1.Expression and function of tumor susceptibility gene 101 in multidrug-resistant gastric cancer cells
Huiqin SHEN ; Zheyi HAN ; Yan XUE
Chinese Journal of Digestion 2001;0(09):-
Objective To investigate the expression and possible function of tumor susceptibility gene 101 (TSG101) in multidrug-resistant cell lines of gastric cancer.Methods The expression of TSG101 was examined in gastric cancer cell line SGC7901 and its vincristine(VCR)-resistant subline SGC7901/VCR with semi -quantitative RT-PCR and Western blot. TSG101 eukaryotic expression vector was transfected into SGC7901 cells by lipofectamine TM 2000. The expression levels of TSG101 in SGC7901 and the transfectants were detected with Western blot. Fluorescence-activated cell sorting was applied to examine the cell cycle alteration and the intracellular mean fluorescence intensity of adriamycin (ADR). Growth curve and drug sensitivity of cells to VCR and ADR were analyzed by MTT assay.Results TSG101 was highly expressed in VCR resistant gastric cancer cells. The expression of TSG101 in TSG101 transfectants was up-regulated compared with that in SGC7901 transfected with empty vector or in SGC7901 cells. TSG101 transfectants were accumulated in S phase, with a concomitant decrease of cell population in G 1 phase. MTT assay showed that TSG101 transfectants proliferated rapidly and were more resistant to VCR and ADR than control cells. Conclusions The over-expression of TSG101 could promote the multidrug resistance phenotype of sGC7901 cells. TSG101 may play a certain role in multidrug resistance of gastric cancer.
2.Expression of IGF-Ⅱ and IGFbP-6 in colorectal cancer and their significance
Jing XUE ; Yang ZHAO ; Jianwei ZHANG ; Yan JIAO ; Xingbin SHEN
Chinese Journal of Clinical and Experimental Pathology 2015;(6):628-631,632
Purpose To investigate the expression of IGF-Ⅱand IGFBP-6 in co1orecta1 cancer,and to exp1ore the c1inica1 significance in co1orecta1 cancer. Methods IGF-Ⅱand IGFBP-6 were detected by immunohistochemistry and RT-PCR in 50 cases of co1orecta1 cancer(experimenta1 group)and 50 cases of the adjacent mucosa(contro1 group). Results (1)In the experimenta1 group,the ex-pression 1eve1 of IGF-Ⅱprotein and mRNA was significant1y higher than the contro1 group. The expression 1eve1s of IGF-Ⅱprotein and mRNA in co1orecta1 cancer were significant1y 1ower than the contro1 group.(2)The expression 1eve1s of IGF-Ⅱand IGFBP-6 were sig-nificant1y different between different tumor infi1tration depth,1ymph node metastasis,invasion depth and Duke’s stages( P<0. 05), but no difference between genders,age and the degree of tumor differentiation( P>0. 05). Conclusions There is a obvious corre1a-tion between of IGF-Ⅱ and IGFBP-6 in c1inica1 patho1ogica1 parameters in co1orecta1 cancer. Combined detection of the two markers may be the bio1ogica1 indicators of occurrence and prognosis of co1orecta1 cancer,and provide a new scheme for diagnosis and treatment of co1orecta1 cancer.
3.Expression and clinic significance of IGFBP-2 and IGFBP-6 in colorectal adenomas
Jing XUE ; Yang ZHAO ; Jianwei ZHANG ; Yan JIAO ; Xingbin SHEN
Chongqing Medicine 2015;(29):4112-4115
Objective To study the insulin‐like growth factor binding protein (IGFBP)‐2 and IGFBP‐6 expression and clini‐cal significance in colorectal adenomas .Methods A collection of Chengde Medical College Hospital from July 2012 to March 2013 after surgical treatment of colorectal cancer confirmed by pathology (colorectal cancer ,CRC) tissue samples of 50 patients ,colorec‐tal adenomas (colorectal adenoma ,CRA) 50 cases ,20 cases of colorectal normal mucosa .Immunohistochemistry and RT‐PCR were used to detect the expression of IGFBP‐2 and IGFBP‐6 protein and mRNA ,combined with clinical and pathological data were statis‐tically analyzed .Results IGFBP‐2 protein positive expression and the amount of mRNA expression in the CRA group compared with normal colorectal mucosa had a rising trend;While compared with CRC group had a tendency to reduce ,and the differences are obvious statistical significance (P<0 .05) ,and IGFBP‐6 protein positive expression in the CRA group compared with normal color‐ectal mucosa had a lower trend;While compared with CRC group amount IGFBP‐6 mRNA expression in the CRA group compared with normal colorectal mucosa had a rising trend;While compared with CRC group had a tendency to reduce ,and the differences were obvious statistical significance (P<0 .05) .In the CRA group ,IGFBP‐2 ,IGFBP‐6 positive expression and the patient′s age , sex ,tumor and the number of parts were no significant statistical difference (P>0 .05) ,but with the degree of hyperplasia had sig‐nificant statistical difference(P<0 .05);In the CRA group ,IGFBP‐2 and IGFBP‐6 expression were negatively related to each other , and the difference was statistically significant (P<0 .01) .Conclusion Colorectal adenomas in normal colorectal mucosa of colorec‐tal cancer to the middle part of the transformation process ,and in its occurrence and development process ,insulin‐like growth factor family (IGFs) and IGFS‐R axis plays an important and irreplaceable role ,so IGFBP‐2 ,IGFBP‐6 may be used as diagnostic colorectal adenomas and early predictors of prognosis ,clinical studies on colorectal adenomas is important .
4.Protective effects of vascular endothelial growth factor on cerebral ischemia
Xue SHEN ; Lihui XUAN ; Rongyin QING ; Yan ZHU ; Yingzhu CHEN
International Journal of Cerebrovascular Diseases 2014;22(9):704-708
Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen.Studies have shown that VEGF is closely associated with ischemic stroke,and this makes it possible to intervene in ischemic stroke from the level of VEGF and its receptor.This article reviews the biological effect of VEGF and its receptor,mechanism of action involving in various stages of ischemic stroke,and the therapeutic prospect in ischemic stroke.
5.Diagnostic value of reflectance confocal microscopy combined with dermoscopy for melanocytic nevus
Yajie ZHENG ; Xue SHEN ; Yan JING ; Yatong WU ; Ruixing YU ; Lei WANG ; Ke XUE ; Yong CUI
Chinese Journal of Dermatology 2017;50(7):517-520
Objective To evaluate the diagnostic value of dermoscopy and reflectance confocal microscopy (RCM) alone or in combination for melanocytic nevus.Methods A total of 37 patients with clinically diagnosed melanocytic nevus were collected.Skin lesions were firstly examined by dermoscopy and RCM,then were resected to be subjected to histopathological examination for final diagnosis.The imaging features of melanocytic nevus were summarized.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of different skin imaging techniques were calculated,and the consistency was analyzed between skin imaging techniques and histopathological examination.Results Based on the dermoscopic and RCM findings,2 kinds of nevus cells with different morphological features were observed in the dermis of intradermal nevus.One kind of nevus cells was characterized by a nonfusional,highly-refractive round structure in the papillary dermis under RCM,and by a brown or light brown homogenous pattern under dermoscopy,which was observed in 5 skin lesions.The other kind of nevus cells appeared as irregular,highly-refractive cell clumps in the papillary dermis under RCM,and by a cobblestone or globular pattern under dermoscopy,which was observed in 31 skin lesions.For the diagnosis of melanocytic nevus,the sensitivity,specificity,accuracy,positive predictive value and negative predictive value of RCM combined with dermoscopy were 91.7%,87.5%,90.9%,97.1% and 70% respectively,those of RCM were 86.1%,75%,84%,93.9% and 54.5% respectively,and those of dermoscopy were 77.8%,87.5%,75%,96.3% and 41.2% respectively.All the diagnostic indices of RCM combined with dermoscopy were higher than those of RCM or dermoscopy alone,except that the specificity was equal to that of dermoscopy alone.RCM showed higher sensitivity,accuracy and negative predictive value,but lower specificity and positive predictive value compared with dermoscopy.There were no significant differences in the diagnostic yield in melanocytic nevus between RCM combined with dermoscopy or RCM alone and histopathological examination (x2 =0.25,0.57,P =0.63,0.45,Kappa value =0.72,0.53,respectively).However,a significant difference in the diagnostic yield in melanocytic nevus was observed between dermoscopy and histopathological examination (x2 =5.81,P =0.012).Conclusion RCM combined with dermoscopy shows higher diagnostic accuracy for melanocytic nevus compared with RCM or dermoscopy alone.
6.Optmization for cutting procedure of astragali radix with Box-Behnken design and response surface method.
Xiu-Juan SHEN ; Qian ZHOU ; Li-Li SUN ; Yan-Peng DAI ; Xue-Sheng YAN
China Journal of Chinese Materia Medica 2014;39(13):2498-2503
Astragali Radix was firstly recorded in the "Shen Nong's Herbal Classic" as a top-grade and commonly used traditional Chinese medicine. Its frequently used slices include raw Astragali Radix and honey-processed products. In current studies, many reports were made on honey-processed Astragali Radix, whereas fewer study reports were made on the cutting process of Astragali Radix. Currently, because Astragali Radix is primarily cut by drug workers according to their operating experience, but with out specific cutting parameters, it is easy to cause the loss or mildew of active ingredients. As a result, the quality of Astragali Radix circulated in the market is not guaranteed, and the quality of their slices and preparations are hard to be controlled, which seriously impact the clinical efficacy. In response, this experiment was performed, in which the optimum cutting process of Astragali Radix was taken as the study objective, the Box-Benhnken central composite design in the response surface analysis was adopted, and the content and appearance character of astragaloside and calycosin-7-glucoside were regarded as the study indicators. Three factors, namely the softening time, the drying temperature and the drying time, were selected to optimize the cutting process of Astragali Radix and obtain the optimum cutting process parameters as follows: the softening time was 3 hours, the drying temperature was 50 degrees C, and the drying time was 4 hours. According to the verification test, the Astragali Radix cutting process is steady and feasible, which has certain significance for normalizing the cutting process of Astragali Radix.
Astragalus Plant
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chemistry
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Chemistry, Pharmaceutical
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methods
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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Glucosides
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chemistry
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Plant Roots
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chemistry
7.Isolation and identification of placental exosomes from maternal serum
Yujing LI ; Zhenyu DIAO ; Pingping XUE ; Li SHEN ; Ping GONG ; Guijun YAN ; Yali HU
Journal of Medical Postgraduates 2015;(6):632-636
Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.
8.Preparation and in vitro evaluation of doxorubicin-loaded magnetic iron oxide nanoparticles.
Song SHEN ; Lin WU ; Cheng-Run WANG ; Xue-Yong QI ; Yan-Ru GE ; Yi JIN
Acta Pharmaceutica Sinica 2013;48(12):1844-1849
PEG-modified magnetic Fe3O4 (Fe3O4-PEG) nanoparticles were sythesized using a solvothermal reaction and characterized with transmission electron microscopy (TEM) and thermo gravimetric analysis (TGA). The photothermal effect and photothermal destruction of cancer cells were evaluated. Then the doxorubicin loaded Fe3O4-PEG (DOX-Fe3O4-PEG) nanoparticles were prepared. The cytotoxicity and combined chemotherapy/photothermal therapy (PTT) effect were investigated. Uniform PEG coated Fe3O4 nanoparticles with particle size of 155 nm were obtained in the experiment. The loading and release of doxorubicin on Fe3O4-PEG were pH-dependent. The drug loading capacity in water was 21%. The results of MTT indicated a good biocompatiblity of Fe3O4-PEG nanoparticles and high cytotoxicity of DOX-Fe3O4-PEG. In combined therapy experiment, photothermal therapy demonstrated unambiguously enhanced chemotherapy efficacy. In conclusion, the obtained Fe3O4-PEG nanoparticles which exhibit good photothermal effect and drug loading capacity can be used for chemotherapy and photothermal therapy. The synergetic anti-tumor activity indicates the potential for the combined application of chemotherapy and photothermal therapy in cancer treatment.
Antibiotics, Antineoplastic
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administration & dosage
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pharmacology
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Cell Survival
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drug effects
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Doxorubicin
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administration & dosage
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pharmacology
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Drug Carriers
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Ferrosoferric Oxide
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chemistry
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Humans
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Hyperthermia, Induced
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MCF-7 Cells
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Magnetite Nanoparticles
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chemistry
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Particle Size
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Polyethylene Glycols
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chemistry
9.Effects of phosphatidylinositol-3 kinase-protein kinase B signaling pathway on iodine-intake ability of lactating rats
Da WEN ; Liang XUE ; Lixiang LIU ; Dandan WANG ; Yingnan YAN ; Hongmei SHEN
Chinese Journal of Endemiology 2016;35(6):395-400
Objective To observe the variation of phosphatidylinositol-3 kinase (PI3K),protein kinase B (AKT),sodium iodide symporter (NIS) mRNA and protein expression in rat mammary tissues and serum insulin growth factor Ⅰ (IGF-1) under different iodine nutrition levels,and to study the role of PI3K-AKT signaling pathway in the process of mammary gland intaking iodine during lactation period.Methods Totally 130 Wistar rats (100 female rats,30 male rats) were randomly divided into five groups with 20 female rats in each group:①control group (NI):was feed with normal diet and iodine content 50 μg/L in deionized water;②low iodine group 1 (LI1 group):was feed with low iodine diet and deionized water;③low iodine group 2 (LI2):was feed with low iodine diet and iodine content 5 μg/L in deionized water;④high iodine group 1 (HI1 group):was feed with normal diet and iodine content 3 000 μg/L in deionized water;⑤high iodine group 2 (HI2):was feed with normal diet and iodine content 10 000 μg/L in deionized water.After feeding for 3 months,females were mated with male rats,then male rats were taken out and every female rat was feed individually.Urinary iodine level of rats in lactation period 10 days after giving birth was tested.Blood and mammary tissue samples of rats in lactation period were taken after killing them.Enzyme linked immunosorbent assay (ELISA) was used to detect serum IGF-1 level,real-time fluorescence quantification PCR to detect the mRNA expression of mammary gland PI3K,AKT and NIS,Western blotting to detect mammary gland PI3K,total AKT,phosphorylation AKT (p-AKT) and NIS protein expression.Results The medians urinary iodine of lactation period rats in LI1 and LI2 (3.16,6.36 μg/L) were significantly lower than that in NI group (162.59 μg/L),and were significantly higher in HI1 and HI2 (2 356.27,11 507.29 μg/L) than that in NI group.The differences were statistically significant (all P < 0.01).Compared with control group [(8.84 ± 2.12) μg/L],the content of serum IGF-1 increased significantly in lactation period rats in LI1 and LI2 groups [(13.30 ± 2.37) and (10.90 ± 1.92) μg/L,all P< 0.01].The real-time fluorescence quantification PCR detection results indicated that the differences were statistically significant by comparing NIS,AKT,PI3K mRNA expression of the mammary tissues of lactation period rats in the five groups (F=14.916,36.477,14.994,all P< 0.01).Among them,NIS mRNA expression quantities in LI1 and LI2 groups (0.75 ± 0.40,0.89 ± 0.51) were significantly higher than that in NI group (0.53 ± 0.31),and significantly lower in HI2 group (0.30 ± 0.24) than that in NI group (P < 0.05 or < 0.01).AKT mRNA expression quantities in LI1 and LI2 groups (0.90 ± 0.19,0.64 ± 0.22) were significantly higher than that in NI group (0.43 ± 0.22),and significantly lower in HI2 group (0.29 ± 0.15) than that in NI group (P < 0.05 or < 0.01).PI3K mRNA expression quantity in LI1 group (0.85 ± 0.42) was significantly higher than that in NI group (0.50 ± 0.24),and significantly lower in HI2 group (0.28 ± 0.10) than that in NI group (all P < 0.01).Western blot detection results indicated that the differences were statistically significant by comparing mammary gland NIS protein expression of lactation period rats in the five groups (F=4.510,P< 0.01).Among them,LI1 group (1.67 ± 0.97) was significantly higher than NI group (0.87 ± 0.43,P < 0.05).The differences were statistically significant by comparing the p-AKT protein expression among groups (F =3.528,P < 0.05).Among them,HI2 group (1.10 ± 0.30) was significantly higher than NI group (0.75 ± 0.23,P <0.05).The differences were not statistically significant by comparing total AKT and PI3K protein expression among groups (F =0.558,1.319,all P > 0.05).Conclusion The inhibitory effect of PI3K-AKT signaling pathways on NIS in the mammary gland was weaker than the effect of iodine intake.But the expression of functional p-AKT was gradually increased with the increment of iodine intake,which had been presented inhibit effect on NIS expression in lactating mammary gland.
10.Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity.
Yanbo ZHU ; Qi WANG ; Chengyu WU ; Guoming PANG ; Jianxiong ZHAO ; Shilin SHEN ; Zhongyuan XIA ; Xue YAN
Journal of Integrative Medicine 2010;8(11):1023-8
To explore the relationships between traditional Chinese medicine (TCM) constitutional types and overweight or obesity so as to provide evidence for adjusting constitutional bias and preventing and treating obesity.