1.Analysis of protein expression profiling in a mouse model of oxygen-induced retinopathy
Chen, QI ; Qian, HAN ; Qiyu, BO ; Xun, LIU ; Fei, WANG ; Yan, ZHANG ; Xiaorong, LI
Chinese Journal of Experimental Ophthalmology 2015;33(12):1069-1075
Background Retinal neovascularization (RNV) is one of the major causes of blindness worldwide,but the pathogenic mechanism of this disease remains unclear, and therapeutic modalities need to be improved.Therefore,it is necessary to identify ocular protein markers with significant expression changes during RNV, thereby providing novel therapeutic targets for neovascular retinopathies.Objective This study was to investigate retinal vessel morphological characteristics and protein expression profiling in a mouse model of oxygen-induced retinopathy (OIR).Methods Forty four C57BL/6J mouse pups were randomly divided into normal control group and OIR group at postnatal day 7 (P7).The mice in the normal control group were raised under the normal air for 10 days.The mice of the OIR group were exposed to (75±2)% oxygen for 5 days.The mother mice were alternated between the two groups every day.The mice of the OIR group returned to normal air at P12.Fluorescein isothiocyanate-dextran (FITC-dextran) was retrobulbarly injected at P17 mice from both groups, and the retinal flatmounts were prepared after fixation.The FITC-dextran-labeled retinal vessels were observed and quantified;the paraffin sections of eyeballs were processed for hematoxylin and eosin staining,and the number of pre-retinal vascular cell nuclei was quantified.The total proteins were extracted from the eyes, and the expression profiling was analyzed by a customized protein array and verified by Western blot and ELISA.Results The retinal flatmounts labeled with FITC-dextran showed that the peripheral retinal microvessels in the OIR group were tortuous, disorganized with neovascular buds,and the vascular obliteration was prominent in the center of retina.Contrastly,the vessels were smooth,organized, and evenly distributed in the normal control group.The percentage of vascular obliteration area in the OIR group was (25.53±2.16)% ,which was significantly higher than (0.66±0.36)% in the normal control group (t=-27.61 ,P< 0.01).The number of pre-retinal vascular cell nuclei,as revealed by hematoxylin and eosin staining, was (28.41 ± 3.97)/slide in the OIR group, which was substantially higher than (0.16±0.31)/slide in the normal control group (t =-54.42,P<0.001).Protein array showed that 10 out of the 62 examined pro-inflammatory, pro-angiogenic cytokines exhibited more than 1.5-fold expression changes,including 3 up-regulated cytokines and 7 down-regulated cytokines;4 cytokines showed more than 2-fold expression changes,in which 3 cytokines were down-regulated and 1 cytokine was up-regulated.The differential expressions were verified by Western blot and ELISA.The expression trends of platelet factor 4 (PF-4), vascular endothelial growth factor A (VEGF-A), selectin P (SELP), vascular cell adhesion molecule 1 (VCAM-1) ,soluble tumor necrosis factor receptor Ⅱ (sTNF-RⅡ) and chemokine C-X-C motif ligand 16 (CXCL16) were consistent with those revealed by protein array.PF-4,VEGF-A and SELP were up-regulated,and the other 3 were significantly down-regulated (all at P<0.05).Conclusions Differential expression patterns of the cytokines,including PF-4,VEGF-A, SELP, VCAM-1, sTNF-RⅡ and CXCL16, are identified between normal and OIR mouse eyes.These differential expression patterns suggest that under the condition of OIR,the platelet system is activated,and proinflammatory factors are down-regulated.PF-4 might become a new target for VEGF-independent therapeutic strategy against RNV.
2.Synthesis of 3-mercaptopropionic acid-stabilized CdTe quantum dots and its application for the detection of Cu2+
Bo JIANG ; Qi HUANG ; Qiang LU ; Caolong LI ; Fei WANG ; Qingjia REN ; Zeng REN
Journal of China Pharmaceutical University 2017;48(3):311-316
The water-soluble 3-mercaptopropionic acid (MPA)-stabilized CdTe (MPA-CdTe) quantum dots (QDs) were synthesized by aqueous suspension.The study showed that the fluorescence quenching process of Cu2+ to MPA-CdTe QDs,whose largest emission peak was 599 nm,could be described well by the theory of fluorescence quenching in competitive absorption systems and its modification of Stern-Volmer equations.By fittings,the results showed a good polynomial relationship between the fluorescence intensity F0/F and the concentration of Cu2+,when the concentration was in the range of 2.28 × 10-6-18.24 × 10-6 mol/L and 4.8 × 10-7-12 × 10-7 mol/L,and two polynomial equations were respectively elucidated based on dynamic and static quenching in competitive-absorption systems:F0/F =7.999-2.470c +0.339 c2,F0/F =3.154-0.160 c +0.049 c2 and degree of fitting are 0.991 and 0.993,respectively.The detection limit was 1.326 × 10-7.
3.Biological markers as predictive factors of response to neoadjuvant taxanes and anthracycline chemotherapy in breast carcinoma.
Bo ZHOU ; De-qi YANG ; Fei XIE
Chinese Medical Journal 2008;121(5):387-391
BACKGROUNDNeoadjuvant chemotherapy provides an excellent model for evaluation of potential predictive factors. The objective of this study was to evaluate the predictive value of different biological factors in breast cancer patients treated with neoadjuvant taxane and anthracycline chemotherapy.
METHODSOne hundred and thirty-five patients treated with 4 cycles of neoadjuvant taxanes and anthracycline were included in this retrospective study. Using pretreatment biopsy materials, immunohistochemical studies were performed for estrogen receptor (ER), progesterone receptor (PgR), HER-2, Ki-67 and p53 protein expression. The associations among biological markers and clinical and pathological complete response (pCR) were analyzed.
RESULTSThe overall clinical response was 86%, including 33% clinical complete response (cCR) and 53% clinical partial response. The pCR was just 17%. In the univariate analysis, only HER-2 overexpression was predictive of cCR to neoadjuvant chemotherapy (P=0.018). No significant associations between other biological factors and cCR were found. Absence of ER, PgR expression and overexpression of HER-2 were predictive of the pCR (P=0.002, 0.001, 0.01, respectively). Ki-67 and p53 failed to show an association with pCR. In multivariate analysis, overexpression of HER-2 remained as an independent variable in predicting the cCR (P=0.021). However, negative ER was the only parameter that maintained statistical significance in predicting the pCR (P=0.001).
CONCLUSIONSPatients with overexpression of HER-2 and negative hormonal receptor status are much more likely to respond to neoadjuvant taxane and anthracycline chemotherapy than those with the opposite characteristics. These factors could serve as predictive markers for this regimen.
Adult ; Aged ; Anthracyclines ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; analysis ; Breast Neoplasms ; chemistry ; drug therapy ; Chemotherapy, Adjuvant ; Female ; Humans ; Ki-67 Antigen ; analysis ; Middle Aged ; Receptor, ErbB-2 ; analysis ; Receptors, Estrogen ; analysis ; Receptors, Progesterone ; analysis ; Taxoids ; administration & dosage ; Tumor Suppressor Protein p53 ; analysis
4.Evaluation of iodine supplementation on improvement of developmental quotient at the critical period of infant brain development
Yan-ling, WANG ; Peng-fei, GE ; Yong-qin, CAO ; Jing, ZHENG ; Wei, SUN ; Hong-bo, LI ; Qi-yi, MA
Chinese Journal of Endemiology 2013;32(4):400-403
Objective To evaluate the effect of iodine supplementation on improvement of developmental quotient (DQ) at the critical period of infant brain development.Methods Pregnant,lactating women and infants less than 3 years old were supplemented with iodized oil in Linxia Hui Autonomous Prefecture(Linxia Prefecture) Gansu Province in 2006-2010.Before and after the intervention(2006,2007-2010),five townships were randomly selected in the north,the south,the east,the west and the center of eight counties(cities) of Linxia.One village was chosen from each of those townships and 20 infants,20 pregnant women and 20 lactating women were randomly selected in each village(insufficient was made up from the neighboring villages).Urinary iodine(UI) level of the infants,pregnant and lactating women were determined by arsenic cerium catalytic spectrophotometry.DQ value of infants was measured before and after supplementation of iodized oil in 2006 and 2010.UI value of pregnant,lactating women and infants was monitored every year after iodine supplementation.Results Before iodine supplementation(2006),the median UI level of pregnant,lactating women and infants was 89.28,84.85,107.3 μg/L,respectively.After iodine supplementation,the medians UI level in 2007,2008,2009 and 2010 were,respectively,pregnant women:136.0,187.8,118.2,175.8 μg/L; lactating women:135.2,159.8,187.5,163.5 μ g/L; infants:139.6,174.7,190.7,168.4 μg/L.Before iodine supplementation,the DQ value of infants was 92.8 ± 16.3,and the average score of gross motor,fine motor,adaptive capacity,language and social behavior was 93.7 ± 20.0,91.4 ± 20.0,92.4 ± 19.0,90.3 ± 20.0,96.4 ± 22.1,respectively.After iodine supplementation,the DQ value of infants was 104.3 ± 13.8,and the average score of gross motor,fine motor,adaptive capacity,language and social behavior was 104.8 ± 21.5,104.1 ± 17.2,104.8 ± 16.1,99.9 ± 19.1,108.0 ± 22.7,respectively,which were higher than that before iodine supplementation (t =-10.43,-10.77,-13.78,-14.28,-9.96,-15.33,all P < 0.01).Conclusions Iodine deficiency at the critical period of brain development can affect the intellectual development of infants and young children at all functional areas of intelligence.Iodine supplementation at the critical period of brain development can prevent mental retardation caused by iodine deficiency disorders.
6.Management of Fournier's gangrene: a report of 23 cases.
Fei-Bo QI ; Mhetar WUBULI ; Ying-Gang WANG ; Maimaitijiang DAWUTI
National Journal of Andrology 2010;16(12):1098-1100
OBJECTIVETo study the clinical characteristics and treatment of Fournier's gangrene.
METHODSWe retrospectively analyzed the clinical data of 23 cases of Fournier's gangrene, all with flare and pains in the scrotum and penis and different degrees of involvement of the scrotum, penis, perianal area, perineum, inguinal and lower extremities. The patients were treated by early debridement, incision-drainage, anti-infection and hyperbaric oxygen therapy, respectively. Scrotoplasty was performed for 11 of the cases, penile and scrotal dermatoplasty for 7, and penile amputation and urethral fistulation for 2 with penile necrosis. One of the cases underwent suprapubic cystostomy, and another 1 received colostomy.
RESULTSTwenty of the patients were recovered and 3 (1 with diabetes and 1 with AIDS) died after surgery.
CONCLUSIONFournier's gangrene is a fatal disease. Early diagnosis and timely surgical intervention are essential for the management of the disease.
Adult ; Aged ; Fournier Gangrene ; diagnosis ; surgery ; therapy ; Humans ; Male ; Middle Aged ; Penis ; pathology ; surgery ; Retrospective Studies ; Scrotum ; pathology ; surgery
7.Evaluation of kidney oxygen bioavailability in acute renal failure by blood oxygen level dependent magnetic resonance imaging.
Wen-bo XIAO ; Qi-dong WANG ; Jing-jing XU ; Fei HAN ; Min-ming ZHANG
Journal of Zhejiang University. Medical sciences 2010;39(2):157-162
OBJECTIVETo assess the kidney oxygen bioavailability in acute renal failure using blood oxygen level dependent (BOLD) magnetic resonance (MR) imaging.
METHODSTwenty-one patients with acute renal failure, including 18 patients with oliguric renal failure, 1 nonoliguric acute renal failure and 2 functional renal failure were enrolled in the study; 20 healthy subjects served as controls. All subjects received renal functional MR examination. BOLD MR imaging with 16 gradient-recalled-echoes on a 1.5-T scanner were performed. R2(*)(1/sec) values of the cortex and medulla and R2(*) ratio of the medulla to cortex (R2(*) ratio of M/C) of the renal were recorded respectively.
RESULTSThe R2(*) values of the medulla was higher than those of the cortex in controls (17.64 +/-1.86/sec vs 13.73 +/-0.49/sec, P<0.00). The R2(*) ratio of M/C in controls was 1.28 +/-0.06. The R2(*) values of the medulla (13.31 +/-4.28/sec) and cortex (12.25 +/-2.41/sec) and the R2(* ) ratio of M/C (1.01 +/-0.25) in oliguric renal failure were lower than those in controls (P <0.05). Patients with functional renal failure and nonoliguric acute renal failure had higher R2(*) values in cortex and medulla and higher R2(*) ratio of M/C than those of controls.
CONCLUSIONBOLD MRI demonstrates that decreased R2(*) values of cortex and medulla suggest lower oxygen bioavailability in acute renal failure and decreased R2(*)ratio of M/C suggests the disappearance of a steep cortico-medullary gradient of oxygen.
Acute Kidney Injury ; diagnosis ; metabolism ; physiopathology ; Adult ; Biological Availability ; Female ; Humans ; Kidney ; metabolism ; physiopathology ; Kidney Function Tests ; methods ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Oxygen ; blood ; metabolism
8.Protein and hordein fraction content in barley seeds as affected by sowing date and their relations to malting quality.
Jun-cong QI ; Jin-xin CHEN ; Jun-mei WANG ; Fei-bo WU ; Lian-pu CAO ; Guo-ping ZHANG
Journal of Zhejiang University. Science. B 2005;6(11):1069-1075
The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relationships among these traits were examined. The results showed that grain protein content and B hordein content increased as the sowing date postponed and were significantly affected by sowing date, while C and D hordein contents were less influenced by sowing date. There were significant differences in grain protein and hordein fraction content among the ten cultivars. The coefficient of variation of D hordein content was much larger than that of B and C hordein contents, suggesting its greater variation caused by different sowing dates. Beta-amylase activity and diastatic power were also significantly affected by sowing date, with malt extract being less affected. Significant differences in measured malt quality were found among the ten cultivars. Grain protein was significantly correlated with B hordein and malt extract positively and negatively, respectively. There was no significant correlation between beta-amylase activity or diastatic power and grain protein content. B hordein was negatively and significantly correlated with malt extract, but no significant correlations between C hordein, D hordein and malting quality traits.
Edible Grain
;
chemistry
;
Food Analysis
;
Food Technology
;
Glutens
;
Hordeum
;
growth & development
;
metabolism
;
Plant Proteins
;
metabolism
;
Seeds
;
growth & development
;
metabolism
9.Regulation of proliferation and apoptosis of human vascular endothelial cell by Acheron.
Rong-ju SUN ; Qi-ying WANG ; Jian-bo ZHANG ; Ying-fei GUO ; Xiao-dong ZHAO
Chinese Journal of Burns 2011;27(2):156-160
OBJECTIVETo investigate regulatory effect of Acheron (Achn) on proliferation and apoptosis of human vascular endothelial cell.
METHODS(1) Eahy926 cells were cultured in serum-free DMEM medium (96-well plates) and were divided into Achn inhibition group (transfected with plasmid psi-Achn), psi4.1 group (transfected with psi4.1 empty vector), Achn induction group (transfected with pcDNA-Achn), pcDNA3.1 group (transfected with pcDNA3.1 empty vector), cotransfection group [cotransfected with pcDNA-Achn + psi-calcium/calmodulin-dependent serine protein kinase (CASK)], blank control group (treated with PBS) according to the random number table (the same method below). The cell proliferation was determined by MTT assay at post transfection hour (PTH) 1, 24, 48, 72, with expression of absorbance value. (2) Total protein of Eahy926 cells were extracted and quantitated by BCA assay, and then they were divided into Achn antibody precipitation group (100 µg protein), CASK antibody precipitation group (100 µg protein), IgG antibody group (100 µg protein), Western blot group (20 µg protein). Achn and CASK protein levels were determined by immunoprecipitation and Western blot. (3) Synchronously cultured Eahy926 cells were divided into LPS induction group (treated with 5 mol/L LPS), Achn transfection group (transfected with pcDNA-Achn), cotransfection group (cotransfected with psi-CASK and pcDNA-Achn), KCl group (treated with 5 mol/L KCl), and blank control group (treated with 5 mol/L PBS). Cells in transfection groups were stimulated by LPS for 12 hours after PTH 24. Caspase-3 protein level was detected by immunohistochemistry. (4) Synchronously cultured Eahy926 cells were divided into Achn inhibition group (transfected with psi-Achn vector), Achn induction group (transfected with pcDNA-Achn vector), and blank control group (treated with PBS). Apoptosis rate was determined by FITC/PI with flow cytometry. Data were processed with one-way analysis of variance and t test.
RESULTS(1) The cell proliferation in Achn inhibition group was lower than that in psi4.1 group from PTH 24, and the differences were statistically significant at PTH 48, 72 (with t value respectively 10.777, 6.112, P values all below 0.05). The cell proliferation in Achn induction group during PTH 24-72 were higher that in pcDNA3.1 group (with t value respectively 5.367, 6.053, 9.831, P values all below 0.05). The cell proliferation in cotransfection group at PTH 48, 72 were significantly lower than that in Achn induction group (with t value respectively 5.481, 9.517, P values all below 0.05). (2) Achn protein was detected in CASK antibody precipitation group while CASK protein was also detected in Achn antibody precipitation group. (3) Caspase-3 level in Achn transfection group was lower [(15.6 ± 0.5)%] as compared with that in LPS induction group [(32.8 ± 2.6)%, t = 10.083, P < 0.05], and that in cotransfection group showed further inhibition [(7.0 ± 2.0)%, t = 9.827, P < 0.01]. (4) Apoptosis rate in Achn inhibition group [(45.6 ± 10.9)%] was higher than that in blank control group [(13.2 ± 4.3) %, t = 7.043, P < 0.05]; while that in Achn induction group [(5.3 ± 2.9)%] was lower than that in blank control group (t = 6.499, P < 0.05).
CONCLUSIONSAchn can promote human vascular endothelial cell proliferation, and inhibit its apoptosis induced by LPS or burn serum, and the effect is related to CASK.
Apoptosis ; Autoantigens ; genetics ; metabolism ; Cell Line ; Cell Proliferation ; Endothelial Cells ; cytology ; Guanylate Kinases ; metabolism ; Humans ; Ribonucleoproteins ; genetics ; metabolism ; Transfection
10.Cloning, Sequence Analysis and Expression of dhaT Gene from Citrobacter freundii and Purification and Property of Corresponding Recombinant Enzyme
Xiang-Hui QI ; Zhao-Fei LUO ; Yu-Tuo WEI ; Fa-Zhong CHEN ; Shan-Shan WANG ; Shou-Hai HOU ; Dong-Qing LIAO ; Ri-Bo HUANG ;
China Biotechnology 2006;0(07):-
1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.