1.Role of miR-124 in breast cancer and its underlying mechanism
Hui REN ; Jianfeng OU ; Qingli ZHAO
Chinese Journal of Clinical Oncology 2015;(20):1012-1017
Objective:To evaluate the role of miR-124 in breast cancer and its underlying mechanism. Methods:Quantitative re-verse transcription-polymerase chain reaction (qRT-PCR) was employed to quantify the expression level of miR-124 in the breast can-cer cell lines and matched tissues of 52 patients. Cell proliferation, invasion, and migration of MDA-MB-231 and T-47D were deter-mined by miR-124 overexpression in vitro. Luciferase vectors (pMIR-SP1 3'UTR) were also constructed. The predicted target gene of miR-124 was identified via luciferase activation assay. The mRNA and protein expression of SP1 was detected via qRT-PCR and West-ern blot, respectively. Results:MiR-124 was decreased in breast cancer tissues and cell lines. This result is correlated with metastatic capacity, TMN stages, and prognosis in breast cancer tissues. In breast cancer cell lines, ectopic overexpression of miR-124 inhibited cell proliferation, invasion, and migration in vitro. MiR-124 mimics significantly inhibited luciferase activation (P<0.05) in HEK293 cells and could significantly decrease the mRNA (P<0.05) and protein expression levels of SP1 in MDA-MB-231 and T-47D cells. Con-clusion:MiR-124 could be inhibited in breast cancer. The low miR-124 expression is associated with poor prognosis. In addition, miR-124 could inhibit cell proliferation, invasion, and migration by targeting SP1. These findings confirm that miR-124 downregulation may be a key mechanism for breast cancer carcinogenesis.
2.Ways for the supervision of department directors
Qilin JIN ; Bin ZHOU ; Jianfeng OU
Chinese Journal of Hospital Administration 1996;0(01):-
Objective To explore ways for the supervision of department directors. Methods Job responsibility agreements with department directors were signed and a series of assessment standards were established. Results The methods adopted were standardized, individualized and easy to use. Conclusion Job responsibility agreements with department directors and a standard assessment system are currently effective measures for the supervision of department directors.
3.The study of isolation and culture in vitro of human umbilical cord mesenchymal stem cells and their biological properties
Hai BAI ; Ke YANG ; Jianfeng OU
Chongqing Medicine 2016;45(7):876-879
Objective To identify a detailed biological characterization of mesenchymal stem cells (MSCs) isolated from hu‐man umbilical cord(UC) tissue regarding their morphology ,immunophenotype ,purity and proliferative capacity and establish a rea‐sonably cultured and amplified system .Methods After stripping off arteries and veins ,the remaining parts of umbilical cord were cut into 1 mm3 small sections and cultured with DMEM/F12 containing 10% fetal bovine serum .Adhere cells were obtained and the morphology of the cells was observed under inverted phase contrast microscope .The growth curves of them were drawn by CCK‐8 and the cell cycle and surface antigens (CD29 ,CD73 ,CD90 ,CD105 ,CD31 ,CD14 ,CD34 ,CD45 ,CD11b ,HLA‐DR) were detected by flow cytometry .Results Seven to ten days after primary culture ,adhere cells came out of fragments .The MSCs harvested were a high purity and mainly presented as a fibroblast‐like morphology .UC‐MSCs had a strong ability of proliferation through the cell growth curve .The special surface antigens CD29 ,CD73 ,CD90 ,CD105 were positive expression ,while CD31 ,CD14 ,CD34 ,CD45 , CD11b ,HLA‐DR were negative .More than 80% cells of MSCs were found at G0/G1 phase .Conclusion Human UC‐MSCs could be cultured and proliferated in vitro .
4.Observation on Dynamic Changes of Flavonoids Contents of Citrus grandis(L.)Osbeck var.tomentosa Hort.
Li LIN ; Lanzhen HUANG ; Jianfeng OU ; Kang CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2001;0(03):-
【Objective】To observe the dynamic changes of flavonoids contents of Citrus grandis(L.)Osbeck var.tomentosa Hort..【Methods】The total flavonoid content was determined by spectral photometric analysis and naringin content by HPLC,and the fingerprints of Citrus grandis(L.)Osbeck var.tomentosa Hort.were studied by HPLC.【Results】With the increase of the fruit age,total flavonoid contents in the peel and the leaves of Citrus grandis(L.)Osbeck var.tomentosa Hort.decreased obviously.Fingerprints results showed that at the early fruit age the rhoifolin content in the peel increased with the fruit age and began to decrease 62 days later,while the rhoifolin content in the leaves showed no changes.【Conclusion】In terms of active components contents and economic value,the optimal collecting time for Citrus grandis(L.)Osbeck var.tomentosa Hort.is:young fruit,34 days,and immature fruit,55 days.
5.Immunocompetent cells and immune regulator improve apopotosis of mesenchymal stem cells by down-regulating heme oxygenase-1
Ling ZHANG ; Hai BAI ; Cunbang WANG ; Jianfeng OU ; Guorong YANG
Journal of Third Military Medical University 2003;0(21):-
Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P
6.Random Amplified Polymorphic DNA Analysis of Germplasm Resources of Exocarpium Citri Grandis
Li LIN ; Jianfeng OU ; Fengxia XIAO ; Xiaohan LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2000;0(04):-
Objective To investigate the influence of producing areas and sexual reproduction on the genetic features of Citrus grandis‘Tomentosa’. Methods The genetic distance in different species was counted,and the DNA fingerprint of germplasm resources of Citrus grandis ‘Tomentosa’ was established after random amplified polymorphic DNA analysis of Citrus grandis ‘Tomentosa’ and Citrus grandis (L.) Osbeck of various species from different producing areas. Results The genetic distance index D of Citrus grandis ‘Tomentosa’ and Citrus grandis (L.) Osbeck of various species from different producing areas was in the range of 0.01~0.64. Conclusion Sexual reproduction and the changes of producing areas can result in the genetic diversity of Citrus grandis ‘Tomentosa’.
7.Diagnostic Significance of Detecting Peripheral Blood Granulocyte CD55/CD59 and FLAER in Anemia and PNH
Ke YANG ; Xiaoyu GUO ; Jianfeng OU ; Hai BAI ; Yaozhu PAN
Journal of Modern Laboratory Medicine 2017;32(3):6-10
Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019<0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163~17.890,P=0.000~0.038<0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ~ 6.308,P=0.000 ~0.016< 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.
8.Influence of Different Processing Technique on the Content of Naringin in Exocarpium Citri Grandis
Lanzhen HUANG ; Zhaoheng LIANG ; Li LIN ; Daoxia GAO ; Jianfeng OU
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To assess the influence of different processing technique on the quality of Exocarpium citri grandis.Methods The RP-HPLC method was adopted to determine the naringin content in Exocarpium citri grandis.The chromatographic conditions were: Kromasil ODS column(250 mm?4.6 mm),mobile phase being methanol-water-acetic acid(35∶61∶4),flow rate 1.0 mL/min,detection wavelength at 283nm and column temperature at 25 ℃.Results There were obvious differences in the naringin content of Exocarpium citri grandis which was processed by different methods: the naringin content processed by electro static drying method was much higher than that by bake drying method, natural drying method and vacuum drying method.Conclusion The electro static drying method is the best way for drying Exocarpium citri grandis.
9.Dynamic changes of CD34~+ cell and T lymphocyte subset from different populations after G-CSF treatment
Cunbang WANG ; Xinghua CHEN ; Yingxian OU ; Hai BAI ; Yaming WEI ; Jianfeng OU ; Haixia MU ; Xiaojin WANG
Journal of Third Military Medical University 2002;0(12):-
Objective To explore the dynamic changes of CD34 + cells and T lymphocyte subsets and best time of harvesting peripheral blood stem cell when G-CSF was used for peripheral blood stem cell mobilization in donors and patients. Methods A total of 12 donors and 16 patients who received chemotherapy for 7 d were injected G-CSF of 300 ?g/d to mobilize peripheral blood stem cells for 5 d and flow cytometry were used to detect the changes of CD34 + cells and T lymphocyte subsets everyday for 5 d. Results ① Before G-CSF treatment, there were obvious differences in bone marrow CD34 + cells between patients and donors (P
10.Autologous bone marrow-derived mesenchymal stem cells and peripheral blood stem cells cotransplantation in treatment of hematological malignant diseases
Tao WU ; Hai BAI ; Cunbang WANG ; Qian ZHANG ; Linfang TANTAI ; Xiaojing WANG ; Jianfeng OU ; Qiang ZHAO
Chinese Journal of Internal Medicine 2009;48(5):392-395
Objective To explore the feasibility and safety of cotransplantation of autologous bone marrow-derived mesenehymal stem cells (MSCs) and peripheral blood stem cells in hematological malignant diseases and to observe its effect on hematopoietic reconstruction after cotransplantation. Methods Adult human MSCs were isolated from the healthy bone marrow of the patient himself with Percoll (1. 073 g/ml) and cultured in Dulbecco's modified Eagle's medium with low glucose containing 10% AB type human serum. After conditioning regimen of high-dose chemotherapy and radiotherapy, cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells was done in five patients with hematological malignant diseases. Results The process of the infusion was safe and there were no adverse reactions or other toxicities related to the infustion of MSCs. The median time to achieve neutrophil counts greater than 0. 5 × 109/L was 9.4 days ( ranging from 8 to 11 days) after cotransplantation and platelet counts greater than 20 × 109/L 12. 2 days (ranging from 10 to 14 days). Conclusion Cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells in hematological malignant diseases is feasible and safe. The rapid hematopoietic reconstruction after cotransplantation shows that MSCs have an effect on hematopeiesis, but the mechanism is still to be investigated.