1.Effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis by inhibiting AKT, ERK and upstream signal SRC activity.
Shi-mei QI ; Jun LV ; Yu MENG ; Zhi-lin QI ; Lie-feng LING
China Journal of Chinese Materia Medica 2015;40(16):3267-3272
To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic
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pharmacology
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Apoptosis
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drug effects
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Breast Neoplasms
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drug therapy
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enzymology
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genetics
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physiopathology
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Down-Regulation
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Extracellular Signal-Regulated MAP Kinases
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genetics
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metabolism
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Female
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Humans
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MCF-7 Cells
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Proto-Oncogene Proteins c-akt
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genetics
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metabolism
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Saponins
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pharmacology
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Signal Transduction
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drug effects
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Triterpenes
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pharmacology
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src-Family Kinases
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genetics
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metabolism
2.Experimental study on cervical lymph nodes removal enhance allograft survival in alkali-burned cornea
Shi-Qi, LING ; Dao-Man, XIANG ; Shi-Yi, XIAO ; Hui, ZHANG
International Eye Science 2007;7(2):287-290
AIM:To explore the inhibitive effects of cervical lymphadenectomy on kerstoplasy after alkaline burns.METHODS:The Wistar rats' corneas were transplanted into Sprague-Dawley(SD)rats' eyes which were randomly divided into 4 groups:group A(control group);group B,the cervicallymphadenectomy group;group C,corneal transplantation after the alkali burn injury;group D,cervical lymphadenectomy following group C.Out of 6 rats in each group,the cornea of one rat Was used for mawophage immunohistochemistry at day 14 after the transplantstion,and the remaining 5 rats were used for studying corneal immune rejection with a slit lamp.The time when allograft rejection occurred was recorded and mean survival times(MST)were compared among the groups.RESULTS:Compared with the MST of group A(10.40±1.14 days),the MST of group B(46.30±9.46 days)Was significantly longer(P<0.05).MST of grafts between group C(7.00± 1.58 days)and group D(15.00±3.39 days)was also significant (P<0.05).At 14th day after the transplantation,there was no CD68 immunoreactivity in the graft of group B,and CD68 proteins were expressed to some extent in the grafts of group A and D.However,in the graft of group C,the expression of proteins Was dramatically up-regulated.CONCLUSION:Cervical lymphadenectomy therapy has a significant effect in preventing corneal allograft rejection in normal and alkali burned oorneai beds.
3.Clinical outcome of patients with follicular development retardation by prolonged duration of gonadotropin administration for in vitro fertilization
Shi-Ling CHEN ; Ling SUN ; Linghong KONG ; Li LI ; Jin LI ; Liang ZHU ; Tian-Ming GAO ; Fu-Qi XING ;
Chinese Journal of Obstetrics and Gynecology 2001;0(08):-
0.05).There were more polycystic ovary (PCO) and (or) polycystic ovarian syndrome (PCOS) patients,more basal antra] follicles,longer duration of Gn stimulation (range 16-33 days),higher Gn dose,lower serum peak estradiol (E_2) level,fewer oocytes,fewer embryos transferred,in group 1 compared with group 2 (P
4.A High-throughput and Quantitative Assay Based on Fluorescence Intensity for Detection of Apoptosis
Ling-Ling YE ; Hong LIU ; Xing-Mao LIU ; Shi-Chong LI ; Ben-Chuan WU ; Qi-Wei WANG ; Zhao-Lie CHEN ;
China Biotechnology 2006;0(06):-
Based on the different permeability of DNA-intercalant dyes YO-PRO-1(YP) and propidium iodide (PI) to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4?mol/L YP and 4?g/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively. The correlation between YP fluorescence intensity and the apoptotic cell number was confirmed by fluorescence microscope and linear regression(r=0.999,P
5.Diallyl disulfide inhibits migration and invasion in human colon cancer SW480 cells through Rac1-ADF/cofilin1 pathway
Jian SU ; Ling SHI ; Yujuan ZHOU ; Hong XIA ; Qianjin LIAO ; Lin DONG ; Shulin XIANG ; Qi SU
Chinese Journal of Clinical Oncology 2013;(14):815-820
Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.
6.CT findings of primary malignant fibrous histiocytoma of the liver
Peihong QI ; Dapeng SHI ; Hongwei ZHENG ; Ling WANG ; Lifu WANG ; Peng XUE
Chinese Journal of Hepatobiliary Surgery 2015;21(10):654-657
Objective To investigate the CT imaging characteristics of primary malignant fibrous histiocytoma (MFH) of the liver.Methods The CT, clinical and histopathologic data of 6 patients with MFH were analyzed retrospectively.All the cases were confirmed by surgery and histopathological examinations.Results The tumors were located in the right liver in 4 patients and the left liver in 2 patients.The maximum diameter of the tumors ranged from 5 to 20 cm.All the tumors were unevenly hypodense on plain CT scan.Four tumors had relatively well-defined margins, while 2 tumors had ill-defined margins.After contrast enhancement, the peripheral and internal divisions of the three storiform-pleomorptic MFH showed progressive enhancement.The peripheral and internal divisions of the two inflammatory MFH were obviously enhanced in the arterial phase, and the enhancement decreased in the portal and delayed phases.The hypodense areas of the one myxiod MFH were not enhanced, but the internal floc and divisions were progressive enhanced.On gross pathology, the tumors were round or irregular in shape, most had a complete capsule with multiple surrounding vessels.These findings correlated well with the CT findings.Conclusions CT findings of primary Hepatic MFH have some characteristics.The enhancement pattern is manifested in a fast wash-in and slow wash-out pattern.However, the final diagnosis still relies on histopathological examination.
7.Corneal lymphangiogenesis correlates closely with hemangiogenesis after keratoplasty
Shi-Qi, LING ; Can, LIU ; Wei-Hua, LI ; Jian-Gang, XU ; Wen-Hui, KUANG
International Eye Science 2010;10(3):405-408
AIM: To examine the relationship between corneal lymphangiogenesis and hemangiogenesis after keratoplasty.METHODS: Nineteen human corneas were obtained from 19 patients undergoing a second corneal transplantation in Zhongshan Ophthalmic Center in 2005. Blood and lymphatic vessels in human transplanted corneas were identified by lymphatic vessel endothelial receptor(LYVE-1) and platelet endothelial cell adhesion modecule-1(PECAM-1) immunohistochemistry, and double enzyme-histochemistry; then the association of corneal blood vessel counting (BVC) with lymphatic vessel counting (LVC) was examined.RESULTS: Corneal hemangiogenesis was present in 12 cases (63%), and lymphangiogenesis occurred in 5 cases (26%) of human transplanted corneas. In addition, corneal lymphangiogenesis was only present in vascularized corneas. LVC was strongly and positively correlated with BVC(r=0.725;P<0.01).CONCLUSION: Corneal lymphangiogenesis develops after keratoplasty, and strongly associates with hemangiogenesis.
8.Molecular Cloning and Sequencing of HCV E2 Gene
Dong-wei, ZHANG ; Bu-feng, LIANG ; Zi-bai, QI ; Shi-Gan, LING
Virologica Sinica 2001;16(1):40-44
HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.
9.CENP-A Regulates Chromosome Segregation during the First Meiosis of Mouse Oocytes
LI LI ; QI SHU-TAO ; SUN QING-YUAN ; CHEN SHI-LING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(3):313-318
Proper chromosome separation in both mitosis and meiosis depends on the correct connection between kinetochores of chromosomes and spindle microtubules.Kinetochore dysfunction can lead to unequal distribution of chromosomes during cell division and result in aneuploidy,thus kinetochores are critical for faithful segregation of chromosomes.Centromere protein A (CENP-A) is an important component of the inner kinetochore·plate.Multiple studies in mitosis have found that deficiencies in CENP-A could result in structural and functional changes of kinetochores,leading to abnormal chromosome segregation,aneuploidy and apoptosis in cells.Here we report the expression and function of CENP-A during mouse oocyte meiosis.Our study found that microinjection of CENP-A blocking antibody resulted in errors of homologous chromosome segregation and caused aneuploidy in eggs.Thus,our findings provide evidence that CENP-A is critical for the faithful chromosome segregation during mammalian oocyte meiosis.
10.Study on standardization of thread-embedding technique: elucidation on the establishment of the national standard standardized manipulation of acupuncture and moxibustion, part X, thread-embedding.
Ling GUAN ; Fang ZUO ; Qi SONG ; Xian SHI
Chinese Acupuncture & Moxibustion 2009;29(5):401-405
To facilitate actualizing the national standard Standardized Manipulation of Acupuncture and Moxibustion, Part X, Thread-Embedding and answer some questions that may arise during practice, overall ideas and principles, the process and basis for its establishment, the main contents and their related research results, especially some disputed questions and the way to solve, as well as the end-results are explained in detail in this paper.
Acupuncture Therapy
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methods
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standards
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China
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Humans
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Moxibustion
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methods
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standards