1.Preparation and Characterization of Polyclonal Antibody Against Xenopus PAPC
Ruiying HU ; Peng XU ; Yuelei CHEN ; Xin LOU ; Xiaoyan DING
Progress in Biochemistry and Biophysics 2007;34(2):222-228
Xenopus Paraxial Protocadherin (PAPC), which was initially identified in a screen for genes present in the Spemann organizer of Xenopus embryos, is required for gastrulation, somitogenesis and otic vesicle formation. In order to investigate its function in various developmental events, an antibody was prepared which could specifically recognize Xenopus PAPC. Glutathione S transferase (GST) expression system was used to express the fusion protein GST-PAPC. Rabbits were immunized with GST-PAPC Western blotting analysis of FL-PAPC transfected HEK 293T cells lysates, which could be specifically blocked by pre-adsorption of prokaryotic expressed GST-PAPC fusion protein. Furthermore, by using immunofluorescence analysis the polyclonal antibody recognized membrane-bound PAPC in FL-PAPC transfected 293T cells and Xenopus animal cap cells. By Western blotting analysis,the endogenous 150 ku PAPC protein was detected in Xenopus embryos using the anti-PAPC antibody. Take together it could be concluded that a polyclonal antibody specifically against Xenopus PAPC was developed.
2.Autogenous iliac bone implantation after core decompression for the treatment of ischemic necrosis of the femoral head in dogs
Ruiying WANG ; Guodong CHEN ; Chao SUN ; Yan GAO
Chinese Journal of Tissue Engineering Research 2009;13(53):10509-10512
BACKGROUND: The pathological change of femoral head was observed after treating necrosis with core decompression and autogenous iliac bone implantation, expected to obtain a better understanding of the prognosis of autogenous iliac bone implantation method.OBJECTIVE: To explore the curative effect of the ischemic necrosis of femoral head filling the hole of the core decompression with autogenous iliac bone implantation in animal models. DESIGN, TIME AND SETTING: Animal trial observation. The experiment was performed at the Central Laboratory of the Guilin Medical University from April 2006 to September 2007.MATERIALS: Nine hybrid adult dogs were prepared for necrosis of femoral head models.METHODS: The ischemic necrosis of femur head was made with the liquid nitrogen in dogs. The rod-shape autogenous cancellous bone that comes from the posterior superior iliac spine was driven into the full depth of the core r decompression tunnel to the subchondral bone. The dogs were divided into 3 groups, 3 dogs in each groups were sacrificed respectively at 3, 6, 12 weeks after operation. Radiograph and histological examination were used to evaluate the results.MAIN OUTCOME MEASURES: Histological changes of the femoral head necrosis and bone graft at the weeks 3, 6 and 12.RESULTS: No collapse of femoral head was observed at the 3 weeks after operation. The graft as well as perivascular tissues showed necrosis with empty lacune. Fiber granulation tissues could be seen ingrowth of capillaries and perivascular tissues into the bone graft. At 6 weeks after operation, basophilia blue area and new born osteoblast, cartilage, as well as chondroblasts zone could be found at the surface of bone trabecula. After 12 weeks of operation, new bone formation, representing typical process of "creeping substitution". And the cancellous bone of necrotic femoral head was rebuilt gradually. CONCLUSION: Core decompression combined with implantation of autogenous iliac bone implantation can accelerate femoral head repairing, delay or prevent femoral head from necrosis.
3.The role of adiponectin postconditioning in myocardial ischemia reperfusion injury in 3-week old rats
Linjing ZHAO ; Ruiying BAI ; Zhengyue CHEN ; Baosheng YANG ; Jinying ZHANG
Chinese Journal of Applied Clinical Pediatrics 2015;30(9):718-720
Objective To study the role of adiponectin (ADP)post-conditioning in alleviating myocardial ischemia reperfusion injury (MIRI) in 3-week old rats.Methods SD rats (3-week old) were randomly divided into 4 groups by random digits table (n =6):sham group was processed identically except that the left anterior descending coronary artery(LAD) was not ligated;MIRI group was subjected to occlusion of the LAD followed by reperfusion,occlusion for 30 min and reperfusion for 120 min;ADP group was subjected to injection of ADP into femoral vein before reperfusion;LY294002 group (ADP post-conditioning and LY294002) was subjected to injection of ADP and LY294002 into femoral vein before reperfusion.The titer of lactate dehydrogenase(LDH),cardiac troponin I(cTnl) and malondialdehyde(MDA) in plasma were observed.The levels of superoxide dismutase (SOD)and MDA in myocardial tissue were observed.Results The levels of LDH,cTnI and MDA in plasma of MIRI group was (732.11 ± 111.29) IU/L,(38.05 ± 6.17) g/L and (4.49 ± 0.17) nmol/L,respectively,compared with sham group,the levels of LDH,cTnI and MDA in plasma of MIRI group increased obviously(all P < 0.05);the levels of LDH,cTnI and MDA in plasma of ADP group were (581.72 ± 68.41) IU/L,(20.70 ± 5.43) g/L and (3.10 ± 0.20) nmol/L,respectively;compared with MIRI group,the levels of LDH,cTnⅠ and MDA in plasma of ADP group declined significantly (all P <0.05);the levels of LDH,cTnI and MDA in plasma of LY294002 group were (701.10 ±99.59) IU/L,(33.13 ± 4.32) g/L and (4.19 ± 0.46) nmol/L,compared with ADP group,the levels of LDH,cTnI and MDA in plasma of group LY294002 increased remarkably (t =2.477,1.804,2.961,all P < 0.05).Compared with sham group,the SOD level decreased and the MDA level increased in myocardial tissue of MIRI group(all P < 0.05);compared with MIRI group,the SOD level increased and the MDA level decreased in myocardial tissue of ADP group (all P < 0.05);compared with ADP group,the SOD level decreased and the MDA level increased in myocardial tissue of LY294002 group (all P < 0.05).Conclusions ADP postconditioning is perhaps a protective factor for alleviating myocardial ischemia reperfusion injury in 3-week-old rats,the protective effect is perherps related to alleviating the damage of the lipid peroxidation by signaling pathway of adiponectin/phosphoinositide-3 kinase/protein kinase B.
4.Effect of Zusanli and Guanyuan Electroacupuncture on Inflammatory Reaction in Sepsis Patients
Guang YANG ; Ruiying HU ; Ming CHEN ; Jian LI
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(3):430-433
Objective To study the effect of Zusanli and Guanyuan electroacupuncture ( EA) on inflammatory reaction in patients with sepsis. Methods A prospective and randomized trial was carried out in 58 sepsis patients, and the patients were randomly divided into EA group and control group. Patients in both groups were given conventional western medicine treatment, and EA group received EA at acupoints of Zusanli and Guanyuan additionally. The hospitalization time in intensive care unit ( ICU) of the two groups was compared. The changes of serum lactic acid (LA) level and inflammatory indexes including white blood cell and neutrophil count, C- reactive protein (CRP), erythrocyte sedimentation rate (ESR) and procalcitonin (PCT) of the two groups were observed before and after treatment. Results The hospitalization time in ICU of EA group was less than that of the control group, and the difference was statistically significant ( P<0.05) . On the seventh day after treatment, the serum LA level of EA group was significantly lower than that of the control group ( P<0.05) . Inflammatory indexes of EA group were decreased after treatment compared with those before treatment ( P<0.05) , and on seventh day after treatment, the inflammatory indexes of EA group were significantly lower than those of the control group ( P<0.05) . Conclusion EA at acupoints of Zusanli and Guanyuan is effective for the treatment of sepsis through reducing the inflammatory reaction of sepsis patients.
5.Study on a new urine analysis core module based on semi-reflection mirror.
Longcong CHEN ; Gaiqin LIU ; Nan HU ; Ruiying ZHANG ; Qifeng JIANG ; Bin GAO ; Xingliang XIONG
Journal of Biomedical Engineering 2014;31(6):1288-1293
A new urine analysis core module based on high performance 32-bit microprocessor and high precision color sensor was presented. A novel optical structure and a specific circuit were applied to improve measurement precision and temperature was used to compensate for results in this core module. The information of urine test peice, such as all original data and color RGB value, reflectivity, semi-quantitative level, etc. can be output. The results showed that the measuring precision was about 95% or above with ideal stability and reliability using this presented core module, which can be conveniently applied in various urine analyzers, and can greatly decrease the cost of urine analyzers in development and production.
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6.CHOP expression and its correlation with proliferative/apoptotic ratio in colorectal adenoma-carcinoma sequence under same genetic background
Yiqian LI ; Junrong CHEN ; Chujun LI ; Ruiying ZHAO ; Huiling YANG ; Yingyu ZHU ; Qinghong CAI ; Siyuan HU
Chinese Journal of Pathophysiology 2014;(6):1004-1009
AIM:To investigate the expression of C/EBP homologous protein (CHOP) and its correlation with proliferative/apoptotic ratio (PAR) in colorectal adenoma-carcinoma sequence under the same genetic background .ME-THODS:Four kinds of tissue samples under the same genetic background from 23 patients, including normal colorectal tissue, adenoma with low-grade intraepithelial neoplasia , adenoma with high-grade intraepithelial neoplasia and colorectal adenocarcinoma samples , were collected .TUNEL method and Ki-67 immunohistochemistry were applied to determine the PAR.The expression of CHOP was detected by immunohistochemistry SABC method .RESULTS: ( 1 ) Under the same genetic background , the level of CHOP expression is significantly higher in colorectal adenocarcinoma than that in the ade -noma with high-grade intraepithelial neoplasia , the adenoma with low-grade intraepithelial neoplasia and the normal muco-sa.The level of CHOP expression was significantly higher in the adenoma with high -grade intraepithelial neoplasia than that in the adenoma with low-grade intraepithelial neoplasia and the normal mucosa .The level of CHOP expression was signifi-cantly higher in the adenoma with low-grade intraepithelial neoplasia than that in normal mucosa .(2) Under the same ge-netic background , PAR was significantly higher in the colorectal adenocarcinoma than that in the adenoma with high -grade intraepithelial neoplasia , the adenoma with low-grade intraepithelial neoplasia and the normal mucosa .PAR was significant-ly higher in the adenoma with high-grade intraepithelial neoplasia than that in the adenoma with low-grade intraepithelial neoplasia and the normal mucosa .PAR was significantly higher in the adenoma with low-grade intraepithelial neoplasia than that in the normal mucosa.(3) CHOP levels were positively correlated with PAR in the adenoma with low-grade intraepi-thelial neoplasia , adenoma with high-grade intraepithelial neoplasia and colorectal adenocarcinoma .CONCLUSION:CHOP expression and PAR continuously increased and positively correlated along the adenoma -carcinoma sequence , indica-ting that endoplasmic reticulum stress mediates the carcinogenesis of colorectal adenomas .
7.Application of standard treatment protocol for type 2 diabetes patients with obstructive sleep apnea-hypopnea syndrome in the elderly
Xiao ZHANG ; Songyun OUYANG ; Peizong SUN ; Ruiying CHEN ; Liping DAI ; Xialian LI ; Lijuan WANG
Chinese Journal of Geriatrics 2013;32(11):1141-1144
Objective To evaluate the efficacy of continuous positive airway pressure (CPAP)-based standard treatment on obstructive sleep apnea-hypopnea syndrome (OSAHS) patients with type 2 diabetes and insulin resistance (IR) in the elderly.Methods 63 elderly type 2 diabetes patients diagnosed as OSAHS were randomly divided into control group (n =31) and CPAP group (n =32).Patients in control group were treated with conventional therapy including diet control,exercise and antidiabetic drugs,and patients in CPAP group were treated with CPAP treatment combined with conventional therapy.Fasting glucose (FBG),oral glucose tolerance test (OGTT),homeostasis model assessment of IR index (HOMA-IR),body mass index (BMI),apnea-hypopnea index (AHI),lowest oxygen saturation (L-SaO2) and the dosage of insulin application were observed after 7 days,1 month,3 months of the treatment.Glycosylated hemoglobin (HbA1c) was determined before and after 3 months of the treatment.Results In CPAP group,levels of FBG and 2 h OGTT glucose,HOMA-IR,AHI,daily insulin dosage were significantly reduced and L-SaO2 was significantly increased after 7 days of the treatment; BMI was significantly reduced after 1 month of the treatment;HbA1c level was significantly reduced after 3 months of the treatment.In control group,L-SaO2 was significantly reduced along with the multiple time points; daily insulin dosage was significantly increased after 3 months of the treatment.There were no significant differences in all observed indicators between groups before treatment.There were significant differences in the observed indicators between groups after treatment except for BMI after 7 days of the treatment.Conclusions Compared with conventional therapy,CPAP-based standard treatment has more efficacy on increasing insulin sensitivity and improving insulin resistance in elderly patients with OSAHS and type 2 diabetes.
8.Comparison of genotype distribution of patients with acute hepatitis B infection or chronic hepatitis B infection in Shanghai
Xuesong LIANG ; Mobin WAN ; Chengzhong LI ; Hao XU ; Jianya XUE ; Ruiying ZHENG ; Jixiu CHEN
Chinese Journal of Infectious Diseases 2009;27(1):23-26
Objective To investigate the distribution of genotypes in chronic HBV infection (CHB) and acute HBV infection (AHB) patients in Shanghai. Methods Sixty-two patients with AHB and 73 patients with CHB admitted to ('hanghai Hospital of Shanghai between 2003 and 2007 were studied. Viral genotypes of all the patients were determined by direct gene sequencing.Meanwhile, epidemiological, clinical and biochemical parameters of all patients were collected. Mean values of different groups were compared by t test while frequency was compared by chi square test. Results The major prevalent genotypes in both AHB and CHB patients were genotype B and C (48.4% vs 51.6% in AHB patients and 26.0% vs 74.0% in CHB patients). The proportion of genotype B was higher in AHB patients compared to CHB patients (P= 0.02). Epidemiological factors and clinical outcomes were not statistically different among patients with different viral genotypes. The proportion of genotype C was much higher in CHB patients compared to AHB patients (P=0.006). The main transmission route of AHB was heterosexual interaction which was 18 out of 62 (29.0%), but in CHB patients, it was prenatal transmission which was 38 out of 73 (52.1%). Conclusions In shanghai, the main HBV genotypes in both AHB and CHB patients are genotype B and C. The proportion of genotype B is relatively high in AHB patients while proportion of genotype C is more common in CHB patients. There is no significant relationship between genotypes and the clinical outcomes of AI-IB patients.
9.Clinical significance of serum levels of growth factors in different kinds of inflammatory response
Xiuhua DAI ; Baochun ZHAO ; Ruiying CHEN ; Suxiang LI ; Yuemin BAI ; Jin MENG
Clinical Medicine of China 2009;25(11):1215-1217
Objective To evaluate the changes of platelet-derived growth factor(PDGF) and vascular endo-thelial growth factor(VEGF) in infectious and autoimmunity diseaases. Methods The growth factors were measured by ELISA in 149 patients for seven times in different periods,and the levels and duration were compared. Results PDGF was (6.32±2.54) μg/ml and (2.57±1.65) μg/ml (P<0.05), and VEGF was (179.5±53.30) ng/ml and (221.4±70.04) ng/ml(P<0.05) in the two groups before treatment;There was statistical significance in dif-ferent time points between groups and within groups for PDGF(P<0.05) and significant difference for time chan-ging to the peak and changed duration between two groups for VEGF(P<0.05). For the two groups,the peak time of PDGF were (2.6±1.1) and (5.4±3.3) days;The peak time of VEGF were (2.4±0.7) and (7.2±3.3) days respectively(P<0.05 for each). However in the two groups, the change duration were (6.7±3.1) and (15.4±6.1) days for PDGF and (8.1±3.4) and (16.7±7.2) days for VEGF (P<0.05 for each). Conclu-sions There is significant difference for serum levels of PDGF and VEGF and the level change duration as well in the two groups, which maybe correlated with inflammatory nature and outcome.
10.Comparative study on drug resistance of pseudomonas aeruginosa isolated from different sites of patients
Ruiying LI ; Xiaosong XU ; Sicong CHEN ; Jinghui LU ; Xin LI ; Tao ZENG
International Journal of Laboratory Medicine 2015;(14):2023-2024
Objective To analyze the infection sites of pseudomonas aeruginosa (PAE) isolated from patients in hospital ,and in‐vestigate their drug resistance situation ,in order to provide reference information for clinical use of antibiotics rationally .Methods The sample distribution of PAE between January 2012 and December 2013 were retrospectively analyzed .And the resistance rates of PAE to antibacterial drugs from different sites of patients were statistically compared .Results The isolation rate of PAE in re‐spiratory tract was the highest ,accounting for 74 .1% ,closely followed by isolation rate in urine and wound secretion .The resist‐ance rate of PAE to antibacterial drugs in these three kinds of specimen is statistically different (P<0 .05) .The resistance rate of PAE is high in respiratory tract ,and low in wound secretion .Conclusion The pseudomonas aeruginosa infection is mostly common‐ly found in respiratory tract ,and has the highest drug resistance rate .The choice of antibacterial drug should be made according to the infection sites of patients ,because the resistance rate of PAE in different sites of patients is significantly different .