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Credible and Facile Fluorometric Detection of Soybean Trypsin Inhibitor Activity with a Water-Soluble Poly(diphenylacetylene) Derivative
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作者 Jiao Tang Xiao Tang +7 位作者 Jiaxiang Tang Wei Qi Qianwei Pan Jinhong Zeng Housheng Xia Jianping Zhou Zhongyi Sheng Junfeng Niu 《Transactions of Tianjin University》 EI CAS 2024年第4期305-313,共9页
Herein,a novel method for fl uorometric detection of soybean trypsin inhibitor(SBTI)activity based on a water-soluble poly(diphenylacetylene)derivative was reported.Fluorescence quenching of the polymer via p-nitroani... Herein,a novel method for fl uorometric detection of soybean trypsin inhibitor(SBTI)activity based on a water-soluble poly(diphenylacetylene)derivative was reported.Fluorescence quenching of the polymer via p-nitroaniline,produced from the trypsin-catalyzed decomposition of N-benzoyl-DL-arginine-4-nitroanilide hydrochloride(L-BAPA),was well described using the Stern-Volmer equation.SBTI activity was quantitatively assessed based on changes in the fl uorescence intensity of the polymer.This strategy has several advantages,such as high sensitivity and ease of operation.Moreover,its applicability to other biochemical analyses is promising. 展开更多
关键词 Fluorometric detection Soybean trypsin inhibitor Inhibition activity Fluorescence quenching
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A Proteinase from Mung Bean Sprouts That Inactivaties Soybean Trypsin Inhibitor 被引量:2
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作者 陈中 杨晓泉 赵谋明 《Acta Botanica Sinica》 CSCD 2001年第11期1150-1153,共4页
By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE... By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE 15HR), a proteinase which can inactivate soybean trypsin inhibitor (STI) was purified from mung bean ( Vigna rabiata (L.) Wilezek) sprouts. Its molecular weight was estimated to be 29.8 kD by SDS-PAGE, and its K-m and V-max for STI were 769.2 N-alpha -benzoyl-L-arginine ethyl ester BAEE/mL and 115.3 BAEE . mL(-1) . min(-1) respectively. This proteinase was stable at temperatures lower than 50 degreesC and pH 6.5 - 8.5, and 90.91% STI activity of defatted soybean powder was inactivated by this preparation, with proteolytic activity 5 000 BAEE/mL at 50 degreesC and pH 8.0 in 4 h. 展开更多
关键词 PROTEINASE inactivate soybean trypsin inhibitor
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血清LncRNA FAF、ITIH4在慢性心力衰竭患者中的表达意义及对预后的预测价值
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作者 江洋 刘泽岩 +3 位作者 叶丽 汪栋林 程景林 万俊 《疑难病杂志》 CAS 2024年第4期418-422,428,共6页
目的探讨血清长链非编码RNA(LncRNA)FAF、间α胰蛋白酶抑制因子重链4(ITIH4)在慢性心力衰竭(CHF)患者中的表达意义及对预后的预测价值。方法选择2020年1月—2022年1月安徽医科大学第二附属医院急诊内科收治的CHF患者187例为CHF组,再根据... 目的探讨血清长链非编码RNA(LncRNA)FAF、间α胰蛋白酶抑制因子重链4(ITIH4)在慢性心力衰竭(CHF)患者中的表达意义及对预后的预测价值。方法选择2020年1月—2022年1月安徽医科大学第二附属医院急诊内科收治的CHF患者187例为CHF组,再根据NYHA心功能分级分为Ⅱ级亚组65例,Ⅲ级亚组77例,Ⅳ级亚组45例。于同期招募健康志愿者103例为健康对照组。2组受试者均检测血清LncRNA FAF表达和ITIH4水平,CHF患者出院后随访12个月,统计随访期间主要不良心血管事件(MACE)发生率;多因素Logistic回归分析影响CHF患者发生MACE的因素;受试者工作特征(ROC)曲线分析LncRNA FAF、ITIH4预测CHF患者发生MACE的价值。结果CHF组血清LncRNA FAF表达、ITIH4水平低于健康对照组(t/P=24.469/<0.001、35.196/<0.001)。血清LncRNA FAF表达、ITIH4水平Ⅳ级亚组低于Ⅲ级亚组低于Ⅱ级亚组(F/P=91.653/<0.001、102.345/<0.001)。MACE亚组血清LncRNA FAF表达,ITIH4水平低于非MACE亚组(t/P=13.556/<0.001、6.293/<0.001)。NYHAⅣ级、高水平NT-proBNP是CHF患者发生MACE的危险因素[OR(95%CI)=4.627(2.245~9.538)、2.284(1.505~3.468)],高表达LncRNA FAF、高水平ITIH4是保护因素[OR(95%CI)=0.599(0.425~0.844)、0.666(0.478~0.928)]。LncRNA FAF、ITIH4及二者联合预测CHF患者发生MACE的曲线下面积为0.796、0.801、0.896,二者联合预测曲线下面积高于单独预测(Z=2.453、2.404,均P<0.001)。结论CHF患者血清LncRNA FAF表达和ITIH4水平均降低,且与不良预后有关,联合LncRNA FAF和ITIH4可预测CHF患者预后不良风险。 展开更多
关键词 慢性心力衰竭 成纤维细胞生长因子相关因子 间α胰蛋白酶抑制因子重链4 预后
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Optimization of Solid-State Fermentation with Lactobacillus brevis and Aspergillus oryzae for Trypsin Inhibitor Degradation in Soybean Meal 被引量:19
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作者 GAO You-ling WANG Cai-sheng +1 位作者 ZHU Qiu-hua QIAN Guo-ying 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第5期869-876,共8页
The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation (SSF) with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on ph... The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation (SSF) with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on phytic acid, crude protein, crude fat, and amino acid profile. Response surface methodology (RSM) with Box-Behnken design was used to optimize SSF. The optimal conditions derived from RSM for L. brevis fermentation were: pH=5. 1; inoculum size=10%; duration=72 h; substrate to water ratio=1.5. The minimum content of trypsin inhibitors was 6.4 mg g^-1 dry matter. The optimal conditions derived from RSM for A. oryzae fermentation were: substrate to water ratio= 0.8 1; inoculum size=4%; duration=120 h. The minimum content of trypsin inhibitors was 1.6 mg g^-1 dry matter. Both L. brevis and A. oryzae decreased trypsin inhibitors dramatically (57.1 and 89.2% respectively). L. brevis fermentation did not affect phytic acid (0.4%) and crude fat (5.2%) considerably, whereas A. oryzae fermentation degraded phytic acid (34.8%) and crude fat (22.0%) contents to a certain extent. Crude protein content was increased after both fermentation (6.4 and 12.9% for L. brevis and A. oryzae respectively). Urease activity was reduced greatly (83.3 and 58.3% for L. brevis and A. oryzae respectively). In conclusion, SSF with A. oryzae and L. brevis reduced trypsin inhibitor content and modified major macronutrients in soybean meal. 展开更多
关键词 Aspergillus oryzae Lactobacillus brevis response surface methodology solid-state fermentation soybean meal trypsin inhibitors
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Urinary trypsin inhibitor attenuates hepatic ischemia-reperfusion injury by reducing nuclear factor-kappa B activation 被引量:28
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作者 Wu, Yi-Jun Ling, Qi +4 位作者 Zhou, Xin-Hui Wang, Yan Xie, Hai-Yang Yu, Ji-Ren Zheng, Shu-Sen 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2009年第1期53-58,共6页
BACKGROUND: Urinary trypsin inhibitor (UTI) inhibits the inflammatory response and protects against ischemia-reperfusion (I/R) injury. The inflammatory response is mediated by nuclear factor-kappa B (NF-kappa B) and i... BACKGROUND: Urinary trypsin inhibitor (UTI) inhibits the inflammatory response and protects against ischemia-reperfusion (I/R) injury. The inflammatory response is mediated by nuclear factor-kappa B (NF-kappa B) and its related target genes and products such as vascular endothelial cell adhesion molecule and CXC chemokines. We aimed to assess the roles of those mediators in a UTI-treated mouse model of hepatic I/R injury. METHODS: Treatment group 1 (UTI given 5 minutes prior to liver ischemia), treatment group 2 (UTI given 5 minutes after the anhepatic phase) and a control group were investigated. Blood and liver samples were obtained and compared at 1, 3, 6 and 24 hours after reperfusion. RESULTS: Attenuation of pathological hepatocellular damage was greater in the treatment groups than in the control group (P < 0.05). Compared with the control group, the UTI treatment groups showed significantly lower serum alanine aminotransferase and aspartate aminotransferase levels, decreased myeloperoxidase activity, and reduced NF-kappa B activation. Also downregulated was the expression of tumor necrosis factor-alpha, cytokine-induced neutrophil chemoattractant, and macrophage inflammatory protein-2 at the mRNA level. P-selectin protein and intercellular adhesion molecule-1 protein expression were also downregulated. In addition, the treatment group I showed a better protective effect against I/R injury than the treatment group 2. CONCLUSIONS: UTI reduces NF-kappa B activation and downregulates the expression of its related mediators, followed by the inhibition of neutrophil aggregation and infiltration in hepatic I/R injury. The protective role of UTI is more effective in prevention than in treatment. 展开更多
关键词 ischemia-reperfusion injury nuclear factor-kappa B tumor necrosis factor-alpha urinary trypsin inhibitor
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Purification, characterization and evaluation of insecticidal activity of trypsin inhibitor from Albizia lebbeck seeds 被引量:1
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作者 Pratima Sharma Amarjit K Nath +1 位作者 Reena Kumari SV Bhardwaj 《Journal of Forestry Research》 CAS CSCD 2012年第1期131-137,共7页
A Bowman-Birk inhibitor with activity against gut proteases of Helicoverpa armigera was extracted in 0.I M sodium phosphate buffer from defatted seed flour of Albizia lebbeck. It was purified to 29.62 folds with 51.43... A Bowman-Birk inhibitor with activity against gut proteases of Helicoverpa armigera was extracted in 0.I M sodium phosphate buffer from defatted seed flour of Albizia lebbeck. It was purified to 29.62 folds with 51.43% recovery using ammonium sulfate precipitation, gel filtration chromatography on Sephadex G-100 column and ion ex- change chromatography on DEAE-Sephadex As0. The purified protein had a molecular weight of 12,303 daltons as determined by SDS-PAGE. It was found to be heat stable up to 60~C and had two pH optima of 7.5 and 9.0. The inhibitor exhibited non-competitive pattern of inhibition with a low Ki value of 0.2 ~tM. The inhibitoi- was found to be susceptible to varying concentrations of reducing agents like DTT and 2- mercaptoethanol, thereby indicating the role of disulphide bridges in maintaining its three dimensional structure and stability. The purified inhibitor caused mortality and suppressed larval growth ofPieris brassi- cae larvae. It was also found to be effective against gut trypsin extracted from Spodoptera littoralis. The sequence of the genes encoding for such inhibitors can be determined and the genes expressing protease inhibitors can be used in vegetable crops to confer resistance against insect pests and other plant pathogens. 展开更多
关键词 Albizia lebbeck trypsin inhibitor insecticidal activity purifi-cation Pieris brassicae
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Purification and Trypsin Inhibitor Activity of a Sporamin B from Sweet Potato (Ipomoea batatas Lam. 55-2) 被引量:1
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作者 SUN Yan-li SUN Jun-mao LI Qing-peng 《Agricultural Sciences in China》 CAS CSCD 2009年第7期808-820,共13页
Sporamin is a soluble protein in sweet potato, and falls into two distinct homology groups, subfamilies A and B. In this research, a sporamin B was purified and its amino acid sequences, trypsin inhibitor activity (T... Sporamin is a soluble protein in sweet potato, and falls into two distinct homology groups, subfamilies A and B. In this research, a sporamin B was purified and its amino acid sequences, trypsin inhibitor activity (Ti activity) were analyzed. This sporamin B was isolated from sweet potato tubers [Ipomoea batatas (L.) Lam cv. 55-2] through extraction of the water-soluble fraction, dialysis, ultrafiltration and ion-exchange chromatography. Homology determined by polyacrylamide gel electrophoresis showed that mainly one bond appeared in gel after being reduced by SDS (sodium dodecyl sulfate), or by SDS and 2-mercaptoethanol, or in native situation. By comparing the data of the polypeptide mass Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry with those of the mass of the theoretical amino acid sequences from NCBI protein database, it was revealed that it was Q40091|Q40091_IPOBA, sweet potato sporamin B - Ipomoea Batatas (sweet potato) (Batate). The sequence coverage was 70.6%. N-terminal sequence was SETPV (Ser-Glu-Thr-Pro-Val). There is a linear relationship between trypsin inhibitor activity (Ti activity) and amounts of this sporamin B (3-18 μg mL-1). The equation of linear regression was y = 2.5809x + 17.049 (r2 = 0.9966). There was a curvilinear relationship between Ti activity and amounts of this sporamin B (21-150 μg mL-1). The equation of curvilinear regression is y = 14.417ln(x) + 23.26 (r2 = 0.9924). The concentration of sporamin B with Ti activity after heating at 40°C may induce part denature of this sporamin B, and there was no statistic difference after heating at 40, 50, 60°C for 20 min. Heat treatment at more than 90°C leads to a dramatic decrease of trypsin inhibitor efficiency. The results suggested that Q40091|Q40091_IPOBA was the major sporamin B in sweet potato tubers [Ipomoea Batatas (L.) Lam cv. 55-2], which had strong Ti activity, and was stable to both thermal and DTT (DL-dithiothreitol) relatively. 展开更多
关键词 sweet potato sporamin B IDENtiFICAtiON trypsin inhibitor activity
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Storage Proteins and Trypsin Inhibitors of an Underutilized Legume, <i>Mucuna</i>: Variability and Their Stability during Germination 被引量:1
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作者 Kempohalli S. Chandrashekharaiah 《American Journal of Plant Sciences》 2013年第4期910-916,共7页
The proteins and trypsin inhibitors were isolated from the seeds of different varieties/accessions of an underutilized legume, Mucuna. The crude protein content of all the germplasms of Mucuna is varied from 15% - 26%... The proteins and trypsin inhibitors were isolated from the seeds of different varieties/accessions of an underutilized legume, Mucuna. The crude protein content of all the germplasms of Mucuna is varied from 15% - 26%, showed little variation and contain higher crude protein when compared with other Mucuna species reported earlier and the pulse crops commonly consumed in India. The seeds of all the varieties of Mucuna exhibited trypsin inhibitor activity. The trypsin inhibitor activity varied from 11 - 14 TIA/mg of protein. Not much variation was observed in trypsin inhibitory activities in soaked seeds compared to dry seeds. Germination of Mucuna pruriens has been carried out and the change in the protein content and trypsin inhibitors were monitored. The protein content of the endosperm increased up to 72 hrs of germination and then decreased. The trypsin inhibitory activity decreased with increase in germination time. The trypsin inhibitor activity was decreased from 14.81 TIA/mg to 2.62 TIA/mg (82% reduction in the trypsin inhibitor activity) after 144 hrs germination. 展开更多
关键词 MUCUNA Seeds Proteins trypsin inhibitors GERMINAtiON Variation
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Pancreatic secretory trypsin inhibitor:More than a trypsin inhibitor 被引量:2
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作者 Gai-Ping Wang,Cun-Shuan Xu,College of Life Science,Henan Normal University,Xinxiang 453007,Henan Province,China Co-construction Key Laboratory for Cell Differentiation and Regulation,Henan Normal University,Xinxiang 453007,Henan Province,China 《World Journal of Gastrointestinal Pathophysiology》 CAS 2010年第2期85-90,共6页
Kazal-type serine protease inhibitor is one of the most important and widely distributed protease inhibitor families. Pancreatic secretory trypsin inhibitor (PSTI), also known as serine protease inhibitor Kazal type I... Kazal-type serine protease inhibitor is one of the most important and widely distributed protease inhibitor families. Pancreatic secretory trypsin inhibitor (PSTI), also known as serine protease inhibitor Kazal type I(SPINK1), binds rapidly to trypsin, inhibits its activity and is likely to protect the pancreas from prematurely activated trypsinogen. Therefore, it is an important factor in the onset of pancreatitis. Recent studies found that PSTI/SPINK1 is also involved in self-regulation of acinar cell phagocytosis, proliferation and growth of a variety of cell lines. In addition, it takes part in the response to inflammatory factor or injury and is highly related to adult type II citrullinemia. 展开更多
关键词 Pancreatic secretory trypsin inhibitor/serine protease inhibitor Kazal type I PANCREAtitiS AUTOPHAGY Cell proliferation Inflammatory factor Adult-II citrullinemia.
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Relationship between inter-α-trypsin inhibitor heavy chain 4 and ovarian cancer 被引量:2
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作者 Min Huang Wei Zhang +1 位作者 Bingbing Zhao Li Li 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2019年第6期955-964,共10页
Objective:The inter-α-trypsin inhibitor heavy chain 4(ITIH4)protein is involved in the development of tumors.However,the relationship between ITIH4 and ovarian cancer(OC)has not been extensively examined.This study a... Objective:The inter-α-trypsin inhibitor heavy chain 4(ITIH4)protein is involved in the development of tumors.However,the relationship between ITIH4 and ovarian cancer(OC)has not been extensively examined.This study aimed to explore the effect of ITIH4 on OC and to identify its underlying mechanism.Methods:Expressions of ITIH4 in OC tissues and cells were determined using quantitative reverse transcription polymerase chain reaction(RT-qPCR)and western blots.The function of ITIH4 in the OC cell line HO8910 pm was tested via ITIH4 knockdown.The cell growth rate was measured using MTT and colony formation assays.Flow cytometry was performed to evaluate cell cycle progression.Cell migration and invasion abilities were observed using the transwell migration assay.Results:ITIH4 was downregulated in OC tissues and cells.ITIH4 knockdown promoted cell growth and cell cycle progression.Consistent with these results,inhibition of ITIH4 in OC cells significantly increased cell migration and invasion abilities.Cox regression analysis suggests that ITIH4 expression alone is not a good predictor of the prognosis of malignant ovarian tumors in patients.Conclusions:ITIH4 inhibits the progression of OC,suggesting that ITIH4 may be a useful biomarker for OC.This study may provide a potential novel target for the treatment of OC. 展开更多
关键词 Ovarian cancer inter-α-trypsin inhibitor H4 protein quantitative reverse transcription PCR RNAI
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Silica-supported Macroporous Chitosan Bead for Affinity Purification of Trypsin Inhibitor
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作者 Feng Na XI Jian Min WU Ming Ming LUAN 《Chinese Chemical Letters》 SCIE CAS CSCD 2005年第8期1089-1092,共4页
Macroporous cross-linking chitosan layer coated on silica gel (CTS-SiO2) was prepared by phase inversion and polyethylene glycol (PEG)molecular imprinting methods. Formation of macroporous surtace was investigated... Macroporous cross-linking chitosan layer coated on silica gel (CTS-SiO2) was prepared by phase inversion and polyethylene glycol (PEG)molecular imprinting methods. Formation of macroporous surtace was investigated by scanning electron microscopy (SEM) and BET analysis.The prepared bead was activated by reacting with 1,2-ethylene digiycidyl ether for introducing epoxy groups, and trypsin could be efficiently immobilized on the bead as a biospecific ligand.The bead bearing trypsin was employed to purify trypsin inhibitor (TIs) from egg white as affinity adsorbent. 展开更多
关键词 CHITOSAN SILICA trypsin inhibitor trypsin affinity purification.
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Polymorphism of Kunitz Trypsin Inhibitor Protein in Natural Populations of Wild Soybean in Hebei
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作者 WANG Ke-jing WANG Ying-dian HAI Lin DAI Xin LI Fu-shan 《Agricultural Sciences in China》 CAS CSCD 2001年第1期40-44,共5页
Hebei Province is one of the main distribution areas growing wild soybean( Glycine soja ) in China. In this study, 461 seed samples,collected from 18 natural populations in this province, were used to electrophoretica... Hebei Province is one of the main distribution areas growing wild soybean( Glycine soja ) in China. In this study, 461 seed samples,collected from 18 natural populations in this province, were used to electrophoretically observe the change in forms and their frequencies of the Kunitz trypsin inhibitor protein (KTI)in individual populations and geographical areas. Allelic frequencies accounted for 85% for Tia and 15% for Tib in the total samples. Twelve populations examined were polymorphic at the KTI locus, accounting for over 50% in the populations investigated. Four populations, 22% of all the populations, were found to have natural cross-pollination with varied heterozygote rates of 3% -5.5%, and the average was 1% in the total sampies. Geographically, the mean Tib frequency in the north areas was higher than in the south, and higher in the mountainous area than in the plain areas. The populations in a lake ecological environment (Baiyangdian Lake) were almost monomorphic. No obvious relationship between the frequency and the geographical distance was observed. In addition, we first found a mutation for the absence of the KTI in wild soybean. 展开更多
关键词 KUNITZ trypsin inhibitor GEOGRAPHICAL DIVERGENCE Isozymic frequency WILD soybean
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A Kunitz trypsin inhibitor from chickpea (<i>Cicer arietinum</i>L.) that exerts an antimicrobial effect on Fusarium oxysporum f.sp. ciceris
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作者 Meera Nair Sardul Singh Sandhu 《Agricultural Sciences》 2013年第11期585-594,共10页
Fusarium oxysporum f.sp. ciceris (Foc) is one of the most important fungal pathogens of chickpea and is regarded as a constant threat in tropical and subtropical countries. In order to correlate Fusarium wilt resistan... Fusarium oxysporum f.sp. ciceris (Foc) is one of the most important fungal pathogens of chickpea and is regarded as a constant threat in tropical and subtropical countries. In order to correlate Fusarium wilt resistance/susceptibility in Cicer arietinum to the presence or absence of trypsin inhibitor (TI) in the crude extract, trypsin inhibitory assay (TIA) and in vitro activity of TI against Foc were studied. In the present study, a 20 kDa trypsin inhibitor was purified from Fusarium wilt resistant cultivar (viz. JG 2001-12) by ammonium sulfate precipitation, dialysis and chromatographies with Sephadex G-100 and Diethyl aminoethyl cellulose (DEAE-cellulose-52) ion-exchange column. Results of pathogenecity assay were found to be in correlation to the trypsin inhibitor assay where the Fusarium wilt resistant cultivar showed high trypsin inhibitory activity (99%) in the presence of trypsin enzyme using both natural and synthetic substrates. Preliminary studies using crude extracts of JG 2001-12 showed a decrease in radial growth of Foc. A 45%-82% reduction in conidium germination at 20 μg&middotmL-1?Cicer arietinum trypsin inhibitor (CaTI) concentration was observed, thereby, indicating the use of CaTI in suppression of pathogen and in its deployment through transgenic plants for the management of Fusarium wilt. 展开更多
关键词 CICER arietinum Fusarium Wilt KUNITZ PROTEINASE inhibitor trypsin inhibitor
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A Novel Trypsin Inhibitor-Like Cysteine-Rich Peptide from the Frog Lepidobatrachus laevis Containing ProteinaseInhibiting Activity
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作者 Yu-Wei Wang Ji-Min Tan +4 位作者 Can-Wei Du Ning Luan Xiu-Wen Yan Ren Lai Qiu-Min Lu 《Natural Products and Bioprospecting》 CAS 2015年第4期209-214,共6页
Various bio-active substances in amphibian skins play important roles in survival of the amphibians.Many protease inhibitor peptides have been identified from amphibian skins,which are supposed to negatively modulate ... Various bio-active substances in amphibian skins play important roles in survival of the amphibians.Many protease inhibitor peptides have been identified from amphibian skins,which are supposed to negatively modulate the activity of proteases to avoid premature degradation or release of skin peptides,or to inhibit extracellular proteases produced by invading bacteria.However,there is no information on the proteinase inhibitors from the frog Lepidobatrachus laevis which is unique in South America.In this work,a cDNA encoding a novel trypsin inhibitor-like(TIL)cysteine-rich peptide was identified from the skin cDNA library of L.laevis.The 240-bp coding region encodes an 80-amino acid residue precursor protein containing 10 half-cysteines.By sequence comparison and signal peptide prediction,the precursor was predicted to release a 55-amino acid mature peptide with amino acid sequence,IRCPKDKIYKFCGSPCPPSCKDLTPNCIAVCKKGCFCRDGTVDNNHGKCVKKENC.The mature peptide was named LL-TIL.LL-TIL shares significant domain similarity with the peptides from the TIL supper family.Antimicrobial and trypsin-inhibitory abilities of recombinant LL-TIL were tested.Recombinant LL-TIL showed no antimicrobial activity,while it had trypsin-inhibiting activity with aKi of 16.5178 lM.These results suggested there was TIL peptide with proteinase-inhibiting activity in the skin of frog L.laevis.To the best of our knowledge,this is the first report of TIL peptide from frog skin. 展开更多
关键词 trypsin inhibitor Cysteine-rich peptide AMPHIBIAN Skin
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CRYSTAL STRUCTURE OF THE COMPLEX OF MUNG BEAN TRYPSIN INHIBITOR LYSINE ACTIVE FRAGMENT WITH BOVINE TRYPSIN AT 1.8 A RESOLUTION
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作者 You Qi TANG Gen Pei LI Zhong Guo CHEN Jie ZENG(Institute of Physical Chemistry,Peking University,Beijing 100871)Tien Chin TSAO Guang Da LIN Rong Guang ZHANG Zheng Wu CHI(Institute of Biochemistry,Academic Sinica,Shanghai 200031) 《Chinese Chemical Letters》 SCIE CAS CSCD 1990年第1期61-64,共4页
The structure of the complex of mung bean trypsin inhibitor lysine active fragment with bovine trypsin has been determined at a resolution of 1.8 A by A-ray crystallographic analysis and the complex model refined by r... The structure of the complex of mung bean trypsin inhibitor lysine active fragment with bovine trypsin has been determined at a resolution of 1.8 A by A-ray crystallographic analysis and the complex model refined by restrained least-squares minimization with the data between 10 and 1.8 resolution.The current conventional R factor is 17.3%,and the model con- tains 1648 protein atoms,219 inhibitor atoms and 126 water molecules.The most prominent feature of the inhibitor fragment is that it does not contain any alpha-helices.Most of the chain fold in an irregular fashion.The seven residues of the binding segment of the inhibitor lysine active frag- ment are in specific contact with bovine trypsin.The binding interaction and geometry around the reactive site are similar to that observed in other studies of trypsin-inhibitor complexes. 展开更多
关键词 maps CRYSTAL STRUCTURE OF THE COMPLEX OF MUNG BEAN trypsin inhibitor LYSINE ACtiVE FRAGMENT WITH BOVINE trypsin AT 1.8 A RESOLUtiON AT
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血清ITIH4和MCL-1水平与急性缺血性脑卒中患者病情程度及预后的关系
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作者 席俊男 冯松松 +2 位作者 薛惠元 腾东 陈丽薇 《疑难病杂志》 CAS 2024年第4期429-434,共6页
目的探究急性缺血性脑卒中(AIS)患者血清间α胰蛋白酶抑制因子重链4(ITIH4)、髓样细胞白血病因子-1(MCL-1)表达与病情程度及预后的关系。方法纳入2019年7月—2022年7月河南科技大学附属黄河医院神经内科诊治AIS患者128例为AIS组。根据... 目的探究急性缺血性脑卒中(AIS)患者血清间α胰蛋白酶抑制因子重链4(ITIH4)、髓样细胞白血病因子-1(MCL-1)表达与病情程度及预后的关系。方法纳入2019年7月—2022年7月河南科技大学附属黄河医院神经内科诊治AIS患者128例为AIS组。根据入院时美国国立卫生研究院卒中量表(NIHSS)评分,分为轻度亚组(NIHSS<6分,n=42)、中度亚组(NIHSS 6~<14分,n=52)和重度亚组(NIHSS≥14分,n=34)。根据出院3个月时AIS患者改良Rankins评分,分为预后不良亚组(mRS评分>2分,30例)和预后良好亚组(mRS评分≤2分,98例)。另选取同期医院体检的健康人70例为健康对照组。酶联免疫吸附试验检测血清ITIH4、MCL-1水平。Pearson相关分析血清ITIH4、MCL-1水平与病情程度及预后的相关性;多因素Logistic回归分析影响AIS患者预后的因素;受试者工作特征曲线分析血清ITIH4、MCL-1对AIS患者预后的预测价值。结果AIS组患者血清ITIH4、MCL-1水平显著低于健康对照组(t/P=43.211/<0.001,43.191/<0.001);病情程度越重,AIS患者血清ITIH4/MCL-1水平越低(F/P=107.796/<0.001,297.976/<0.001);预后不良亚组梗死面积、入院24 h NIHSS评分高于预后良好亚组(t/P=9.637/<0.001,9.752/<0.001),血清ITIH4、MCL-1水平及出院3个月简易智能状态量表(MMSE)评分、蒙特利尔认知评估量表(MoCA)评分低于预后良好亚组(t/P=26.723/<0.001,11.709/<0.001,13.674/<0.001,10.782/<0.001);AIS患者血清ITIH4、MCL-1与梗死面积、入院24 h NIHSS评分呈负相关(r/P=-0.705/<0.001,-0.685/<0.001;-0.761/<0.001,-0.619/<0.001),与出院3个月MMSE评分、MoCA评分呈正相关(r/P=0.656/<0.001,0.632/<0.001;0.751/<0.001,0.789/<0.001);出院3个月MMSE评分高、出院3个月MoCA评分高是影响AIS患者预后不良的独立保护因素[0.622(0.446~0.868),0.606(0.427~0.861)],血清ITIH4低、MCL-1低、梗死面积大、入院24 h NIHSS评分高是危险因素[OR(95%CI)=1.467(1.150~1.870),1.415(1.094~1.829),1.605(1.168~2.205),1.765(1.233~2.526)];血清ITIH4、MCL-1及两项联合预测AIS预后不良的AUC分别为0.811、0.835、0.923,两项联合预测AIS预后不良的AUC大于单一指标,差异具有统计学意义(Z=4.258、4.119,P均<0.001)。结论AIS患者血清ITIH4、MCL-1表达下调,两者表达水平与病情严重程度有关,两者联合对AIS患者预后具有较高的预测价值。 展开更多
关键词 急性缺血性脑卒中 间α胰蛋白酶抑制因子重链4 髓样细胞白血病因子-1 病情程度 预后
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Low trypsinogen-1 expression in pediatric ulcerative colitis patients who undergo surgery 被引量:1
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作者 Maija Piekkala Jaana Hagstrm +3 位作者 Maarit Tanskanen Risto Rintala Caj Haglund Kaija-Leena Kolho 《World Journal of Gastroenterology》 SCIE CAS 2013年第21期3272-3280,共9页
AIM: To investigate whether matrix metalloproteinases-9 (MMP-9) or trypsinogens could serve as histological markers for an aggressive disease course in pediatric ulcerative colitis (UC). METHODS: We identified 24 pati... AIM: To investigate whether matrix metalloproteinases-9 (MMP-9) or trypsinogens could serve as histological markers for an aggressive disease course in pediatric ulcerative colitis (UC). METHODS: We identified 24 patients with pediatric onset (≤ 16 years) UC who had undergone surgery during childhood/adolescence a median of 2.1 years (range 0.1-7.4 years) after the diagnosis (between 1990 and 2008) in Children's Hospital, Helsinki, Finland. We also identified 27 conservatively treated UC patients and matched them based on their age at the time of diagnosis and follow-up at a median of 6 years (range 3-11 years) to serve as disease controls. Twenty children for whom inflammatory bowel disease (IBD) had been excluded as a result of endoscopy served as non-IBD controls. Colon biopsies taken by diagnostic endoscopy before the onset of therapy were stained using immunohistochemistry to study the expression of MMP-9, trypsinogen-1 (Tryp-1), Tryp-2, and a trypsin inhibitor (TATI). The profiles of these proteases and inhibitor at diagnosis were compared between the surgery group, the conservatively treated UC patients and the non-IBD controls. RESULTS: The proportions of Tryp-1 and Tryp-2 positive samples in the colon epithelium and in the inflammatory cells of the colon stroma were comparable between the studied groups at diagnosis. Interestingly, the immunopositivity of Tryp-1 (median 1; range 0-3) was significantly lower in the epithelium of the colon in the pediatric UC patients undergoing surgery when compared to that of the conservatively treated UC patients (median 2; range 0-3; P = 0.03) and non-IBD controls (median 2; range 0-3; P = 0.04). For Tryp-2, there was no such difference. In the inflammatory cells of the colon stroma, the immunopositivities of Tryp-1 and Tryp-2 were comparable between the studied groups at diagnosis. Also, the proportion of samples positive for TATI, as well as the immunopositivity, was comparable between the studied groups in the colon epithelium. In the stromal inflammatory cells of the colon, TATI was not detected. In UC patients, there were significantly more MMP-9 positive samples and a higher immunopositivity in the stromal inflammatory cells of the colon when compared to the samples from the non-IBD patients (P = 0.006 and P = 0.002, respectively); the immunopositivity correlated with the histological grade of inflammation (95%CI: 0.22-0.62; P = 0.0002), but not with the other markers of active disease. There were no differences in the immunopositivity or in the proportions of MMP-9 positive samples when examined by epithelial staining. The staining profiles in the ileal biopsies were comparable between the studied groups for all of the studied markers.CONCLUSION: For pediatric UC patients who require surgery, the immunopositivity of Tryp-1 at diagnosis is lower when compared to that of patients with a more benign disease course. 展开更多
关键词 Children ULCERAtiVE COLItiS Inflammatory BOWEL disease Matrix metalloproteinase-9 resection trypsin inhibitor
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Corrosion resistance improvement of Ti-6Al-4V alloy by anodization in the presence of inhibitor ions 被引量:2
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作者 A.L.MARTINEZ D.O.FLAMINI S.B.SAIDMAN 《Transactions of Nonferrous Metals Society of China》 SCIE EI CAS CSCD 2022年第6期1896-1909,共14页
Colorful thin oxide films were synthesized by galvanostatic anodization on Ti−6Al−4V alloy.Three different aqueous solutions containing corrosion inorganic inhibitors(Na_(2)MoO_(4),NaH_(2)PO_(4) and NH4VO3)were employ... Colorful thin oxide films were synthesized by galvanostatic anodization on Ti−6Al−4V alloy.Three different aqueous solutions containing corrosion inorganic inhibitors(Na_(2)MoO_(4),NaH_(2)PO_(4) and NH4VO3)were employed for the anodization treatment.The effect of inhibitor anions on the corrosion behavior of the alloy in Ringer solution was studied.Open circuit potential(OCP),Tafel polarization,linear sweep voltammetry(LSV)and chronoamperometry(CA)were performed to evaluate the corrosion performance of the treated electrodes.The incorporation of the inhibitor ions was detected by the release of Mo,V and P through ICP-AES technique.The formed oxides were characterized by scanning electron microscopy(SEM),X-ray diffraction(XRD)and X-ray photoelectron spectroscopy(XPS).The results show that compact,amorphous oxides without pores or cracks were obtained independently of the solution used.The sample anodized in Na_(2)MoO_(4) solution registered the lowest corrosion current density(0.11μA/cm^(2)),and it was able to protect the alloy even after 168 h of immersion in Ringer solution.No cracks or corrosion products were detected.The XPS analysis reveals the incorporation of molybdenum to the oxide film in the form of Mo^(6+) and Mo^(4+). 展开更多
关键词 ti−6Al−4V alloy anodization inorganic inhibitors corrosion protection
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miR-184通过TIMP-2/MMP-14促进PNX模型中代偿性肺生长 被引量:1
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作者 彭静 向旭东 +5 位作者 王忠慧 王琼川 邵世豪 马维浩 朱波波 赵力 《中国病理生理杂志》 CAS CSCD 北大核心 2023年第12期2214-2222,共9页
目的:探究微小RNA-184(miR-184)对多原发肺癌(MPLC)肺叶切除术后代偿性肺生长(CLG)的影响和作用机制。方法:(1)收集MPLC患者和肺叶切除术后恢复较好患者(CLG)的肺组织样本(n=16)。RT-qPCR检测miR-184的表达。(2)将人肺泡上皮细胞根据转... 目的:探究微小RNA-184(miR-184)对多原发肺癌(MPLC)肺叶切除术后代偿性肺生长(CLG)的影响和作用机制。方法:(1)收集MPLC患者和肺叶切除术后恢复较好患者(CLG)的肺组织样本(n=16)。RT-qPCR检测miR-184的表达。(2)将人肺泡上皮细胞根据转染物分为NC-mimic组、miR-184 mimic组、OE-NC组、金属蛋白酶组织抑制物2(TIMP-2)过表达(OE-TIMP-2)组和miR-184 mimic+OE-TIMP-2组(n=3)。RT-qPCR检测细胞中miR-184、TIMP-2 mRMA和基质金属蛋白酶14(MMP-14)mRNA表达;Western blot检测TIMP-2和MMP-14蛋白表达;CCK-8和集落形成实验检测细胞增殖能力。(3)将C57BL/6J小鼠分为肺切除术(PNX)组和PNX+miR-184 mimic组(n=5)。用flexiVent肺功能检测系统检测小鼠肺活量和肺顺应性;水置换法检测肺容量;HE染色观察肺组织变化。结果:miR-184在肺叶切除术后恢复较好患者中的表达显著升高(P<0.01)。过表达miR-184能促进人肺泡上皮细胞增殖和PNX后小鼠肺功能恢复。miR-184与TIMP-2存在靶向结合关系;过表达miR-184通过抑制TIMP-2促进MMP-14的表达,进而促进人肺泡上皮细胞增殖和PNX后小鼠肺功能恢复。结论:miR-184通过TIMP-2/MMP-14轴促进PNX后的CLG。 展开更多
关键词 多原发肺癌 代偿性肺生长 肺切除术 微小RNA-184 金属蛋白酶组织抑制物2 基质金属蛋白酶14
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On the Activities of Pancreatic Proteases and Alpha-1 Proteinase Inhibitor in Meat-Type Chicken
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作者 Vladimir G. Vertiprakhov Alena A. Grozina +3 位作者 Ivan A. Egorov Tatiana N. Lenkova Vardges A. Manukyan Tatiana A. Egorova 《Open Journal of Animal Sciences》 2017年第3期289-296,共8页
The study was aimed at the evaluation of the effects of breed, age, different digestion stimulators, and dietary crude protein (CP) level on the activities of proteolytic enzymes in pancreatic tissue and duodenal chym... The study was aimed at the evaluation of the effects of breed, age, different digestion stimulators, and dietary crude protein (CP) level on the activities of proteolytic enzymes in pancreatic tissue and duodenal chymus (in vivo), serum trypsin and α1-proteinase inhibitor (A1PI) concentrations in meat-type chicks. The study of age dynamics of trypsin and A1PI concentrations was performed on the chicks of hybrid cross “Smena-8”and two parental lines (Plymouth Rock and Cornish) fed standard commercial corn-wheat-SBM diets. Twenty birds per breed were euthanized at 1, 7, 14, 21, 28 and 35 days of age to obtain blood samples and pancreatic homogenate. Experiments on the effects of different digestion promotors (probiotic, acidifier, phytobiotic, enzymatic preparation) and different CP levels (finisher diet, CP 20%, vs. ground corn, CP 8.5%) were performed on 12 hybrid chicks with fistulated duodenum from 14 to 50 days of age. The following conclusions were made: 1) At 1 day of age high proteolytic activity in pancreatic tissue and maximal serum concentrations of trypsin and A1PI were found in both hybrid and parental lines. Since 7 to 35 days of age A1PI concentration was nearly constant, serum trypsin concentration decreased while proteolytic activity in pancreatic tissue exhibited undulate increase;2) Proteolytic activity in pancreatic tissue was higher in hybrids compared to the parental lines from 7 to 35 days of age (p 0.05);3) Supplementation of diet with exogenous enzymes stimulated the digestion due to the increase in protease activity in duodenal chymus by 9.1% compared to unsupplemented control (p 0.05);4) Proteolytic activity in duodenal chymus significantly responded to the substitution of ground corn for the complete diet by 2-fold decrease while serum trypsin concentration responded by 2.5-fold increase (p 0.001). This fact can indicate that physiological functions of digestive proteases are not confined to the digestive processes. 展开更多
关键词 CHICKS Pancreas trypsin Alpha-1 PROTEINASE inhibitor (Antitrypsin) Serum DUODENAL Fluid
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