2.Expression and tyrosine phosphorylation of insulin receptor substrate-1 in adipose tissue from patients with gestational diabetes mellitns
Jianwen ZHU ; Ming HE ; Yan ZHOU ; Han XIE ; Juan WANG ; Huan WANG ; Li ZOU
Chinese Journal of Endocrinology and Metabolism 2008;24(4):397-399
Objective To investigate the expression and tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and in the adipose tissue of patients with gestational diabetes mellitus (GDM), and to explore molecular mechanisms of insulin resistance in GDM. Methods The serum and adipose tissue were sampled from patients with GDM (GDM group, n = 20) and normal pregnant women (control group, n = 20). Fasting plasma glucose was measured by glucose oxidase assay. The expressions of IRS-1 protein and mRNA were determined by Western blot and semi-quantitative RT-PCR. The tyrosine phosphorylation of IRS-1 was measured by immunoprecipitation. Results Compared with control group, in GDM group, the expression of IRS-1 mRNA was markedly decreased (0.61 ±0.06 vs 1.12 ± 0.17, P < 0.01), the expression of IRS-1 protein was significantly decreased (0.57 ±0.08 vs O. 83 ±0.07, P <0.01) and tyrosine phosphorylation was significantly reduced (0.23 ± O. 06 vs O. 62 ±0.04, P < 0.01) in the adipose tissue. Conclusion The decline of protein expression and tyrosine phosphorylation of IRS-1 in the adipose tissue of gestational diabetes appears to be one of the moleculemechanisms of insulin resistance in patients with GDM.
3.Relationship between variation of phosphatidyl inositol-3 kinase expression and activity and insulin resistance in pregnant women with gestational diabetes mellitus
Jianwen ZHU ; Ming HE ; Yan ZHOU ; Han XIE ; Juan WANG ; Huan WANG ; Li ZOU
Chinese Journal of Perinatal Medicine 2009;12(1):6-9
Objective To investigate the variations of the expression and activity of phosphatidyl inositol-3 kinase(PI-3K)in the adipose tissue of pregnant women with gestational diabetes mellitus(GDM)and to explore its relationship with insulin resistance(IR)in these women.Methods The expression of PI-3K p85aprotein and mRNA in adipose tissue were detected by Western blot and Semi quantitative reverse transcription polymerase chain reaction(RT-PCR)in 20 GDM women(GDM group)and 20 normal pregnant women(normal group).The activity of PI-3K was determined by ELISA and IR index was calculated using homeostasis model assessment(HOMA)according to the results of fasting insulin(FINS)and fasting plasma glucose(FPG),which measured by oxidase assay and immunoradioassay,respectively.Results The expressions of PI-3K p85αmRNA and PI-3K p85a protein were markedly increased in the adipose tissue of GDM group than in the normal group(mRNA:0.83±0.03 vs 0.53±0.07,P<0.01;protein:0.93±0.04 vs 0.71±0.06,P<0.01).However,the PI-3K activity in the GDM groups was significantly down-regulated compared with the normal group(1.7±0.6 vs 5.2±0.5,P<0.01).The levels of FPG and FINS (HOMA-IR)in the GDM group were all significantly higher than the normal group[FPG:(5.8±0.2)mmol/L vs(4.7±0.3)mmol/L;FINS:(14.8±0.2)mmol/L vs(11.2±0.3)mmol/L;HOMA-IR:1.3±0.4 vs 0.9±0.3,a11 P<0.01]. Conclusions The decreased activity of PI-3K in the adipose tissue may be one of the molecular mechanisms for IR in GDM.
4.Influence of Pranlukast on Expression of Eotaxin-3 in Bronchial Epithelia Cells
wei-lin, OU ; huan, WEI ; zhi-ying, FENG ; chun-jiang, ZHU ; ming-ci, ZHENG
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To investigate whether leukotriene D4 (LTD4) regulates eotaxin-3 (Eot-3) expression in bronchial epithelial cells, and study effect of pranlukst on the regulation.Methods BEAS-2B cells and normal human bronchial epithelia cells were pre- treated with LTD4 for 1 hour,stimulated with interleukin-4, the cells were incubated for 24 hours. Eot-3 protein in supernatant were measured by enzyme linked immunosorbent assay(ELISA). The cells were pretreated with pranlukast in different concentration, then the above procedure was repeated. Results The untreated bronchial epithelial cell expressed Eot-3 protein on a very low level. After stimulating with IL-4 and incubating for 24 hours, Eot-3 production increased significantly. Pretreating the cells with LTD4 enhanced the inducing effect of IL-4. Pranlukast inverted the upregulation of LTD4. Conclusions Upregulating the expression of Eot-3 induced by IL-4 on bronchial epithelial cells may explain partially the mechanism of leukotrienes involving airway allergic inflammation of asthma. The invertion impact on upregulation of LTD4 by pranlukast may be one of mechanisms that leukotrienes receptor antagonist cure asthma.
5.Effects of fuzheng jianpi decoction combined chemotherapy on the quality of life and the survival time of children with solid tumor.
Xue SHI ; Xiu-Dan ZHU ; Huan-Ming WANG
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(9):1175-1179
OBJECTIVETo study the effects of Fuzheng Jianpi Decoction (FJD) combined chemotherapy on the quality of life (QOL) and the survival time of children with solid tumor.
METHODSRecruited were 167 solid tumor children patients at Department of Tumor, Beijing Children's Hospital from Jan. 2005 to Jan. 2008. They were randomly assigned to the treatment group (83 cases) and the control group (84 cases) according to the random digit table. All had chemotherapy. Those in the treatment group additionally took FJD, 50 -100 mL each time, twice daily. After chemotherapy those in the treatment group took modified FJD. The WBC, Hb, and PLT were detected in all patients before treatment, 6 months after treatment, and 1 year after treatment. The 1-, 2-, and 3-year QOL, 3-year survival rate, and the survival life of dead children patients were observed.
RESULTSCompared with the control group of the same period, the 6-month and 1-year WBC and Hb increased, the 1-year PLT increased in the treatment group, showing statistical difference (P<0.05, P<0.01). Compared with 1-year treatment in the treatment group, the 2-and 3-year psychological functions and the general symptoms scores decreased (P<0.05, P<0.01). Compared with the control group of the same period, the 2-and 3-year somatic functions, psychological functions, and the general symptoms scores decreased in the treatment group; the 2-year somatic functions and psychological functions decreased in the treatment group; the 3-year psychological functions and the general symptoms scores decreased in the treatment group, all with statistical difference (all P<0.05). Compared with the control group, the death number decreased, the survival rate increased, the life span of dead children was prolonged in the treatment group, showing statistical difference (P<0.05).
CONCLUSIONFJD combined chemotherapy could effectively improve the QOL of solid tumor children patients, elevate their survival rate, and prolong their life spans.
Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Infant ; Integrative Medicine ; Male ; Neoplasms ; drug therapy ; mortality ; Phytotherapy ; Quality of Life ; Survival Rate ; Treatment Outcome
6.Establishment and application of screening methods for non-agonist PPARγ ligand.
Yi HUAN ; Jun PENG ; Yue WANG ; Chun-Ming JIA ; Ke WANG ; Ke-Hua WANG ; Zhi-Qiang FENG ; Zhu-Fang SHEN
Acta Pharmaceutica Sinica 2014;49(12):1658-1664
In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator
Genes, Reporter
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Hepatocytes
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Hypoglycemic Agents
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chemistry
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Ligands
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PPAR gamma
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agonists
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chemistry
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Plasmids
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Response Elements
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Sulfonamides
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chemistry
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Transcriptional Activation
7.Glycated Hemoglobin and Blood Pressure Related to Carotid Artery Atherosclerosis in Continuum in Non-diabetic Persons
Wen ZHU ; Yong LI ; Huan YU ; Hai-Ming SHI ; Jian LI ; Jun ZHU ; Wei-Lin QI ; Xin-Ping LUO ; Feng-Xia ZHANG
Chinese Journal of Hypertension 2007;0(03):-
Objective To analyze the relationship between glyeated hemoglobin,blood pressure and carotid ar- tery atheroselerosis (AS) in non-diabetic patients.Methods To recruite 300 non-diabetic patients and retrospec tively study the relationship between glycated hemoglobin HbAlc,blood pressure and carotid artery AS.Carotid AS was determined by carotid uhrasound.Carotid AS was defined as intima-media thickness (IMT) of common ca- rotid artery≥0.9 mm,or occurrence of carotid plaque.Results Compared with the carotid AS negative subjects, the carotid AS positive patients had significant higher fasting glucose,2 h glucose,HbAlc and systolic blood pres- sure (SBP) level,without difference in DBP.Higher HbAlc or SBP levels were associated with increased incidence of carotid AS.For patients with similar SBP levels,higher HbAlc was associated with more prevalence of carotid AS.Logistic regression analysis shows HbAlc (OR=4.1,P=0.009) was an independent predictor of carotid AS,after adjusting for sex,BMI,SBP,DBP,LDL-C,HDL-C,hypersensitive C-reactive protein (hsCRP),and fasting and 2 h glucose.Conclusion These results suggests that even a slight increase of HbAlc may be independ- ently associated with carotid AS in non-diabetic subjects;and the coexistence of elevated systolic blood pressure syn- ergetically enhance the carotid atherosclerosis.
8.Construction and application of a multiplex PCR system for genotyping of human red blood cell antigens.
Huan-huan GAO ; Yun-lei HE ; Lu-yi YE ; Pan WANG ; Zhong-hui GUO ; Zi-yan ZHU ; Yong-ming ZHU
Chinese Journal of Medical Genetics 2012;29(5):547-552
OBJECTIVEA reliable method for genotyping blood group antigens Dib, k, Jsb1910 and Jsb2019 was developed. Through screening for rare blood types, the National Rare Blood Bank of China may be enriched.
METHODSThe controls for allele detection of blood groups Dib, k, Jsb1910 and Jsb2019 were prepared via polymerase chain reaction (PCR)-mediated gene site-directed mutagenesis (SDM) technique. Sequence-specific primers were designed according to known single nucleotide polymorphism (SNP) sites of alleles of blood groups antigens Dib, k, Jsb1910 and Jsb2019, a multiplex PCR system was developed by optimizing PCR reaction system. And 4190 random healthy donors samples were screened for the blood group antigens.
RESULTSUsing SDM technique, controls for alleles in blood group Dib, k, Jsb1910 and Jsb2019 were successfully generated. And a multiplex PCR system for genotyping above blood groups was developed. After verification, the system has performed with good stability and reproducibility. Two Di (b-) samples have been discovered from 4190 samples, no k- and Js(b-) sample was found.
CONCLUSIONMultiplex PCR features rapid detection, high throughput and low cost, and can be used for screening for donors of rare blood types. Information of donors may be registered in a database, which in turn can help those with rare blood types or require long-term blood transfusion to obtain matched blood, thereby reduce the adverse reactions of blood transfusion.
Blood Group Antigens ; genetics ; Erythrocytes ; immunology ; Genotyping Techniques ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Mutagenesis, Site-Directed ; Polymorphism, Single Nucleotide
9.Comparison of the early control process of H7N9 avian influenza in Shanghai and COVID-19 in Wuhan
Li LUO ; Ge BAI ; Rui-ming DAI ; Qian WANG ; Huan-huan ZHU ; Xin LIU ; Ping WU ; Ying WANG ; Tian-tian ZHANG
Shanghai Journal of Preventive Medicine 2020;32(6):478-
Objective To analyze and estimate the possibility of early control in Shanghai if COVID-19 had begun in Shanghai. Methods Comparison was made in the processes of early control between H7N9 avian influenza in Shanghai in 2013 and COVID-19 in Wuhan in 2019.The early incidence data of Korean COVID-19 was simulated and analyzed to predict whether the medical resources needed in Shanghai were available. Results If it had occurred in Shanghai, it would have taken 22 days from the first case to the government′s emergency response.It was estimated that there would have been 602-763 patients with cumulative onset and onset after incubation period.At least 500 beds of infectious diseases could have been allocated in Shanghai in case of emergency.Through adding beds and resources reallocation in the whole city, patients could have been fully admitted and treated. Conclusion If COVID-19 epidemic had occurred in Shanghai, it′s early control would have been possible though there might have difficulties.
10.Immune regulatory effect of human bone marrow mesenchymal stem cells on T lymphocyte.
Xiao-Xi LU ; Ting LIU ; Wen-Tong MENG ; Huan-Ling ZHU ; Ya-Ming XI ; Yong-Mei LIU
Journal of Experimental Hematology 2005;13(4):651-655
To investigate the immune regulatory effects of human bone marrow mesenchymal stem cells on alloantigen T lymphocyte in vitro, human MSCs were isolated and expanded from bone marrow cells, and identified with cell morphology, and the phenotypes were assessed by immunohistochemistry and flow cytometry. As the stimulation factor of T lymphocytes proliferation, either PHA or dendritic cells isolated from cord blood were cocultured with CD2(+) T lymphocytes from peripheral blood mononuclear cells by magnetic beads with or without MSC in 96-well plats for seven days. T cell proliferation was assessed by [(3)H]-thymidine incorporation using a liquid scintillation counter. T cell subsets, Th1, Th2, Tc1 and Tc2 were analyzed by flow cytometry after co-culture of CD2(+) T cells with MSCs for 10 days. The results showed that a significant decrease of CD2(+) T cell proliferation was evident when MSC were added back to T cells stimulated by DC or PHA, and an increase of Th2 and Tc2 subsets were observed after co-culture of MSC with T lymphocytes. It is suggested that allogeneic MSC can suppress T cell proliferation in vitro and the cause of that was partly depend on interaction of cells and the alteration of T cell subsets.
Bone Marrow Cells
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cytology
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immunology
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CD2 Antigens
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immunology
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Cell Communication
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immunology
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Cell Proliferation
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Cells, Cultured
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Coculture Techniques
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Flow Cytometry
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Humans
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Immunohistochemistry
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Mesenchymal Stromal Cells
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cytology
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immunology
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T-Lymphocyte Subsets
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cytology
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immunology
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T-Lymphocytes
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cytology
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immunology