan GUO ; Tiecheng ZHANG ; Shun YUE ; Mingde HUANG ; Rong YAO
Chinese Journal of Tissue Engineering Research 2013;(49):8596-8601
BACKGROUND:To study the phenotypes of side population cells in leukemia is important for understanding the heterogeneity and origin of tumor cells, molecular markers and targeted therapy.
OBJECTIVE:To identify whether the human chronic myeloid leukemia cellline-K562 contains side population cells or not, and to further observe the differences in expressions of leukocyte differentiation antigens from side population cellsubset and non-side population cells subset.
METHODS:Flow cytometry was used to detect whether there were side population cells in the K562 celllines. Then, the expression of CD34+, CD34+CD38-, CD34+CD38+, HLA-DR+cells in the side population subsets and non-side population subsets.
RESULTS AND CONCLUSION:Flow cytometry results showed that the K562 cellline contained side population cells, and the proportion of side population cells was much lower. The side population cells accounted for (2.7±0.5)%of viable cells in K562. The expressions of CD34+cells and CD34+CD38-cells in the side population subset were significantly higher than those in the non-side population subsets. The expressions of CD34+CD38+cells and HLA-DR+cells in the side population subset and non-side population subset did not have a significant difference. Heterogeneity was found in the differentiation antigen expression between the side population subset and non-side population subset.
8.IE-HPLC analysis of trigonelline in Fructus Quisqualis.
China Journal of Chinese Materia Medica 2004;29(2):135-187
OBJECTIVETo establish a IE-HPLC method for the determination of trigonelline in the fruit and seed of Quisqualis indica.
METHODThe method was carried out by using Sperisorb SCX column with a mobile phase consisting of 20 mmol x L(-1) phorsphate (pH3.2, titrate a 20 mmol x L(-1) monsodium salt with a 20 mmol x L(-1) solution of phosphoric acid), flow rate of 1.0 mL x min(-1), and the detection at 265 nm.
RESULTBy using SCX-modified silica column, trigonelline in Fructus Quisqualis can be baseline separated and determinated. The retention mechanisms proved to be contributed mainly by ion-exchange with the SCX moieties. The validity of methods was demonstrated with respect to linearity, precision, reproducibility and recovery. It was found that there was no trigonelline occurring in the peel of Fructus Quisqualis.
CONCLUSIONThis assay method can be used for the quality control of Fructus Quisqualis.
Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Combretaceae ; chemistry ; Fruit ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Seeds ; chemistry
9.The practical value of a Logistic regression model of serum indexes in differentiating between geriatric depression and depressive state
Yuhao XU ; Ming YU ; Yuefeng LI ; Shun YAO
Chinese Journal of Geriatrics 2018;37(9):984-987
Objective To investigate the practical value of a logistic regression model of serum indexes in distinguishing between geriatric depression and geriatric depressive state. Methods A total of 160 patients were recruited from the outpatient department from January 2013 to January 2016 ,and were divided into a depression group (n= 80)and a depressive state group (n= 80) ,with retrospective diagnoses based on the Chinese Classification of Mental Disorders-Third-Edition(CCMD-3).Serum samples were collected and enzyme linked immunosorbent assays (ELISA)were used to determine the concentrations of brain-derived neurotrophic factor (BDNF ) ,glial cell line-derived neurotrophic factor(GDNF) ,fibroblast growth factor 2(FGF-2) ,vascular endothelial growth factor (VEGF) ,interleukin-1β(IL-1β) ,interleukin-6 (IL-6) ,interleukin-10 (IL-10) ,tumor necrosis factor-α(TNF-α) ,cortisol (CORT ) ,and platelet-derived growth factor (PDGF).Binary logistic regression analysis was used to establish the regression model ,and the ROC curve was drawn to explore its value of differentiating geriatric depression from depressive state. Results The levels of serum BDNF , GDNF and VEGF in the depression group were lower than those in the depressive state group (BDNF :208.7 ± 41.4 vs.262.9 ± 84.6 ng/L ,GDNF :92.3 ± 18.6 vs. 101.4 ± 30.9 ng/L ,VEGF :223.1 ± 98.2 vs. 257.8 ± 77.2ng/L) ,while the levels of IL-1βand CORT in the depression group were higher than in the depressive group(IL-1β:27.0 ± 4.9 vs.19.6 ± 5.7 μg/L ,CORT :96.3 ± 16.7 vs.83.0 ± 17.3 nng/L).In multivariate logistic regression analysis ,BDNF(OR = 0.987 ,P = 0.001) ,IL-1β(OR =1.29 ,P = 0.000)and CORT (OR = 1.065 ,P = 0.000)were selected to build the regression model. The regression equation was P=1/[1+e-(- 8.546 - 0.013(BDNF)+ 0.258(IL -1β)+ 0.063(CORT)) ]and the area under the ROC curve was 0.966.Compared with retrospective diagnoses made two weeks later ,the correct diagnosis rate of the logistic model was 90.47%. Conclusions The Logistic regression model of serum indexes can further differentiate between geriatric depression and depressive state which also offers additional benefits for the diagnosis ,differential diagnosis ,and treatment of depression.
10.Effect of annonaceous acetogenin mimic AA005 on proliferative inhibition of leukemia cells in vitro and its possible mechanisms.
Bing HAN ; Zhu-Jun YAO ; Li-Shun WANG
Journal of Experimental Hematology 2012;20(3):549-553
This study was aimed to investigate the biological behavior of annonaceous acetogenin mimic AA005 in various kinds of leukemia cells and further elaborated its possible mechanisms in acute promyelocytic leukemia (APL) cell line NB4. The proliferative inhibition of leukemia cells was measured by CCK-8 method. Cell death was determined by trypan blue. Cell morphological features of NB4 treated with AA005 were examined by microscopy after Wright's staining. The form of cell death was measured by flow cytometry. Proteins PARP-1 and caspase-3 were detected by Western blot. Flow cytometry was used to detect the cell cycle arrest induced by AA005 of low concentration. The results showed that AA005 (> 200 nmol/L) significantly inhibited proliferation of all tested leukemia cell lines in a concentration-dependent manner. The vast majority of cells went to die after leukemia cell lines of NB4, U937 and K562 were treated with different concentration of AA005 for 48 h. Typical morphologic changes significantly appeared in NB4 cells after AA005 treatment. AA005 almost simultaneously induced early apoptosis and late apoptosis. The little cleavage of PARP-1 and activation of caspase-3 happened in AA005-induced cell death, and caspase-3 inhibitor Z-VAD-fmk could not block the cell death. The non-toxic concentrations of AA005 (< 200 nmol/L) caused NB4 cells G(2)/M-phase arrest. It is concluded that annonaceous acetogenin mimic AA005 induces significant proliferative inhibition of various leukemia cell lines in a concentration-dependent manner, which may be associated with cell death and G(2)/M-phase arrest induced by AA005.
Acetogenins
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pharmacology
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Apoptosis
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drug effects
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Caspase 3
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metabolism
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Cell Cycle Checkpoints
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Cell Proliferation
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drug effects
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Fatty Alcohols
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pharmacology
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HL-60 Cells
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Humans
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K562 Cells
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Lactones
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pharmacology
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Poly (ADP-Ribose) Polymerase-1
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Poly(ADP-ribose) Polymerases
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metabolism