1.An Experimental Investigation on Correlations of Left Atrial Size with Left Ventricular Diastolic Dyfunction
Qinyun RUAN ; Shusheng LIAO ; Lei YAN
Chinese Journal of Hypertension 2007;0(01):-
Background Left atrium size was reported as a marker for ventricular diastolic dysfunction and predictive risk factors for cardiovascular disease.Objective To analyze the relations between left atrial(LA) size and left ventricular(LV) diastolic function echocardiographically in rabbit model with LV dysfunction while with normal LVEF.Methods LV pressure overloaded animal model was established by abdominal aorta constriction in 18 New-Zealand rabbits(model group) which developed LVH with normal EF with 8 healthy rabbits as normal controls.LA dimension and volume(LAD,LAV),left ventricular dimension(LVD) and wall thickness(IVST,PWT),Transmitral inflow E and A,and mitral annulus velocities Ea and Aa were determined by echocardiography.LVEDP was measured within 24 hours after echo examination by catheterization.Results ①Increased LVD,IVST,PWT and LA size were found in model group(P
2.Dot-immunobinding assay for the activity of egg yolk anibody
Guangping RUAN ; Mei AN ; Guihua WANG ; Lei YE ; Shufen DENG
Chinese Journal of Tissue Engineering Research 2007;11(12):2397-2400
BACKGROUND: There are differences in physical and biological activity between the antibody from mammals and egg yolk antibody (IgY) from chicken. IgY is acid- and heat-resistant, and can prevent and cure the infectious diseases in animals and human being, which is also benefit to develop routine diagnostic immunoassays. Conventional ELISA assay for IgY takes much more time than dot-immunobinding assay.OBJECTIVE: To detect the IgY stability byusing dot-immunobinding assay.DESIGN: Open trail.SETTING: Department of Transfusion, Kunming General Hospital of Chinese PLA.MATERTALS: The experiment was completed in the Kunming General Hospital of Chengdu Military Area Command of Chinese PLA from January to June 2006. Two White Leghorn hens (30 weeks old) were selected. HLA-A*0201 α chain served as the antigen. The total protein concentration of the purified antigen was 0.04 g/L with the molecular mass of 32 000(self-prepared); nitrocellulose filter (NC, import and divided); nonfat dry milk (Anyi Corp. No. 20051220); DAB (Boshide Corp.);caprylic acid (made by Shanghai Xinghuo Chemical Factory); ammonium sulfate (Shantou Guanghua Chemical product).METHODS: ①HLA-A*0201 α chain with the total protein concentration of 0.04 g/L was purified with egg yolk antibody,and identified by SDS-PAGE. ②1 μL antigen was spotted into the center of NC membrane and dried in the incubator at 37 ℃. Then the NC membrane was blocked in 1 mL PBST and put in the incubator at 37℃ with shaking in 90 r per minute for 15 minutes. Then the liquid was exchanged with 1 mL PBST and added the primary antibody at a final concentration of 10 mg/L. After 30 minutes shaking in the incubator at 37 ℃, the NC membrane was washed in PBST for three times. The second antibody, mouse anti-chicken IgY conjugated to horseradish peroxidase (HRP) was added and after 30 minutes incubation, the NC membrane was washed three times in PBST. Binding was revealed by incubation with a DAB reagent. A positive reaction was represented by adeep brown spot,Irdlcating that IgY had better activity; if the spot became lighter IgY lost part activity, and when the spot disappeared, the IgY lost a the activty.According to intensity (gray degree)of the dot compared tothe standard, the remained percent of activity of the IgY was calculated. ③IgY was adjusted to three different protein concentrations with PBS: 1, 0.1, 0.01 g/L and stayed at room temperature for four months. 10 μg lgY was taken out from each concentration sample every month to detect the activity by dot-immunobinding assay. ④IgY was put into seven EP tubes with 100 μL per tube and numbered 1-7. Number 1 to 3 was adjusted pH to 5, 3 and 2, respectively with 1 mol/L HCI; Number 4 to 6 was to 9, 11 and 12, respectively with 1 mol/L NaOH. The pH of number 7 was neutral without adding acid or base. The samples were stayed in incubator at 37 ℃ for 3 hours. 10 μg IgY from each tube was taken every hour to detect the stability at different pH by dot-immunobinding assay. ⑤IgY was added to six EP tubes (10 μL per tube) and numbered 1-6. Number 1-6 was put into waterbath at 30, 40, 50, 60, 70 and 80 ℃ for 15 minutes. After cooled in refrigerator at 4 ℃, 10 mg samples from each tube and standard sample (untreated sample) taken to check the thermal stability by dot-immunobinding assay.MAIN OUTCOME MEASURES: ①SDS-PAGE of IgY. ②IgY stability at room temperature. ③IgY stability at different pH. ④ Detection of IgY thermal stability.RESULTS: ①Purified IgY after SDS-PAGE had two major binds, the molecular mass of the heavy chain was 66.000,and the light chain was 25 000. ②1, 0.1, 0.01 g/L IgY still had partial activity after staying at room temperature for four months. ③When pH ranged from 5 to 9, IgY still had partial activity after staying in 37 ℃ for 3 hours. If pH was lower than 5 or higher than 9, it lost the whole activity in above condition. ④Purified IgY was added to six EP tubes, the number 1-4 still had partial activity, but number 5 and 6 showed some white precipitate, which was caused by protein denaturation at higher temperature.CONCLUSION: IgY stability is higher than others. The dot-immunobinding assay described a rapid and simple method for the demonstration and characterization of functional activity of egg yolk antibody. With only small volume antigen and antibody, and specific dot, the dot-immunobinding assay method could process many samples at the same time.
3.Comparison of references for assessment of overweight and obesity in children of Shanghai
Lei CHEN ; Qingya TANG ; Huijuan RUAN ; Xiuhua SHEN
Journal of Shanghai Jiaotong University(medical Science) 2010;30(1):36-38
Objective To compare and analyse four references for assessment of obesity in Chinese children. Methods The height and weight of 2347 children(1175 boys and 1172 girls) aged 7 to 8 years in Shanghai were measured,and their body mass indexes (BMI) were calculated.The prevalences of overweight and obesity were evaluated and compared among reference of Weight for Height 1985(WFH 1985 reference),BMI reference of Working Group on Obesity in China (WGOC reference),BMI reference of Europe International Obesity Task Force(IOTF reference) and BMI reference of Centers for Disease Control and Prevention of American 2000 (CDC reference). Results The prevalence of overweiight in boys evaluated by IOTF reference was significantly higher than those by the other three references(P<0.05),and the prevalence of overweight in girls evaluated by IOTF reference was significantly higher than those by WGOC and CDC references (P<0.05).The prevalence of obesity in boys evaluated by IOTF reference was significantly lower than those by the other three references (P<0.01),and the prevalence of obesity in girls evaluated by IOTF reference was significantly lower than those by WGOC and WFH 1985 references(P<0.01).There was no significant difference in the evaluation findings of obesity and overweight between WFH 1985 and WGOC references(P>0.05). Conclusion WFH 1985 and WGOC references are more suitable than IOTF and CDC references for assessment of overweight and obesity in Chinese children.
4.Analysis of an outbreak of nosocomial infection caused by Burkholderia cepacla using Sau-PCR method
Xi LU ; Weimei RUAN ; Xin XU ; Shinji YAMASAKI ; Lei SHI
Chinese Journal of Laboratory Medicine 2008;31(9):1047-1050
Objective San-PCR was used to analyze an outbreak of nosocomial infection caused by Brukholderia cepacia.Meanwhile a new DNA amplification technique for genetic fingerprinting-San-PCR was introduced,which owns high sensitivity and facility for homology analysis.Methods The proposed technique was based on the digestion of genomic DNA with the restriction endonuclease Sau3AI and subsequent amplification with primers which carried San3AI recognition site.Finally the homology among the DNA samples was analyzed on the basis of the profiles of agarose gel electrophoresis.Results All of the 11 strains isolated from the patients shclwed the homology except one.The results were confirmed by using PFGE and the results showed consistence with PFGE results.Condusion Sau-PCR is simple,robust,rapid method for DNA fingerprinting with broad perspective.
5.Role of FAT/CD36 in high-fat diet-induced adipose tissue inflammation
Yanyan ZHANG ; Lei ZHAO ; Yunxia XIE ; Yaxi CHEN ; Xiongzhong RUAN
Chinese Journal of Pathophysiology 2015;(3):463-467
AIM: To investigate the role of fatty acid translocase/CD36 (FAT/CD36) in adipose tissue in-flammation induced by a high-fat diet.METHODS:C57BL/6J mice were fed with a normal-chow diet ( NCD) or a high-fat diet ( HFD) for 14 weeks.The content of free fatty acid ( FFA) in the serum was measured by ELISA.The expression of CD36, cytokines and chemokines at mRNA and protein levels in the adipose tissues was determined by real-time poly-merase chain reaction and Western blotting.Immunohistochemical staining was used to examine the macrophages infiltration in the adipose tissues.The inflammatory responses in CD36 knockout mice and wild type mice with high-fat diet were ana-lyzed.RESULTS:The levels of FAT/CD36 were higher in HFD group than that in NCD group.HFD feeding enhanced the mRNA and protein expression of IL-1β, IL-6, TNF-α, MCP-1 and MIP-1, as well as promoted macrophage infiltration in the adipose tissues.Interestingly, as fed with HFD, the expression of cytokines/chemokines and macrophage infiltration were significantly reduced in adipose tissues of the CD36 knockout mice, compared with the wild type mice.CONCLU-SION:High-fat diet promotes adipose tissue inflammation in the mice in a FAT/CD36-dependent manner.
6.The practice of flipped classroom teaching based on WeChat platform in geriatrics teaching
Lei RUAN ; Xiaofen WU ; Kai ZHENG ; Cuntai ZHANG
Chinese Journal of Medical Education Research 2017;16(7):697-701
The clinical teaching of geriatrics is difficult to arouse the students' interest. The flipped classroom teaching method can improve it through the interaction. We used WeChat as a platform for flipped classes and designed a variety of micro lessons. The participation and study interests of students were obvi-ously increased through exchanging teaching, answering questions and discussing. The teaching efficiency was also improved after strengthening practice. The preliminary practice shows that this teaching method can provide students a virtual teaching platform. It not only can enhance the teaching effect, but also can share teaching resources in network even for the long-term cross hospital and cross school exchange promotion.
7.Primary cardiac lymphoma of non-Hodgkin's lymphoma located in the right atrium: report of a case.
Yong-li GAN ; Xiang-lei HE ; Ya-jun RUAN
Chinese Journal of Pathology 2007;36(5):355-356
Antigens, CD20
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metabolism
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CD79 Antigens
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metabolism
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Heart Atria
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Heart Neoplasms
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diagnostic imaging
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metabolism
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pathology
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surgery
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Humans
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Immunohistochemistry
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Lymphoma, Large B-Cell, Diffuse
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diagnostic imaging
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metabolism
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pathology
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surgery
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Male
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Middle Aged
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Tomography, X-Ray Computed
8.Advance of study on Mer function.
Journal of Experimental Hematology 2007;15(4):892-895
Mer is one member of Axl receptor tyrosine kinase family, and its ligand Gas6 can stimulate activity of Mer receptor tyrosine kinase after binding it, and then activate the downstream signal transduction pathway, Mer participates in cell inflammation, apoptosis, tumorigenesis, thrombosis and hemostasis. Rencet advances of study on Mer function were reviewed, and its potential prospects of antithrombosis and antitumor were discussed in this article.
Animals
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Fibrinolytic Agents
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Humans
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Intercellular Signaling Peptides and Proteins
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metabolism
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Proto-Oncogene Proteins
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genetics
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metabolism
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Receptor Protein-Tyrosine Kinases
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genetics
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metabolism
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Signal Transduction
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c-Mer Tyrosine Kinase
9.Umbilical cord mesenchymal stem cells with bone marrow stem cells in the treatment of decompensated cirrhosis:a 1-year follow-up study
Zhiyu ZENG ; Dongliang LI ; Jian FANG ; Mei RUAN ; Lei XIA ; Zhiqiang ZHANG ; Shian ZHANG
Chinese Journal of Tissue Engineering Research 2015;(10):1533-1538
BACKGROUND:There are most single-center studies about bone marrow stem cels applied to treat decompensated cirrhosis, but the therapeutic results are not ideal. It is possibly related to aging, physical weakness, poor bone marrow hematopoietic function, less available number of stem cels and feeble ability of regeneration and proliferation in liver cirrhosis patients. Umbilical cord mesenchymal stem cels are characterized of easy to obtain, wide source and weak immunogenicity. Co-transplantation of bone marrow stem cels and umbilical cord mesenchymal stem cels may improve the therapeutic effects on decompensated cirrhosis patients. OBJECTIVE:To investigate the efficacy and safety of co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels on decompensated cirrhosis.METHODS:Thirty-two decompensated cirrhosis patients were randomly divided into two groups: in stem cel group, 13 patients received co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels based on regular medical treatment; in control group, 19 patients only underwent the regular medical treatment. Al the patients were folow-up for 1 year. Alanine aminotransferase, albumin, total bilirubin, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score, 1-year survival rate, Quality of Life score and adverse reactions related to stem cel therapy were observed and recorded in the two groups at 4, 12, 52 weeks after treatment. RESULTS AND CONCLUSION:At 4, 12, 52 weeks after treatment, improvements in the liver function, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score were found in the two groups, but there was no difference between the two groups (P > 0.05). At 4 weeks after transplantation, the clinical symptoms and Quality of Life score in the stem cel group were significantly improved, which were better than those in the control group (P < 0.05). But at 12 and 52 weeks after treatment, no difference was found between the two groups (P > 0.05). In addition, the 1-year survival rate showed no difference between the two groups, and no severe adverse reactions related to stem cel therapy occurred during the folow-up. Co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels is safe and effective to improve the clinical symptoms of decompensated cirrhosis patients. However, further studies with larger samples are warranted to better clarify the co-transplantation effects.
10.Expansion of cytomegalovirus-specific cytotoxic T lymphocytes after peptides stimulation
Guang-Ping RUAN ; Xiang YAO ; Mei AN ; Shu-Fen DENG ; Gui-Hua WANG ; Lei YE ;
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.