1.Sorting methods of breast cancer stem cells
Chongyong GUO ; Keying SONG ; Ke LI
Journal of International Oncology 2011;38(5):358-361
In recent years, the theory of cancer stem cells has provided a new perspective on the treatment of cancers including breast cancer. The accurate sorting of breast cancer stem cells is critical. The sorting procedure is consist of 4 steps: isolation of side population, serum-free suspension culture,determination of specific cell surface markers and of the activity of aldehyde dehydrogenase 1 ( ALDH1 ) enzymatic through the ALDEFLUOR assay. Some studies choose drug resistance as an additional method for sorting cancer stem cells because of the enrichment of cancer stem cells after chemotherapy . The controversy about the sorting outcome of breast cancer stem cell mainly focus on molecular markers like CD44+ CD24- and ALDH1+ . The problem needed to be settled is to identify which sorting method and markers are appropriate.
2.High-risk Human Papillomavirus and the Detection of Viral Load in the Diagnosis of CINⅡ and Ⅲ or Cervical Cancer
Chinese Journal of Minimally Invasive Surgery 2005;0(11):-
Objective To explore the role of high-risk human papillomavirus(Hr-HPV) DNA testing in the detection of cervical intraepithelial lesions(CIN) Ⅱ and Ⅲ,or cervical cancers for patients with abnormal cervical cytology.Methods A total of 2197 patients with abnormal cervical liquid-based cytology,who had received Hr-HPV detection and colposcopy,were enrolled into this study.The data were analyzed using ?2-test and Logistic regression.Among the patients,1013(46.5%) had atypical squamous cells undetermined significance(ASC-US),89(4.1%) had atypical squamous cell which cannot exclude high grade squamous intraepithelial lesion(ASC-H),757(34.7%) low squamous intraepithelial lesion(LSIL),253(11.6%) high-grade squamous intraepithelial lesion(HSIL),and 67(3.1%) atypical gland cells(AGC).Results Pathological examination showed CINⅡ in 224 patients(10.3%),CIN Ⅲ in 140(6.4%),cervical cancer in 27(1.2%),and adenocarcinoma in 9(0.4%).The other patients(1779/2179,81.6%) had CINⅠ,condyloma,or chronic cervicitis.The detection rate of CIN Ⅱ and Ⅲ,and cervical cancer was increased in the patients with Hr-HPV.The differences in the rate among ASC-US,ASC-H,LSIL,and HSIL patients were significant.The incidence rate of CINⅡ and Ⅲ,or cervical cancer was 2.2%(13/588),9.7%(21/216),24.9%(97/389),29.8%(182/611),and 23.2%(87/375) for the patients with a Hr-HPV load of 0-0.9,1.0-9.9,10.0-99.9,100.0-999.9,and ≥1000.0.Logistic regression showed that the risk of CINⅡ and Ⅲ,and cervical cancer increased by 1.325 folds with the log of the HPV load.Conclusions Hr-HPV DNA test is helpful for the detection of CIN Ⅱ and Ⅲ or cervical cancer in patients with abnormal cervical cytology.
3.Effect of LY294002 on protein kinase B activation in human lens epithelial cell
Rui, GUO ; Hai-ke, GUO ; Qian-li, MENG
Chinese Journal of Experimental Ophthalmology 2013;32(11):1026-1030
Background Protein kinase B (Akt) is the center of multiple cellular signaling pathways,and it participates in the regulation of cell function,such as proliferation,migration,and metabolism of cells.Phosphoinositide 3-kinase (PI3K) promotes activation of Akt and therefore triggers many signal pathways.PI3K inhibitor can silent Akt,but whether it can affect the biological behavior of lens epithelial cells (LECs) during the posterior capsular opacity (PCO) is worthy of investigation.Objective This study was to explore the effect of LY294002,a PI3K inhibitor,on Akt activation in human LECs.Methods Human LECs strain,HLEC-B3 cells,were cultured and passaged.The cells were incubated to 96-well plate for 24 hours,then LY294002 was added with the final concentration at 0,10,20,30,40,50,60,70,80 μmol/L,respectively.After incubation for 24 hours,cell counting kit-8 (CCK-8) was used to detect the inhibitory rate of cell proliferation.Transforming growth factor-β2 (TGF-β2) of 10 μg/L was added in the medium of cells as TGF-β2 group,TGF-β2 + LY294002 (20 μmol/L) was used to coculture the other group of cells,and the cells without TGF-β2 and LY294002 served as the control group.Phosphorylation of Akt (p-Akt) in the cells was detected by laser scan confocal immunofluorescence microscope.The expression level of p-Akt was evaluated using Western blot.Results CCK-8 assay showed that the A value of the cells was gradually reduced with the increase of LY294002 concentration (Fgroup =9.72,P =0.00),but the A value was significant raised along with the lapse of time (Ftime =1737.54,P=0.00).Confocal immunofluorescence revealed that a little of p-Akt was expressed in the control cells.After induced by TGF-β2,lots of red fluorescence of p-Akt was seen in cells around the cell membranes.But in the TGF-β2+LY294002 co-culture cells,the fluorescence of Akt was much weaker.Western blot showed that the expression level of p-Akt was 0.91±0.08,1.48±0.13 and 0.95±0.19 in the control group,TGF-β2 group and TGF-β2 +LY294002 group,respectively,with a significant difference among the three groups (F =15.04,P =0.00).Conclusions LY294002 can inhibit the activation of Akt induced by TGF-β2.LY294002 may have utility in the prevention and treatment of PCO.
4.Clinical assessment of chronic prostatitis based on infrared imaging technology.
An-guo LIU ; Xing-ke YAN ; Li-li KAN
National Journal of Andrology 2016;22(1):22-27
OBJECTIVETo investigate the characteristic changes in the infrared thermogram of chronic prostatitis (CP) patients and find some evidence for the auxiliary diagnosis and therapeutic evaluation of the disease.
METHODSFifty CP patients and 20 healthy male volunteers were included in this clinical trial. The infrared thermograms of the subjects were compared between the two groups for characteristic changes. The values obtained were used for the auxiliary diagnosis and therapeutic evaluation of the disease.
RESULTSCompared with the healthy males in the same age group, the CP patients showed extremely significant abnormal changes in the average temperature value in the hypogastrium (H), pubis (P), scrotum (S), and groin (G) (P < 0.01). The average H temperature value of the CP patients was correlated negatively with the CP symptom index (CPSI) (P < 0.01, Pearsons correlation coefficient = -0.519), while the S temperature positively with CPSI (P < 0.01, Pearsons correlation coefficient = 0.446). In addition to the H value, the P, S, and G values were all correlated in different degrees with CPSI (P < 0.01), which the S value exhibited the most significantly negative correlation (Pearson's correlation coefficient = -0.898).
CONCLUSIONThere are some characteristic changes in the hypogastrium temperature of CP patients in the infrared thermogram, which has a potential application value for the auxiliary diagnosis, symptom assessment, and therapeutic evaluation of CP.
Case-Control Studies ; Humans ; Infrared Rays ; Male ; Prostatitis ; diagnosis ; Temperature ; Thermography
5.Anti-tumor Effect of Phellinus Linteus and Coriolus Versicolor Capsule on S180 sarcoma and H22 hepatoma in mice
Kunxing LI ; Xueping ZHU ; Hailin ZHANG ; Ke NIE ; Wei GUO
Chinese Journal of Experimental Traditional Medical Formulae 2009;(7):83-86
Objective: To observe the anti-tumor effect of Phellinus Linteus and Coriolus Versicolor Capsules (PLCVC) in S180 sarcoma and H22 hepatoma animal models in mice. Methods: The sarcoma S1180 and hepatoma H22 models were established in mice. After 12 days of treatment, the animals were killed, and the subcutaneous sarcoma were separated and weighted. The levels of vascular endothelial growth factor(VEGF), CD4 and CD8 of S180 tumor tissue were investigated by immunohistochemical method. KM mice were intraperitoneal injected with H22 hepatoma cells, and treated with different experimental drugs. The survival time was observed and recorded, and life-prolongation rate was calculated. Result: PLCVC could inhibit the growth of S180 and H22 tumor, and inhibit the expression of VEGF, improve the expression of CD4 and CD8. The survival time of the mice treated by PLCVC were significantly longer than the untreated group. Conclusion: PLCVC can inhibit the growth of tumour, the mechanism is partially related to inhibiting angiogenesis and improving the immunological function.
6.Research Progress of Silk Fibroin As a Drug Delivery Materials.
Lijing JI ; Jing KE ; Lan JIA ; Li GUO ; Jingxin ZHU
Journal of Biomedical Engineering 2015;32(6):1364-1368
Recently, drug delivery materials have become the hotspot of medical study. Suitable delivery material plays an important role in constructing an excellent drug delivery system. Silk fibroin is a naturally occurring protein polymer with excellent biocompatibility, remarkable mechanical properties, biodegradability and outstanding processability. Due to its unique properties, silk fibroin has become a favorable carrier material for the incorporation and delivery of a range of therapeutic agents. Based on the structure and characteristics of silk fibroin, this article provides an overview of the recent research progress of silk fibroin used as drug delivery materials.
Biocompatible Materials
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Drug Delivery Systems
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Fibroins
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chemistry
7.Expression of focal adhesion kinase in human lens epithelial cells induced by hydrogen peroxide
Yu, HUANG ; Qian-li, MENG ; Hai-ke, GUO
Chinese Journal of Experimental Ophthalmology 2013;31(10):914-918
Background The pathogenesis of age-related cataract is associated with the apoptosis of lens epithelial cells (LECs) caused by oxidative stress.Previous studies showed that intracellular focal adhesion kinase (FAK) pathway can be activated by H2O2 in vitro,which induced apoptosis of cells.To investigate the effect of oxidative on FAK expression in LECs is one of important studies in the prevention of age-related cataract.Objective This study was to investigate the expression and function of FAK in human LECs treated by H2O2.Methods Human LECs strain (HLECs-B3) were cultured in vitro in the low glucose DMEM with 10% fetal bovine serum.Different concentrations (0,30,50,70,100,300,500,700,1000 μmol/L) of H2O2 were added into the culture medium for 24 hours.The survival rate of the cells was detected by Cell Counting Kit-8 (CCK-8) assay.Cell morphology as well as the expression and distribution of FAK in the cells were observed by immunofluorescent staining under the laser confocal microscope.Apoptosis was observed by hoechst33258 staining,and Western blot assay was used to quantitatively detect the expression and phosphorylation of FAK.Results The survival rate of the cells was (1.00±0.03) %,(1.24±0.03)%,(1.36±0.24) %,(0.93±0.02)%,(1.75±0.19)%,(1.37±0.18) %,(0.64±0.01)%,(0.59±0.11)%,(0.14±0.05)% in 0,30,50,70,100,300,500,700,1000 μmol/L H2O2 groups,with a significant difference among them (F =95.30,P =0.00).The survival rates of the cells in the below 300 μmol/L H2O2 groups were significantly higher than those in the 0 μmol/L H2O2 group,and survival rates of the cells in the above 500 μmol/L H2O2 groups were significantly lower than those in the 0 μmol/L H2O2 group(all at P<0.05).After H2 O2 treatment for 24 hours,HLECs-B3 cells transformed from polygon shape to spindle shape and extended pseudopodiums,meanwhile the green fluorescence for FAK exhibited in the cytoplasm.Cell apoptosis was found in the 1000 μ mol/L H2O2 group.Western blot assay revealed that the expressing levels (grey scale) were significantly different among the various groups (F=28.08,P=0.00),and FAK expressing levels in the below 300 μmol/L H2O2 groups were significantly higher than those of the 0 μmol/L H2O2 group; while the expressing levels in the above 500 μmol/L H2O2 groups were lower than those of the control 0 μmol/L H202 group (all at P<0.05).After treated by different concentrations of H2O2,the phosphorylation level of intracellular FAK (p-FAK) was significantly higher in 3 hours group than that in 30 minutes group (all at P<0.05).Conclusions H2 O2 can affect the survival,proliferation and morphology of human LECs by activating the intracellular FAK pathway,indicating that FAK may play roles in the regulation process of cell biological behavior.
8.Evaluation of HR-HPV testing in diagnosis of cervical intraepithelial neoplasia and cervical cancer
Yanli GUO ; Li GENG ; Ke YOU ; Jie QIAO
Chinese Journal of Clinical Laboratory Science 2006;0(06):-
Objective To evaluate the role of high-risk human papillomavirus (HR-HPV) testing in diagnosis of high-grade cervical intraepithelial neoplasia (CIN2/3+) and cervical cancer.Methods Total 3426 women with known cytologic abnormality tests were subjected to detect high-risk HPV DNA by hybrid capture II and to have established diagnoses by cervical coloposcopy-directed biopsy test.Results Among 3426 women,45 were cervical cancer,670 were CIN 2/3,and 2711 were CIN 1 or other benign lesions based on the cervical histologic results. Compared with the women with CIN 1 or other benign lesions,the women with CIN 2/3+ had higher positive rates of HR-HPV. Higher load of HR-HPV was more common among women with cervical lesions than those without. Fifty-seven women who were transiently HR-HPV positive did not develop CIN2/3+ during follow-up.Conclusions Our data suggest that a high viral load could be used as a short-term marker of CIN 2/3+,and women repeatedly tested positive for HR-HPV are at risk of developing CIN2/3+. Detecting HR-HPV and follow up the positive women would be useful in diagnosis of CIN2/3+.
9.Preliminary study of vulvar and vaginal intraepithelial neoplasias
Yanli GUO ; Li GENG ; Ke YOU ; Jie QIAO ; Congrong LIU
Journal of Peking University(Health Sciences) 2003;0(05):-
Objective:To detect the clinical features of vulvar and vaginal intraepithelial neoplasias(VIN and VAIN,respectively).Methods:Total 148 women were performed vulvar or vaginal coloposcopy-directed biopsy pathology tests,from Sep.2004 to Dec.2007.Results:Among 148 women,vulvar or vaginal histologic results were vulvar cancer for 1,VIN or VAIN 2,3 for 23,VIN or VAIN for 16,condyloma for 61,vulvitis and vaginitis for 47.Eighty-five percent(33/39) women with VIN or VAIN 2,3 were more than 30 years old.Compared to women with cervical intraepithelial neoplasia(CIN),women with VIN or VAIN were older.The rate of high-risk HPV DNA in women with vulvar or vaginal lesions was 84%(84/100).VAIN occurred mainly in the upper vagina(90%,69/75).VIN or VAIN often accompanied or followed CIN or cervical cancer(79%,31/39),and VIN or VAIN 2,3 often accompanied or followed CIN 2,3 or cervical cancer(70%,16/23).Conclusion:Our data suggest that women with high-risk HPV infection are at risk of developing VIN or VAIN 2,3.The vulva and vagina should be carefully inspected by colposcopic examination at the time of colposcopy for any abnormal findings.
10.Chemical constituents from Polygonum hydropiper
Mengyun LI ; Yangmin MA ; Ke QIAO ; Linxin GUO
Chinese Traditional Patent Medicine 2017;39(4):769-773
AIM To study the chemical constituents from Polygonum hydropiper L..METHODS The petroleum ether,ethyl acetate and n-butanol fractions of 95% ethanol extract from P.hydropiper were isolated and purified by silica,Sephadex LH-20 and recrystallization,then the structures of obtained compounds were identified by spectral data.RESULTS Thirteen compounds were isolated and identified as β-sitosterol (1),aniba dimer A (2),succinic acid (3),quercetin (4),gallic acid (5),daucosterol (6),quercetin-3-O-β-D-glucoside (7),kaempferol-3-O-β-D-glucoside (8),quercetin-3-O-β-galactoside (9),kaempferol-3-O-β-galactoside (10),stigmast-4-ene-3β,6a-diol (11),fumaric acid (12),ellagic acid (13).CONCLUSION Compounds 11 and 12 are isolated from genus Polygonum for the first time,compounds 2,8,10 and 13 are first obtained from this plant.