Correlation analysis of the expression of LACTB and GST-π in focal tissues of endometrialcarcinoma with their survival and estrogen and progesterone receptors / 中华内分泌外科杂志

Objective:To investigate the expression of LACTB and GST-π in endometrial carcinoma and their correlation with estrogen and progesterone receptors.Methods:A total of 165 patients with endometrial cancer admitted to our hospital from Jan. 2015 to Oct. 2020 were selected to observe the expression of LACTB, GST-π, estrogen and progesterone receptors in patients with different tumor stages, degrees of differentiation, muscular infiltration, tissue type, tumor diameter and whether there was lymph node metastasis. The correlation between the expression of LACTB and GST-π and the expression of female and progesterone receptors and the survival of patients with different expressions of LACTB and GST-π were analyzed.Results:The LATCB positive rate decreased in patients with tumor stage III to IV, high differentiation, myometrial infiltration ≥1/2, tissue type I, tumor diameter <2 cm and no lymph node metastasis ( P<0.05), and the GST-π positive rate was not statistically significant ( P>0.05). The positive rate of GST-π in patients with chemotherapy resistance was higher than that in patients with chemotherapy sensitivity, and the difference was statistically significant ( P<0.05). The positive rate of estrogen receptor decreased in patients with tumor stage III to IV, high differentiation, tissue type II, no lymph node metastasis, and chemotherapy resistance, and the difference was statistically significant ( P<0.05). The positive rate of progesterone receptor decreased in patients with tumor stage III to IV, low differentiation, tissue type I and no lymph node metastasis, and the difference was statistically significant ( P<0.05). In this study, 110 patients with LACTB positive expression were detected, with an average LACTB positive staining intensity (25.92±4.77) %, and 99 patients with GST-π positive expression were detected, with an average GST-π positive staining intensity (27.46±4.83). A total of 50 patients with LACTB positive and GST-π negative had an average survival time of (41.48±5.52) months, a total of 39 patients with LACTB negative and GST-π positive had an average survival time of (21.25±3.15) months, and a total of 60 patients with LACTB positive and GST-π positive had an average survival time of (21.25±3.15) months. The mean survival time of 16 patients with both LACTB negative and GST-π negative was 29.31±3.77 months. The mean survival time was 31.54±4.61 months. Pearson correlation test showed that the staining intensity of LACTB positive staining was positively correlated with the survival time of patients. The positive staining intensity of GST-π was negatively correlated with survival (correlation coefficient =0.392, -0.284, P<0.01). Pearson correlation analysis showed that LACTB was positively correlated with estrogen and progesterone receptors. GST-π was negatively correlated with estrogen receptor ( P<0.05) . Conclusions:The expression of LACTB in patients with endometrial cancer is associated with the disease of the patients. The survival of patients with high expression is longer. The expression of GST-π is associated with the chemotherapy resistance of patients, both of which can be used as indicators to evaluate the prognosis of patients. Moreover, the expressions of LACTB and GST-π are correlated with the expression of female and progesterone receptors, which may be regulated by the expression of female and progesterone receptors.

Construction, identification and expression of recombinant Schistosoma japonicum vaccine mediated by Enterococcus faecalis / 中华地方病学杂志

Objective:To construct a recombinant vaccine of Schistosoma japonicum (Sj) mediated by Enterococcus faecalis (Efs, rEfs-Sj26GST vaccine), and to study the expression of Sj26GST-GST fusion protein in the recombinant vaccine. Methods:The recombinant plasmid pGEX-Sj26GST was transformed into the susceptible strain Efs ATCC47077 by electroporation to construct rEfs-Sj26GST vaccine, and the plasmid was extracted for PCR identification. After induction of expression with isopropyl-beta-D-thiogalactopyranoside (IPTG), the products were analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot.Results:After PCR identification, a 676 bp fragment was amplified, which was consistent with the length of Sj26GST amplification fragment. SDS-PAGE analysis showed that the relative molecular mass was 52 × 10 3, which was consistent with the band of Sj26GST-GST fusion protein. Western blot results showed that the Sj26GST-GST fusion protein expressed by rEfs-Sj26GST vaccine could be specifically recognized by the serum of Sj infected patients. Conclusion:The rEfs-Sj26GST vaccine is successfully constructed, and the Sj26GST-GST fusion protein expressed by recombinant vaccine can be specifically recognized by the serum of Sj infected patients.

Prokaryotic expression and functional identification of androgen receptor functional domains / 安徽医科大学学报

Objective@#To construct the full-length prokaryotic expression plasmid of the wild type of androgen receptor (AR) and the truncated body of four functional domains，and to identify the fusion protein by Western blot and electrophoretic mobility shift assay ( EMSA) ．@*Methods@#Based on the pGEX-4T-1 vector ，the recombinant plasmids were constructed to express the full-length and functional domains of AR. IPTG was used to induce the expression of the recombinant proteins，which were isolated and purified by glutathione sepharose 4B beads under the optimized condition．The specific protein expression in the bacterial lysate and the purified protein isolated with glutathione sepharose 4B beads was identified by Western blot with AR antibody and GST labeled antibody．The purified protein was incubated with a fluorescent probe of the virus，and the complex was detected by electrophoresis in a non-denaturing gel. @*Results @# The prokaryotic recombinant plasmids of full length and three functional domain truncated AR were successfully constructed．The recombinant clones were identified by using bacterial culture as a template，and further verified by double enzyme digestion．It showed that there were identical bands in the same sizes as the inserted fragments．The nucleotide and the amino acid sequences were aligned to the reference sequence in NCBI GenBank．The GST fusion protein，GST-AR-NTD + DBD (96 ku) and GST-AR-NTD (86 ku) were successfully induced and verified． The purified protein could be directly combined with the viral genome DNA．@*Conclusion@#The prokaryotic expression conditions of truncated AR plasmid from the same gene sequence are different．The purified AR protein can be used to understand the direct interaction mechanism between functional domains of AR and other molecules.

CAUSES AND CONSEQUENCES OF GOODS & SERVICE TAX (GST) FRAMEWORK IN INDIA

The giant indirect tax framework in India known as the Goods and Services Tax, is meant to promote and bolster the economic growth of a country. GST has been introduced in more than 150 countries. The Vajpayee government came up with the concept of GST in 2000. Growth and development may be hindered by an aggressive performance of GST. For this study, the primary goal is for researchers to get a better understanding of the in?uence of the Goods and Services Tax (GST) on the Indian economy, as well as the many features of the Indian GST system. Using data from periodicals, newspapers, and approved websites, the article is based on secondary data sources. In conclusion, I believe GST has certain bene?cial effects, such as helping to create a tax administration that is open and devoid of corruption. Small ?rms with annual revenue of between INR 20 lakhs and INR 50 lakhs might elect to pay lesser taxes under the new GST system. What it is referred to as is known as the compositional scheme. A hike to Rs. 75 lakhs has been proposed, however there are also drawbacks, such as the fact that some retail items presently only carry a 4% tax, and that after GST, clothing and apparel may become more expensive. The GST does not apply to three key tax income generators: petroleum goods, alcoholic beverages, and electricity. The tax will be levied by the state government on certain industries. Tax income generated by GST would be a major factor in the inclusion of these sectors in the GST. A higher tax rate of 18 percent from the current 15 percent would have a signi?cant impact on the services industry, such as communications and restaurants. In India, the introduction of the Goods and Services Tax (GST) has had mixed results

REVOLUTIONARY IMPACT OF GST ON INDIAN INDUSTRIAL SECTORS

In this paper I illustrated the economic structure in India is the basis of such an amazing withstand that is to say, the introduction of Goods and Services Tax (GST) as a multipoint tax system is an another milestone in the economic growth by enveloping all indirect taxes and avoiding cascading effect in taxation to create India as a single uni?ed common market all over the world. The implementation of GST will reduce tax burden on manufacturers and thus encourages for the higher production. This process will increase the export of India and it will increase the total GNP. Avoidance of cascading effect empowers the manufacturers to produce to their optimum capacity and retards growth.

Effect and Mechanism of Banxia Xiexintang on Rats with Chronic Atrophic Gastritis Based on Keap1/Nrf2/ARE Signaling Pathway / 中国实验方剂学杂志

Objective:To explore the mechanism of Banxia Xiexintang （BXXX） in preventing and treating chronic atrophic gastritis （CAG） through Kelch-like ECH-associated protein 1 （Keap1）/nuclear factor erythroid 2-related factor 2 （Nrf2）/antioxidant response element （ARE） signaling pathway. Method:SD rats were divided into a normal group （<italic>n</italic>=12） and an experimental group for CAG model induction. The model rats were then randomly divided into a model group， a vatacoenayme （VG） group （60 mg·kg^-1）， and high- （280 mg·kg^-1）， medium- （140 mg·kg^-1）， and low-dose （70 mg·kg^-1） BXXX groups. The doses in the BXXX groups were equivalent to 28， 14， and 7 g·kg^-1 crude drugs. The rats in the normal group and the model group received distilled water at an equal volume， and those in the VG group and the BXXX groups were treated correspondingly by gavage. After 12 weeks of treatment， hematoxylin-eosin （HE） staining was carried out to observe pathological changes in the gastric mucosa of CAG rats. Western blot and real-time fluorescence-based quantitative PCR was used to detect the protein and mRNA expression levels of Nrf2， glutathione S-transferase （GST）， and NAD （P）H：quinone oxidoreductase 1 （NQO1） in the gastric mucosa of CAG rats. Result:Compared with the normal group， the model group showed increased protein and mRNA expression levels of Nrf2， NQO1， and GST in the gastric mucosa of the rats （<italic>P</italic><0.05）， atrophic gastric mucosa， and even intestinal metaplasia. The protein and mRNA expression levels of Nrf2， NQO1， and GST in the VG group and the high- and medium-dose BXXX groups were lower than those in the model group （<italic>P</italic><0.05）， and gastric mucosa atrophy and intestinal metaplasia were significantly improved， especially in the high-dose BXXX group. However， the effect in the low-dose BXXX group was not significant. Conclusion:BXXX can blunt the transcriptional activity of Nrf2， shut down Nrf2 signaling pathway， and reduce the expression levels of NQO1 and GST to achieve normal oxidation-anti-oxidation balance， which may be one of its action mechanisms in the treatment of CAG.

Resveratrol elongates the lifespan and improves antioxidant activity in the silkworm Bombyx mori / 药物分析学报

A number of research has shown that the plant polyphenol resveratrol,one of the most prominent small molecules,has beneficial protective effects in multiple organisms,including worms,flies,and killifish.To understand the effects of resveratrol on lifespan,we evaluated its effects in the silkworm Bombyx mori.In this study,we found that lifespan was significantly prolonged in both female and male silkworms treated with resveratrol.Silkworm larval weight was significantly increased from day 3 of the 5th larval instar(L5D3) to day 7 of the 5th larval instar (L5D7).However,the weight of the pupa,cocoon,and total cocoon was not significantly different in female silkworms with resveratrol treatment than that in controls.Meanwhile,resveratrol significantly improved the thermotolerance of the silkworms,which enhanced their survival rate.Moreover,antioxidant activity was increased by resveratrol in both female and male silkworms.Furthermore,an antioxidant-related signalling pathway,SIRT7-FoxO-GST,was activated in silkworms with resveratrol treatment.Collectively,these results help us to understand the molecular pathways underlying resveratrol induced pro-longevity effects and indicate that silkworm is a promising animal model for evaluating the effects of lifespan-extending drugs.

Identification of phosphatidic acid interacting proteins in Ganoderma lingzhi / 生物工程学报

Ganoderma lingzhi is widely recognized as a medicinal basidiomycetes. Triterpene acids (TAs) are the key bioactive medicinal components of G. lingzhi. Our previous studies have shown that phospholipid acid (PA) produced by phospholipase D (PLD) plays a regulatory role in TA synthesis. In order to further elucidate the molecular mechanism how PA regulates TA synthesis in G. lingzhi, PA beads enrichment combined with LC-MS/MS technology was used to identify PA interacting proteins in G. lingzhi. A total of 19 PA interacting proteins were identified, including cytochrome P450 monooxygenase (GL22084), specific protein kinase MAPK (GL23765), catalase and cell surface hydrophobicity-associated protein. GST tagged GL22084 and GL23765 proteins were obtained through gene cloning, heterologous expression, and purification. The interactions between GL22084/GL23765 and PA were verified by GST pull down assay. The identification of PA interacting proteins provides a basis for further understanding the molecular mechanism how PLD-mediated PA signaling molecules regulates the TA synthesis in G. lingzhi. Moreover, the PA interacting proteins identified in this study can also provide clues for the research of PLD/PA signaling pathway in other species.

Relationship of anti-tuberculosis drug-induced liver injury and genetic polymorphisms in CYP2E1 and GST

ABSTRACT Setting: Treatment of tuberculosis (TB) can result in Drug-Induced Liver Injury (DILI) since hepatotoxic metabolites are formed during the biotransformation of isoniazid (INH).DILI can be related to the genetic profile of the patient. Single nucleotide polymorphisms in the CYP2E1 gene and GSTM1 and GSTT1 deletion polymorphisms have been associated with adverse events caused by INH. Objective: To characterize the genetic polymorphisms of CYP2E1, GSTT1 and GSTM1 in TB carriers. Design: This is an observational prospective cohort study of 45 patients undergoing treatment of TB. PCR-RFLP and multiplex-PCR were used. Results: The distribution of genotypic frequency in the promoter region (CYP2E1 gene) was: 98% wild genotype and 2% heterozygous. Intronic region: 78% wild genotype; 20% heterozygous and 2% homozygous variant. GST enzyme genes: 24% Null GSTM1 and 22% Null GSTT1. Patients with any variant allele of the CYP2E1 gene were grouped in the statistical analyses. Conclusion: Patients with the CYP2E1 variant genotype or Null GSTT1 showed higher risk of presenting DILI (p = 0.09; OR: 4.57; 95% CI: 0.75-27.6). Individuals with both genotypes had no increased risk compared to individuals with one genotype.

Expression and receptor binding characteristics of porcine P6 genotype rotavirus GST-VP8*-Z84 protein / 中华实验和临床病毒学杂志

Objective@#The gene engineering technique was used to express the P[6] genotype rotavirus (rotaviruses, RVs) GST-VP8*-Z84 protein from pigs, and the binding characteristics of the protein to oligosaccharide and salivary receptor were studied.@*Methods@#The GST-VP8*-Z84 protein was purified by GST Escherichia coli expression system and affinity chromatography using porcine P[6]. Enzyme-linked immunosorbent assay (ELISA) saliva binding test and oligosaccharide binding test were used to analyze the binding characteristics of the genotype to receptors.@*Results@#Porcine P[6] GST-VP8*-Z84 protein bound well to mucin core 2.@*Conclusions@#The potential receptor of P[6] RV may be the core of mucin, which may provide the experimental basis and theoretical basis for the mechanism of rotavirus and receptor interaction and the development of RV vaccine and highly effective therapeutic drugs.

Detection of Human Anti-Trypanosoma cruzi Antibody with Recombinant Fragmented Ribosomal P Protein

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and is endemic in many Latin American countries. Diagnosis is based on serologic testing and the WHO recommends two or more serological tests for confirmation. Acidic ribosomal P protein of T. cruzi showed strong reactivity against positive sera of patients, and we cloned the protein after fragmenting it to enhance its antigenicity and solubility. Twelve positive sera of Chagas disease patients were reacted with the fragmented ribosomal P protein using western blot. Detection rate and density for each fragment were determined. Fragments F1R1, F1R2, and F2R1 showed 100% rate of detection, and average density scoring of 2.00, 1.67, and 2.42 from a maximum of 3.0, respectively. Therefore, the F2R1 fragment of the ribosomal P protein of T. cruzi could be a promising antigen to use in the diagnosis of Chagas disease in endemic regions with high specificity and sensitivity.

Combination of mass spectrometry and GST pull-down techniques to study potential interacting protein of PCV2 ORF4 / 生物工程学报

A novel protein encoded by the open reading frame 4 (ORF4) was recently discovered in porcine circovirus type 2 (PCV2). However, little is known about the interaction proteins of ORF4 which hindered better understanding the biological functions of ORF4 in the life cycle of PCV2. In the present study, the ORF4 was inserted into the multiple cloning site of pCMV-N-Flag-GST, yielding recombinant plasmid pCMV-N-Flag-GST-ORF4. The recombinant plasmid was transfected into 293T cells and the intracellular interaction complex of ORF4 were enriched and separated by GST pull-down and SDS-PAGE, sequentially. The potential interacting proteins of PCV2 ORF4 were stained with silver and identified by mass spectrometry (MS). Finally, five candidate ORF4-interacting proteins, including Serine/threonine-protein phosphatase 6 catalytic subunit, alpha cardiac muscle 1, actin, SEC14-like protein 5 and myosin 9 were identified. These results would benefit a better understanding of the biological function of ORF4 in PCV2 infected cells.

Water pH and hardness alter ATPases and oxidative stress in the gills and kidney of pacu ( Piaractus mesopotamicus )

This study aimed to assess the effects of low and high water hardness in interaction with different water pH in pacu (Piaractus mesopotamicus). Pacu juveniles were subjected to low (50 mg CaCO3 L-1 - LWH) or high water hardness (120 mg CaCO3 L-1 - HWH) at water pH of 5.5 (acidic), 7.5 (circumneutral) or 9.0 (alkaline) for 15 days. Gills and kidneys were collected (days 1, 5 and 15). Gill Na+/K+-ATPase (NKA) and vacuolar-type H+-ATPase (V-ATPase) activities were higher in alkaline pH with HWH on day 1. Gill and kidney NKA and V-ATPase activities were higher in acidic pH with LWH on day 15. Gill NKA activity of pacus under alkaline pH with LWH was higher than those exposed to HWH. Reduced antioxidant capacity in the gills and kidney and enhanced thiobarbituric acid reactive substances (TBARS) levels were demonstrated in fish exposed to acidic or alkaline pH, mainly with LWH. HWH increased glutathione-S-transferase (GST) activity and reduced TBARS levels in the gills and kidney. On day 15, GST activity was increased at acidic pH with LWH. In conclusion, circumneutral pH presents less oxidative stress and fewer variations in ATPases and HWH reduced deleterious effects in fish exposed to acidic or alkaline pH.(AU)

Este estudo objetivou analisar o efeito de baixa e alta dureza da água em interação com diferentes pH da água em pacu (Piaractus mesopotamicus). Juvenis de pacu foram submetidos a baixa (50 mg CaCO3 L-1 - BDA) ou alta dureza da água (120 mg CaCO3 L-1 - ADA) em pH da água de 5.5 ácido), 7.5 (circum-neutro) ou 9.0 (alcalino) por 15 dias. Foram coletados brânquias e rim (dias 1, 5 e 15). Atividade de Na+/K+-ATPase (NKA) e H+-ATPase do tipo vacuolar (V-ATPase) branquial foram maiores em pH alcalino com ADA no dia 1. Atividade de NKA e V-ATPase branquial e renal foram maiores em pH ácido com BDA no dia 15. Atividade de NKA branquial de pacus submetidos a pH alcalino com BDA foi maior que aqueles expostos para ADA. Em peixes expostos a pH ácido ou alcalino com BDA houve redução da capacidade antioxidante nas brânquias e rim e aumento dos níveis de "substâncias reativas ao ácido tiobarbitúrico" (TBARS). Em ADA aumentou a atividade da "glutationa-S-transferase" (GST) e reduziu níveis de TBARS nas brânquias e rim. No dia 15, a atividade da GST foi maior em pH ácido com BDA. Em conclusão, pH circum-neutro apresentou menor estresse oxidativo e poucas variações na atividade de ATPases e ADA reduziu efeitos deletérios em peixes expostos a pH ácido ou alcalino.(AU)

Association Of Angiotensin-Converting Enzyme (ACE) And Glutathione S-Transferase (GST) Gene Polymorphisms With Diabetic Nephropathy

Diabetic nephropathy accounts for the most serious microvascular complication of diabetes mellitus. It is suggested that the prevalence of diabetic nephropathy will continue to increase in future pretense a major challenge to the healthcare system resulting in increased morbidity and mortality. It occurs as a result of interaction between both genetic and environmental factors in individuals with T2DM-Type 2 diabetes mellitus. Genetic susceptibility has been offered as an important factor for the development of diabetic nephropathy, and various research efforts are being executed worldwide to identify the susceptibility gene for diabetic nephropathy. Several single nucleotide polymorphisms have been found in various genes giving rise to various gene variants which have been found to play a role in genetic susceptibility to diabetic nephropathy. The risk of developing diabetic nephropathy is increased several times by inheriting risk alleles at susceptibility loci of various genes like ACE, GST, TNF-α, COL4A1, eNOS, GLUT, etc. The identification of these genetic variants at a biomarker level could thus, let the detection of those individuals at high risk for diabetic nephropathy which could thus help in the treatment, diagnosis and early prevention of the disease. The present review discusses about the ACE-Angiotensin Converting Enzymeand GST-Glutathione S Transferase gene variants associated with diabetic nephropathy.

Herbal extracts differentially inhibit oxidative effects caused by the biotransformation of nifurtimox, nitrofurantoin and acetaminophen on rat liver microsomes / Extractos herbales inhiben diferencialmente los efectos oxidativos causados por la biotransformación de nifurtimox, nitrofurantoína y acetaminofeno en microsomas hepáticos de rata]

Inflammation is a cellular defensive mechanism associated to oxidative stress. The administration of nitrofurantoin, nifurtimox and acetaminophen generates oxidative stress by their biotransformation through CYP450 system. The main adverse effect described for the first two drugs is gastrointestinal inflammation and that of the last, hepatitis. Therefore, standardised dry extracts from Rosmarinus officinalis, Buddleja globosa Hope, Cynara scolymus L., Echinacea purpurea and Hedera helix were tested to evaluate their capacity to decrease drug-induced oxidative stress. For that, rat liver microsomes were incubated with drugs in the presence of NADPH (specific CYP450 system cofactor) to test oxidative damage on microsomal lipids, thiols, and GST activity. All drugs tested induced oxidation of microsomal lipids and thiols, and inhibition of GST activity. Herbal extracts prevented these phenomena in different extension. These results show that antioxidant phytodrugs previously evaluated could alleviate drugs adverse effects associated to oxidative stress.

Inflamación es un mecanismo de defensa el cual está asociado a estrés oxidativo. La administración de nitrofurantoína, nifurtimox y paracetamol genera estrés oxidativo al metabolizarse a través del sistema CYP450. El principal efecto adverso de los dos primeros fármacos es inflamación gastrointestinal y del tercero, hepatitis. Por lo tanto, utilizamos diversos extractos herbales para disminuir el estrés oxidativo inducido por estos fármacos. Para esto se incubaron microsomas hepáticos de rata con dichos fármacos en presencia de NADPH (cofactor específico del sistema CYP450) y se evaluó el daño oxidativo generado sobre los lípidos, los tioles y la actividad GST microsómica. Todos los fármacos indujeron oxidación de los lípidos y los tioles microsómicos e inhibieron la actividad GST. Los extractos herbales previnieron estos fenómenos oxidativos en diferente extensión. Estos resultados indican que fitofármacos antioxidantes previamente evaluados, podrían aliviar los efectos adversos asociados a estrés oxidativo de los fármacos.

Expression, purification, stability and transduction efficiency of GST-SOD1-R9 fusion protein / 生物工程学报

The fusion of cell permeable peptide TAT and bifunctional antioxidant enzymes, GST (Glutathione sulfur transferase)-TAT-SOD1 (Cu, Zn superoxide dismutase), is an intracellular superoxide scavenger. Compared with SOD1-TAT, GST-TAT-SOD1 has better protective effect on oxidative damage but less transduction efficiency. A novel cell permeable bifunctional antioxidant enzymes with the fusion of GST, SOD1 and polyarginine R9 was constructed for higher transduction efficiency. The full nucleotide sequence of SOD1-R9 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1 with the GST tag. After the successful construction of the prokaryotic expression vectors of GST-SOD1-R9, the recombinant vector was then transformed into Escherichia coli BL21 (DE3) and the GST-SOD1-R9 fusion protein was produced with the induction of IPTG. The soluble expression of GST-SOD1-R9 fusion protein was combining with the induction temperature and time. The best soluble expression was obtained with the induction temperature of 25 ℃ and the induction time of 11 h. The fusion protein was purified through the combination of 80% ammonium sulfate precipitation and affinity chromatography using glutathione agarose, and verified by SDS-PAGE and special enzymatic activity. The thermal and pH stability of GST-SOD1-R9 fusion protein were analyzed and the SOD and GST activity of fusion protein were proved to be well maintained under physiological conditions. Finally, the transduction efficiency of GST-SOD1-R9 fusion protein was proved to be better than GST-TAT-SOD1 fusion protein (P<0.05). These works establish a foundation for further study of the protective effect of GST-SOD1-R9 fusion protein against oxidative damage.

Effects of lymphocyte DNA damage levels in Korean plant food groups and Korean diet regarding to glutathione S-transferase M1 and T1 polymorphisms / 한국영양학회지

PURPOSE: GST (glutathione S-transferase) M1 and T1 gene polymorphisms are known to affect antioxidant levels. This study was carried out to evaluate genetic susceptibility by measuring the effect of DNA damage reduction in the Korean diet by vegetable food according to GST gene polymorphisms using the ex vivo method with human lymphocytes. METHODS: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups. A total of 84 foods, which constituted more than 1% of the total intake in each food group, were finally designated as a vegetable food in the Korean diet. The Korean diet applied in this study is the standard one-week meals for Koreans (2,000 Kcal/day) suggested by the 2010 Dietary Reference Intakes for Koreans. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. RESULTS: In the Korean food group, the DNA damage protective effect of GSTM1 and GSTT1 was found to be greater in mutant type and wild-type, respectively. and the DNA damage protective effect according to the combined genotype of GSTM1 and GSTT1 was different depending on the food group. On the other hand, in Korean Diet, the DNA damage protective effect appeared to be larger in GSTM1 wild-type than in mutant type and was found to not be affected by GSTT1 genotype. CONCLUSION: These results can be used as basic data to demonstrate the superiority of the antioxidant function of Korean dietary patterns and food groups. Furthermore, it may be a starting point to begin research on customized antioxidant nutrition according to individual genes.

Expression and Purification of an N?terminal Fragment of the Cav1.2 Calcium Channel and Characterization of Its Interaction with Calmodulin / 中国医科大学学报

Objective To investigate a method for the purification of the N?terminal peptide fragment(NT)of the myocardial calcium channel Cav1.2,and characterize its interaction with calmodulin(CaM). Methods EscherichiacoliBL?21 cells were transformed with plasmid pGEX?6p?3/NT harboring the NT?GST fusion gene. The cells harboring pGEX?6p?3/NT were cultured and protein expression was induced with isopropyl?β?D?thiogalactoside(IPTG). Then,the GST?NT fusion protein was purified by using glutathione Sepharose 4B(GS?4B)beads. GST was cleaved off with the PreScission protease,and SDS?PAGE was performed to detect the purity and relative molecular weight of the purified peptide. Further, GST pull?down assay was performed to characterize the interaction of the NT peptide with CaM. Results SDS?PAGE analysis showed that the NT peptide was successfully purified,with high purity. Results of the GST pull?down assay showed that the NT peptide could interact with CaM. Conclusion This study establishes a method for the purification of the NT peptide and lays the foundation for further research on the interaction partners and biological functions of NT.

Oxidative stress responses of juvenile tambaqui Colossoma macropomum after short-term anesthesia with benzocaine and MS-222

ABSTRACT The present study aimed to evaluate the effects of benzocaine and tricaine methanesulfonate on oxidative stress parameters of juvenile tambaqui tissues. Fish (n=80) were anesthetized with benzocaine (100 mg L-1) or tricaine (240 mg L-1) and two control groups were used (non-anesthetized fish and fish exposed to ethanol-only). After anesthetic induction 10 fish/anesthetic were euthanized after 3, 12 and 24 hours post-anesthesia and tissue samplings (gills, liver and brain) were performed. Samples were submitted to analyses of enzyme activity glutathione-S-transferase (GST), cellular lipid peroxidation (TBARS) and total antioxidant capacity (ACAP). ACAP increased in gills of benzocaine treatment after 12 hours. The liver showed a reduction in ACAP of tricaine treatment after 12 hours. Both anesthetic treatments showed an increase of ACAP at 24 hours compared to control group. The activity of the GST enzyme increased in the gills for treatments benzocaine and tricaine after 3 and 12 hours. Liver showed increased GST activity (benzocaine after 24 hours and tricaine after 3 and 24 hours). Lipid damage decreased in gills (both anesthetics) and brain (tricaine) after 24 hours. The results demonstrate that benzocaine and tricaine did not cause oxidative damage in juvenile tambaqui under the experimental conditions herein established.

Association of GSTT1 and GSTM1 polymorphisms in South Indian Epilepsy Patients.

Experimental studies suggest that oxidative stress is one of the contributing factors in the onset of epileptic seizures. Glutathione S-transferases (GSTs) are able to conjugate electrophilic compounds, and thus possess neuroprotective role by removing exogenous and endogenous oxidants, detoxifying therapeutic drugs, environmental toxins through conjugation with glutathione (GSH). Several studies from different ethnic groups showed that polymorphisms of the GST gene have been associated with Epilepsy. In the present study, we investigated the association of GST polymorphism in the South Indian epilepsy patients population. A total 371 samples (110 cases and 261 controls) were genotyped for the GST1 and GSTM1 polymorphism by multiplex PCR method. We observed a significant association of GSTT1 null polymorphism in patients with epilepsy. The frequency of the GSTT1 null genotype was found to be significantly higher in cases (35.45 %) than the controls (18.39 %) (OR: 2.44, 95%CI: 1.4-4.02, P <0.0001). In contrast, the frequency of the GSTM1 null variant was significantly lower in cases (11.81%) than controls (32.95%) (OR: 0.27, 95%CI: 0.14-0.51, P <0.001) indicating a protective role. These results indicated that individuals who have GSTT1 null variant are at higher risk for developing seizure than those of GSTT1 wild genotype. On the other hand, individuals carrying GSTM1 null variant showed protective role against seizure. Further, these two null variants did not show any significant association with antiepileptic drug-induced skin rash.