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Objective:To investigate whether endoplasmic reticulum aminopeptidase 1 ( ERAP1) is a susceptible gene for pre-eclampsia (PE) and the possible mechanism in the pathogenesis. Methods:This retrospective study included 990 PE patients (case group) and 1 240 healthy pregnant women (control group) in six prefecture-level tertiary hospitals in Shandong Province, including the Affiliated Hospital of Qingdao University and Zaozhuang Maternal and Child Health Hospital, from September 2018 to April 2021. Peripheral blood were collected for DNA extraction. Single-nucleotide polymorphisms in the ERAP1 gene (rs30187, rs27044, and rs469783 loci) were analyzed by Taqman probe polymerase chain reaction (PCR). Two missense mutant plasmids, rs30187(c.1583A>G) and rs27044(c.2188C>G), were constructed by point mutation induction based on wild-type plasmids. Six groups (knock-down control, knock-down, over-expression control, over-expression, variant 1 and 2 groups) were set up in this study. After transfecting Htr8 cells with different transfection molecules, the expression of ERAP1 at mRNA and protein levels were detected. Besides, the effects of different transfections on cell function were detected using Transwell migration assay, Transwell invasion assay, cell scratch assay, and CCK-8 assay. Statistical analysis was performed using two independent samples t-test, rank sum test, and Chi-square test. Results:(1) There were significant differences in the genetic distribution of rs30187 (Genotype: χ2=29.25, Allele: χ2=4.68) and rs469783 (Genotype: χ2=7.01, Allele: χ2=6.45) as well as the genotype distribution of rs27044 ( χ2=28.95) between the case group and the control group (all P<0.05). Statistical analysis of the genetic model revealed that rs30187 and rs27044, both recessive models, were statistically different between the two groups with a higher frequency of CC genotypes in the case group ( χ2=20.82 and 19.97, both P<0.05), but a lower frequency in CC dominant gene pattern for rs469783 ( χ2=5.82, P=0.016). (2) Compared with the knock-down control group, the knock-down group showed significantly inhibited expression of ERAP1 (mRNA: 0.5±0.1 vs 1.0±0.0, t=7.49; protein: 0.4±0.1 vs 0.7±0.1, t=2.81; both P<0.05), reduced cell migration rate after 48 h of scratching [(16.5%±1.8%) vs (23.8%±2.4%), t=3.33, P=0.031] and decreased number of cells crossing Transwell chambers after 24 h of culture (423.7±21.3 vs 499.0±24.6, t=3.29, P=0.031). Compared with the over-expression group, variant 1 group and variant 2 group showed significantly inhibited expression of ERAP1 at mRNA (both P<0.001) and protein ( P=0.003 and 0.006) levels after transfection, decreased number of cells crossing Transwell chambers ( P=0.001 and 0.032) and down-regulated cell migration rate after 48 h of scratching [variant 1: P=0.004; variant 2: (21.1±4.6)% vs (28.3±1.1)%, t=2.10, P=0.099]. ERAP1 expression at both mRNA ( P<0.001) and protein ( P=0.008) levels, as well as cell proliferation ( P<0.001) and invasion ability ( P<0.001), were all enhanced in the over-expression group than those in the over-expression control group. Moreover, the migration rate of cells after 48 h of scratching ( P=0.002) and the number of cells crossing Transwell chambers after 24 h of culture ( P=0.001) were also increased. Conclusions:The rs30187, rs27044, and rs46978 on ERAP1 gene were all associated with PE susceptibility, with more carriers of the CC genotype in PE patients at rs30187 and rs27044 loci and more carriers of the CC genotype in healthy gravida at rs469783 locus. ERAP1 may be involved in the pathogenesis of PE by affecting the migratory and invasive ability of trophoblast cells.
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La insuficiencia cardíaca (IC) se define como la incapacidad del corazón de satisfacer los requerimientos metabólicos de los tejidos en reposo o durante el ejercicio ligero. Esta incapacidad motiva una respuesta neurohormonal que se interrelaciona con las alteraciones hemodinámicas vinculadas a las cargas ventriculares, más los problemas funcionales y estructurales del miocardio que puedan existir. En la fisiopatología cardiovascular (de la IC, de la hipertensión arterial, de las valvulopatías, de la enfermedad coronaria, etc.), tiene participación clave el sistema renina angiotensina (SRA), cuyas acciones principales incluyen la regulación de la presión arterial, el tono vascular, la volemia y facilitar la transmisión simpática. El SRA participa en la remodelación ventricular del infartado y del hipertenso, así como en la remodelación vascular. Esta revisión sobre algunos nuevos aspectos del SRA, publicados recientemente en la literatura médica, se acompaña de un resumen de los conceptos clásicos, para ver cómo los nuevos se instalan en ellos, se abren nuevos caminos en la investigación fisiológica y farmacológica, al ampliarse significativamente el espectro de acción del SRA, eje de la fisiopatología de la hipertensión arterial, de la insuficiencia cardíaca y de otras patologías.
Heart failure (HF) is defined as the inability of the heart to satisfy the metabolic requirements of the tissues at rest or during light exercise. This failure motivates neurohormonal response that interrelates with the hemodynamic changes related to ventricular loads more functional and myocardial structural problems that may exist. In cardiovascular pathophysiology (HF, arterial hypertension, valvular heart disease, coronary disease, etc.) has key role the renin angiotensin system (RAS), whose main activities include to regulate blood pressure, vascular tone, volemia and to facilitate sympathetic transmission. The RAS participates in ventricular remodeling of the infarcted and of the hypertensive as well as vascular remodeling. This review of some new aspects of the RAS, recently published in the medical literature, is accompanied by a summary of the classical concepts, to see how new they are installed, opening new pathways in physiological and pharmacological research, to significantly expand the RAS spectrum of action, axis of the pathophysiology of hypertension, heart failure and other diseases.
A insuficiência cardíaca (IC) é definida como a incapacidade do coração para atender às necessidades metabólicas dos tecidos em repouso ou durante exercícios leves. Esta falha motiva resposta neuro-hormonal que interage com as alterações hemodinâmicas relacionadas às cargas ventriculares mais os problemas funcionais e estruturais do miocárdio que possam existir. Na fisiopatologia cardiovascular (IC, hipertensão arterial, doença cardíaca valvular, doença coronariana, etc.) tem papel fundamental no sistema renina angiotensina (SRA), cujas ações principais incluem a regulação da pressão arterial, do tônus vascular, da volemia e facilitar a transmissão simpática. O SRA participa de remodelação ventricular do infartado e do hipertenso assim como a remodelação vascular. Esta revisão de alguns novos aspectos da SRA, recentemente publicado na literatura médica, é acompanhada por um resumo dos conceitos clássicos, para ver como novo eles estão instalados, abrindo novos caminhos na pesquisa fisiológica e farmacológica, para expandir de forma significativa a espectro de ação da SRA, eixo da fisiopatologia da hipertensão, insuficiência cardíaca e outras doenças.
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The proteolytic system of lactic acid bacteria contribute to the development of flavor during the ripening of cheese through the generation of short peptides and free amino acids, which directly or indirectly act as flavor precursors. Newly isolated lactic acid bacteria (LAB) as well as those procured from culture collection centers were screened for the production of various substrate specific aminopeptidases. Among all the strains screened, L. brevis (NRRL B-1836) was found to produce quantifiable amount of intracellular arginine specific aminopeptidase (EC 3.4.11.6). The productivity of arginine aminopeptidase in 5 L fermentor was 36 IU/L/h. The Luedeking and Piret model was tested for intracellular production of aminopeptidase and the data seemed to fit well, as the correlation coefficient was 0.9964 for MRS. The αAP and βAP was 0.4865 and 0.0046, respectively in MRS medium indicating that the yield was predominantly depended on growth. The culture produced lactic acid and also tolerated pH 2.0-3.0 and 0.3-0.5 percent bile salts, the most important probiotic features.
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The proteolytic system of lactic acid bacteria contribute to the development of flavor during the ripening of cheese through the generation of short peptides and free amino acids, which directly or indirectly act as flavor precursors. Newly isolated lactic acid bacteria (LAB) as well as those procured from culture collection centers were screened for the production of various substrate specific aminopeptidases. Among all the strains screened, L. brevis (NRRL B-1836) was found to produce quantifiable amount of intracellular arginine specific aminopeptidase (EC 3.4.11.6). The productivity of arginine aminopeptidase in 5 L fermentor was 36 IU/L/h. The Luedeking and Piret model was tested for intracellular production of aminopeptidase and the data seemed to fit well, as the correlation coefficient was 0.9964 for MRS. The αAP and βAP was 0.4865 and 0.0046, respectively in MRS medium indicating that the yield was predominantly depended on growth. The culture produced lactic acid and also tolerated pH 2.0-3.0 and 0.3-0.5 percent bile salts, the most important probiotic features.
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Objective To detect the genetic transcription and protein expression level of endoplasmic reticulum aminopeptidase 1 (ERAP1) in human ovarian cancer cells and tissue and study their relationship with directional lymphatic metastasis. Methods Real-time fluorescent quantitative PCR and western blot were used to determine the expressions of ERAP1 gene and protein in the human ovarian cancer cell lines between non-directional (SKOV3) and directional highly lymphatic metastasis (SKOV3-pm2, SKOV3-pm3, SKOV3-pm4 ). Immunohistochemistry method was used to further validate the ERAP1 expressions of the transplanted ovarian tumor primarily focus and the lesions of lymph node metastasis from nude mice and the human ovarian cancer primarily focus and the lesions of lymph node metastasis. Results The expression level of ERAP1 gene and protein were down-regulated in SKOV3, SKOV3-pm2, SKOV3-pro3, SKOV3-pm4 cell sublines (0.118±0.012, 0.031±0.003,0.028±0.003, 0.016±0.005 ; 0.91± 0.33, 0.09±0.03, 0.10±0.04, 0.05+0.04; respectively), and the level of ERAP1 in SKOV3 cell was higher than those in the other three kinds of cell lines (P<0.05 ). The results showed that there were significant declining trend of expression of ERAP1 in the human ovarian cancer cell lines between non-directional and directional highly lymphatic metastasis; the transplanted ovarian tumor primarily focus and the metastasis lesions of lymph node from nude mice (143±22 vs. 97±12, P<0.05), the primarily focus (184±14) and the lesions of lymph node metastasis from human ovarian tumors (P<0.05). Conclusion The absence or down-regulated expression of ERAP1 is closely related to the metastasis and invasion of lymph node in ovarian carcinoma.
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Objective To study the activity of aminopeptidase N (APN) in the bile of patients with cholesterol gallstones (CGS) and rabbits fed with lithogenic diets (LD) to explore the lithogenic role of APN in the formation of gallstones. Methods Enzymatic activity of APN and lipid concentrations in bile of investigated CGS patients and LD fed rabbits were measured. The relationship between the activity of APN and CSI was statistically analyzed.Results APN activity in CGS patients′ gallbladder (GB) and bile duct (BD) biles 〔(66 0?24 1)*!U/ml and (38 4?9 4)*!U/ml〕 as well as in LD fed rabbits′ GB and BD biles 〔(14 5?4 08)*!U/ml and (9 13?1 67)*!U/ml〕 were significantly higher than that in the corresponding biles of their clinical and experimental control (CC and EC) groups 〔(23 0?7 7)*!U/ml〕 in GB bile of CC group and 〔(6 05?0 95)*!U/ml and (3 09?0 92)*!U/ml〕 in EC group. All P values were less than 0 05. A statistical positive correlation was found between the APN and CSI value in each of the groups.Conclusions It is more likely that APN might play certain role in the early neucleating period of the pathogenesis of gallstones based on its proneucleative potential and measurement of APN activity may serve as a predictive marker among patients with high risk of gallstone formation.