1.Screening and Analysis of Proteins Interacting with HCMV UL145 from a Human Fetal Brain cDNA Library
Fei ZOU ; Shuang WANG ; Si WU ; Zhengrong SUN
Journal of China Medical University 2017;46(4):309-312
Objective To screen a human fetal brain cDNA library for proteins that can interact with HCMV UL145 using a yeast two-hybrid system.Methods A bait plasmid (pGBKT7-UL145) was constructed.Using HCMV UL145 as bait,a human fetal brain cDNA library was screened and proteins interacting with UL145 were identified using bioinformatic methods to sequence and analyze the positive clones.Results Three clones interacting with HCMV UL145 were found,and identified as FOXG1.Conclusion Several proteins interacting with HCMV UL145 in the human fetal brain cDNA library were identified as FOXG1,indicating that this protein may play an important role in the course of HCMV infection.
2.The variables in normalizing glomerular filtration rate
Fei LI ; Hongwei SI ; Jianhua GENG ; Shengzu CHEN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2013;(3):238-240
Normalizing GFR with variables is important for non-cancer patients,especially for kidney donors.The most frequently evaluated variables are body surface area (BSA),extracellular fluid volume (ECV) and lean body mass (LBM).It is difficult to accurately quantify BSA and the power of BSA normalization decreases in children and obesity population.The ECV normalization is suitable for healthy children,but its clinical value decreases in patients with damaged renal function.The LBM can be accurately measured and has a larger serviceable range in the normalization.Although the influence factors of LBM should be extensively evaluated,the available data indicate that LBM is more suitable in the normalization of GFR than BSA and ECV.
3. Study on quality standard improving of Shenmai Dihuang Pills
Chinese Traditional and Herbal Drugs 2017;48(2):299-304
Objective: To develop the quality standard for Shenmai Dihuang Pills (SDP). Methods: The TLC methods applied to identify MoutanCortex, CorniFructus, Alismatis Rhizoma, and GlehniaeRadix in the formulation were carried out according to the methods recorded in general rules of Chinese Pharmacopeia (2015 edition, volume 4). The contents of psoralen, iso psoralen, and paeonol were analyzed by high performance liquid chromatography on a Diamonsil C18 column (250 mm ×4.6 mm, 5μm) with the mobile phase of acetonitrile (A)-0.1% formic acid (B) by gradient elution at a flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm and the column temperature was controlled at 30℃. Results: There were good specificities of TLC for MoutanCortex, CorniFructu, Alismatis Rhizoma, and Glehniae Radix. The methodology validation for the assay of psoralen, iso psoralen, and paeonol presented that they were in good linear correlation in the ranges of 6.531-58.78, 6.115-55.04 and 20.40-183.6 μg/mL, with the regression equations of Y =1.523×104X-1.209×104 (r = 0.999 9), Y =1.809×104 X-1.523×104 (r = 0.999 9), and Y =2.760 ×104 X-1.683 ×104 (r = 0.999 9), respectively. The average recoveries were 100.4% (RSD 1.1%), 101.5% (1.3%), and 101.5% (0.79%).The content ranges of psoralen, isopsoralen, and paeonol from five different batches of SDP were 322.1-331.0, 298.0-350.1, and 929.5-982.7 μg/g, respectively. Conclusion: The established quality of TLC and HPLC methods are specific, reliable, accurate, and suitable, which can be successfully applied to the quality control for the preparation.
4.Screening and identification of proteins interacting with HCMV UL132 protein
Fei ZOU ; Shuang WANG ; Si WU ; Zhengrong SUN
Journal of Jilin University(Medicine Edition) 2017;43(4):690-693,前插1
Objective:To screen the proteins interacting with the human cytomegalovirus(HCMV)UL132 protein from the human fetus brain cDNA library by using Yeast Two-Hybrid System, and to elucidate the possible mechanism of UL132 protein in congenital cytomegalovirus infection.Methods:The HCMV UL132 fragment was amplified by polymerase chain reaction,the amplified HCMV UL132 fragment and expression vector pGBKT7 were digested and purified,and the HCMV UL132 fragment was linked to the vector pGBKT7.The pGBKT7-UL132 was constructed and transformed to yeast AH109, then the Human Fetal Brain DNA Library DNA was transformed into AH109 yeast.Using HCMV UL132 as abait, a human fetus brain cDNA was screened and the proteins interacting with UL132 protein were searched, the positive clone was sequenced and analyzed by bioinformatics methods.Results:The bait expression vector pGBKT7-UL132 was successfully constructed.The results of double enzyme digestion showed that there were two visible bands of 800 and 7 000 bp, respectively.After transformation of library plasmid, the transformation efficiency was calculated, and the transformation efficiency was 6.6×103 cfu· μg-1.There were 95 blue clones by X-gal coloration reactionsequencing and there were 10 clones interacting with the protein encoded by UL141 protein.The BLAST analysis showed that 7 of them were highly homologous with CAML.Conclusion:CAML might be one interaction protein with HCMV UL132 in Human Fetus Brain cDNA Library,suggesting that the interaction may be associated with the invasion and proliferation of the HCMV.
5.Relationship between interleukin-6 and serum polyunsaturated fatty acids in patients with metabolic syndrome
Dajun LOU ; Qiqian ZHU ; Fei YE ; Haiyan DONG ; Dihua HUANG ; Xuwei SI
Chinese Journal of Clinical Nutrition 2017;25(1):36-39
Objective To investigate the relationship between serum phospholipid polyunsaturated fatty acids (PUFA) and interleukin-6 (IL-6) in patients with metabolic syndrome.Methods Serum phospholipid PUFA profiles of 61 patients with metabolic syndrome and 55 healthy controls was analyzed with high performance gas chromatography.Enzyme-linked immunosorbent assay was used to measure serum IL-6 levels.Results The IL-6 concentration was significantly higher in the MS group than in the control group [(16.0 ± 4.7) pg/ml vs.(9.2 ± 2.6) pg/ml,P =0.000].The total PUFA percentage was lower in the MS group than in the control group [(47.2±2.4)% vs.(50.8 ±2.5)%,P=0.001].The serum IL-6 concentration was negatively correlated with n-3 PUFA,n-6 PUFA,and total PUFA (r =-0.51,P <0.01;r=-0.27,P<0.01;r=-0.38,P<0.01).Conclusions Serum PUFA in patients with metabolic syndrome is lower than in healthy people.Serum IL-6 concentration may be negatively correlated with total PUFA concentration.
7.Association of triglyceride with serum polyunsaturated fatty acid in patients with metabolic syndrome
Dajun LOU ; Qiqian ZHU ; Fei YE ; Duo LI ; Dihua HUANG ; Haiyan DONG ; Xuwei SI
Chinese Journal of Clinical Nutrition 2013;21(5):263-266
Objective To investigate the relationship between triglyceride and serum polyunsaturated fatty acid (PUFA) in patients with metabolic syndrome (MS).Methods Totally 74 MS patients (MS group) and 62 healthy subjects (normal control group,NC group) were recruited from June 2011 to June 2012 in Shaoxing People's Hospital.Serum phospholipid fatty acid profiles were analyzed with capillary gas chromatography.Serum lipids were measured by enzymatic assay.Results The triglyceride concentration was significantly higher in MS group than that in NC group [(2.0 ±0.8) mmol/L vs.(1.3 ±0.5) mmol/L,P=0.000].The levels of n-3 PUFA,n-6 PUFA,and PUFA were significantly lower in MS group than those in NC group (9.8% ±2.2% vs.11.1% ±2.4%,P=0.002; 35.4% ±6.7% vs.39.5% ±7.8%,P=0.009; 45.2% ±8.9% vs.50.6% ±10.1%,P =0.000).Triglyceride concentration was inversely correlated with n-3 PUFA and PUFA levels (r =-0.42,P =0.008 ; r =-0.23,P =0.013).Conclusions n-3 PUFA,n-6 PUFA,and PUFA decrease in MS patients.Triglyceride concentration is inversely correlated with PUFA.
8.The experimental study of histology and apoptosis after the bone cement leakage into the intervertebral disc
Hui ZHAO ; Cai-Fang NI ; Long CHEN ; Tian-Si TANG ; Hui-Lin YANG ; Jian-Fei HUANG ;
Chinese Journal of Radiology 2000;0(12):-
Objective To discuss the histology change,apoptosis state and Bcl-2,Bax expression after the bone cement leakage into the intervertebral disc in vertebroplasty.Methods Eight majority canis familiaris were studied.Three lumbar intervertebral discs(L2 to L5)in each dog were randomly classified into three groups(control group,PMMA group,and CPC group),the canine discs were stabbed by 18-gauge needle,and 0.1 ml cement was injected into them.Control discs were only stabbed and injected with nothing.Histology of all discs was studied 24 weeks after the operation.Terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)and immunohistochemisty were used to detect apoptosis and Bcl-2,Bax expression in the discs.The data were statistically analyzed by SPSS 12.0.Results Intervertebral disc degeneration was not found in control groups.In bone cement groups,however,ruptured or serpentine patterned fibers,decreased cellularity of the nucleus pulposus and condensed matrix of the nucleus pulposus were found in histologic results.The Bax protein decreased in the order of control group, CPC group,and PMMA group.However,the Bcl-2 protein increased in the order of control group,CPC group,and PMMA group.The histology grade was significantly different among the three groups under ANOVA analysis(P
9.Degradation of a magnesium alloy stent in the rabbit abdominal aorta
Qian AN ; Wenjun CUI ; Jiangtao SI ; Ying WANG ; Fei WU ; Yu DING ; Yang LI ; Bing WANG
Chinese Journal of Tissue Engineering Research 2017;21(18):2864-2869
BACKGROUND:Foreign studies have found that the magnesium alloy stent is safe and effective, but there are few studies on the degradation performance of magnesium alloy stents in China.OBJECTIVE:To investigate the degradation of degradable AZ31 magnesium alloy stent in the rabbit abdominal aorta and the effect of degradation process on vascularization.METHODS:Twenty-eight rabbits were enrolled, and the degradable AZ31 magnesium alloy stent was implanted into the rabbit abdominal aorta. Postoperative abdominal aortic X-ray examination and histological observation were done at 30, 60, 90, 120 days after implantation.RESULTS AND CONCLUSION:(1) X-ray examination: 30 days after implantation, the stent expanded completely with structural integrity; 60 days after implantation, the stent deformation, partial stent fracture, and lose of support were found; 90 days after implantation, only a small amount of support rod residues were found, and the majority of the stent was degraded; and 120 days after implantation, there was no support rod residual, and the stent was degraded completely. (2) Histological observation: 60 days after implantation, the number of residual support rods was less than that 30 days after implantation (P< 0.05), the number value at 90 days after implantation was lower than that at 30 and 60 days after implantation (P< 0.05), and the number value at 120 days after implantation was lower than that at 30, 60, 90 days after implantation (P < 0.05), indicating that the number of residual support rods was negatively correlated with post-implantation days. The time for complete stent degradation was 124.8 days. The intimal area at 90 days after implantation was higher than that at 30, 60, 120 days after implantation (P < 0.05), while the lumen area was smaler than that at 30, 60, 120 days after implantation (P < 0.05). There was no significant difference in the intimal area and lumen area at latter three time points after implantation. To conclude, the degradation of the degradable AZ31 magnesium alloy stent in the rabbit abdominal aorta can be completed within 124.8 days, and at 90 days after the stent is implanted, vascular intimal hyperplasia and lumen stenosis are most serious, and then gradualy reduced.
10.Characteristics of serum free fatty acid and high-sensitivity C-reactive protein in patients with type 2 diabetes mellitus and metabolic syndrome
Dajun LOU ; Qiqian ZHU ; Weikun GONG ; Fei YE ; Haiyan DONG ; Dihua HUANG ; Xuwei SI
Chinese Journal of Clinical Nutrition 2015;23(6):351-355
Objective To investigate characteristics of free fatty acid (FFA) and high-sensitivity Creactive protein (hs-CRP) in patients with type 2 diabetes mellitus and metabolic syndrome (MS) compared to those with type 2 diabetes mellitus without MS.Methods Totally 120 patients with type 2 diabetes mellitus who received blood glucose control in Shaoxing People's Hospital from January to June 2013 were recruited and divided into MS group (n =56) and non-MS group (n =64).The serum FFA profile of the patients was measured by enzymatic assay, and the serum hs-CRP level measured by particle-enhanced immuno-precipitation assay.Results The level of FFA was higher in the MS group than that in the non-MS group [(0.60 ±0.25) mmoL/L vs.(0.45 ±0.21) mmol/L, P =0.033].The level of hs-CRP was higher in the MS group than that in the non-MS group [5.29 (4.69-5.82) mg/L vs.0.73 (0.42-1.26) mg/L, P =0.000].The level of hs-CRP was higher in the patients with central obesity than that in those without central obesity [4.34 (0.91-5.46) mg/L vs.1.80 (0.82-3.27) mg/L, P=0.014], also higher in hypertensive patients than in non-hypertensive patients [5.21 (3.18-5.96) mg/L vs.2.93 (0.89-4.98) mg/L, P =0.012].Conclusions Serum FFA and hs-CRP concentrations may be significantly higher in type 2 diabetes mellitus patients with MS.The increased levels of FFA and hs-CRP may play an important role in the pathogenesis of type 2 diabetes mellitus and MS.