1.Expression and significance of Oct4 B1 in colorectal cancer stem cells
Jiaping CHENG ; Jiang LI ; Xian SU ; Yilin CHEN ; Qingliang ZENG ; Kunming WEN
Chinese Journal of Immunology 2016;32(6):794-797
Objective:To investigate the expression and its possible role of Oct4B1 subtype of Embryonic stem cell transcription factor Oct4 in colorectal cancer stem cells. Methods: 3D microspheres were cultured by suspension culture to human colorectal cancer cell line SW480 cells. The 3D microspheres and SW480 cells were used as the research objects. Whether 3D microspheres were enriched cancer stem cells,we used the methods of cell differentiation experiments,soft agar cloning experiments,and the expression levels of cancer stem cells markers CD133,CD44 detected by flow cytometry. The expression levels of Oct4B1 mRNA were detected by RT-qPCR. Results:3D microspheres could differentiate into normal cancer cells. Compared with the parental SW480 cells,in vitro colony formation was significantly enhanced(P<0. 01),the percentage of positive cells of CD133 and CD44 were significantly increased ( P < 0. 01 ), the expression levels of Oct4B1 mRNA were obviously higher ( P < 0. 01 ) in 3D microspheres. Conclusion: Oct4B1 subtype of Embryonic stem cell transcription factor Oct4 in 3D microspheres enriched human colorectal cancer stem cells,which may be involved in the regulation of colorectal cancer stem cells.
2.Locally administered lentivirus-mediated siRNA inhibits wear debris-induced inflammation.
Xiao-chun PENG ; Xian-long ZHANG ; Kun TAO ; Tao CHENG ; Jun-feng ZHU ; Bing-fang ZENG
Chinese Journal of Surgery 2009;47(5):377-380
OBJECTIVETo determine the safety and efficacy of local administration of lentivirus-mediated small interfering RNA (siRNA) targeting tumor necrosis factor-alpha (TNF-alpha) in murine air pouch model.
METHODSFrom May 2007 to April 2008 a siRNA targeting TNF-alpha and a missense siRNA were designed, and recombine lentivirus which coexpressed the green fluorescent protein (GFP) as a marker gene was constructed. Air pouches were established and stimulated by Ti-6Al-4V particles. Pouches were divided into 3 groups randomly. Lentivirus-mediated siRNA targeting TNF-alpha (TNF-alpha group) or lentivirus-mediated missense siRNA (MS group), or virus-free saline (control group) were injected into pouches respectively. Pouch membrane, peripheral blood, heart, liver, spleen, kidney, lung and brain were harvested at 28 d after transfection, and assayed for markers of inflammation using histological, molecular, immunological techniques and Xenogen in vivo imaging system (IVIS) 50 vivo bioluminescent assay system.
RESULTSXenogen IVIS 50 vivo image revealed strong expression of GFP localized in pouch areas and no expression in other parts of mice both in TNF-alpha group and MS group at 4 weeks after transfection, while no expression of GFP was found in control group. By RT-PCR and ELISA, the mRNA and protein levels of TNF-alpha in TNF-alpha group decreased by 81.6% and 82.6% respectively compared to control group (P < 0.01), and decreased by 78.9% and 84.0% respectively compared to MS group (P < 0.01), whereas TNF-alpha level in peripheral blood, heart, liver, spleen, kidney, lung and brain remained invariant (P > 0.05). Less inflammatory responses (thinner pouch membrane and decreased cellular infiltration) were observed in TNF-alpha group.
CONCLUSIONEfficient local delivery of lentivirus-mediated siRNA targeting TNF-alpha into modified murine air pouch can inhibit debris-induced inflammation effectively, with no systemic adverse effects.
Animals ; Disease Models, Animal ; Genetic Therapy ; Genetic Vectors ; genetics ; Inflammation ; therapy ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; genetics ; Random Allocation ; Transfection ; Tumor Necrosis Factor-alpha ; genetics
3.Curative effect analysis on closed reduction and external fixator under local anesthesia for the treatment of intertrochanteric fracture in elderly patients with high-risk.
Zeng-ping WANG ; Lin LIU ; Wen XUE ; Hui-ru ZHOU ; Yu-xin SONG ; Li-yang CAI ; Xian-tang CHENG ; Yao-wen QIAN
China Journal of Orthopaedics and Traumatology 2016;29(6):502-504
OBJECTIVETo explore clinical efficacy of closed reduction and external fixation under local anesthesia for the treatment of high-risk elderly patients with intertrochanteric fracture.
METHODSFrom March 2013 to March 2015, 10 patients with intertrochanteric fractures treated with closing reduction and external fixator under local anesthesia were analyszed, including 4 males and 6 females, aged from 69 to 88 years old with an average of 75.2 years old. All fractures were caused by injury and classified to type I (5 cases), II (3 cases), and V (2 cases) according to Evans classification. According to American Society of Anesthesiologists (ASA), 6 cases were type III and 4 cases were type IV. Blood loss,operative time,hospital stays, postoperative complications, ambulation time and fracture healing time were observed, and Harris scoring were used to evaluate hip joint function.
RESULTSAll patients were followed up from 3 to 23 months with an average of 13.1 months. One patient with chronic obstructive pulmonary disease died for non-operation reason at 4 months after operation, the other fractures were healed at stage I, the mean fracture healing time was 5.6 months. There were no coxa vara, lower limb venous thrombosis, loosen and remove of needle passage. The average operative time was 46 min, blood loss was (35.00 ± 8.46) ml without blood transfusion. One patient was occurred pulmonary infection and stent-tract infection on the 2 nd and 3 rd day after operation, and improved with active anti-infection and dressing change; the other patients gone to ground activity at 4.2 d after operation. The patients stayed hospital for 10.6 d on average. According to Harris scoring at final following-up, the total score was 83.42 ± 3.27, 3 cases obtained excellent results, 5 cases good and 1 case poor.
CONCLUSIONClosed reduction and external fixation under local anesthesia in treating high-risk elderly patients with intertrochanteric fracture,which has advantages of shorter operative time, less blood loss, good recovery of postoperative function, is a safe, stable and economic method.
Aged ; Aged, 80 and over ; Anesthesia, Local ; Bone Nails ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Fracture Fixation, Intramedullary ; Fractures, Closed ; surgery ; Hip Fractures ; surgery ; Humans ; Male ; Treatment Outcome
4.Cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.
Ying-Xian DENG ; Zeng-Chun MA ; Hong-Ling TAN ; Cheng-Rong XIAO ; Yu-Guang WANG ; Yue GAO
China Journal of Chinese Materia Medica 2006;31(24):2070-2073
OBJECTIVETo explore the cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.
METHODRats were ovariectomized and administered drugs for 3 monthes. Bone mineral density and biomechanics properties, histomorphometric analysis and biochemical index such as calcium, phosphorus, alkaline phosphatase were detected.
RESULTJiangu Fufang could significantly increase bone density and biomechanics properties. The level of calcium, phosphorus and alkaline phosphatase were restored by Jiangu Fufang. Jiangu Fufang could significantly increase area of bone trabecula, thickness of cortical bone and bone trabecula.
CONCLUSIONJiangu Fufang could cure osteoporosis through increasing bone mineral density, improving bone biomechanics properties, and effecting bone metabolism.
Alkaline Phosphatase ; blood ; Animals ; Biomechanical Phenomena ; Bone Density ; drug effects ; Calcium ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Female ; Femur ; physiopathology ; Lumbar Vertebrae ; drug effects ; pathology ; physiopathology ; Osteoporosis ; blood ; drug therapy ; physiopathology ; Ovariectomy ; Phosphorus ; blood ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley
5.Predictive role of diagnostic information in treatment efficacy of rheumatoid arthritis based on neural network model analysis
Qinglin ZHA ; Yiting HE ; Xiaoping YAN ; Li SU ; Yuejin SONG ; Shengping ZENG ; Wei LIU ; Xinghua FENG ; Xian QIAN ; Wanhua ZHU ; Seqi LIN ; Cheng Lü ; Aiping Lü
Journal of Integrative Medicine 2007;5(1):32-8
OBJECTIVE: To analyze the indications of the therapies for rheumatoid arthritis (RA) with neural network model analysis. METHODS: Three hundred and ninety-seven patients were included in the clinical trial from 9 clinical centers. They were randomly divided into Western medicine (WM) treated group, 194 cases; and traditional Chinese herbal medicine (CM) treated group, 203 cases. A complete physical examination and 18 common clinical manifestations were prepared before the randomization and after the treatment. The WM therapy included voltaren extended action tablet, methotrexate and sulfasalazine. The CM therapy included Glucosidorum Tripterygii Totorum Tablet and syndrome differentiation treatment. The American College of Rheumatology 20 (ACR20) was taken as efficacy evaluation. All data were analyzed on SAS 8.2 statistical package. The relationships between each variable and efficacy were analyzed, and the variables with P<0.2 were included for the data mining analysis with neural network model. All data were classified into training set (75%) and verification set (25%) for further verification on the data-mining model. RESULTS: Eighteen variables in CM and 24 variables in WM were included in the data-mining model. In CM, morning stiffness, swollen joint number, peripheral immunoglobulin M (IgM) level, tenderness joint number, tenderness, rheumatoid factor (RF), C-reactive protein (CRP) and joint pain were positively related to the efficacy, and disease duration and more urination at night negatively related to the efficacy. In WM, erythrocyte sedimentation rate (ESR), weak waist, white fur in tongue, joint pain, joint stiffness and swollen joint were positively related to the efficacy, and yellow fur in tongue, red tongue, white blood negatively related to the efficacy. In the analysis with the neural network model in the patients of verification set, the predictive response rates of 20% patients would be 100% and 90% in the treatment with CM and WM, respectively. CONCLUSION: Neural network model analysis, based on the full clinical trial data with collection of both traditional Chinese medicine and modern medicine diagnostic information, shows a good predictive role for the information in the efficacy in rheumatoid arthritis.
6.Association of single nucleotide polymorphism on chromosome 12q24.31 with susceptibility to coronary artery disease.
Wen-wen XIAO ; Chuan-wei LI ; Zhi ZENG ; Rui LIU ; Xian LI ; Yu-cheng CHEN
Chinese Journal of Medical Genetics 2011;28(4):455-459
OBJECTIVETo determine whether the single nucleotide polymorphism (SNP) on chromosome 12q24.31(rs2259816) is associated with coronary artery disease (CAD) in Han population of southwest China.
METHODSA case-control association study with 592 unrelated patients with coronary artery disease and 463 normal controls from Chinese Han population was performed. Genotype for the SNP on chromosome 12q24.31 (rs2259816) was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).
RESULTSThe genotypes of AA, AC, CC were both detected in the coronary artery disease group and the control group. The frequencies of A allele were 49.5% in case group and 43.8% in control group, showing statistically significant difference(OR=1.129, 95%CI:1.029-1.239, P=0.010).
CONCLUSIONThe replication study showed that the genetic polymorphism in rs2259816 is associated with coronary artery disease in Han population of southwest China.
Base Sequence ; Case-Control Studies ; Chromosomes, Human, Pair 12 ; genetics ; Coronary Artery Disease ; blood ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Lipids ; blood ; Male ; Polymorphism, Single Nucleotide ; genetics
7.Effect of Siwu decoction on function and expression of P-glycoprotein in Caco-2 cells.
Yi JIANG ; Zeng-chun MA ; Xian-ju HUANG ; Qing YOU ; Hong-ling TAN ; Yu-guang WANG ; Qian-de LIANG ; Xiang-lin TANG ; Cheng-rong XIAO ; Yue GAO
China Journal of Chinese Materia Medica 2015;40(5):933-937
To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B
;
genetics
;
metabolism
;
ATP-Binding Cassette, Sub-Family B, Member 1
;
genetics
;
metabolism
;
Caco-2 Cells
;
Drugs, Chinese Herbal
;
pharmacology
;
Humans
;
Up-Regulation
;
drug effects
8.Health literacy level and health related behavior of high school students in Xuhui District of Shanghai
Zi-yi CUI ; Xian WANG ; Ze-liang XUAN ; Xiao-lei JIN ; Shu-jun ZENG ; Cheng-gang ZHANG
Shanghai Journal of Preventive Medicine 2020;32(6):468-
Objective To investigate the health literacy level and health related behavior of high school students in Xuhui District of Shanghai, and to provide basis for health literacy intervention. Methods Using qualitative and quantitative methods, the quantitative part adopted the multi-stage stratified sampling method.A total of 355 aged 15-18 from senior high schools in Xuhui District were selected for face-to-face survey.The qualitative part adopted the form of group interviews, and 20 students from two senior high schools in Xuhui District were selected for interviews. Results The health literacy level of high school students in Xuhui District was 27.32%, which was lower than that of adults in the same period.Among the three dimensions, the level of basic knowledge and concept literacy was the highest, followed by the level of health basic skills literacy, the level of healthy lifestyle and behavior literacy was the lowest; among the six types of health problems, the level of infectious disease prevention literacy and basic medical literacy was at a low level.Statistical analysis of the correct answer rate showed that the answer rate of various questions in the health literacy survey was quite different.Further analysis of the health-related behavioral characteristics of high school students showed that most students had correct health concept and awareness of health change, but they encountered resistance in the process of knowledge transformation into action, which mainly came from academic burden, school education, peer influence and so on.The main channels for high school students to obtain health-related information were classroom, parents, school and various media.They had a high degree of trust in medical professionals and encountered difficulties in seeking health information. Conclusion The health literacy level of senior high school students in Xuhui District of Shanghai needs to be improved.We should explore a health intervention model more suitable for the behavioral characteristics of this group weak links in their health literacy.
9.In vitro effect of all-trans retinoic acid on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells in patients received peripheral blood stem cell transplantation.
Cheng CHANG ; Xing-Hua CHEN ; Pei-Yan KONG ; Xian-Gui PENG ; Dong-Feng ZENG ; Wen-Bo YANG ; Xue LIANG ; Lin LIU ; Hong LIU ; Qing-Yu WANG
Journal of Experimental Hematology 2006;14(4):768-772
The aim of this study was to evaluate the effect of all-trans retinoic acid (ATRA) on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells (BMSC) in patients after conditioning treatment for peripheral blood stem cell transplantation (PBSCT). BMSC of 27 patients before and after conditioning treatment for PBSCT were cultured in vitro. After treated with ATRA at 0.01, 0.1, or 1 micromol/L, expression of intercellular adhesion molecule-1 (ICAM-1) protein and vascular adhesion molecule-1 (VCAM-1) protein were detected by flow cytometry, and soluble ICAM-1 (sICAM-1) protein was determined by using radioimmunoassay. Then BMSC was co-cultured with CD34+ cells, and adhesion rate of BMSC to CD34+ cells was measured. The results showed that after pretreatment with conditioning regimen for PBSCT, the expressions of ICAM-1 and VCAM-1 proteins in BMSC and the expression level of sICAM-1 protein in supernatant of BMSC culture were down-regulated, and the adhesion rate of BMSC to CD34+ cells was decreased, after administration of ATRA, the expression of ICAM-1 protein in BMSC, sICAM-1 protein in culture medium and adhesion rate of BMSC to CD34+ cells all increased significantly, but expression of VCAM-1 protein changed no significantly. It is concluded that the ATRA can partly restore adhesion function of BMSC injured by pretreatment for PBSCT and contribute to hematopoietic reconstitution.
Adolescent
;
Adult
;
Antigens, CD34
;
Antineoplastic Agents
;
pharmacology
;
Bone Marrow Cells
;
metabolism
;
pathology
;
Cell Adhesion
;
drug effects
;
Cell Adhesion Molecules
;
biosynthesis
;
genetics
;
Child
;
Coculture Techniques
;
Hematologic Neoplasms
;
metabolism
;
pathology
;
therapy
;
Humans
;
Middle Aged
;
Peripheral Blood Stem Cell Transplantation
;
Stromal Cells
;
metabolism
;
pathology
;
Tretinoin
;
pharmacology
;
Tumor Cells, Cultured
;
Vascular Cell Adhesion Molecule-1
;
biosynthesis
;
genetics
10.Effect of the NHE-1-specific inhibitor DMA on pHi, proliferation and apoptosis of HL-60/ADM cells in vitro.
Cheng CHANG ; Pei-Yan KONG ; Xing-Hua CHEN ; Xian-Gui PENG ; Lin LIU ; Hong LIU ; Dong-Feng ZENG ; Xue LIANG ; Qing-Yu WANG
Journal of Experimental Hematology 2006;14(3):488-491
The aim of this study was to evaluate the effect of dimethyl amiloride (DMA), a specific inhibitor of Na(+)/H(+) exchanger-1 (NHE-1), on intracellular pH value (pHi), proliferation and apoptosis of HL-60/ADM cells in vitro. After treatment with DMA at different doses, pHi of HL-60 and HL-60/ADM cell lines were determined by using pH-sensitive fluorescence dye BECEF-AM; the rate of growth inhibition of cells was detected with MTT assay; cell cycle was detected by flow cytometric DNA analysis; cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The results showed that pHi in HL-60/ADM cells was higher than that in HL-60 cells. After treatment with DMA at different doses, pHi decreased, the rate of growth inhibition and the rate of apoptotic cells in HL-60/ADM cells were all higher than those in HL-60 cells. Meanwhile, after treatment with DMA during 100 micromol/L to 150 micromol/L, the increase amplitude of G(0)/G(1) phase cells and the decrease amplitude of S + G(2)/M cells in HL-60/ADM cells were higher than those in HL-60 cells. It is concluded that by causing intracellular acidification, the NHE-1-specific inhibitor DMA inhibits proliferation of HL-60/ADM cells and induces apoptosis of HL-60/ADM cells, and the degree of this growth inhibition of HL-60/ADM cells is higher than that of HL-60 cells.
Amiloride
;
analogs & derivatives
;
pharmacology
;
Apoptosis
;
drug effects
;
Cell Cycle
;
drug effects
;
Cell Proliferation
;
drug effects
;
Doxorubicin
;
pharmacology
;
Drug Resistance, Neoplasm
;
HL-60 Cells
;
Humans
;
Hydrogen-Ion Concentration
;
Sodium-Hydrogen Exchangers
;
antagonists & inhibitors