1.Increased risk of metabolic disease in offspring rats due to maternal high fat diet
Fang FANG ; Yuanwen CHEN ; Ning LIN ; Xiaoyu WAN ; Chongxiao LIU ; Yan DONG
Journal of Clinical Pediatrics 2013;(7):655-659
Objectives To examine the association of the maternal high-fat (HF) diet with increased susceptibility to obe-sity and the development of metabolic diseases in their offspring, and observe difference in the effect of maternal vs. acquired high fat diet on metabolic state in their offspring. Methods A total of 15 SD female rats were divided into HF diet group (group H, n=9) and control diet group (group C, n=6). After fed on different diet for seven weeks, they were mated at the age of ten weeks and became pregnant. Their offspring were then divided to groups CH and HH fed HF diet and groups CC and HC fed control diet. At the age of 3 and 8 weeks, the metabolic markers and the liver pathohistological evidences of their offspring were obtained. Results The body weight, area under curve (AUC) of glucose tolerance, cholesterol and triglyceride were all higher in group H than those in group C (P<0.05) before pregnancy. The offspring of group H had a higher body weight than the offspring of group C at the age of 3 weeks (P=0.002), and no difference in AUC was found between two groups (P>0.05). At the age of 8 weeks, there was no difference in fasting glucose and insulin levels among the four offspring groups. The AUC and body weight were higher in group H than in group C (main effect of maternal diet, P=0.024, P=0.013). The AUCs were also higher in groups CH and HH than groups CC and HC respectively (main effect of acquired diet, P=0.041). The levels of total cholesterol, triglycerides and LDL at the age of 8 weeks were all higher in HH and CH groups than those in HC and CC groups (main effect of acquired diet, P=0.008, 0.007, 0.000, respectively). Their histological analysis at 8 weeks showed different degrees of fatty liver in HH, HC and CH groups, and normal liver in CC group. Conclusions Maternal HF diet may result in increased body weight, fatty liver and impaired glucose tolerance in their adult offspring, and thus increase the risk of developing metabolic diseases at their later age. .
2.Shoulder back lumbar pain treated with application with argy wormwood feeleaf volatile oil.
Wan-Ning LIU ; Hong-Ren GAN ; Cun-Zhong FANG
Chinese Acupuncture & Moxibustion 2013;33(2):171-172
Adult
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Aged
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Artemisia
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chemistry
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Female
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Humans
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Male
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Massage
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Middle Aged
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Oils, Volatile
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therapeutic use
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Plant Oils
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therapeutic use
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Shoulder Pain
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drug therapy
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therapy
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Young Adult
3.Inducing differentiation of human amnion-derived mesenchymal stem cells into insulin-secreting cells in vitro
Yujie ZHAO ; Ning FANG ; Daixiong CHEN ; Limei YU ; Huangfei YU ; Weihong WAN ; Chunhua ZHAO
Chinese Journal of Endocrinology and Metabolism 2011;27(12):1013-1016
ObjectiveTo investigate the potential of human amnion-derived mesenchymal stem cells to differentiate into insulin secreting cells in vitro.MethodsThe hAD-MSCs were isolated from human amnion by trypsin-collagenase digestion.The phenotype of the isolated cells was identified by flow cytometry and immunocytochemical staining.The 3rd generation cells were inoculated at density of 2.5 × 106 unit/ml or 5 × 105 unit/ml in 6 well plates or preset coverslip 24 well plates.Induced groups were treated in serum-free HG-DMEM with 10 mmol/L nicotinamide and N2 supplement.The cells in the non-induced groups were incubated in LG-DMEM supplemented with 10% fetal bovine serum.At days 7,14 and 21 after induction,insulin and β2 microglobulin was determined by immunocytochemical stain,the content of insulin in the culture supernatant was assayed by radioimmunoassay,and insulin mRNA and PDX-1 mRNA were detected by reverse transcriptase-polymerase chain reaction.Results( 1 ) The hAD-MSCs highly expressed CD29,CD44,CD73,CD166,and vimentin.( 2 ) At 7,14,and 21 days,the percentages of insulin-positive cells in hAD-MSCs induced groups were 74.67% ± 1.53%,75.00%:1.00%,and 74.33% ±1.53%,respectively.Contents of insulin in the supematant of hAD-MSCs induced groups were ( 331.62 ± 1.76 ),( 330.50 ± 1.22 ),and ( 331.65 ± 0.48 ) μIU/ml,respectively,but non-induced groups were negative.(3) PDX-1 mRNA and PDX-1 protein were expressed before and after the induction of hADMSCs,but insulin mRNA was expressed only in the induced groups.( 4 ) Both hAD-MSCs induced groups and non-induced groups expressed β2 microglobulin ( all P > 0.05 ).ConclusionThe hAD-MSCs have a potential of differentiating into ISCs and thus may become a new cell source of therapy for type 1 diabetes.
4.Significance of NF-?B in immunopathogenesis of Graves disease
Xun MIN ; Daixiong CHEN ; Yonglun WANG ; Ning FANG ; Weihong WAN ; Jinwei LIU ; Yu XIAO ; Zhenzhong ZHOU
Chinese Journal of Pathophysiology 1986;0(03):-
AIM:To investigate the activity of NF-?B in peripheral blood mononuclear cells (PBMCs) from patients with Graves disease (GD) and the significance in immunopathogenesis of GD.METHODS:Peripheral blood was collected from 22 untreated GD,20 treated GD with tapazole more than 1 year,and 25 healthy volunteers. PBMCs were isolated from the blood by histopaque-1077 density-gradient centrifugation. The activity of NF-?B in PBMCs was analyzed using gel electrophoretic mobility shift assay (EMSA). The contents of IL-1?,IL-6 and TNF-? were tested by radioimmunoassay.RESULTS:The activity of NF-?B in PBMCs of untreated GD group was increased remarkably,compared with that in the treated group and control (P
5.Properties of hematopoietic stem/progenitor cells derived from human placenta tissues
Tao ZHANG ; Ning FANG ; Daixiong CHEN ; Zulin LIU ; Jinwei LIU ; Weihong WAN ; Ying QI ; Jianhui XIAO ; Yu XIAO
Chinese Journal of Tissue Engineering Research 2008;12(21):4172-4176
BACKGROUND: As is well known that hematopoietic stem and progenitor cells (HSPCs) contain in bone marrow, peripheral blood, and cord blood. Recent studies found that human placenta tissue (PT) also exists in HSPCs. But so far the property and differentiation capacity of human PT-HSPCs is not yet known. Furthermore the composition of lymphocyte subpopulations and immunogenicity regarded to human PT-HSPCs are also unclear.OBJECTIVE: To verify whether there are more HSPCs in human PT than those in human umbilical cord blood (UCB), to investigate their capacities of proliferation and differentiation, and to analyze the phenotypes of lymphocyte subpopulations in human PT.DESIGN, TIME AND SETTING: Open eXperiments were performed at the Key Laboratory of Cell Engineering of Guizhou Provinee from January 2004 to December 2006.SETTING: Key Laboratory of Cell Engineering of Guizhou Province, the Affiliated Hospital of Zunyi Medical College.MATERIALS: Twelve human placenta and UCB samples through cesarean delivery were collected aseptically with the informed consents of parturients derived from Maternity Department of the Affiliated Hospital of Zunyi Medical College. The main reagents were detailed as follows: lymphocyte subpopulations analysis reagents Simultest IMK-lymphocyte Kit, CD34 absolute counting reagents Kit (Becton Dickinson); CD34 Multisort Kit, FITC conjugated CD38 monoclonal antibody, anti-FITC microbeads and MS/LS mini MACS segregating, columns (Miltenyi Biotec).METHODS: UCB samples were 1:1 diluted with RPMI-1640 containing 0.1 volume fraction of fetal bovine serum and the mononuclear cells (MNCs) were isolated on Ficoll-Histopaque by centrifugation for 30 minutes. The MNCs at the interface were collected and washed with PBS. Single cells suspension liquid of human PT was prepared by mechanical method combined with 0.25g/L collagenase digestion. After that, the placenta samples underwent the same protocol as used in UCB to isolate MNCs. The percentage of CD34+CD38-, CD34+CD38+ HSPCs and the phenotype of lymphocyte subpopulations derived from human PT-MNCs were analyzed by flow cytometry (FCM). CD34+CD38-, CD34+CD38- cell subsets isolated by magnetic-activated cell sorting (MACS) from human PT were used to carry out colony-forming culture including granulocyte/macrophage colony-forming unit (CFU-GM), burst forming unit-erythroid (BFU-E) and mixed colony-forming unit (CFU-Mix) in order to assess their capacities of hematopoietic progenitor cells' proliferation and differentiation. In parallel, UCB samples underwent the same protocols for comparison.MAIN 0UTCOME MEASURES: Percent compositions of CD34+ HSPCs, hematopoietic progenitors' lineage colony-forming capacities of CD34+ HSPCs, phenotypes and compositions of lymphocyte subpopulations both in PT and UCB.RESULTS: The percentage of CD34+ cells contained in human PT was 8.8 times higher than that of in UCB (P<0.01). The total number of lymphocytes, T cells (CD3+CD2+), B cells (CD19+), Th (CD3+CD4+) and Th/Ts ratio were apparently lower in human placenta, while the number of CD8+CD28- T suppressor cells were higher compared to UCB samples (P<0.01). Among PT, CFU-GM, BFU-E and CFU-Mix frequencies of CD34+CD38- cells subset were much higher than that of CD34+CD38- (P<0.01). Within the same phenotype of cell subsets, however, the number of each colony-forming unit was similar between PT and UCB (P 0.05).CONCLUSION: Human PT is richer in CD34+CD38-, CD34+CD38+ HSPCs and both of them have the abilities of proliferating and differentiating into CFU-GM, BFU-E and CFU-Mix. Considering that human PT have a lower lymphocyte subpopulations and higher Ts cells, human PT might be a alternative and suitable source of HSPCs for clinical transplantation.
6.Research situation of effects of sulfur fumigation on quality of traditional Chinese medicine.
Tu-Lin LU ; Zi-Wan NING ; Xin SHAN ; Lin LI ; Chun-Qin MAO ; De JI ; Fang-Zhou YIN
China Journal of Chinese Materia Medica 2014;39(15):2796-2800
Sulfur fumigation (SF) is a universal phenomenon in primary processing of Traditional Chinese Medicine (TCM) in modern times. In the process, fumigation, sulfur or both of them act on the TCMs. Some active components of TCMs change quantitatively or qualitatively during the processing. At the same time, the sulfur dioxide and heavy metal would remain and cause a serious influence on quality and future development of TCM. This article reviews the chemical compositions change after SF to study the change law and their influence on quality. This article provide references for SF in TCMs' processing for a better and safer quality.
Drug Contamination
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Fumigation
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methods
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Medicine, Chinese Traditional
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methods
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Quality Control
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Sulfur
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chemistry
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Technology, Pharmaceutical
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methods
7.Differentiation of Human Amniotic Mesenchymal Cells into Cardiomyocyte-like Cells in Vitro
Lu ZHANG ; Ning FANG ; Dai-Xiong CHEN ; Zu-Lin LIU ; Jin-Wei LIU ; Wei-Hong WAN ; Tao ZHANG ;
China Biotechnology 2006;0(12):-
To explore the plasticity of human amniotic mesenchymal cells(hAMCs) into cardiomyocyte-like cells,hAMCs were isolated from human amnion with collagenase digestion.Phenotype of the isolated cells was analyzed by flow cytometry(FCM).hAMCs were treated with 5-azacytidine and basic fibroblast growth factor(bFGF) to investigate their ability of differentiation into cardiomyocytes.The induced differentiated cells were evaluated by immunofluorescence for desmin and ?-actin expression and by RT-PCR for Nkx2.5,GATA-4 and alpha-myosin heavy chain(?-MHC) mRNA expression.The results showed that,after primary culture,hAMCs could reach a confluence of 80% with swirl like growth at 6 days.The cells proliferated rapidly after passages with a 100% confluence at 3~4 d.hAMCs were positive expression of CD44 and vimentin,but negative for CK19.After induced differentiation at 8~10d,the differentiated cells have close-up arranged with long spindle-shape.At 2 weeks and 4 weeks,induced cells expressed ?-actinin and cardiac-specific transcription factor Nkx2.5.Expressions of GATA-4 and desmin can be detected but ?-MHC can not in the hAMCs both before and after the induction.In conclusion,hAMCs have the ability of differentiation into cardiomyocyte-like cells,which means that hAMCs can be regarded as candidate cells for cellular cardiomyoplasty(CCM).
8.Phenotypic characteristics of peripheral blood lymphocytes and its clinical significance in patients with coal arsenic poisoning
Ning, FANG ; Tao, ZHANG ; Wei-hong, WAN ; Xi-jie, HU ; Ying, QI ; Xin-jiang, ZHANG ; Dai-xiong, CHEN
Chinese Journal of Endemiology 2008;27(5):561-565
Objective To study the change and rule of immunological function among the patients with coal arsenic poisoning in order to provide a basis for tumor risk evaluation and monitoring.Methods Seventy patients with coal arsenic poisoning aged from 24 to 71 years old(44 men,26 women,averaging 41 years old)were divided into 4 groups including 23 cases having a course less than 10 years,21 case8 lasting for 10~19 years,20 cages for more than 20 years,6 cases of cancer,and 26 healthy normal controls.Flow cytometer(FCM)was used to analyze the frequency of CD3+(total T cell),CD3+CD4+(inducer/helper T cell),CD3+CD8+(suppressor/cytotoxic T cell),CD19+(B lymphocyte),and CD56+CD16+(natural killer cell)lymphocyte subsets in the peripheral blood of the subjects and the expression rates of lymphocytic membrane surface molecules of human leucocvte antigen (HLA)-DR,CD25,CD38 were also determined by FCM.Results The pmportions of CD3+cells in periDheral blood of less than 10 years,10~19 years,more than 20 years and cancer groups were (63.76±9.32)%。(55.63± 12.97)%,(51.00±12.23)%and(49.83±,9.89)%respectively,which were significantly lower than that in control group[(68.10±8.62)%],and there was a significant difference between different groups(F=12.862,P<0.05). In less than 10 years,10~19 years,more than 20 years and cancer groups,the proportion of CD3+CD4+cells cells was (31.35±6.62)%,(28.38±8,66)%,(24.13±6.46)%and(19.17±4.96)%respectively,which wag significantly lowerthan that in control group[(34.28±7.32)%],and significant in a-group difference was found(F=10.455, P<0.05).The percentages of CD19+cells in more than 20 yeats and cancer groups[(9.00±5.32)%,(9.00± 3.29)%]were lower than that in control group and less than 10 years group[(11.80±3.43)%,(12.35±4.53)%] (P<0.05),while no statistical difference was found between other groups.The expression rates of CD25 and CD38 in lymphocytes of cancer group[(17.96 ±4.98)%,(41.38±8.54)%]were obviously higher than those in control group[(13.10±338)%,(28.60±5.51)%]and there were statistical differences between the experimental groups(P<0.05).The expression rate of HLA-DR in 10~19 years groups[(18.20±6.25)%]was significantly higher than that in control group[(10.72±7.06)%]and less than 10 years group[(11.78±5.13)%],while it was the same in more than 20 years and cancer group[(20.30±8.01)%,(21.82±10.97)%].Conclusions Reduction of cellular immune function caused by coal arsenic poisoning may be an important mechanism of skin cancer.CelMar immune function may be used as a warning signal of skin cancerization of patients with coal arsenic poisoning.
9.The effects of CT120B over-expression on growth suppression and changes of gene expression profiles in lung adenocarcinoma cells.
Dong-ning PAN ; Lin WEI ; Ming YAO ; Da-fang WAN ; Jian-ren GU
Chinese Journal of Oncology 2006;28(5):321-325
OBJECTIVECT120B gene is a splicing variant of CT120A, which deletes 96 nucleotides and leads to an in-frame loss of 32 amino acids between the codon 136 and 167 as compared with CT120A. This study was undertaken to assess the effects of CT120B expression on lung cancer cell growth and to explore the gene expression profiles.
METHODSCT120B cDNA was transfected into the human lung adenocarcinoma SPC-A-1 cells, and stable cell lines overexpressing CT120B were established. CCK-8 assay and tumorigenecity in a xenograft model were performed to analyze cell proliferation in vitro and in vivo. The differential gene expression induced by overexpressed CT120B was investigated using Atlas cDNA expression array. Flow cytometry was performed to analyze cell cycle and cell apoptosis.
RESULTSOverexpression of CT120B in SPC-A-1 cells resulted in a reduced cell growth rate in vitro, and decrease of the tumorigenicity in nude mice. A total of 38 genes were identified as differential expressions with more than a 2.0-fold change by Atlas cDNA expression array analysis, including downregulated cyclin E1, cdk 2, c-kit, CXCR4 and upregulated caspase 8 gene. Overexpression of CT120B also induced G1 phase arrest, but had no effect on cell apoptosis.
CONCLUSIONThe G1 cell cycle arrest, but not apoptosis, underlay the growth inhibitory activities of CT120B. The down-regulation of c-kit and CXCR4 expression might also contribute to the suppressive effects on cell growth of CT120B.
Adenocarcinoma ; metabolism ; pathology ; Animals ; Cell Line, Tumor ; Cell Proliferation ; DNA, Complementary ; genetics ; G1 Phase ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Proteins ; genetics ; metabolism ; Neoplasm Transplantation ; Oligonucleotide Array Sequence Analysis ; Proto-Oncogene Proteins c-kit ; metabolism ; Receptors, CXCR4 ; metabolism ; Transfection