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.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.
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.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.
6.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
7.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.
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.Effect of aging-related changes in gap junction protein on arrhythmia in rats
Xingfen WANG ; Cuntai ZHANG ; Rende XU ; Lei RUAN ; Xiaoqing QUAN ; Ming CAO ; Xueying TONG ; Yi YANG
Chinese Journal of Geriatrics 2011;30(5):427-430
Objective To investigate the aging-related changes in gap junction protein-connexin 43 (Cx 43) in rats and their effect on the high incidence rate of ventricular arrhythmia in aged rats. Methods The 64 healthy male Fischer 344 (F344) rats were randomly divided into four age groups (n=16,each): 3-6 months (juvenile), 9-12 months (young-adult), 18-21 months (middle-aged) and 24-26 months (aged). The incidence rate of ventricular arrhythmia was recorded by monitoring their limb-lead Electroa rdiogram(ECG). Morphological changes of ventricular myocardium were observed under optical microscope in Hematoxylin (HE) and Masson's stain. The distribution of connexins 43 (Cx43) and deophosphatase (NP) Cx43 was observed by confocal immunofluorescence microscopy and the Cx43 and NP-Cx43 protein expression was assessed by Western-Blot. Results The incidence rate of ventricular arrhythmia was much higher in aged group (75.0%) than in other three groups (0%,0%,12.5%), all P<0.05, and the aged group showed that ventricular muscle cells were hypertrophy and arrayed sparsely and disorderly with hyperplasia of connective tissues. The distribution of Cx43 changed from end-to-end to disordered arrangement and the total expression amount of Cx43 decreased as age increased (P<0.05). The expression amount of NP-Cx43 in middle-aged rats was notably decreased than in juvenile and aged rats (P<0.05). Conclusions For aged rats, the high incidence rate of ventricular arrhythmia may be associated with ventricular myocardium reconstruction, disarrangement of ventricular muscle cells and gap junction proteins, decreased expression amount of Cx43 and relatively increased NP-Cx43.
10.Combining plates and nails in the treatment of ipsilateral unicondylar tibial plateau and shaft fractures
Lei HUANG ; Yanzhao ZHU ; Feng ZHANG ; Penghan YE ; Xianfeng HE ; Yongping RUAN ; Rongming XU
Chinese Journal of Orthopaedics 2012;32(8):739-744
Objective To analyze the feasibility and effect of combining intramedullary nails and plates in the treatment of ipsilateral unicondylar tibial plateau fractures with non.contiguous tibial shaft fractures.Methods From May 2003 to November 2005,21 patients with ipsilateral unicondylar tibial plateau fracture and non-contiguous tibial shaft fracture were treated with combining intramedullary nails and plates,including 15 males and 6 females,with the mean age of 34 years (range,20-55 years).The tibial plateau fractures were classified according to Schatzker et al.; 4 cases were type Ⅰ,11 cases type Ⅱ,6 cases type Ⅲ and 1 case type Ⅵ.Middle third tibial shaft fractures were in 13 patients,and distal third tibial shaft fractures were in 8 patients.Once the reduction was obtained,we first fixed tibial plateau with plate and then tibial shaft with medullary nail in 19 cases; in 2 cases,we first fixed the tibial shaft with medullary nail and then tibial plateau with plate.Results 1All patients were followed up for 0.9 to 4 years (average,2.2 years).Bone union was obtained in all patients.The tibial plateau fractures united after an average of 12 weeks,and the tibial shaft fractures united after an average of 29 weeks.Delayed union of the tibial shaft fracture occurred in 3 patients,and the fracture healed finally by removing the proximal locking.Tibial plateau malunion occurred in one patient due to malreduction.According to HSS scores,excellent result was obtained in 17 patients (80.95%),good in 3 patients (14.29%),fair in 1 patient (4.76%).Conclusion With careful attention to some techniques,ipsilateral unicondylar tibial plateau fractures with non-contiguous tibial shaft fractures can be treated successfully by combining medullary nail and plate.