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.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.
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.Study of Electroacupuncture at Ganshu (BL18) and Zusanli (ST36) on Interaction between Brain and Gut in Rats with Depressive Functional Dyspepsia
Kaiyan JIANG ; Haibo LI ; Ke ZHANG ; Xiaowei GUO ; Lu REN
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(3):62-66
Objective To observe the effects of electroacupuncture at Ganshu (BL18) and Zusanli (ST36) on depression state and gastrointestinal motility and relevant protein expressions in brain of rats with stress-induced depressive functional dyspepsia. Methods Totally 48 Wistar rats were randomly divided into normal, model, electroacupuncture (EA) and sham-operation group, with 12 rats in each group. Depressive functional dyspepsia model was established by chronic unpredictable mild stress protocols. From day 15 to day 28, the EA group was given electroacupuncture at BL18 and ST36, and the sham-operation group was stimulated by electroacupuncture at 1/3 and 2/3 of the tail. Body weight test, open field test and sucrose preference test were measured on day 0, day 14 and day 28. After 14-day administration, gastric residual volume and intestinal propulsion rate of semisolid paste were detected. The expression of βCaMK inⅡ lateral habenulas and BDNF in hippocampus were measured by Western blot. Results Compared with the normal group, the weight, the sucrose preference rate, the behavior score, the small intestinal propulsion rate of semisolid paste and the expression of BDNF in hippocampus decreased significantly both in the model group and sham-operation group (P<0.01), and the gastric residual volume of semisolid paste and the expression of βCaMK inⅡ lateral habenulas increased significantly (P<0.01). Compared with the model group, the weight, the sucrose preference rate, the behavior score, the small intestinal propulsion rate of semisolid paste and the expression of BDNF in hippocampus increased significantly in EA group (P<0.01), and the gastric residual volume of semisolid paste and the expression of βCaMK inⅡ lateral habenulas decreased significantly (P<0.01). Compared with the sham-operation group, the expression of βCaMKⅡ in lateral habenulas decreased significantly, the expression of BDNF in hippocampus increased significantly in EA group (P<0.01). Conclusion Electroacupuncture at Ganshu (BL18) and Zusanli (ST36) can improve depression and gastrointestinal motility of rats with depressive functional dyspepsia, which may be related to the inhibition of βCaMK inⅡ lateral habenulas and the activation of BDNF in hippocampus dyspepsia.
8.Efficacy and complication of caffeine citrate and aminophylline in primary apnea of premature infants:a comparative analysis
Hua KE ; Zhankui LI ; Jinzhen GUO ; Yunfan YANG
Chinese Pediatric Emergency Medicine 2017;24(1):56-59
Objective To investigate the effects of caffeine citrate and aminophylline in treatment of primary apnea of premature infants and its related complications. Methods A total of 152 preterm infants who were diagnosed primary apnea within 10 days after birth were randomly divided into caffeine citrate group(n=77) and aminophylline group(n =75). The changes in the time of the apnea disappeared after treating,needing oxygen,non-invasive and invasive mechanical ventilation,and the incidence of bronchopul-monary dysplasia ( BPD ) , necrotizing enterocolitis, retinopathy of prematurity, patent ductus arteriosus (PDA),intercranial hemorrhage(ICH)were compared between the two groups. Results The time of the apnea disappeared after treating[(47. 4 ± 5. 3) h],needing oxygen[(20. 5 ± 7. 6) d],non-invasive mechani-cal ventilation[(8. 7 ± 4. 2) d] and invasive mechanical ventilation[(1. 0 ± 1. 3) d] in the caffeine citrate group were significantly lower than those in the aminophylline group [ ( 54. 8 ± 6. 2 ) h, ( 24. 4 ± 8. 5 ) d, (10.4±5.3)d,(10.4±5.3)d,respectively](P<0.05).TheincidenceofBPD[16.9%(13/77)],PDA [13. 0%(10/77)] and ICH[16. 9%(13/77)] in the caffeine citrate group were significantly lower than those in the aminophylline group[33.3%(25/75),26.7%(20/75),22.7%(17/75),respectively](P <0. 05),but there were no differences between the two group about the incidence of necrotizing enterocolitis [caffeine group 7. 8%(6/77),aminophylline group 9. 3%(7/75)] and retinopathy of prematurity[caffeine group 6. 5%(5/77),aminophylline group 9. 3%(7/75)](P>0. 05). Conclusion The caffeine citrate has a better efficacy in the treating primary apnea of preterm infants than aminophylline. It can also decrease the incidence of BPD,PDA and ICH in premature infants.
9.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.
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.