Efeito Anti-Hipertensivo de Novos Agonistas do Receptor de Adenosina em Ratos Espontaneamente Hipertensos / Antihypertensive Effect of New Agonist of Adenosine Receptor in Spontaneously Hypertensive Rats

Resumo Fundamento A hipertensão arterial sistêmica é um fator de risco para disfunções cardíacas, renais e metabólicas. A busca por novas estratégias para prevenir e tratar doenças cardiovasculares levou à síntese de novas N-acilidrazonas para produzir efeito anti-hipertensivo. Os receptores de adenosina são um alvo alternativo para reduzir a pressão arterial devido à sua ação vasodilatadora e propriedades antioxidantes, que podem reduzir o estresse oxidativo característico da hipertensão arterial sistêmica. Objetivo Avaliar o perfil anti-hipertensivo de novos compostos contendo selênio desenvolvidos para melhorar sua interação com os receptores de adenosina. Métodos Foi avaliada a reatividade vascular, registrando-se a tensão isométrica da aorta torácica pré-contraída de ratos Wistar machos após exposição a concentrações crescentes de cada derivado (0,1 a 100 μM). Para investigar o efeito anti-hipertensivo em ratos espontaneamente hipertensos, foram determinadas a pressão arterial sistólica, pressão arterial diastólica, pressão arterial média e a frequência cardíaca após administração intravenosa de 10 e 30 μmol/kg do composto selecionado LASSBio-2062. Resultados Os compostos denominados LASSBio-2062, LASSBio-2063, LASSBio-2075, LASSBio-2076, LASSBio-2084, LASSBio-430, LASSBio-2092 e LASSBio-2093 promoveram vasodilatação com concentrações efetivas médias de 15,5 ± 6,5; 14,6 ± 2,9; 18,7 ± 9,6; 6,7 ± 4,1; > 100; 6,0 ± 3,6; 37,8 ± 11,8; e 15,9 ± 5,7 μM, respectivamente. O LASSBio-2062 (30 μmol/kg) reduziu a pressão arterial média em ratos espontaneamente hipertensos de 124,6 ± 8,6 para 72,0 ± 12,3 mmHg (p < 0,05). A ativação do receptor de adenosina subtipo A3 e dos canais de potássio parece estar envolvida no efeito anti-hipertensivo do LASSBio-2062. Conclusões O novo agonista do receptor de adenosina e ativador dos canais de potássio é um potencial agente terapêutico para o tratamento da hipertensão arterial sistêmica.

Abstract Background Systemic arterial hypertension is a risk factor for cardiac, renal, and metabolic dysfunction. The search for new strategies to prevent and treat cardiovascular diseases led to the synthesis of new N-acylhydrazones to produce antihypertensive effect. Adenosine receptors are an alternative target to reduce blood pressure because of their vasodilatory action and antioxidant properties, which may reduce oxidative stress characteristic of systemic arterial hypertension. Objective To evaluate the antihypertensive profile of novel selenium-containing compounds designed to improve their interaction with adenosine receptors. Methods Vascular reactivity was evaluated by recording the isometric tension of pre-contracted thoracic aorta of male Wistar rats after exposure to increasing concentrations of each derivative (0.1 to 100 μM). To investigate the antihypertensive effect in spontaneously hypertensive rats, systolic, diastolic, and mean arterial pressure and heart rate were determined after intravenous administration of 10 and 30 μmol/kg of the selected compound LASSBio-2062. Results Compounds named LASSBio-2062, LASSBio-2063, LASSBio-2075, LASSBio-2076, LASSBio-2084, LASSBio-430, LASSBio-2092, and LASSBio-2093 promoted vasodilation with mean effective concentrations of 15.5 ± 6.5; 14.6 ± 2.9; 18.7 ± 9.6; 6.7 ± 4.1; > 100; 6.0 ± 3.6; 37.8 ± 11.8; and 15.9 ± 5.7 μM, respectively. LASSBio-2062 (30 μmol/kg) reduced mean arterial pressure in spontaneously hypertensive rats from 124.6 ± 8.6 to 72.0 ± 12.3 mmHg (p < 0.05). Activation of adenosine receptor subtype A3 and potassium channels seem to be involved in the antihypertensive effect of LASSBio-2062. Conclusions The new agonist of adenosine receptor and activator of potassium channels is a potential therapeutic agent to treat systemic arterial hypertension.

Characteristics of tumor patients with anti-P1 antibody: a retrospective study / 中国输血杂志

【Objective】 To evaluate the detection and distribution characteristics of anti-P1 in tumor patients, so as to aid in blood screening and transfusion safety. 【Methods】 The clinical data of 112 658 tumor patients who underwent blood preparation and transfusion in our hospital from January 2014 to December 2021 were retrospectively analyzed, and column agglutination technique was used to perform transfusion compatibility test. 【Results】 A total of 1 079 (0.96%, 1 079/112 658) cases were detected with unexpected antibodies, of which 71 (6.58%, 71/1 079) were identified as anti-P1. In anti-P1 cases, 59.15% (42/71) were males; 60.56% had no pregnancy history (P<0.01); 29.58% (21/71), 52.11%(37/71), 12.68%(9/71) and 5.63%(4/71) of anti-P1 patients were with type A, B, O and AB, respectively. 57 cases of anti-P1 patients (80.28%) had difficulty in ABO blood group identification. The incidence of interfering in patients with type B was higher than that of other blood types (P<0.05), as the frequency of w+ in reverse blood typing was higher than other reactive patterns (P<0.05). The incidence of gastric tumor and brain space-occupying lesion in patients with anti-P1 was higher than that in patients with other alloantibodies, while the incidence of gynecological tumors was lower (P<0.05). 【Conclusion】 Anti-P1 affects the ABO blood group identification of tumor patients, and most of them had difficulty in ABO blood group identification. Compared with patients with other alloantibodies, patients with anti-P1 are more likely to be male and suffer from gastric and brain tumors, but less likely from gynecological tumors.

Dickkopf-1 inhibits the secretion of MUC5AC induced by Mycoplasma pneumoniae P1-C in mouse lung epithelial cells / 生物工程学报

Mycoplasma pneumoniae is the most common pathogen of respiratory tract infection in children and adults. Clinical observation shows that M. pneumoniae infection can cause massive mucus secretion in the respiratory tract, which makes the breathing of patients difficult. Studies have shown that M. pneumoniae infection can cause massive secretion of mucin 5AC (MUC5AC). Adhesin P1 plays an important role in the pathogenesis of M. pneumoniae infection by mediating the adhesion of pathogens to host cells, and the C-terminal residues of P1 (P1-C) are immunogenic. This study investigated the molecular mechanism of Wnt/β-catenin signaling pathway inhibitor Dickkopf-1 (DKK1) in the secretion of MUC5AC in mouse airway epithelial cells (MAECs) induced by P1-C. Scanning electron microscope and hematoxylin-eosin staining were used to observe the effect of P1-C on mucus secretion of MAECs. Protein chip was used to detect the secretion of cytokines and analyse the enrichment of related signaling pathways induced by P1-C in MAECs. Periodic acid schiff stain (PAS) staining, Tunel staining and Masson staining were used to detect the damage of the lungs of mouse exposed to P1-C. Immunohistochemistry was used to detect the secretion of MUC5AC expression, and Western blotting was used to reveal the molecular mechanism of DKK1-regulated secretion of MUC5AC induced by P1-C protein in MACES. The results showed that P1-C induced the massive secretion of mucus and inflammatory factors in MAECs. During P1-C infection, DKK1 down-regulated janus kinase 2 (JAK2), phosphorylation signaling and transcription activator 1 (p-STAT1) and phosphorylation signaling and activator of transcription 3 (p-STAT3) expression. Overexpression of DKK1 significantly up-regulated the expression of MUC5AC repressor transcription factor fork-head box protein A2 (FOXA2). At the same time, the expression of MUC5AC induced by P1-C was inhibited significantly. It is speculated that DKK1 can effectively reduce the secretion of MUC5AC in MAECs induced by P1-C by inhibiting the JAK/STAT1-STAT3 signaling pathway and up-regulating the expression of FOXA2.

Galvanic vestibular evoked myogenic potentials: normative data and the effect of age / Potenciais evocados miogênicos vestibulares galvânicos: dados normativos e o efeito da idade

Abstract Introduction: Galvanic vestibular evoked myogenic potentials evaluate vestibular nerve responses using electric stimulation by records collected from the sternocleidomastoid muscle. A normal vestibular evoked myogenic potential response consists of the first positive, P1, and negative, N1, peaks. The response can be affected by factors such as age and gender and is also consequential in the diagnosis of pathologies. Objectives: The present study was performed to obtain normative data on healthy adults, to help in diagnosis by establishing clinical norms as well as to investigate changing test parameters with age in galvanic vestibular evoked myogenic potentials. Methods: A total of 100 healthy participants were included in the study. Galvanic vestibular evoked myogenic potential (current 3 mA, duration 1ms) was performed randomly on both ears of each participant. The participants between the ages of 18-65 (mean age 39.7 ± 13.9) were divided into 5 groups according to their ages. Normative data of galvanic vestibular evoked myogenic potentials parameters were calculated in groups and in total, and age-related changes were examined. Results: The galvanic vestibular evoked myogenic potential waveform was elicited from all participants (200 ears). The latency of P1 and N1 was 7.82 ± 3.29ms and 22.06 ± 3.95 ms, respectively. The P1-N1 amplitude value was 66.64 ± 24.5 μV. The percentage of vestibular asymmetry was 16.29 ±11.99%. The latencies of P1 and N1 and P1-N1 amplitude values demonstrated significant differences among different age groups (p < 0.01). Conclusions: The results of this study show that as age increased, latencies were prolonged, and amplitudes gradually decreased. The normative data aids in the diagnosis of retrolabyrinthine lesions and the increase in the clinical use of galvanic vestibular evoked myogenic potentials.

Resumo Introdução: Os potenciais evocados miogênicos vestibulares galvânicos avaliam as respostas do nervo vestibular com estimulação elétrica por meio de registros coletados do músculo esternocleidomastóideo. Uma resposta normal de potenciais evocados miogênicos vestibulares consiste nos primeiros picos positivo, P1, e negativo, N1. A resposta pode ser afetada por fatores como idade e sexo e também tem importância no diagnóstico de doenças. Objetivos: Obter dados normativos em adultos saudáveis, para ajudar no diagnóstico através do estabelecimento de normas clínicas, e investigar a alteração dos parâmetros de teste com a idade em potenciais evocados miogênicos vestibulares galvânicos. Método: Foram incluídos no estudo 100 participantes saudáveis. O potencial evocado miogênico vestibular galvânico (corrente 3mA, duração 1ms) foi realizado de forma aleatória nas duas orelhas de cada participante. Os participantes entre 18 e 65 anos (média de 39,7 ±13,9) foram divididos em 5 grupos de acordo com a idade. Os dados normativos dos parâmetros dos potenciais evocados miogênicos vestibulares galvânicos foram calculados nos grupos e no total e as alterações relacionadas à idade foram examinadas. Resultados: A forma de onda do potencial evocado miogênico vestibular galvânico foi obtida de todos os participantes (200 orelhas). A latência de P1 e N1 foi de 7,82±3,29ms e 22,06 ±3,95 ms, respectivamente. O valor da amplitude P1-N1 foi de 66,64 ±24,5 μV. O percentual de assimetria vestibular foi de 16,29± 11,99%. Os valores das latências de P1 e N1 e da amplitude P1-N1 mostraram diferenças significantes entre os diferentes grupos etários (p < 0,01). Conclusão: Os resultados deste estudo mostram que à medida que a idade aumentou as latências foram prolongadas e as amplitudes diminuíram gradualmente. Os dados normativos auxiliam no diagnóstico de lesões retrolabirínticas e na disseminação do uso clínico dos potenciais evocados miogênicos vestibulares galvânicos.

Clinicopathological and prognostic significance of expression of forkhead box P1 in patients with intrahepatic cholangiocarcinoma / 中华肝胆外科杂志

Objective:To study the expression of forkhead box P1 (FOXP1) in intrahepatic cholangiocarcinoma (ICC), and its clinicopathological and prognostic significance.Methods:The clinical data of ICC patients treated with radical resection at Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 1, 2013 to December 12, 2019 were retrospectively analysed. Of 48 ICC patients, there were 24 males and 24 females, with age of (59.1±10.1) years old (range 42 to 83 years old). Their clinicopathological data, including age, gender, tumor size, degree of differentiation, and staging were recorded. Immunohistochemical method was used to detect the expression of FOXP1 protein in ICC cancer tissues and the corresponding adjacent normal tissues. Kaplan-Meier method was used to calculate survival rates and to construct survival curves of patients. Cox regression model was used to analyze factors affecting prognosis of patients.Results:Forty-eight ICC cancer tissues and 40 corresponding paracancerous tissues were collected. The positive rates of FOXP1 proteins in ICC were significantly lower than the adjacent normal tissues [54.2%(26/48) vs. 92.5%(37/40), χ 2=15.76, P<0.05]. The degrees of tumor differentiation, lymph node metastasis, organ invasion and TNM staging were related to expression of FOXP1 ( P<0.05). Forty-two patients were followed-up with a median follow-up time of 11.5 (7.75, 19.25) months. Cox multivariate analysis revealed that invasion to adjacent organs, lymph node metastasis, high TNM staging (stage Ⅲ) and negative expression of FOXP1 were independent risk factors affecting overall survival of ICC patients. The overall survival and recurrence-free survival of FOXP1-positive ICC patients were 17.5 months and 15.5 months, which were significantly higher than the 14.0 months and 11.1 months, respectively, in FOXP1-negative patients. Conclusion:Negative FOXP1 expression was closely correlated with aggressive biological behavior and poor prognosis of ICC. FOXP1 may be used as new diagnostic and therapeutic targets.

Efficacy of anti-CD 20 monoclonal antibody in combination with Gemcitabine-based chemotherapy regimen in children with recurrent diffuse large B-cell lymphoma / 中华实用儿科临床杂志

Objective:To investigate the efficacy of anti-CD 20 monoclonal antibody in combination with Gemcitabine-based chemotherapy regimen in children with recurrent diffuse large B-cell lymphoma (DLBCL). Methods:The clinical data of 62 children with DLBCL admitted to the Department of Pediatrics, Yantai Mountain Hospital of Yantai City and the Department of Pediatrics, Muping District Traditional Chinese Medicine Hospital in Yantai City from January 2010 to January 2018 were analyzed retrospectively.Different treatment options were selected according to the children′s stage and the presence of risk factors such as huge tumors.Among them, 32 cases in the control group were treated with the Gemcitabine-based treatment plan (Gemcitabine+ Cisplatin+ Dexamethasone treatment). Thirty patients in the study group were treated with anti-CD 20 monoclonal antibody on the basis of the control group for a total of 4 cycles (21 d per cycle). After 4 cycles of treatment, the clinical efficacy, the positive expression of forkhead box protein P1 (FOXP1) and B-cell lymphoma factor-6 (Bcl-6) before and after treatment, the occurrence of adverse reactions and survival[3-year progression-free survival (PFS), 3-year overall survival (OS)] were evaluated.The measurement data that meets the normal distribution is expressed that the t test is used, and the counting data is represented by (%), and the χ2 test is used.Level data is compared with ranking and inspection. Results:All patients were followed up to March 2021.The total response rate (RR) and disease control rate (DCR) of the study group were 93.33% (28/30 cases) and 96.67% (29/30 cases), respectively.The RR and DCR of the control group were 68.75% (22/32 cases) and 81.25% (26/32 cases), respectively.The RR of the study group was higher than that of the control group ( χ2=5.995, P<0.05), and there was no statistical difference in DCR between the two groups ( χ2=3.674, P>0.05). After treatment, the positive expression rate of FOXP1 in the study group and the control group was lower than before treatment[23.33% (7/30 cases) vs. 76.67% (23/30 cases); 50.00% (16/32 cases) vs. 75.00% (24/32 cases), χ2=17.067, 4.267, all P<0.05], and the positive expression rate of the study group was lower than that of the control group ( χ2=4.179, P<0.05). After treatment, the positive expression rate of Bcl-6 in the study group and the control group was higher than before treatment[86.67% (26/30 cases) vs. 26.67% (8/30 cases); 62.50% (20/32 cases) vs. 31.25% (10/32 cases), χ2=21.991, 6.275, all P<0.05], and the positive expression rate of the study group was higher than that of the control group ( χ2=4.723, P<0.05). There was no significant difference between the study group and the control group in the level of gastrointestinal reactions, elevated transa-minase, and decreased white blood cell ( Z=-1.074, -1.078, -0.834, all P>0.05). There was a difference between the 3-year PFS survival curve and the 3-year OS survival curve between the two groups ( χ2=3.997, 4.723, all P<0.05). Conclusions:Anti-CD 20 monoclonal antibody combined with Gemcitabine-based chemotherapy is effective for children with DLBCL, and will not significantly increase the adverse reactions in children.

Exploration of IRES Elements within the ORF of the Coxsackievirus B3 Genome / 生物医学与环境科学（英文）

Objective@#This study aimed to identify internal ribosome entry sites (IRESs) in the open reading frame (ORF) of the Coxsackievirus B3 (CVB3) genome.@*Methods@#The sequences of P1, P2, or P3 of the CVB3 genome or the truncated sequences from each antithymocyte globulin (ATG) to the end of the P1, P2, or P3 gene were inserted into the pEGFP-N1 vector. After transfection, possible IRES-dependent green fluorescent protein (GFP)-fused proteins were detected by anti-GFP western blotting. The sequences of possible IRESs were inserted into specific Fluc/Rluc bicistronic vectors, in which the potential IRESs were determined according to the Fluc/Rluc activity ratio. Expression of Fluc and Rluc mRNA of the bicistronic vector was detected by RT-qPCR.@*Results@#After transfection of full length or truncated sequences of the P1, P2, or P3 plasmids, six GFP-fused protein bands in P1, six bands in P2 and nine bands in P3 were detected through western blotting. Two IRESs in VP2 (1461-1646 nt) and VP1 (2784-2983 nt) of P1; one IRES in 2C (4119-4564 nt) of P2; and two IRESs in 3C (5634-5834 nt) and 3D (6870-7087 nt) of P3 were identified according to Fluc/Rluc activity ratio. The cryptic promoter was also excluded by RT-qPCR.@*Conclusion@#Five IRESs are present in the CVB3 coding region.

Distribution of ABO, Rh, Mn, P, h blood groups and gene frequency in Lahu ethnic minority in Yunnan / 中国输血杂志

【Objective】 To understand the distribution and gene frequency of main red blood cell blood groups in Lahu ethnic minority and analyze the genetic characteristics of Lahu people. 【Methods】 1) ABO forward and reverse typing had been performed by microplate method; 2) Rh, MN, H, P1Pk and Mur antigen were tested by the tube method. If the ABO forward and reverse typing were incompatible, the tube method was used for confirmation. 【Results】 The distribution characteristics of blood group and gene frequency in Lahu ethnic minority were as follows: B>O>A>AB for ABO, with genotype frequency as p 11.1%, q 27.5% and r 61.4%; the frequency of Rh genotype was CDe 83.3%, cDE 12.0%, cDe 2.42%, CDE 2.32%, CdE 0%, Cde 0%, cdE 0% and cde 0%; M > MN>N for MN blood group, with genotype frequency as M 75.26% and N 24.74%; P1

Expression of antimicrobial peptide Cecropin P1 in Saccharomyces cerevisiae and its antibacterial and antiviral activity in vitro

BACKGROUND: Cecropin P1, acting as an antimicrobial, has a broad-spectrum antibacterial activity with some antiviral and antifungal properties. It is a promising natural alternative to antibiotics which is originally isolated from the pig intestinal parasitic nematode Ascaris suum. Many studies have shown that Cecropin P1 is helpful for the prevention or treatment of clinical diseases. Therefore, it is very necessary to establish a safe, nontoxic, and efficient expression method of Cecropin P1. RESULTS: The results indicated that the recombinant protein was about 5.5 kDa showed by Tricine­SDS­ PAGE and Western blot. And Cecropin P1 was efficiently secreted and expressed after 12 h of induction, with an increasing yield over the course of the induction. Its maximum concentration was 7.83 mg/L after concentration and purification. In addition, in vitro experiments demonstrated that Cecropin P1 not only exerted a strong inhibitory effect on Escherichia coli, Salmonella sp., Shigella sp., and Pasteurella sp., but also displayed an antiviral activity against PRRSV NADC30-Like strain. CONCLUSIONS: Collectively, the strategy of expressing Cecropin P1 in Saccharomyces cerevisiae is harmless, efficient, and safe for cells. In addition, the expressed Cecropin P1 has antiviral and antibacterial properties concurrently.

Progress of molecular epidemiology of mycoplasma pneumoniae in children / 国际儿科学杂志

Mycoplasma pneumoniae(MP)is the main pathogen causing community-acquired pneumonia in children, usually treated with macrolide drugs.The type of MP genes is mainly based on PCR-RFLP(P1-restriction fragment length polymorphism analysis)and MLVA(multiple locus variable number tandem repeat analysis)typing methods.During epidemics, MP subtypes will undergo certain transformation.Studying and mastering the prevalence characteristics and transformation laws of MP subtypes can deeper understand the distribution area, prevalence year and clinical relevance of each subtype of MP.Due to the extensive use of antibiotics, the resistance of mycoplasma pneumoniae to macrolides has increased , which has become a global public health concern.Studies have shown that MP resistance is related to mutations in the V region of 23S rRNA gene domain.The improvement of typing technology also guides significance for the rational selection of antibiotics.It is imperative to carry out systematic and comprehensive molecular epidemiological studies of MP genotypes and its resistance mutations, and reveal the distribution characteristics, epidemic trends and resistance mutations of MP subtypes at the molecular level.This paper reviews the research progress of molecular epidemiology of mycoplasma pneumoniae in children, and provides ideas for the monitoring, prevention and clinical treatment of MP infection.

Advances in plant heavy metal transporter P1B-ATPases / 生物工程学报

P1B-ATPases are a group of proteins that can transport heavy metal ions across membranes by hydrolyzing ATP and they are a subclass of the P-type ATPase family. It was found that P1B-ATPases are mainly responsible for the active transport of heavy metal ions in plants and play an important role in the regulation of heavy metal homeostasis in plants. In this paper, we dissusses the mechanism of P1B-ATPases from the structure and classification of P1B-ATPases, and review the current research progress in the function of P1B-ATPases, in order to provide reference for future research and application of P1B-ATPases in improving crop quality and ecological environment management.

hsa-miR-124-3p.1 inhibits the migration and invasion of human gastric cancer cells by targeting TRAF6 / 西安交通大学学报(医学版)

【Objective】 To observe the effects of hsa-miR-124-3p.1 in inhibiting epithelial-mesenchymal transition （EMT）， migration and invasion of human gastric cancer cells induced by transforming growth factor β1 （TGF-β1） by targeting tumor necrosis factor receptor-associated factor 6 （TRAF6）. 【Methods】 A total of 43 gastric cancer tissues and 43 normal para-carcinoma tissues were collected. The human gastric mucosal epithelial cells GES-1 and gastric cancer cells （NCI-N87， MGC-803， BGC-823， SGC-7901， and MKN-45） were cultured. The expressions of miR-124-3p.1 and TRAF6 in tissues and cells were detected by fluorescent quantitative PCR and Western blotting. The targeted relationship between miR-124-3p.1 and TRAF6 was verified by dual-luciferase reporter gene system assay. SGC-7901 cell lines with miR-124-3p.1 and TRAF6 overexpression were constructed. The cells were induced by TGF-β1. The invasion and migration abilities of the cells were evaluated by Transwell chamber assay and scratch test. 【Results】 Compared with normal para-carcinoma tissues and normal gastric mucosal cells， the expression of miR-124-3p.1 was downregulated， while the expressions of TRAF6 mRNA and protein were upregulated in gastric cancer tissues and cells （P<0.05）. Compared with control group， expression of E-cadherin in cells was downregulated， expressions of N-cadherin and Vimentin were upregulated， invasion and migration rates of cells were increased in TGF-β1 group （P<0.05）. Compared with TGF-β1 group， after cells were transfected with miR-124-3p.1 mimic， the expression of E-cadherin was upregulated， the expressions of N-cadherin and Vimentin were down-regulated， and invasion and migration rates of cells were decreased （P<0.05）. Compared with miR-124-3p.1 mimic group， invasion and migration rates of cells were increased in TGF-β1+mimic+TRAF6 group， expressions of TRAF6， N-cadherin and Vimentin were up-regulated， and the expression of E-cadherin was down-regulated （P<0.05）. 【Conclusion】 hsa-miR-124-3p.1 is lowly expressed in gastric cancer. Overexpression of miR-124-33p.1 can inhibit EMT， cell invasion and migration induced by TGF-β1. And the action mechanism may be related to the downregulated expression of TRAF6.

Toxicological effects of bioactive peptide fractions obtained from Bothrops jararaca snake venom on the structure and function of mouse seminiferous epithelium

Pathogenesis of Bothrops envenomations is complex and despite numerous studies on the effects of this snake venom on various biological systems, relatively little is known about such effects on the male reproductive system. In the present study, the toxicological outcomes of the low molecular weight fraction (LMWF) of B. jararaca snake venom - containing a range of bioactive peptides - were investigated on the dynamics and structure of the seminiferous epithelium and 15P-1 Sertoli cells viability. Methods: LMWF (5 µg/dose per testis) venom was administered in male Swiss mice by intratesticular (i.t.) injection. Seven days after this procedure, the testes were collected for morphological and morphometric evaluation, distribution of claudin-1 in the seminiferous epithelium by immunohistochemical analyses of testes, and the nitric oxide (NO) levels were evaluated in the total extract of the testis protein. In addition, the toxicological effects of LMWF and crude venom (CV) were analyzed on the 15P-1 Sertoli cell culture. Results: LMWF induced changes in the structure and function of the seminiferous epithelium without altering claudin-1 distribution. LMWF effects were characterized especially by lost cells in the adluminal compartment of epithelium (spermatocytes in pachytene, preleptotene spermatocytes, zygotene spermatocytes, and round spermatid) and different stages of the seminiferous epithelium cycle. LMWF also increased the NO levels in the total extract of the testis protein and was not cytotoxic in concentrations and time tested in the present study. However, CV showed cytotoxicity at 10 μg/mL from 6 to 48 h of treatment. Conclusions: The major finding of the present study was that the LMWF inhibited spermatozoa production; principally in the spermiogenesis stage without altering claudin-1 distribution in the basal compartment. Moreover, NO increased by LMWF induce open of complexes junctions and release the germ cells of the adluminal compartment to the seminiferous tubule.(AU)

Quantitation of DNA by nuclease P1 digestion and UPLC-MS/MS to assess binding efficiency of pyrrolobenzodiazepine / 药物分析学报

Accurate DNA quantitation is a prerequisite in many biomedical and pharmaceutical studies. Here we established a new DNA quantitation method by nuclease P1 digestion and UPLC-MS/MS analysis. DNA fragments can be efficiently hydrolyzed to single deoxyribonucleotides by nuclease P1 in a short time. The decent stabilities of all the four deoxyribonucleotides were confirmed under different conditions. Deoxyadenosine monophosphate (dAMP) was selected as the surrogate for DNA quantitation because dAMP showed the highest sensitivity among the four deoxyribonucleotides in the UPLC-MS/MS analysis. The linear range in DNA quantitation by this method is 1.2-5000 ng/mL. In the validation, the inter-day and intra-day accuracies were within 90％-110％, and the inter-day and intra-day precision were acceptable (RSD<10％). The validated method was successfully applied to quantitate DNA isolated from tumors and organs of a mouse xenograft model. Compared to the quantitation methods using UV absorbance, the reported method provides an enhanced sensitivity, and it allows for the accurate quantitation of isolated DNA with contamination of RNA and ribonucleotide.

EffectofGSTP1geneticvariationontherecurrenceriskandprognosisofcolorectal cancerpatients received postoperative adjuvant chemotherapy / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the influence of glutathione S-transferase P-1 (GSTP1) genetic variation on the recurrence risk and prognosis of colorectal cancer (CRC) patients received postoperative adjuvant chemotherapy. Methods: The clinical data of 195 CRC patients, who received postoperative adjuvant chemotherapy in the Department of Gastroenterology of the FirstAffiliated Hospital of Zhengzhou University from January 2010 to December 2018, were collected for this study. 5-fluorouracil (5-FU) based adjuvant chemotherapy was given after surgical resection. The recurrence status of the patients was assessed during hospitalization period, and the long-term survival data of patients were obtained by telephone follow-up after finishing the scheduled adjuvant chemotherapy. GSTP1 genotyping was performed with the DNA extracted from peripheral blood specimens, and its correlation with patients’clinical characteristics wasanalyzed.Additionally, RNAwasextractedfrom peripheral blood mononuclear cell (PBMC) specimens of some CRC patients that prior to chemotherapy for GSTP1 mRNA expression analysis. The univariate analysis of genotypes and prognosis was carried out by Kaplan-Meier survival analysis, and adjusted by multivariate Cox regression model. Results: The median disease-free survival (DFS) of the 195 CRC patients was 4.8 years, and the median overall survival (OS) was 6.2 years. Polymorphism analysis indicated that the I105VlocusofGSTP-1coding region was correlated with prognosis. The prevalence of I105V in the study population: AA genotype of 135 cases (69.23%), AG genotype of 56 cases (28.72%) and GG genotype of 4 cases (2.05%), the minor allele frequency of I105V was 0.16. The genotype distribution was in accordance with Hardy-Weinberg equilibrium (P>0.05). The analysis of recurrence risk and prognosis found that the median DFS of patients with AA genotype and AG/GG genotype was 5.7 and 3.9 years respectively (P<0.01), while the median OS of two groups of patients was 7.0 and 4.5 years respectively (P<0.01). The multivariate Cox regression results indicated that AG/GG genotype was an independent factor for OS (OR=1.54, P<0.05). The mRNA expression of GSTP1 in PBMC of the patients with AG/GG genotypes were significantly higher than those patients with AA genotype (P<0.01). Conclusion: GSTP1 I105V genetic variation influences the recurrence risk and prognosis of CRC patients received postoperative adjuvant chemotherapy possibly via mediating the mRNAexpression of GSTP1.

A novel S1P1 modulator IMMH002 ameliorates psoriasis in multiple animal models

Psoriasis is characterized by abnormal proliferation of keratinocytes, as well as infiltration of immune cells into the dermis and epidermis, causing itchy, scaly and erythematous plaques of skin. The understanding of this chronic inflammatory skin disease remains unclear and all available treatments have their limitations currently. Here, we showed that IMMH002, a novel orally active S1P modulator, desensitized peripheral pathogenic lymphocytes to egress signal from secondary lymphoid organs and thymus. Using different psoriasis animal models, we demonstrated that IMMH002 could significantly relieve skin damage as revealed by PASI score and pathological injure evaluation. Mechanistically, IMMH002 regulated CD3 T lymphocytes re-distribution by inducing lymphocytes' homing, thus decreased T lymphocytes allocation in the peripheral blood and skin but increased in the thymus. Our results suggest that the novel S1P agonist, IMMH002, exert extraordinary capacity to rapidly modulate T lymphocytes distribution, representing a promising drug candidate for psoriasis treatment.

Design and synthesis of selective sphingosine-1-phosphate receptor 1 agonists with increased phosphorylation rates

FTY720 and IMMH002, prodrugs for sphingosine-1-phosphate receptor 1 (S1P) agonists, show inadequate and inconsistent levels of phosphorylation in humans compared to that in rats. In this study, FTY720 or IMMH002 analogues (-) were designed and synthesized with modified head pieces to improve the biotransformation of the prodrugs to the active phosphorylated forms. Target compounds were synthesized a convergent route using the key and optically pure building block , which was first synthesized asymmetrically catalyzed amination. The phosphorylation rates of these analogues in rat or human blood were compared. The new methyl-substituted analogue compound showed higher phosphorylation rates in both rats and humans than the parent compound, whereas compound showed improvements in rats, but not in humans. In pharmacokinetics studies of rats, compounds and both had higher levels of phosphorylation than FTY720 and IMMH002. Thus, our study not only yielded new compounds with therapeutic potential, but also showed species differences between rats and humans in response to the structural modifications, which might be useful for predicting the biotransformation behavior and efficacy of this class of prodrugs in the clinic.

Productive infection and transduction by bacteriophage P1 in the species Salmonella bongori

Background: Horizontal gene transfer (HGT) is the most important mechanism in the evolution of new genetic capabilities in bacteria, including specific degradative pathways, virulence factors, and resistance to antibiotics. Among the processes involved in HGT, transduction is noteworthy. This is a mechanism for gene transmission mediated by a bacteriophage that functions both as a reservoir and as a vector of exogenous genes, which remain protected from environmental effects in the bacteriophage's capsid. Within this context, this investigation aimed to evaluate the ability of the generalized transducing bacteriophage P1 to productively infect and transduce in the bacterial species Salmonella bongori. Results: We could establish that a derivative of bacteriophage P1, P1Cm, infects strains of S. bongori with frequencies of lysogenization in the order of ~10−2 lysogens/UFP. Through thermal induction, infective viral progeny was obtained, and we could show that P1Cm readily formed plaques on S. bongori lawns, a phenomenon thus far not reported for other members of the genus Salmonella. Finally, we showed P1Cm-mediated transduction of the model plasmid RP4 at frequencies of ~10−7 transductants/donor. Conclusion: Therefore, bacteriophage P1 can be used as a tool for the genetic manipulation in the species S. bongori.

S1P₁ Regulates M1/M2 Polarization toward Brain Injury after Transient Focal Cerebral Ischemia

M1/M2 polarization of immune cells including microglia has been well characterized. It mediates detrimental or beneficial roles in neuroinflammatory disorders including cerebral ischemia. We have previously found that sphingosine 1-phospate receptor subtype 1 (S1P₁) in post-ischemic brain following transient middle cerebral artery occlusion (tMCAO) can trigger microglial activation, leading to brain damage. Although the link between S1P₁ and microglial activation as a pathogenesis in cerebral ischemia had been clearly demonstrated, whether the pathogenic role of S1P₁ is associated with its regulation of M1/M2 polarization remains unclear. Thus, this study aimed to determine whether S1P₁ was associated with regulation of M1/M2 polarization in post-ischemic brain. Suppressing S1P₁ activity with its functional antagonist, AUY954 (5 mg/kg, p.o.), attenuated mRNA upregulation of M1 polarization markers in post-ischemic brain at 1 day and 3 days after tMCAO challenge. Similarly, suppressing S1P₁ activity with AUY954 administration inhibited M1-polarizatioin-relevant NF-κB activation in post-ischemic brain. Particularly, NF-κB activation was observed in activated microglia of post-ischemic brain and markedly attenuated by AUY954, indicating that M1 polarization through S1P₁ in post-ischemic brain mainly occurred in activated microglia. Suppressing S1P₁ activity with AUY954 also increased mRNA expression levels of M2 polarization markers in post-ischemic brain, further indicating that S1P₁ could also influence M2 polarization in post-ischemic brain. Finally, suppressing S1P₁ activity decreased phosphorylation of M1-relevant ERK1/2, p38, and JNK MAPKs, but increased phosphorylation of M2-relevant Akt, all of which were downstream pathways following S1P₁ activation. Overall, these results revealed S1P₁-regulated M1/M2 polarization toward brain damage as a pathogenesis of cerebral ischemia.

Serum Procollagen Type I N-Terminal Propeptide and Osteocalcin Levels in Korean Children and Adolescents

PURPOSE: Bone markers can be useful for the diagnosis and treatment of skeletal diseases in children and adolescents. Owing to high skeletal growth velocity and rapid bone turnover, children and adolescents have higher bone marker levels than adults. Thus, a valid age- and sex-specific reference should be established for pediatric populations living in similar environments. We aimed to assess the associations of procollagen type I N-terminal propeptide (P1NP) and osteocalcin with age and sex in a group of healthy Korean children and adolescents. MATERIALS AND METHODS: The participants (290 boys and 290 girls, age range 0–18 years) were Korean outpatients. Serum P1NP and osteocalcin levels were measured in control materials and patient samples by electrochemiluminescence immunoassay using an automated Cobas e411 analyzer. RESULTS: Significant age-dependent variations in bone marker levels were observed in both sexes (p<0.001). The highest P1NP levels were observed during the first year of life; thereafter, levels decreased until puberty. There was no postnatal peak for osteocalcin; however, its levels remained higher than the adult reference range throughout childhood. Significant differences were observed between boys and girls (p<0.05), especially between the ages of 12 and 17 years. Cobas e411 results for P1NP showed satisfactory precision and linearity. CONCLUSION: We established reference data for P1NP and osteocalcin levels in healthy Korean children and adolescents, as the first and only study of these parameters in pre-adulthood in Korea. Cobas e411-quantified bone markers may be useful for determining bone metabolism indices.