1.Association of general level of total anti-oxidation capacity, superoxide dismutase, nitric oxide synthase activity in serum with the lifestyle of residents of Bin County of Heilongjiang Province
Chunmei GUAN ; Yang ZHANG ; Zhenjiang YAO ; Rong LI ; Ping YU
Chinese Journal of Tissue Engineering Research 2005;9(27):198-201
BACKGROUND: When the dynamic equilibrium between oxidation system and anti-oxidation system in the body is upset, the overly produced active oxygen-derived free radicals will attack the target organs, thus resulting in the damage to organs and big molecules, and then diseases. Total anti-oxidation capacity (T-AOC), superoxide dismutase (SOD), and nitric oxide synthase (NOS) activity are the majorindexes for the defense system of the body, and their status is closely related to diseases. OBJECTIVE:To explore the level of T-AOC, SOD and NOS in serum,and find out its relationship with lifestyle. DESIGN:Single sample investigation. SETTING:Department of Health Laboratory Technology, College of Public Health of Harbin Medical University. PARTICIPANTS:Totally 531 residents of Bin County aged 20 to 70years were recruited between September 2000 and May2001. MATERIALS:The reagents kits of T-AOC, SOD and NOS were manufactured by Nanjing Jiancheng Bio-engineering Institute. METHODS:The investigators surveyed the residents with the same questionnaire. Questionnaire was made and the investigators were trained in advance. The questionnaire was filled in item by item as required. Questionnaire items consisted of the general situation, financial status, dietary habit, hobby, status of health care, and mental factor. Finally 95 investigation indexes were completed. The activity of serum T-AOC, SOD and NOS of the 531 residents in Bin County was determined, respectively, with reagent kits produced by Nanjing Jiancheng Bio-engineering Institute. The lifestyle factors were analyzed by multivariate stepwise regression analysis,and informed consent was obtained from the participants. MAIN OUTCOME MEASURFS:① Serum T-AOC, SOD and NOS in the participants; ② lifestyle factors affecting T-AOC, SOD and NOS.RFSULTS:According to actual analysis, the 531 participants entered the results analysis. T-AOC was measured in 489 participants, SOD in 525participants, and NOS in 531 participants. ① Indexes of the participants:T-AOC of the males was obviously higher than that of the females [(20.01±7.21), (15.25±6.22) kU/L, P < 0.05]. SOD of the males was slightly lower than that of the females while NOS of the males was slightly higher than that of the females, but there was no significant difference. ②Multivariate stepwise regression analysis showed that risk factors related to T-AOC were gender difference (OR=2.188), educational level (OR=1.859),and the presence of rheumatism, respectively (OR =1.142). SOD-related risk factors were educational level (OR=1.584), years of spirit drinking (OR =1.048), presence of nephritis (OR=1.093), and irradiation (OR=1.770);frequency of tea drinking was a beneficial factor (OR=0.800). NOS-related risk factors were the average amount of cigarette smoking (OR=1.194) and the times of weekly spirit drinking (OR=1.368). However, the beneficial factor that affected serum NOS was the frequency of weekly mutton eating (OR=0.458) CONCLUSION:This experiment revealed that the ability to clean free radical in the 531 subjects was good and that it was better in males than in females. The increased educational level and presence of rheumatism can decrease T-AOC. Drinking, smoking and irradiation decrease SOD activity while the frequency of tea drinking and mutton eating are beneficial factors.
2.Effect of long non-coding RNA 114227 on gastric cancer cell proliferation and migration.
Haining GAN ; Huiying XIANG ; Yue XI ; Min YAO ; Chen SHAO ; Shihe SHAO
Journal of Central South University(Medical Sciences) 2023;48(2):157-164
OBJECTIVES:
Gastric cancer is a common cancer of the digestive system. Long non-coding RNA (lncRNA) plays an important role in the formation and development of gastric cancer. This study aims to investigate the effect of long non-coding lncRNA 114227 on biologic behaviors in gastric cancer cells.
METHODS:
The experiment was divided into 4 groups: a negative control (NC) group, a lncRNA 114227 small interference (si-lncRNA 114227) group, an empty vector (Vector) group, and an overexpression vector (OE-lncRNA 114227) group. The expressions of lncRNA 114227 in gastric mucosa and gastric cancer tissues, gastric mucosal epithelial cells and different gastric cancer strains were determined by real-time reverse transcription PCR (real-time RT-PCR).The proliferation were detected by CCK-8 assay in gastric cancer cells. The epithelial-mesenchymal transformation (EMT) was utilized by Transwell assay, scratch healing assay, and Western blotting in gastric cancer cells. The effect of lncRNA 114227 on proliferation of gastric cancer cells was detected by tumor bearing experiment in nude mice in vivo.
RESULTS:
The expression level of lncRNA 114227 in the gastric cancer tissues was significantly lower than that in the gastric mucosa tissues, and in 4 kinds of gastric cancer strains was all significantly lower than that in gastric mucosal epithelial cells (all P<0.01). In vitro, the proliferation and migration abilities of gastric cells were significantly reduced after overexpressing lncRNA 114227, and cell proliferation and migration were enhanced after silencing lncRNA 114227 (all P<0.05). The results of in vivo subcutaneous tumorigenesis in nude mice showed that the tumorigenic volume of the tumor-bearing mice in the OE-lncRNA 114227 group was significantly smaller than that of the Vector group, and the tumorigenic quality was lower than that of the Vector group (P<0.05), indicating that lncRNA 114227 inhibited tumorigenesis.
CONCLUSIONS
The expression of lncRNA 114227 is downregulated in gastric cancer gastric cancer tissues and cell lines. LncRNA 114227 may inhibit the proliferation and migration of gastric cancer cells through EMT process.
Animals
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Mice
;
RNA, Long Noncoding/metabolism*
;
Stomach Neoplasms/pathology*
;
Mice, Nude
;
Cell Line, Tumor
;
Cell Proliferation/genetics*
;
Carcinogenesis/genetics*
;
Cell Movement/genetics*
;
Epithelial-Mesenchymal Transition/genetics*
;
Gene Expression Regulation, Neoplastic
;
Apoptosis/genetics*
3.Preparation of thermoresponsive micelles loaded with indocyanine green and doxorubicin for combined therapy in MCF-7 cells
Ling ZONG ; An-ran YUAN ; Yao ZHU ; Yun-shuang GU ; Yan-ru GE ; Song SHEN
Acta Pharmaceutica Sinica 2018;53(7):1169-1176
In this study, the thermoresponsive micelles were synthesized with random copolymerization method and the photosensitizer indocyanine green (ICG) was loaded on micelles through the physical adsorption. The light energy was converted into heat energy to increase the temperature after irradiation with near-infrared light. When the phase transition temperature was reached, the micelle was disassembled and the targeted therapy was achieved. The nanoparticles were characterized with a transmission electron microscopy, Fourier transform infrared spectrometer, nuclear magnetic resonance spectrometer and other characterization were used to investigate. The critical micelle concentration (CMC), upper critical solution temperature, the photothermal properties of the carrier and the release of drug triggered by light were investigated after the doxorubicin (DOX) loaded. The carrier was evaluated for toxicity, cellular uptake, the effect of photothermal, the combination of photothermal and chemotherapy; the p(AAm-co-AN)-g-PEG (PAAP) was spherical in shape with a particle size of about 45 nm and a phase transition temperature was about 43℃. The critical micelle concentration was 24 μg·mL-1. The particle size increased to 88 nm after loaded with ICG and DOX which the photothermal effect was obvious. The cumulative release of the drug under the irradiation of near-infrared light (808 nm, 2 W·cm-2, 2 min·h-1) was increased to 59.4% (pH 5.0) after 5 h. The results of the cell experiment indicated that ICG-PAAP was almost non-toxic and uptaken by the lysosomal pathway. The cell killing effect was stronger with combination of chemotherapy (DOX as 20 μg·mL-1) with more than 70% of the cells killed. The results showed that the prepared micelle with low toxicity was thermoresponsive and could be used in combined therapy of tumor under the irradiation of near-infrared light.
4.Overexpression of LncRNA ITGB2-AS1 Predicts Adverse Prognosis in Acute Myeloid Leukemia.
Ying-Li ZHOU ; Zi-Jun XU ; Jing-Dong ZHOU ; Ting-Juan ZHANG ; Dong-Ming YAO ; Ji-Chun MA ; Jiang LIN ; Jun QIAN
Journal of Experimental Hematology 2021;29(5):1436-1449
OBJECTIVE:
LncRNA ITGB2-AS1 has been found to play important roles in the occurrence and development of human solid tumors. However, its role in hematological diseases, especially acute myeloid leukemia (AML), remains unclear. The aim of this study was to identify the expression pattern of ITGB2-AS1 in AML patients and to further explore its clinical significance.
METHODS:
ITGB2-AS1 expression was analyzed in public datasets (including TCGA and GSE63270) and further validated in a cohort of 109 AML patients by real-time quantitative PCR (RT-qPCR).
RESULTS:
The level of ITGB2-AS1 was up-regulated among two independent cohorts (TCGA, P<0.05; GSE63270, P<0.05), which was confirmed by the data from 109 AML patients enrolled in this study (P<0.05). Clinically, high ITGB2-AS1 expression was associated with older age (P=0.023) and lower complete remission (CR) rate (P=0.005). Multivariate analysis identified that high ITGB2-AS1 expression was an independent prognostic factor not only for CR rate (P=0.027) but also for overall survival (OS) time (P=0.011), and ITGB2-AS1 was positively correlated with ITGB2 expression in both TCGA (r=0.74, P<0.001) and clinical data detected in this study (r=0.881, P<0.001). High ITGB2 expression was also associated with older age (P=0.02) and lower CR rate (P=0.020). Moreover, high ITGB2 expression predicted worse OS (P=0.028).
CONCLUSION
ITGB2-AS1 is overexpressed in AML and predicts poor prognosis in AML patients.
Aged
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Humans
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Leukemia, Myeloid, Acute/genetics*
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Prognosis
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RNA, Long Noncoding/genetics*
5.Preparation and characterization of a gene-activated matrix mimicking extracellular matrix
Hai-jun SHEN ; Zhi-juan WU ; Dong-xia WANG ; Dong-fang HUO ; Yu BAI ; Zhi-han YANG ; Chun-fang YAO
Acta Pharmaceutica Sinica 2017;52(11):1748-1755
This study aims to establish a novel gene-activated matrix that mimics the structure and function of extracellular matrix (ECM-m-GAM). The structure, mechanical property and release profile were also characterized. Firstly, the liposome/DNA lipoplex (LPD) was modified with cell penetrating peptide TAT. The obtained TAT-LPD was then mixed with RGD grafting hyaluronic acid solution. After addition of the matrix metalloproteinase (MMPs) sensitive crosslinker (HS-MMP-SH), hyaluronic acid was crosslinked and TAT-LPD was encapsulated in the subsequently formed hydrogel. As a result, the cell adhesion factor RGD, MMPs sensitive substrate and the efficient gene transfer vector TAT-LPD were all integrated in the hyaluronic acid hydrogel, which was named as ECM-m-GAM. The release profile of DNA from ECM-m-GAM in different release medium was evaluated with PicoGreen kits. The results suggested that the mean diameter of the spherical TAT-LPD was (263.0 ±4.30) nm. TAT-LPD was successfully encapsulated in ECM-m-GAM, which had the typical porous network structure of hydrogels. The mechanical strength of GAM was enhanced with the increasing of hyaluronic acid content. When the content was 4%, the elastic modulus of GAM reached 1 600 Pa. The highly elastic GAM may be suitable for implantation and tissue regeneration. The DNA release showed significant MMPs sensitive property. Especially, the released DNA still existed in form of nanoparticles. Bone marrow mesenchymal stem cells (BMSCs) were successfully transfected with GAM and the green fluorescent protein was expressed. The results have laid a solid foundation for future study of the cell transfection and tissue regeneration.
6.Study on the synchronization of biventricular beats with the control mode of left ventricular assist device.
Fangqun WANG ; Yao ZHANG ; Wanqian HE ; Si CHEN ; Teng JING ; Zhihao ZHANG
Journal of Biomedical Engineering 2021;38(1):72-79
Right ventricular (RV) failure has become a deadly complication of left ventricular assist device (LVAD) implantation, for which desynchrony in bi-ventricular pulse resulting from a LVAD is among the important factor. This paper investigated how different control modes affect the synchronization of pulse between LV (left ventricular) and RV by numerical method. The numerical results showed that the systolic duration between LV and RV did not significantly differ at baseline (LVAD off and cannula clamped) (48.52%
Heart Failure/therapy*
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Heart-Assist Devices
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Humans
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Systole
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Ventricular Dysfunction, Right
;
Ventricular Function, Right
7.CD137 signaling promotes angiogenesis through regulating macrophage M1/M2 polarization.
Tian Xin GENG ; Bo LI ; Yao XU ; Zhong Qun WANG ; Chen SHAO ; Jin Chuan YAN
Chinese Journal of Cardiology 2020;48(6):500-506
Objective: To investigate whether CD137 signaling can promote angiogenesis via regulating macrophage M1/M2 polarization. Methods: (1) The primary peritoneal macrophages in mice induced by 3% thiglycollate broth were divided into three groups: control group, CD137 signaling activated group and CD137 signaling inhibited group. Various specific markers of M1 and M2 macrophages were detected to observe the phenotype change of macrophages, and the macrophages protein expression of CD137, CD86 and CD206 was detected by flow cytometry (FCM). The protein and mRNA expression of induced nitric oxide synthase (iNOS), arginase Ⅰ(Arg-1) was determined by Western blot and RT-PCR, respectively. The secretion levels of IL-12 and IL-10 in culture supernatant of macrophages were detected by ELISA. (2) Macrophages were co-cultured with the endothelial cells (bEnd.3), and macrophages were implanted in the upper chamber, endothelial cells were implanted in stromal glue of the lower chamber. The experiment was divided into three groups: the control group, CD137 signaling activated group and peroxisome proliferator-activated receptor-γ (PPAR-γ) inhibited group, and tube formation ability of endothelial cells in each group was determined. Results: (1) The purity of primary peritoneal macrophages in mice was (97.93±1.31)%. The expression of CD137 on the surface of macrophages was (97.40±2.70)%. (2) Compared with control group, the mRNA and protein expression levels of Arg-1 were significantly increased and the mRNA and protein expression of iNOS were significantly decreased in CD137 signaling activated group (all P<0.05). Compared with CD137 signaling activated group, the mRNA and protein expression of Arg-1 were significantly lower and the mRNA and protein expression levels of iNOS were significantly higher in CD137 signaling inhibited group (all P<0.05). FCM results showed that the average fluorescence intensity of CD206 was higher, while the average fluorescence intensity of CD86 was lower in CD137 signaling activated group than in control group (P<0.05, P<0.01, respectively); the expression of CD206 was significantly lower, while the expression of CD86 was higher, in the CD137 signaling inhibited group than in CD137 signaling activated group (P<0.05, P<0.01, respectively). ELISA results showed that the secretion of IL-10 was higher, and the secretion level of IL-12 was significantly lower in CD137 signaling activated group than in control group (both P<0.01); the secretion of IL-10 was significantly lower and the secretion of IL-12 was significantly higher in CD137 signaling inhibited group than in CD137 signaling activated group (both P<0.05). (3) Values of the formation of tube length and branch number were both longer in CD137 signaling activated group than control group (P<0.05). The formation of the tube length and branch number were less in PPAR-γ inhibited group than in CD137 signaling activated group (P<0.05). Conclusion: CD137 signaling can promote angiogenesis by regulating macrophage M1/M2 polarization.
Animals
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Coculture Techniques
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Endothelial Cells
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Macrophages
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Mice
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Neovascularization, Pathologic
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Signal Transduction
8.Association between CD137 and ischemia-reperfusion injury in patients with acute ST-segment elevation myocardial infarction.
Ying Jie PAN ; Rui CHEN ; Yao XU ; Hao XIA ; Chong XU ; Wei YUAN
Chinese Journal of Cardiology 2021;49(12):1198-1205
Objective: To explore the relationship between the levels of serum soluble CD137 (sCD137) and membrane-bound CD137 (mCD137) and the occurrence of ischemia reperfusion injury (IRI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods: This is a cross-sectional study. Consecutive patients with acute STEMI, who underwent emergency percutaneous coronary intervention (PCI) in the Department of Cardiology, Jiangsu University Affiliated Hospital from May 2019 to September 2020, were enrolled. According to the absence or presence of IRI, patients were divided into IRI group and non-IRI group. Clinical data of the two groups were collected and compared. sCD137 level was detected by enzyme linked immunosorbent assay. Ficoll density gradient centrifugation was used to separate peripheral blood mononuclear cells (PBMC) and RNA was extracted, mCD137 mRNA expression level was detected by PCR. Serum sCD137 levels and the mCD137 mRNA levels of PBMC before, after PCI and 24 hours after PCI were compared. The correlation between serum sCD137 level, PBMC mCD137 mRNA level and clinical indicators was observed. The univariate and multivariate logistic binary regression analyses were performed to evaluate the related risk factors of IRI. ROC curve was used to analyze the predictive value of defined parameters for IRI. Results: A total of 112 STEMI patients were enrolled. There were 42 cases (of which 33 were males (78.6%), mean age was (58.6±12.7) years) in non-IRI group and 70 cases(of which 56 were males (80.0%), mean age was (64.5±11.6) years) in IRI group. Compared with the non-IRI group, patients in the IRI group had longer hospital stays, older age, lower rates of obesity, lower systolic and diastolic blood pressure at admission, higher proportion of the the right coronary artery as culprit vessel, lower rate of the use of angiotensin-converting enzyme inhibitor/angiotensin-Ⅱ receptor blocker/angiotensin receptor neprilysin inhibitor, higher levels of urea nitrogen and creatinine, lower glomerular filtration rate, lower triglycerides, higher D-dimer and B-type natriuretic peptidemax, higher proportion of Killip grade Ⅳ and cardiovascular adverse events (all P<0.05). sCD137 levels at the preoperative, postoperative and 24 hours after surgery were significantly higher in the IRI group than in the non-IRI group, while the mRNA levels of CD137 was similar between the two groups. The level of sCD137 in patients after PCI was lower than that before operation, the level of mCD137 mRNA was higher than that before operation (P<0.05). Serum sCD137 levels were positively correlated with hospitalization days, age, B-type natriuretic peptide, creatinine, ischemic time, C Reactive protein (CRP) and CRP/albumin (P<0.05), and negatively correlated with body mass index, glomerular filtration rate and albumin (P<0.05). The mCD137 mRNA expression level of PBMC was positively correlated with hospital stay, age, B-type natriuretic peptide, ischemic time, CRP and CRP/albumin (P<0.05), and negatively correlated with body mass index, glomerular filtration rate, albumin (P<0.05). Multivariate logistic regression analysis showed that higher sCD137 (OR=1.038, 95%CI: 1.009-1.069), aspartate aminotransferase, (OR=1.029, 95%CI: 1.009-1.050) and lower albumin (OR=0.829, 95%CI: 0.703-0.829) before surgery were independent risk factors of IRI (P<0.05). Receiver operating characteristic curve showed that the area under the curve of sCD137 was 0.672 (95%CI: 0.574-0.770, P=0.002) for the prediction of IRI, the best cut-off value was 28.43×10-3 μg/L with sensitivity of 95.2% and specificity of 48.6%. Conclusion: The significantly increased level of sCD137 in acute STEMI patients is positively correlated with reperfusion injury, which is an independent risk factor of IRI and may be related to the prognosis of patients with IRI.
Aged
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Cross-Sectional Studies
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Humans
;
Leukocytes, Mononuclear
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Male
;
Middle Aged
;
Percutaneous Coronary Intervention
;
Reperfusion Injury
;
ST Elevation Myocardial Infarction
9.Organization and transcription strategy of genome of Bombyx mori bidensovirus (China isolate) VD1.
Yong-Jie WANG ; Qin YAO ; Ke-Ping CHEN ; Xu HAN
Chinese Journal of Biotechnology 2006;22(5):707-712
To better understand the genomic structure and function of Bombyx mori bidensovirus (China isolate) VD1, the VD1 was purified and cloned into the pUC119 vector, and the complete nucleotide sequence of VD1 was determined. Sequence analysis showed that VD1 genome consisted of 6543 nts including inverted terminal repeats (ITRs) of 224 nts. In the viral genome, three major open reading frames (ORF1, ORF2 and ORF3) in the plus strand and one major ORF (ORF4) in the complementary strand were identified. Comparison of the complete genome sequence between Bombyx mori bidensovirus (China isolate) and BmDNV-2 (Yamanashi isolate) showed an identity of 98.4% in VD1, with a total number of 104 bp substitutions and 1 bp insertions found in Bombyx mori bidensovirus (China isolate), the highly variable regions were mainly located in VD1 ORF3 and VD1 ORF4. Northern blotting revealed that VD1 contained 1.1 kb and 1.5 kb transcript in the left-half 'plus' strand, and one transcript about 3.3 kb of 'minus' strand in the right-half. Sequencing of 3' and 5' ends of transcript products showed the 1.1 kb transcript started at nt 290 and ended at nt 1437, the 1.5 kb transcript was found to start nt 1423 and ended at 2931, and the 3.3 kb transcript was found to start nt 6287 and ended at nt 2922. Therefore, the 1.5 kb transcript in the left-half plus' strand and 3.3 kb transcripts of minus' strand in the right-half overlapped for 10 nts at the 3' ends. These results indicate that this virus employs a transcription strategy that is radically different from that of the other reported DNVs.
Amino Acid Sequence
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Animals
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Base Sequence
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Bombyx
;
virology
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Densovirus
;
genetics
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Genome, Viral
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Molecular Sequence Data
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Transcription, Genetic
10.Differential expression of haemolymph proteome of resistant strain and susceptible strain for BmNPV in Bombyx mori L.
Keya CAI ; Keping CHEN ; Xiaoyong LIU ; Qin YAO ; Jun LI
Chinese Journal of Biotechnology 2008;24(2):285-290
Three model silkworms, highly resistant strain, highly susceptible strain and their near isogenic line were established by hybridization and backcross. The resistance of silkworm (Bombyx mori L.) to BmNPV was studied at proteomic level using two-dimensional gel electrophoresis and MALDI TOF/TOF MS. Differential expression profiles of proteome in resistant strain, susceptible strain and near isogenic line were obtained, and 180, 190 and 187 of protein spots were shown, respectively. Among them, 80% were concentrated in pI 5-9. Twelve differential protein spots in total were obtained from 3 gels. Using MALDI TOF/TOF MS, five proteins, including aminoacylase, were identified from these spots. The exclusive expression of aminoacylase in highly resistant strain and near isogenic line and its absence in susceptible strain suggest that it might be a BmNPV-resistance related protein.
Amidohydrolases
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analysis
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genetics
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Animals
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Bombyx
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chemistry
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genetics
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virology
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Electrophoresis, Gel, Two-Dimensional
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Hemolymph
;
chemistry
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Host-Pathogen Interactions
;
genetics
;
Insect Proteins
;
analysis
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Nucleopolyhedrovirus
;
pathogenicity
;
physiology
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Proteome
;
analysis
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Proteomics
;
methods
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Recombination, Genetic
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization