1.Effect of high volume mechanical ventilation on radiation-induced lung toxicity in rats
Sheng JIN ; Jun CHEN ; Fan YE ; Dan FENG
Chinese Journal of Radiological Medicine and Protection 2015;35(5):344-348
Objective To evaluate the effects of mechanical ventilation on radiation induced lung injuries of apoptosis,acute inflammation,and oxidative stress by establishing a rat mechanical ventilation model and animal model.Methods Totally 40 male Sprague-Dawley(SD) rats were randomly divided into 4 groups with 10 rats in each group:control,radiation alone,high tidal volume ventilation,and high tidal volume ventilation following by radiation.After treatment,the pathological changes in lung tissue were observed,NF-κB activity was detected by electrophoretic mobility shift assay (EMSA),the expression of NF-κB subunit P65 protein level in lung cell nucleus was detected by Western blot,and the apoptosis of lung cells was detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) method.The wet to dry weight ratio (W/D) of lung,myeloperoxidase (MPO),malondialdehyde (MDA) and superoxide dismutase (SOD) were detected.In addition,the total protein and white blood cell number in lung lavage fluid were also measured.Results Compared to the control,the acute lung injury (ALI) score,W/D ratio,MPO activity,total protein level,white blood cell number,apoptosis index (AI),lung tissue MDA,NF-κB activity and P65 protein expression were increased significantly (q =0.000 32-0.004 81,P <0.05),while SOD values was decreased significantly (q =0.000 18-0.002 53,P <0.01),in other three groups.Compared with radiation and high tidal volume ventilation group,the above indexes were significantly higher (q =0.004 3-0.022 6,P < 0.05) but the SOD value was significantly lower (q =0.002 9-0.008 3,P < 0.05) than those in the high tidal volume ventilation plus radiation group.Conclusions High tidal volume ventilation delivered to the radiation group produced more transparent ventilator-induced lung injury (VILI) than the high tidal volume ventilation alone induced VILI including permeable pulmonary edema,acute inflammation,oxidative stress and apoptosis in lung.
4.Establishment and application of a TaqMan real-time PCR assay for the detection of encephalomyo-carditis virus
Haixia ZHANG ; Ruofei FENG ; Dan WANG ; Jingjing FAN ; Jingying XIE ; Zhongren MA ; Yuping FENG
Chinese Journal of Microbiology and Immunology 2013;(9):706-710
Objective To establish a TaqMan real-time PCR assay for the detection of encephalo-myocarditis virus ( EMCV) .Methods Based on the conservative region of 3D gene of EMCV published in GenBank , a pair of primers and one TaqMan probe were designed and synthesized .Then a TaqMan real-time PCR assay was set up and the reactive system was optimized .The sensitivity and specificity of the assay was evaluated respectively .The TaqMan real-time PCR assay was then carried out to detect 98 randomly selected swine serum samples and the results were compared with those by using ELISA .Results The Ct value of the templates had a good linear relationship with the log starting quantity , with a correlation coefficient of 0.995.The TaqMan real-time PCR assay was only specific for EMCV and its sensitivity was 100 times higher than that of the ordinary PCR .The coincidence rate between the established assay and the ELISA assay was 98.0%in the detection of 98 blood samples.Conclusion The TaqMan real-time PCR assay for the detec-tion of EMCV was successfully established with advantages of high sensitivity and good specificity .It could be used for detection of EMCV and quantitative analysis .
5.Immunomodulatory effect of oyster peptide on immunosuppressed mice
Dan XU ; Feng LIN ; Xiaoyu ZHU ; Wenying LIU ; Xiaowen CHEN ; Jinqiu FENG ; Aiqin FAN ; Muyi CAI ; Yajun XU
Journal of Peking University(Health Sciences) 2016;48(3):392-397
Objective:To evaluate the immunomodulating effect of oyster peptide on immunosup-pressed mice.Methods:ICR mice injected with cyclophosphamide (CTX)were adopted as the module group,with mice without treatment as the control group,and different dosages of oyster peptide (0.5 g/kg,1 .0 g/kg,and 2.0 g/kg)were given to the low,middle,and high groups for 1 5 days.The body weight,spleen,and thymus weight of the mice,structures under the microscope of the immune organs, numbers of white blood cells,ratios of T lymphocyte subsets,immune cytokines and numbers of nuclear cells,and DNA content in bone marrow were all assessed.Results:Compared with the control group, the structures of thymus and spleen of the mice in the CTX group appeared obscure and shrunk when ob-served under microscope,the number of their white blood cells declined (P =0.04),the proportion of their CD3 +T cells in peripheral blood declined (P =0.003),the proportion of their CD8 +T cells in pe-ripheral blood declined (P =0.002),the concentration of their IL-5 in peripheral blood significantly in-creased (P <0.01 ),the concentration of their nucleated cells and DNA density in bone marrow de-creased (P =0.04,P <0.01 ).Oyster could improve the structures of thymus and spleen of the immuno-suppressed mice.Compared with the CTX group,the number of white blood cells in 2.0 g/kg group in-creased (P =0.003),the proportion of CD3 +T cells in peripheral blood in 1 .0 g/kg group (P =0.04) and 2.0 g/kg group (P =0.02)increased,the proportion of CD8 +T cells in peripheral blood in 2.0 g/kg group increased (P =0.002),the concentration of IL-5 in peripheral blood in all the oyster treated groups increased (P <0.01 in 0.5 g/kg,1 .0 g/kg,and 2.0 g/kg groups),the concentration of IL-1 7 in peripheral blood in 2.0 g/kg group decreased (P =0.03),the concentration of nucleated cells in bone marrow of all the oyster treated groups increased (0.5 g/kg vs.CTX,P =0.04;1 .0 g/kg vs. CTX,P =0.02;2.0 g/kg vs.CTX P =0.01 ),the DNA content in bone marrow of all the oyster treated groups increased (P <0.01 in the 0.5 g/kg,1 .0 g/kg,and 2.0 g/kg groups).Conclusion:Oyster peptide could improve the structures of immune organs of the CTX-induced immunosuppressed mice,re-cover the imbalances of T lymphocyte subsets,improve the immune cytokines and increase numbers of nucleated cells and DNA content in bone marrow,thus improving the immunologic function.
6.Role of Delta-like 1 in differentiation and antigen presentation of mouse bone marrow-derived dendritic cells.
Yun-Ru CHEN ; Fan FENG ; Dan-Dan YIN ; Ying-Min LIANG ; Hua HAN
Journal of Experimental Hematology 2010;18(3):704-708
The aim of this study was to investigate the role of Delta-like 1 (Dll1) in differentiation and antigen pre-sensation of mouse bone marrow-derived dendritic cells (DCs). In the presence of granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin 4 (IL-4), mouse bone marrow cells were co-cultured with OP9-Dll1 and OP9-GFP cell lines respectively. After 8 days, the immature DCs were stimulated with tumor antigen. The surface molecules of the activated DCs including MHC II, CD80 and CD86 were analyzed by flow cytometry. Levels of IL-12 and IL-10 in the culture supernatant were detected by ELISA. In addition, the proliferation of T-cells co-cultured with DCs was analyzed by FACS through mixed T-lymphocyte reaction. The results showed that compared with OP9-GFP, the bone marrow cells co-cultured with OP9-Dll1 produced significantly more CD11c(+) DCs (p < 0.05), and possessed higher levels of surface molecule expression including MHC II, CD80 and CD86 after tumor antigen stimulation. The DCs secreted higher level of IL-12 (p < 0.05) and less IL-10 (p < 0.01). They also resulted in significantly stronger T-cell proliferation response. It is concluded that Dll1 can promote the differentiation of DCs from mouse bone marrow cells and enhance their antigen presentation capacity.
Animals
;
Antigen Presentation
;
immunology
;
Bone Marrow Cells
;
cytology
;
Cell Differentiation
;
immunology
;
Cells, Cultured
;
Dendritic Cells
;
cytology
;
immunology
;
Granulocyte-Macrophage Colony-Stimulating Factor
;
pharmacology
;
Intercellular Signaling Peptides and Proteins
;
metabolism
;
Interleukin-4
;
pharmacology
;
Male
;
Mice
;
Mice, Inbred C57BL
7.Influence of fluoride on osteoprotegerin mRNA and protein expression in rat osteoblasts: an in vitro study
Dan, LI ; Xing-quan, JIANG ; Xiao-feng, JI ; Zhi-tao, ZHAO ; Zhe, FAN ; Ling, JING ; Guang-sheng, LI
Chinese Journal of Endemiology 2009;28(2):134-137
Objective To study the influence of fluoride on the expression of osteoprotegerin(OPG) mRNA and protein in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarial of suckling Wistar rats were cultured in vitro in the media supplemented with NaF at a series of doses[O(control), 1,2 and 4 mg/L groups], and OPG mRNA expression and protein were evaluated by RT-PCR and ELISA methods, respectively. Results OPG mRNA expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h and 72 h(F=333.48,808.34,P<0.05). OPG mRNA expression in suckling rat osteoblasts cultured in vitro after exposure to NaF for 48 h at different doses(0.810±0.003, 0.819±0.031 and 0.870±0.044 for 1,2 and 4 mg/L groups, respectively) compared with that of control (0.800±0.040, all P<0.05). OPG mRNA expression further increased for 72 h exposure to NaF(0.933±0.047,1.031±0.051,1.240±0.062 for 1,2 and 4 mg/L, respectively), significantly higher than that of the control (0.805±0.020,all P<0.05) and corresponding groups at 48 h. NaF doses and time exposure exhibited a significant synergistic effect on OPG mRNA expression(F=2004.16, P<0.05). NaF also enhanced OPG protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed only in 4 mg/L group(0.228±0.014,0.277±0.048) and control(0.205±0.012,0.229±0.010) at 48 h and 72 h (P<0.05). In addition, OPG protein expression at 72 h post-exposure was higher than that at 48 h,but there was no synergistic effect between concentration and time(F=1.21,P>0.05). Conclusions The results suggested that NaF could increase OPG mRNA and protein expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.
8.Mechanical properties of human enamel based on resonant ultrasound spectroscopy
Dan-Dan FENG ; Fan FAN ; Rui WANG ; Qiang ZHANG ; Hai-Jun NIU
Journal of Medical Biomechanics 2017;32(5):448-453
Objective To investigate the mechanical properties of human enamel based on resonant ultrasound spectroscopy (RUS).Methods The rectangular parallelepiped specimens of human enamel were processed.The theoretical resonant frequencies of specimens were estimated and paired with the experimental resonant fre quencies measured from RUS experiments.An iterative procedure was used to adjust elastic constants of enamel until the theoretical frequencies corresponded to the experimental frequencies based on minimum mean-squared error criterion.In addition,elastic modulus,shear modulus and Poisson's ratio were calculated respectively.Results The elastic modulus,shear modulus and Poisson's ratio ranged from 61.52 to 80.46 GPa,21.51 to 51.86 GPa and 0.18 to 0.43,respectively.Eliminating the effect of large specimen variances,the average of elastic modulus,shear modulus and Poisson's ratio was 72.84 GPa,31.94 GPa and 0.27,respectively.Conclu sions RUS performs a feasibility of measuring the mechanical properties of human enamel with repeatable and nondestructive advantages.All the elastic constants and mechanical parameters can be estimated through a sig nal experiment.The results provide references for the development of biomimetic dental restoration materials.
9.Effect of artemether on radiosensitivity in human nasopharyngeal carcinoma cell line CNE-1
Luyu PAN ; Jianping CAO ; Rong JI ; Yang FENG ; Yuanyuan ZHOU ; Xiaomei PENG ; Yang LIU ; Xialin CHEN ; Dan CUI ; Wei ZHU ; Saijun FAN
Chinese Journal of Radiological Medicine and Protection 2010;30(5):550-553
Objective To evaluate the effect of artemether on the cell cycle and the radiosensitivity in human nasopharyngeal carcinoma cell line CNE-1.Methods Cell growth inhibition was assessed with MTT.The method of colony-forming was used to detect the radiation sensitivity.Cell cycle distribution was analyzed by using flow cytometry.The protein expressions of clyclin B1 and Weei were detected by using Western blot.Results The growth of CNE-1 cells was inhibited in a dose-dependent manner.The concentration of 20 μmol/L artemether had radiosensitive effect on CNE-1 cells at 24 h after administration,and SER was 1.481.When CNE-1 cell was irradiated,the G2/M cells increased (t =4.59,P < 0.05).After exposure to combination of artemether and irradiation,the G2/M cells were decreased (t= 10.60,P < 0.05).Western blot showed that artemether increased the level of cyclin B1 expression and inhibited the level of Weel expression.Conclusions The noncytotoxic concentration of artemether could enhance radiosensitization of CNE-1 cells.The radiosensitivity enhancement of artemether might depend on the exposure time.The effect is most obvious when radiation is delivered 24 h after expose to artemetherr.The radiosensitizing effect could be related to apoptosis.
10.Investigation of the characteristics of HCV genotypes of Han and Korean in Yanbian area of Jilin province.
Zhong-Xie LI ; Fan-Ping MENG ; Gang-Tie SHEN ; Dan JIN ; Xiang-Wei FENG
Chinese Journal of Experimental and Clinical Virology 2010;24(2):104-106
OBJECTIVETo investigate the characteristics of HCV genotypes of Han and Korean in Yanbian area of Jilin Province.
METHODSThe HCV RNA load and genotypes of the 119 chronic hepatitis C patients in Yanbian area of Jilin Province were determined by real-time PCR and LiPA. The differences of the HCV genotypes in Han and Korean cases, in severity of the diseases, in HCV-RNA load, and in the relation with type 2 diabetes mellitus were analyzed.
RESULTSThere was no significant difference in the distribution of each HCV genotype between Han and Korean patients (P > 0.05) with chronic hepatitis C. The difference between HCV genotype and HCV-RNA load was not significant (P > 0.05). With and without type 2 diabetes mellitus in these patients. The distribution of HCV genotype was also not significantly different (P > 0.05). The type 1b of HCV genotype in the moderate to severe chronic hepatitis C patients accounted for 58.06%. It was different compared with mild chronic hepatitis C patients (P < 0.05).
CONCLUSION1) The type 1b is the most popular HCV genotype in Yanbian area of Jilin Province, type 2a is the second and there are still a few other genotypes. 2) There is no significant difference in the distribution of HCV genotypes between Han and Korean cases. 3) The HCV genotypes has nothing to do with the load of HCV-RNA. 4) The distribution of HCV genotypes in chronic hepatitis C patients with and without diabetes mellitus is not significantly different. 5) Type 1b of HCV infection is relatively severe.
Adolescent ; Adult ; Aged ; Diabetes Mellitus, Type 2 ; complications ; Female ; Genotype ; Hepacivirus ; genetics ; Hepatitis C ; complications ; epidemiology ; virology ; Humans ; Korea ; epidemiology ; Male ; Middle Aged ; Polymerase Chain Reaction ; Young Adult