1.The relationship between TNF-αγ, IFN-γ and intestinal mucosal permeability in DSS-induced colitis and its effect by balsalazide
Xiaochang LIU ; Qiao MEI ; Jianming XU ; Juan JIN
Chinese Journal of Microbiology and Immunology 2009;29(5):395-399
Objective To investigate the relationship between TNF-α, IFN-γand intestinal muco-sal permeability in a mouse colitis model and its inhibiting effect by balsalazide. Methods Forty-five C57BL/6J mice were divided randomly into five groups. Normal group was only fed with distilled water, DSS group and balsalazide groups at doses of 42, 141,423 mg/kg were both fed with 5% DSS. Balsalazide was given by intragastric administration. At the end of the experiment, colon tissue was collected for assessment of histological index(HI) and the MPO activity. Small intestinal mucosa was collected for assessment of the content of TNF-α and IFN-γ,transmission electron microscope(TEM), and detection of permeability by Ev-arts blue method. Results Compared with normal group, DSS group mice all manifested severe weight loss associated with hematocbezia and diarrhea, HI score, and the colon MPO activity and the content of TNF-α and IFN-γ were increased significantly. Intestinal mucosa showed a thinning of microvillous carpet, with de-curtated and broaden junctional complex and enlarged intercollutar space under TEM observations. The amount of Evans blue permeated into intestinal wall was obvious. Compared with DSS group, the HI score, the MPO activity and the content of TNF-α and IFN-γ were decreased by balsalazide. The amount of Evans blue permeated into intestinal wall was less. Ileal microvillous carpet was ameliorated dose dependently by balsalazide. Conclusion In DSS-induced colitis model, the change of the content of the TNF-α and IFN-γ, was accordance with the increase of intestinal mucosal permeability while balsalazide can significantly amelio-rate intestinal mucosal permeability by its anti-colitis effect.
2.A Survey of the Medical Students' After-class Reading
Qian WANG ; Juan XU ; Juhua MA ; Xiaofei YAO ; Zhongshan JIN
Chinese Journal of Medical Education Research 2002;0(01):-
After-class reading is one of the important activities in the college students' spiritual life.The interests,sorts of books,contents and quantities of their reading can reflect their ideological conditions and their value judgment.Based on the investigation and analysis on their after-class reading activities,we can discover problems and give appropriate guidance in time,hoping to promote the cultivation of good after-class reading atmosphere on campus. The results of this survey can provide reference for the humanistic quality education of medical students.
3.The expression of odontoblast relevant proteins in dental pulp stem cells induced by BMP-7
Juan MA ; Ruiqiao ZHU ; Xiaolei SHI ; Hui XU ; Yan JIN ; Lei JIN
Journal of Practical Stomatology 2015;(6):753-756
Objective:To examine the expression of odontoblast related proteins in dental pulp stem cells(DPSCs)induced by BMP-7.Methods:DPSCs were cultured in the common culture medium or medium supplemented with 1 00 ng/ml BMP-7.Electron microscope,CCK8 and immunohistochemical staining were carried out to estimate the cell morphology and differentiation.Results:In-duced by BMP-7,the morphology of DPSCs was not changed,the proliferation of DPSCs was slower than that of the cells without BMP-7 treatment.DPSCs were negative for the expression of DSPP,DMP-7 and ALP.However,DPSCs were found strongly positive for DSPP,DMP-7 and ALP after the induction of BMP-7.Conclusion:BMP-7 induction may promote the differentiation of DPSCs.
5.Construction and Expression of RNase-Resisting His-Tagged Virus-Like Particles Containing FluA/B mRNA.
Jin ZHANG ; Xiaoning XUE ; Hefei XU ; Ke ZHU ; Xiaoguang CHEN ; Juan ZHANG ; Qi ZHANG ; Yuan LIN
Chinese Journal of Virology 2015;31(6):629-633
To prepare virus-like particles containing FluA/B mRNA as RNA standard and control in Influenza RNA detection, the genes coding the coat protein and maturase of E. coli bacteriophage MS2 were amplified and cloned into D-pET32a vector. Then we inserted 6 histidines to MS2 coat protein by QuikChange Site-Directed Mutagenesis Kit to construct the universal expressing vector D-pET32a-CP-His. In addition, the partial gene fragments of FluA and FluB were cloned to the down-stream of expressing vector. The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. The virus-like particles were purified by Ni+ chromatography. The virus-like particles can be detected by RT-PCR, but not PCR. They can be conserved stably for at least 3 months at both 4 degrees C and -20 degrees C. His-tagged virus-like particles are more stable and easier to purification. It can be used as RNA standard and control in Influenza virus RNA detection.
Escherichia coli
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genetics
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metabolism
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Influenza A virus
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genetics
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metabolism
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Influenza B virus
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genetics
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metabolism
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RNA, Messenger
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genetics
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metabolism
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RNA, Viral
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genetics
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metabolism
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Recombinant Fusion Proteins
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genetics
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metabolism
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Ribonucleases
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chemistry
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Virion
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genetics
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metabolism
6.Impact of fluorine and aluminum and both action combined on the number of rat osteoclasts and bone resorption cultured in vitro
Guang, DU ; Mao-juan, YU ; Xiao-ya, XU ; Wei-fang, JIN ; Jian-jun, GAO
Chinese Journal of Endemiology 2013;32(4):370-373
Objective To determine the impact of fluorine and aluminum,and both action combined on the number of rat osteoclasts and bone resorption cultured in vitro and to explore its mechanisms.Methods The osteoclasts and bone marrow stromal cells (BMSCs) isolated from long bone of new born rats were cultured,respectively,in TC199 medium (containing 10% fetal bovine serum) with fluoride,aluminum and fluoride combined with aluminum.The osteoclasts were inoculated in 96-well culture plate and ivory slice,BMSCs in 6-well culture plate,and culture medium was changed after 2 hours incubation.The cells were divided into control group,fluoride group,aluminum group and fluoride combined with aluminum group; the doses of sodium fluoride were 0,1.0 × 10-4,0,1.0 × 10-4 mol/L and the doses of aluminum chloride were 0,0,1.0 × 10-5,1.0 × 10-5 mol/L,respectively.Tartrate-resistant acid phosphatase (TRAP) staining positive cells were counted under light microscope after TRAP staining on the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue.The expression of osteoprotegerin(OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) was detected by real-time fluorescence quantitative PCR in BMSCs after 8 h treatment.Results ① Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the numbers of osteoclasts (F =7.15,6.56 and 7.98,respectively,all P < 0.05).The numbers of osteoclasts in fluoride group,aluminum group and fluoride combined with aluminum group[(136.9 ± 22.99),(135.4 ± 23.5),(163.0 ± 24.4) per well] were higher than that in the control group[(92.5 ± 22.1) per well,all P < 0.05].② Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the resorption pit area on ivory slices(F =10.47,12.64,14.29,respectively,all P < 0.05).The resorption pit area on ivory slices in fluoride group,aluminum group and fluoride combined with aluminum group[(0.242 ± 0.031),(0.293 ± 0.026),(0.333 ± 0.016)mm2 per slice] was higher than that in the control group [(0.088 ± 0.030)mm2 per slice,all P < 0.05].③Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the expression ratios of RANKL/OPG in BMSCs (F =8.15,15.38,23.59,respectively,all P < 0.05).The expression ratios of RANKL/OPG in BMSCs in fluoride group,aluminum group and fluoride combined with aluminum group [(193.98 ± 137.93)%,(326.11 ± 176.78)%,(599.84 ± 275.82)%] were higher than that in the control group[(100.00 ± 56.02)%,all P < 0.05].Conclusions Both fluoride and aluminum can cause increase in the number of osteoclasts in vitro and promote cell differentiation and bone resorption activity,which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.The stimulating effects of fluoride on osteoclasts differentiation and bone resorption is enhanced by aluminum.
7.Effects of fluorosis on osteoclasts's quantity and bone resorption function in vitro
Guang, DU ; Mao-juan, YU ; Xiao-ya, XU ; Wei-fang, JIN ; Jian-jun, GAO
Chinese Journal of Endemiology 2011;30(3):266-269
Objective To determine the effects of fluoride on osteoclasts's quantity and bone resorption function in vitro and its mechanisms. Methods The osteoclasts and bone marrow stromal cells(BMSCs) isolated from long bone of new born rats were cultured respectively in TC199 medium (containing 10% fetal bovine serum) with fluoride. The osteoclasts were inoculated in 96-well culture plate and ivory slice, BMSCs were inoculated in 6- well culture plate, respectively, medium were changed after 2 hours incubation. They were divided into control group, low-dose fluoride, medium-dose fluoride and high-dose fluoride groups, the doses of sodium fluoride were 0,2.5 × 10-5,5.0 × 10-5,10.0 × 10-5 mol/L, respectively. Tartrate-resistant acid phosphatase(TRAP) staining positive cells were counted under light microscope after TRAP staining on the 2nd and the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue. The expression of receptor activator of NK-κβ ligand(RANKL) and osteoprotegerin(OPC) was detected by real-time fluorescence quantitative (337.5 ± 70.5), (447.5 ± 43.4), (472.9 ± 34.8), (475.3 ± 24.3)/well in the control group, the low-dose, mediumdose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the control group (all P < 0.05). After in vitro culture for 5 days, the numbers of osteoclasts were (92.5 ± 22.1), (123.0 ± 26.4), (135.5 ± 22.2), (136.9 ± 23.0) per well in the control group, the low-dose, medium-dose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the (0.088 ± 0.030), (0.100 ± 0.018), (0.152 ± 0.015), (0.242 ± 0.031 )mm2 per piece in the control group, the lowdose, medium-dose and high-dose fluoride groups, respectively. The values of medium-dose and high-dose fluoride BMSCs in the control group, the low-dose, medium-dose and high-dose fluoride groups were 100.00 ± 56.02, 144.95 ± 97.21,223.25 ± 184.48,193.98 ± 137.93, respectively. The values of medium-dose and high-dose fluoride groups were significantly higher than that of control group (all P < 0.05). Conclusions Fluoride can cause increase in the number of osteoclasts in vitro and promote their cell differentiation and bone resorption activity, which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.
8.An approach to the development of interns' clinical thinking ability by discussing the causes of gastrointestinal hemorrhage
Dong-lin LUO ; Yan XU ; Jin-ping CHEN ; Xiao-juan NG JIA
Chinese Journal of Medical Education Research 2011;10(6):736-738
The article probes into the approach to the interns' capability of clinical thinking and analyzing and solving problems by disscussing causes of gastrointestinal hemorrhage. Correct clinical thinking always comes from solid basis and clinical theory and knowledge, careful history taking, meticulous physical examination, rigorous logical thinking and constant summing up experiences and lessons. As a teacher, we should help interns develop clinical thinking ability during their whole peroid of medical education.Besides, theory should be combined with practice to prevent and correct vicious clinical thinking
9.Exploration of development of the subjective role of students in regional anatomy experiment program
Shengwei GAN ; Jianhua RAN ; Weitian LU ; Juan HUANG ; Shujuan ZHU ; Jin XU ; Ping FAN ; Shanquan SUN
Chinese Journal of Medical Education Research 2015;(7):735-737
During the teaching activities, to stimulate students' subject awareness and encour-age them to play the main role in class activities are inevitable trends in the reform of college educa-tion. Students’ subjective activity is a key to the teaching effect of regional anatomy, a course mainly based on experimental program. Department of Human Anatomy in Chongqing Medical University lay-outs regional anatomy teaching program to develop the students' subjective activities in learning from the course specialty: in preview and review, to train students' image-thinking by drawing; in anatomi-cal operation link, to cultivate students' interest in learning and innovation by discussing the relation-ship of anatomical structure and clinical disease and identifying the variation of structure; in the eval-uation process, taking the formative evaluation system to promote the students' initiatives and ensure the objectiveness and fairness. The implementation of these measures promotes the regional anatomy teaching quality.
10.Combination of MGIT liquid culture and Xpert MTB/RIF test in the rapid diagnosis and drug resistance detection in Mycobacterium tuberculosis
Lifeng XU ; Xuliang YU ; Feng ZHANG ; Ting SHUI ; Juan CHEN ; Jin ZHU
Chinese Journal of Laboratory Medicine 2016;39(4):272-276
Objective To evaluate the feasibility of Mycobacteria growth indicator tube ( MGIT ) liquid culture combined with rifampin resistance test real-time PCR ( Xpert MTB/RIF test) in the diagnosis of tuberculosis.Methods 652 cases of suspected pulmonary tuberculosis from October 2014 to March 2015 in Quzhou People′s Hospital were enrolled. The morning sputum samples were collected for acid-fast staining, L?wenstein-Jensen ( L-J) culture, BACTEC MGIT culture and Xpert MTB/RIF test.Samples with positive results of MGIT liquid culture were subjected to strain identification and drug sensitivity test with fluid method.Methodological comparison between four methods was made and the performance of MGIT liquid culture combined with Xpert MTB/RIF test in the rapid diagnosis and drug resistance detection in Mycobacterium tuberculosis was evaluated by chi-square test. Results Among the samples from 399 confirmed tuberculosis patients, the diagnostic sensitivity of the 4 methods ( acid-fast staining, L-J culture, MGIT culture and Xpert MTB/RIF test) were 17.0%(68/399), 23.8%(95/399), 37.8% (151/399) and 37.3%(149/399) respectively.The sensitivity and specificity of MGIT culture combined with Xpert MTB/RIF test was 39.8%(159/399) and 94.8%(240/253).The sensitivity of MGIT culture combined with Xpert MTB/RIF test was significantly higher than acid-fast staining (χ2 =50.9, P<0.01 ) and L-J culture (χ2 =23.7,P<0.01).The average detection time of MGIT culture was 7.5 days ( smear positive and Xpert MTB/RIF positive), 13.4 days (smear negative and Xpert MTB/RIF positive) and 16.9 days ( smear negative and Xpert MTB/RIF negative) .The sensitivity and specificity of Xpert MTB/RIF test in rifampin resistance detection were 9/9 and 97.3% ( 129/132 ) respectively.The average detection time of fluid method for drug sensitivity test was 8.3 days.Conclusions MGIT liquid culture combined with Xpert MTB/RIF test can detect Mycobacterium tuberculosis and the drug sensitivity rapidly.The method is highly sensitive and specific.It is important for clinical diagnosis and treatment of tuberculosis.