1. Differential proteomic analysis of transgenic Bt rice and its non-transgenic parental rice
Journal of International Pharmaceutical Research 2016;43(4):768-773
Objective: Under the methodology of differential proteomics and bioinformatics, the impact of the exogenous gene on the expression of rice in the proteome is discussed, aiming to explore into the study of genetically modified rice in the proteomics. Methods: The total protein was extracted from genetically modified rice Huahui No.1(HH1) and non-transgenic rice Minghui 63(MH63), the method of two-dimensional gel electrophoresis was applied to generate corresponding proteome two-dimensional polyacrylamide gel(2D-PAGE) electrophoresis spectrum; then, the mass spectrometry and bioinformatics analysis were conducted after the selection of protein spots with significant differences. Results: The comparing and matching of protein spots between transgenic Bt(cry1Ab/1Ac) rice and non-transgenic rice 2D-PAGE profiles identified 28 protein spots with significant differences. With non-transgenic rice as a reference, transgenic Bt rice held 18 relatively high and 10 relatively low expressions; mass spectrometry and bioinformatics retrieval were made on the different protein spots. It was found that the differentiated protein was mainly involved in energy metabolism, protein synthesis, redox stress response and other biological processes. Conclusion: Differences exist between transgenic Bt HH1 and its parental rice MH63 on the expression of proteome; however, there are neither anti-nutritional and allergenic protein, nor new or toxic proteins among these differentiated proteins.
2.Expression of p57kip2 in patients with de novo myelodysplastic syndrome and its relationship with SDF-1/CXCR4 axis.
You-Shan ZHAO ; Juan GUO ; Rui YANG ; Shu-Cheng GU ; Xi ZHANG ; Li-Yu ZHOU ; Xian LI ; Chun-Kang CHANG
Journal of Experimental Hematology 2012;20(2):352-357
This study was purposed to explore the expression of p57kip2 in the bone marrow of patients with de novo myelodysplastic syndrome (MDS) and its role in MDS pathogenesis, as well as the relationship between the expression of p57kip2 and SDF-1/CXCR4 signal. The expression of p57kip2 and CXCR4 in 67 de novo MDS patients was measured by real-time quantitative PCR. The percentage of CD34(+) cells in the bone marrow from MDS patients was measured by flow cytometry. 18 healthy volunteers were recruited for control. The effect of SDF-1 on p57kip2 expression in bone marrow mononuclear cell (BMMNC) from MDS or normal controls was investigated in vitro, and difference between them was compared. The results showed that low-risk MDS and high-risk MDS displayed a significant reduction of p57kip2 mRNA expression in BMMNC compared with that in control group (P < 0.001) and there was a negative correlation between p57kip2 expression and percentage of CD34(+) (r = -0.458, P < 0.001); the patients with abnormal karyotype showed lower expression of p57kip2 gene, compared to patients with normal karyotype (P = 0.045). Although the expression of CXCR4 had no difference between MDS patients and normal controls, a positive correlation between p57kip2 and CXCR4 in MDS patients was still found (r = 0.609, P < 0.001). Moreover, SDF-1 increased p57kip2 expression in normal BMMNC in dose-dependent manner, but BMMNC from MDS patients showed no response to SDF-1. SDF-1-induced p57 expression was blocked by AMD3100. It is concluded that the low expression of p57 gene in MDS may play a role in the pathogenesis of MDS. Furthermore, SDF-1-induced p57kip2 expression in BMMNC, and the decreasing response of BMMNC to SDF-1 may contribute to the low expression of p57kip2 in MDS patients.
Case-Control Studies
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Chemokine CXCL12
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metabolism
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Cyclin-Dependent Kinase Inhibitor p57
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genetics
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metabolism
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Flow Cytometry
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Humans
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Myelodysplastic Syndromes
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genetics
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metabolism
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Receptors, CXCR4
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metabolism
3.Relationship between DNA methylation and expressions of p57kip2 in hepatocellular carcinoma.
Ji-zhi ZHAO ; Zong-ji ZHANG ; Li-juan SHEN ; Ruo-chuan CHENG ; Hua-xian ZHANG ; Zhong-yi QIAN
Chinese Journal of Hepatology 2009;17(9):703-704
Biomarkers, Tumor
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metabolism
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Carcinoma, Hepatocellular
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genetics
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metabolism
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CpG Islands
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Cyclin-Dependent Kinase Inhibitor p57
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genetics
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metabolism
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DNA Methylation
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Gene Expression Regulation, Neoplastic
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Humans
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In Situ Hybridization
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Liver
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metabolism
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Liver Neoplasms
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genetics
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metabolism
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Polymerase Chain Reaction
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methods
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Promoter Regions, Genetic
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RNA, Messenger
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genetics
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metabolism
4.Expression and correlation of NKG2D and sMICA in lung cancer patients
Gang CHEN ; Chun-li WANG ; Shi-ping GUO ; Wen SU ; Li-juan QIAO ; Xian-xia MAI ; Jie MA ; Cheng-guang HU ; Pei-gang ZHANG
Cancer Research and Clinic 2009;21(11):759-761
Objective To investigate the expression and correlation of NKG2D and sMICA in lung cancer patients. Methods By collecting 30 lung cancer patients as the test group,and taking 30 healthy volunteers as the contrast group, the expression of NKG2D and sMICA in the two groups were examined separately by FACS and ELISA method. Results The expressions of NKG2D in the two groups were (81.56±8.78) %, (85.63±6.62) %. The lung cancer patients were high remarkable. There was a significant difference between the two groups (P <0.05). The expression of sMICA in the two groups were (354.13 ±80.575) pg/ml,(216.53±48.175) pg/ml. The lung cancer patients were low remarkable. There was a significant difference between the two groups (P <0.01). There was a significant relation between the two groups (r =-0.349, P =0.006). Conclusion The expression of NKG2D and sMICA may provid one of the immune targets for diagnosing that can forecast the immune state and malignant metastasis of the lung cancer patients. The significant relation between NKG2D and sMICA may take on main role in the immune escaping of tumor. It may provide the suitable target of the patients for tumor organisms and immune treatment.
5.Role of apoptosis of neutrophils and vascular endothelial cells and changes of relative factors in the pathogenesis of traumatic sepsis at the advanced stage
Yu-Miao WEI ; Yu-Hua LIAO ; Shu-Juan CHU ; Quan-Jun ZHOU ; Guo-Rong WANG ; Chao-Hui WANG ; Long-Xian CHENG
Chinese Journal of Trauma 2003;0(09):-
Objective To explore the role of apoptosis of neutrophils and vascular endothelial cells and changes of relative cytokines and thrombotic factors in the pathogenesis of traumatic sepsis at the advanced stage.Methods The venous blood was collected from the patients with traumatic sepsis at advanced stage and traumatic patients without sepsis and healthy subjects.The peripheral hlood mononu- clear cells(PBMC)and neutrophils were isolated and cultured.The apoptosis of neutrophils and vascular endothelial cells was assayed,and the level of IL-4,IL-10 in PBMC culture supernatants were deter- mined,and the tissue factor(TF)and vW factor(vWF)of peripheral plasma were measured.Results The percentages of apoptosis of neutrophils and numbers of apoptotic circulating vascular endothelial cells were higher than that of traumatic patients without sepsis and healthy controls significantly.And the levels of anti-inflammatory cytokines IL-4,IL-10 and thrombotic factors TF,vWF in traumatic sepsis were elevated than that of traumatic patients without sepsis and controls too.Conclusion Immune suppres- sion and abnormal thrombotic state may be one characteristic of traumatic sepsis at advanced stage,which perhaps involveds in the pathogenesis of traumatic sepsis at the advanced stage and multiple organ dys- function syndrome.
6.Function of TTG1A in hepatic stellate cells.
Lin XIAO ; Jun CHENG ; Li-ying ZHANG ; Jiang GUO ; Yuan HONG ; Li-juan ZHANG ; Yong-zhi LUN ; Xian-yong LAN ; Hui-juan WU ; Yue-xin ZHANG ; Jian-long ZHANG ; Yan LI
Chinese Journal of Hepatology 2009;17(12):925-929
OBJECTIVETo investigate the biological functions of TTG1A in liver fibrosis.
METHODSYeast two-hybrid system was used to screen proteins associated with TTG1A. Briefly, the coding sequence of TTG1A was cloned into pGBKT7 vector, and the recombinant plasmid was transformed into yeast cells AH109 ( a type), then these cells were mated with yeast cells Y187 (a type) transformed with human leukocyte cDNA library plasmid pACT2. The obtained diploid yeast cells were plated on synthetic dropout nutrient medium containing X-alpha-gal for double selection. The plasmids from positive colonies were transformed into E.coli and sequenced.
RESULTSThe recombinant yeast expression vector pGBKT7-TTG1A was successfully constructed. Nineteen TTG1A binding proteins, including Homo sapiens major histocompatibility complex, class II DP beta 1 (HLA-DPb1), Homo sapiens ribosomal protein L30 (RPL30), Homo sapiens nucleophosmin Homo sapiens nucleobindin 2 (NUCB2), Homo sapiens ash2, variant Gaucher disease and variant metachromatic leukodystrophy, MORF4L1, Homo sapiens ubiquitin-conjugating enzyme E2L3 (UBE2L3), APOA1, Homo sapiens lectin, and galectin 1, were identified.
CONCLUSIONSThis study may help to elucidate the molecular function of TTG1A.
Carrier Proteins ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Library ; Genes, Regulator ; Genetic Vectors ; Hepatic Stellate Cells ; Humans ; Liver Cirrhosis ; genetics ; Oligonucleotide Array Sequence Analysis ; Plasmids ; genetics ; Ribosomal Proteins ; genetics ; Transcriptional Activation ; Transforming Growth Factor beta1 ; genetics ; Two-Hybrid System Techniques ; Yeasts ; genetics
7.Influence of educational status, burn area and coping behaviors on the complication of psychological disorders in severely burned patients.
Hua CHENG ; Xiao-jian LI ; Wen-juan CAO ; Li-ying CHEN ; Zhi ZHANG ; Zhi-he LIU ; Xian-feng YI ; Wen LAI
Chinese Journal of Burns 2013;29(2):195-200
OBJECTIVETo discuss how the educational status, burn area and coping behaviors influence the psychological disorders in severely burned patients.
METHODSSixty-four severely burned patients hospitalized in Guangzhou Red Cross Hospital, Guangdong Provincial Work Injury Rehabilitation Center, and Guangdong General Hospital were enrolled with cluster random sampling method. Data of their demography and situation of burns were collected. Then their coping behavior, psychological disorders including anxiety, depression and post-traumatic stress disorder (PTSD) plus its core symptoms of flashback, avoidance, and hypervigilance were assessed by medical coping modes questionnaire, self-rating anxiety scale (SAS), self-rating depression scale (SDS), PTSD checklist-civilian version (PCL-C) respectively. Correlation was analyzed between demography, burn area, coping behavior and psychological disorders. The predictive powers of educational status, burn area and coping behaviors on the psychological disorders were analyzed. The qualitative variables were assigned values. Data were processed with t test, Spearman rank correlation analysis, and multiple linear regression analysis.
RESULTS(1) The patients scored (19.0 ± 3.4) points in confrontation coping behavior, which showed no statistically significant difference from the domestic norm score (19.5 ± 3.8) points (t = -1.13, P > 0.05). The patients scored (16.6 ± 2.4) and (11.0 ± 2.2) points in avoidance and resignation coping behaviors, which were significantly higher than the domestic norm score (14.4 ± 3.0), (8.8 ± 3.2) points (with t values respectively 7.06 and 7.76, P values both below 0.01). The patients' standard score of SAS, SDS, PCL-C were (50 ± 11), (54 ± 11), and (38 ± 12) points. Respectively 89.1% (57/64), 60.9% (39/64), 46.9% (30/64) of the patients showed anxiety, depression, and PTSD symptoms. (2) Four independent variables: age, gender, marital status, and time after burns, were correlated with the psychological disorders, but the correlativity was not statistically significant (with rs values from -0.089 to 0.245, P values all above 0.05). Educational status was significantly negatively correlated with anxiety, depression, PTSD and its core symptoms of flashback, avoidance (with rs values from -0.361 to -0.253, P values all below 0.05). Educational status was negatively correlated with hypervigilance, but the correlativity was not statistically significant (rs = -0.187, P > 0.05). Burn area was significantly positively correlated with the psychological disorders (with rs values from 0.306 to 0.478, P values all below 0.05). Confrontation coping behavior was positively correlated with the psychological disorders, but the correlativity was not statistically significant (with rs values from 0.121 to 0.550, P values all above 0.05). Avoidance coping behavior was correlated with the psychological disorders, but the correlativity was not statistically significant (with rs values from -0.144 to 0.193, P values all above 0.05). Resignation coping behavior was significantly positively correlated with the psychological disorder (with rs values from 0.377 to 0.596, P values all below 0.01). (3) Educational status had predictive power on the anxiety, PTSD and flash back symptoms of patients (with t values from -2.19 to -2.02, P values all below 0.05), but not on depression, avoidance and hypervigilance (with t values from -1.95 to -0.99, P values all above 0.05). Burn area had no predictive power on the psychological disorders (with t values from 0.55 to 1.78, P values all above 0.05). Resignation coping behavior had predictive power on the psychological disorders (with t values from 3.10 to 6.46, P values below 0.01). Confrontation and avoidance coping behaviors had no predictive power on the psychological disorders (with t values from 0.46 to 2.32 and -0.89 and 1.75 respectively, P values all above 0.05).
CONCLUSIONSThe severely burned patients with lower educational status, larger burn area, and the more frequently adapted resignation coping behavior are more likely to suffer from anxiety, depression, and PTSD.
Adaptation, Psychological ; Adolescent ; Adult ; Aged ; Burns ; complications ; pathology ; psychology ; Educational Status ; Female ; Humans ; Male ; Mental Disorders ; etiology ; Middle Aged ; Young Adult
8.The nutrition status of children aged 6-23 months after three months of Wenchuan Earthquake in Beichuan and Lixian, Sichuan Province.
Li-juan WANG ; Jun-sheng HUO ; Jing SUN ; Wen-xian LI ; Cheng-yu HUANG ; Shao-lin LAI ; Ji-fang HU ; Chun-ming CHEN ; Yu-ying WANG
Chinese Journal of Preventive Medicine 2010;44(8):696-700
OBJECTIVEThis project aims to investigate the nutrition status of children aged from 6 to 23 months living in Beichuan and Lixian where were seriously affected by the earthquake in 2008.
METHODSThe height, body weight and hemoglobin concentration were measured for 478 children from Leigu township, Qushan township in Beichuan county and Zagunao township, Xuecheng township and Putou township in Lixian county; the number of group 6 - 11, 12 - 17, 18 - 23 months were 190, 144, 144 respectively. The height for age (HAZ), the body weight for age (WAZ) and the weight for height (WHZ) were calculated respectively. The low weight, stunting, wasting prevalence, anemic rate were analyzed.
RESULTSThe WAZ scores of 6 - 23 months males and females in Beichuan county and Lixian county ranged from -0.97 to -0.13 and from -0.67 to -0.23, HAZ scores ranged from -1.23 to -0.31 and from -1.25 to -0.38, respectively. The WAZ (-0.89 ± 1.16) and HAZ (-1.20 ± 1.60) of 18 - 23 months infants in Beichuan county were significantly different from the WAZ and HAZ of 6 - 11 months (WAZ: -0.32 ± 0.92; HAZ: -0.58 ± 0.98) and 12 - 17 months (WAZ: -0.47 ± 1.00; HAZ: -0.68 ± 1.34) infants (F values were 7.161 and 4.584, respectively; all P values < 0.05). The WAZ (-0.63 ± 1.03) and HAZ (-1.11 ± 1.15) of 18 - 23 months in Lixian county infants were significantly different from the WAZ and HAZ of 6 - 11 months (WAZ: -0.23 ± 0.93; HAZ: -0.51 ± 1.55) infants and 12 - 17 months (WAZ: -0.58 ± 0.52; HAZ: -0.80 ± 1.19) (F values were 3.156, 4.345, all P values < 0.05). The prevalence of low weight infants (18 - 23 months) these two counties were 15.6% (12/77) and 9.1% (6/66), respectively. The prevalence of stunting in Beichuan and Lixian have reached 26% (20/77) and 24.2% (16/66), respectively. The wasting rate of 18 - 23 months infants had reached 9.1% (7/77) in Beichuan. The total anemia prevalence among infants reached 49.6% (123/248) and 78.8% (178/226) in Beichuan and Lixian, respectively. The prevalence of moderate anemia have reached 7.7% (19/248) and 19.9% (45/226), respectively.
CONCLUSIONAfter the earthquake in Beichuan, Lixian, the growth of infants aged 6 to 23 months old was below average. The prevalence of anemia was high. The malnutrition status of infant and young children aged from 18 to 23 months in Beichuan and Lixian was more serious, should be targeted of nutrition interventions.
China ; epidemiology ; Disasters ; Earthquakes ; Female ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Male ; Malnutrition ; epidemiology ; Nutritional Status
9.Safety and Efficacy of Etanercept Monotherapy for Moderate-to-severe Plaque Psoriasis: A Prospective 12-week Follow-up Study
XIE FANG ; WANG RUI ; ZHAO ZI-GANG ; MENG XIAN-FU ; LIN BI-WEN ; YANG JIE ; WANG WEN-JUAN ; DING XIANG-YU ; YANG YI ; ZHAO HUA ; LI CHENG-XIN ; LI HENG-JIN ; ZHOU YONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(6):943-947
Etanercept has been shown to be effective for the treatment of moderate-to-severe plaque psoriasis.Since most clinical trials examined etanercept in combination with other drugs,the efficacy and safety of etanercept monotherapy for moderate-to-severe plaque psoriasis have not been well established.This prospective study enrolled 61 Chinese patients with moderate-to-severe plaque psoriasis to explore the efficacy and safety of etanercept monotherapy.These patients were treated with etanercept at a subcutaneous dose of 25 mg,twice a week,for 12 weeks.All the 61 patients completed the treatment and showed significant improvement in psoriasis area and severity index (PASI) scores.At 4,8,and 12 weeks after treatment,the response rates (PASI75) were 0%,21.31%,and 40.98%,respectively.It was concluded that etanercept monotherapy is efficacious and safe for patients with moderate-to-severe plaque psoriasis.
10.Screening and cloning genes transactivated by TGF beta 1 in hepatic stellate cells using suppression subtractive hybridization technique.
Lin XIAO ; Jun CHENG ; Jiang GUO ; Li-ying ZHANG ; Yuan HONG ; Yong-zhi LUN ; Xian-yong LAN ; Hui-juan WU ; Li-juan ZHANG ; Yue-xin ZHANG ; Jian-long ZHANG ; Yan LI
Chinese Journal of Hepatology 2008;16(11):854-857
OBJECTIVESTo construct a cDNA subtractive library of genes transactivated by TGF beta 1 in LX02 hepatic stellate cells (HSC); to screen and to clone the regulated genes transactivated by TGF beta 1; and to elucidate the molecular biological mechanism of hepatic fibrosis mediated by TGF beta 1.
METHODSmRNA was isolated from HSC treated with TGF beta 1 or with PBS (as controls). Suppression subtractive hybridization (SSH) technique was employed to analyze the differentially expressed DNA sequence between the two groups. After restriction enzyme Rsa I digestion, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent polymerase chain reaction twice it then was subcloned into pGEM-Teasy plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5a. The cDNA was sequenced and analyzed in GenBank with Blast search.
RESULTSThe subtractive cDNA library of genes transactivated by TGF beta 1 in HSC was constructed successfully. The amplified library contained 146 positive clones, which contained 200-1000 bp of inserts. Randomly, thirty clones were analyzed by sequencing and bioinformatics, consisting of 28 known genes and 2 unknown genes.
CONCLUSIONSThe subtractive cDNA library of genes transactivated by TGF beta 1 in HSC using SSH technique was constructed successfully. Some gene coding proteins are those involved in cell growth regulation, protein synthesis, signal transduction, extracellular matrix metabolism, and anti-lipid peroxidative, which gives us some new clues for the study of the mechanism of liver fibrosis.
Animals ; Cell Line ; Cloning, Molecular ; Gene Library ; Genetic Vectors ; Hepatic Stellate Cells ; metabolism ; Nucleic Acid Hybridization ; methods ; RNA, Messenger ; genetics ; Rats ; Sequence Homology ; Transforming Growth Factor beta1 ; genetics