2.Notch1 regulates stemness and chemotherapeutic sensitivity of human gli-oma U251 cells
Like ZHANG ; Na XIAN ; Ling LIN ; Yuqing GONG ; Zhiqiang YE ; Zhihong ZHENG
Chinese Journal of Pathophysiology 2015;(11):1943-1949
AIM:To investigate whether Notch1 changes stemness and chemotherapeutic sensitivity in human glioma U251 cells.METHODS: The lentiviral vectors, which expressed Notch1-shRNA or Notch1 intracellular domain ( NICD) , were transfected into U251 cells .Western blot and immunofluorescence staining were applied to monitor the va-lidity of the cells, down-regulation of Notch1 expression or over-expression of NICD.The proportion of CD133 +cells was analyzed by flow cytometry.The expression of nestin and GFAP was identified by immunofluorescence staining.The forma-tion rate of tumor cell spheres and the implanted tumor growth in SCID mice were observed.MTT assay was performed to e-valuate the chemotherapeutic sensitivity to VM-26 and BCNU of the cells with different treatments.RESULTS:Stemness was significantly enhanced in the cells over-expressing NICD.For example, the proportion of CD133 +cells was increased, the expression of nestin was up-regulated, the expression of GFAP was down-regulated, and the formation rate of tumor cell spheres and implanted tumor growth were increased.The chemotherapeutic sensitivity to VM-26 and BCNU of the cells was decreased.In the cells with Notch1 gene down-regulation by RNAi, the stemness was inhibited and chemotherapeutic sensi-tivity was increased.CONCLUSION:Notch1, which leads to the change of stemness and chemotherapeutic sensitivity in human glioma U251 cells, is likely to be a potential molecular target for treatment of glioma.
3.E.max crowns by monolithic technique applied to endodontically treatedmolars
Zhaojie ZHENG ; Wenfang LV ; Na LI ; Peng ZHANG ; Ling SONG ; Shulai LU ; Yang CAO ; Jiangbo YU ; Dawei GUO
Chinese Journal of Tissue Engineering Research 2016;20(21):3124-3130
BACKGROUND:IPS e.max Presshas an excelent biocompati bility and corrosion resistance, which obtains satisfactory clinical outcomes on dental veneers, inlay and onlay restorations. But little is reported on molar monolithic restoration by IPS e.max Presscrown.
OBJECTIVE:To evaluate the clinical effects of IPS e.max Press crown on molar repair after root canal therapy.
METHODS:Totaly 215 patients with 324 affected molars, including 88 males and 127 females, aged 22-58 years old, were enroled for repairing with IPS e.max Presscrown. Then the color, shape, fracture and edge coloring of the restoration, marginal discrepancy, secondary caries and gingival health status were assessed after a 3-year folow-up.
RESULTS AND CONCLUSION:During the folow-up, 324 dental restorations met the class A standards for color, marginal discrepancy, shape as wel as secondary caries. In addition,3restoration swere fractured, 14 restorations had margin coloring, and 8 restorations appeared to have gingival inflammation. More than 95% restorations were scored grade A. These results indicate that IPS e.max Press crown applied to molar repair after root canal therapy can achieve ideal outcomes.
4.Establishment and evaluation of PCR-array for detection of six common pathogens of purulent meningitis in children
Cheng-Na ZHAO ; Chao ZHANG ; Yu-Ling ZHENG ; Peng LIU ; Yong-Qiang JIANG
Military Medical Sciences 2017;41(11):902-905
Objective To develop a PCR-array method for detecting common purulent meningitis pathogens including Streptococcus pneumoniae,Escherichia coli,Haemophilus influenzae type B,Neisseria meningitidis,S.agalactiae and Listeria monocytogenes in children.Methods The amplification efficiency,limit of detection (LOD) and cross-reactivity were validated with individually real-time PCR using genomic DNA of the six pathogenic bacteria.The sensitivity and specificity of the PCR-array method were evaluated using artificial cerebrospinal fluid(CSF),and the consistency between the PCR-array method and the golden method of CSF culture was evaluated using clinical samples.Results The primers and probes of the pathogens in PCR-array had high specificity,and there was no cross reaction between them.The LOD of the PCR-array method was 10 cfu/ml and very sensitive.The sensitivity and specificity of the PCR-array method could reach 95% in the evaluation of artificial CSF,and had a good consistency with the clinical gold standard method.Conclusion The PCR-array method with high sensitivity and specificity can simultaneously detect six common pathogens in children with purulent meningitis at 2.5 h,which could provide reference for the diagnosis of purulent meningitis.
5.Construction of small interfering RNA targeting heparanase gene and its inhibitory effect on the in-vasiveness of human malignant melanoma cell line A375 in vitro
Xiao-Yan LIU ; Hong FANG ; Zheng-Gang YANG ; Li-Ming RUAN ; De-Ren FANG ; Ying-Guo DING ; Yi-Na WANG ; Yu ZHANG ; Xiao-ling JIANG ;
Chinese Journal of Dermatology 2003;0(11):-
Objective To construct the small interfering RNA (siRNA) targeting heparanase gene and its expressing vector,and to observe its interference effect on the expression of heparanase gene and inhibitory effect on the invasive potential of human malignant melanoma A375 cells.Methods Three siRNAs were designed.The recombinant plasmid pRNATU6.1/heparanase-siRNA was designed and constructed. A375 cells were cultured,and transfected with pRNATU6.1/heparanase-siRNA.The cells treated with lipo- fectamine or Opti-MEM served as the controls.Real-time fluorescence quantitative PCR and Western blot were performed to evaluate the expression of heparanase RNA and protein in these treated A375 cells.The in vitro invasive potential of treated A375 cells was assessed by Matrigel gel assay.Results The siRNA targeting heparanase gene was successfully cloned to the eukaryotic expressing vector pRNATU6.1.The expression levels of both heparanase RNA and protein decreased significantly in siRNA-transfected A375 cells than those in the control cells.The in vitro invasive potential of siRNA-transfected cells was also signifi- cantly inhibited as compared with that of the control cells (P
6.Development of time resolved fluorescence microparticle-conjugated anti-human procalcitonin antibody for homogeneous immunoassay
jun Can WU ; yu Qing L(U) ; jie Huai HAO ; na Cheng ZHAO ; ling Yu ZHENG ; qiang Yong JIANG
Military Medical Sciences 2017;41(8):689-693
Objective To develop europium (Ⅲ) [Eu (Ⅲ)] chelated microparticles for homogeneous immunoassay.Methods Anti-human PCT antibodies were labeled with Eu (Ⅲ) chelated nanoscale microparticles as the detection antibody,and another anti-human PCT antibody was labeled with biotin as the solid-phase antibody.Magnetic microspheres labeled with streptavidin were used to separate the complexes of Eu-IgM-PCT-IgM-Biotin.Results In the homogeneous immunoassay,the standard curve fit was not linear.The quadratic curve was Y =19170.12 + 75493.74X-26.00X2(r =0.9986).According to the standard curve,the limit of detection for PCT was 0.04 ng/ml.Conclusion The homogeneous immunoassay which uses Eu (Ⅲ) chelated microparticles is highly sensitive for detection of PCT recombinant antigens and may serve as a promising method to measure serum PCT levels in the future.
7.Ginsenoside Rg3 induces apoptosis of human lung squamous cell carcinoma SK-MES-1 cell line.
Xin WANG ; Yu-ling ZHENG ; Ke LI ; Na LIN ; Qing-xia FAN
Journal of Southern Medical University 2009;29(9):1823-1826
OBJECTIVETo investigate the effect of ginsenoside Rg3 on the apoptosis and survivin expression in human lung squamous cell carcinoma cell line SK-MES-1.
METHODSSK-MES-1 cells were divided into Rg3 treatment group, blank control group and positive control (arsenic trioxide) group. The apoptotic rate of the cells in each group was determined using flow cytometry, and the expression of survivin protein and mRNA was detected by immunocytochemistry and RT-PCR, respectively.
RESULTSA 48-h treatment with Ginsenoside Rg3 induced increased apoptotic rate of SK-MES-1 cells in a dose-dependent manner. Ginsenoside Rg3 significantly downregulated the expressions of survivin protein and mRNA as compared with the expression levels in the blank control group (P<0.05).
CONCLUSIONGinsenoside Rg3 can induce the apoptosis of SK-MES-1 cells, the mechanism of which may involve inhibited survivin expression.
Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Ginsenosides ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; metabolism ; pathology ; Microtubule-Associated Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism
8.Effect of Data Acquisition Parameters on Characterization of Gold Nanoparticles By Single Particle Inductively Coupled Plasma-Mass Spectrometry
Rui-Ping LUO ; Ling-Na ZHENG ; Liang LI ; Juan WANG ; Wei-Yue FENG ; Xiang-Hua YU ; Meng WANG
Chinese Journal of Analytical Chemistry 2018;46(6):925-930
Single particle inductively coupled plasma mass spectrometry (SP-ICP-MS) is a new tool that can provide the element composition, size distribution, and number concentration of nanoparticles. Here we discussed the effects of dwell time and settling time on analysis of nanoparticles by SP-ICP-MS. We analyzed standard materials of gold nanoparticles (30, 40 and 60 nm AuNPs, from NIST and NCNST), showing that better signal-to-noise ratio and higher determination efficiency could be achived when using shorter dwell time and settling time. We utilized a nano mode for SP-ICP-MS, in which the dwell time was set as 0. 05 ms and the settling time was 0. The size of NIST AuNP standard material determined here was in accord with the certified size using the developed method. The detection limits of size and number concentration of AuNPs were 8 nm and 1. 1×105 particle / L, respectively.
9.Effect of fear of disease progression on sleep quality in patients with type 2 diabetes mellitus:the pathway of executive function
Na ZHANG ; Wenkai ZHENG ; Ling LI ; Mengge BAI ; Chunni HENG
Sichuan Mental Health 2024;37(3):249-254
Background Type 2 diabetes mellitus is one of the most prevalent diseases,which imposes a heavy burden on patients' families and the society.Sleep disorders are recognized as risk factors for the development of diabetes,which may affect the onset and development of diabetes through neuro-endocrino-metabolic pathways,so identifying the factors responsible for the sleep quality of diabetic patients is of great importance in improving their sleep quality.Objective To investigate the relationship among fear of disease progression,executive function and sleep quality in patients with type 2 diabetes mellitus,so as to provide references for improvement of sleep quality in patients with type 2 diabetes mellitus.Methods A sample of 197 patients with type 2 diabetes mellitus who were admitted to the Endocrinology Department of the Second Affiliated Hospital of the Air Force Military Medical University from January to May 2023 and met the criteria defined in the Guideline for the Prevention and Treatment of Type 2 Diabetes Mellitus in China(2020 edition)were consecutively selected.All subjects were assessed using Fear of Progression Questionnaire-Short Form(FoP-Q-SF),Behavior Rating Inventory of Executive Function-Adult version(BRIEF-A)and Pittsburgh Sleep Quality Index(PSQI).Then the Process macro for SPSS(Model 4)and Bootstrap technique were applied to examine the mediating effect of executive function on the relationship between fear of disease progression and sleep quality in patients with type 2 diabetes mellitus.Results ①75 patients(38.07%)with type 2 diabetes mellitus were found to have sleep problems.②PSQI score in patients with type 2 diabetes mellitus was positively correlated with FoP-Q-SF score and BRIEF-A score(r=0.159,0.287,P<0.01).③Executive function mediated the relationship between fear of disease progression and sleep quality,the indirect value was 0.076(95%CI:0.022~0.146),accounting for 39.58%of the total effect.Conclusion Sleep disorders are common in patients with type 2 diabetes mellitus,and executive function may play a medicating role in the relationship between fear of disease progression and sleep quality.
10.Immunomodulatory effects of Fomes fomentarius polysaccharides: an experimental study in mice.
Hui-Ling GAO ; Lin-Sheng LEI ; Chuan-Lin YU ; Zheng-Guang ZHU ; Na-Na CHEN ; Shu-Guang WU
Journal of Southern Medical University 2009;29(3):458-461
OBJECTIVETo investigate the immunomodulatory effects of Fomes fomentarius polysaccharides (FFP) in mice.
METHODSMTT assay was employed to evaluate the in vitro metabolic activity of the mouse splenocytes treated with FFP at different concentrations, and the secretion of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and interleukin 2 (IL-2) from the cells were measured by enzyme-linked immunosorbent assay. The changes in the phagocytotic activity of mouse macrophage in response to FFP treatment were evaluated by phagocytosis percentage of chicken red blood cells (CRBCs). The effect of FFP on the humoral immunity was assessed in mice immunized with sheep red blood cells (SRBCs) by measuring the serum levels of specific antibody (hemolysin) against SRBCs.
RESULTSFFP at the concentrations of 25, 50, and 100 microg/ml all significantly enhanced the metabolic activity of mouse splenocytes in vitro and increased the production of TNF-alpha, IFN-gamma and IL-2. FFP treatment also markedly enhanced the metabolic activity of mouse peritoneal exudate cells and TNF-alpha production by the cells. At the doses of 25, 50, and 100 mg/kg, FFP significantly increased serum hemolysin level in mice immunized with SRBCs, and FFP at 50 and 100 mg/kg obviously increased the capacity of mouse peritoneal macrophages in vivo for CRBC phagocytosis.
CONCLUSIONFFP can promote the secretion of TNF-alpha, IFN-gamma and IL-2 by mouse immunocytes and enhance mouse humoral immune response and the phagocytotic activity of the macrophages.
Adjuvants, Immunologic ; pharmacology ; Animals ; Coriolaceae ; chemistry ; Female ; Immunologic Factors ; immunology ; pharmacology ; Interferon-gamma ; secretion ; Interleukin-2 ; secretion ; Macrophages, Peritoneal ; drug effects ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Phagocytosis ; drug effects ; Polysaccharides ; isolation & purification ; pharmacology ; Tumor Necrosis Factor-alpha ; secretion