Transmission of porcine endogenous retrovirus in xenotransplantation / 器官移植

Organ transplantation is the most effective treatment for various types of end-stage diseases. To resolve the problem of donor shortage in organ transplantation, the possibility of xenotransplantation has been gradually explored by surgeons. Pig is one of the common donor sources for xenotransplantation. As a bridge between two species, the viruses carried by pig organs may be transmitted between species and cause the risk of zoonosis. Porcine endogenous retrovirus (PERV) is integrated into the genome, which is a category of retrovirus featuring cross-species transmission. In this article, the influencing factors of transmission characteristics of PERV, the transmission risk of PERV and its recombinant virus, and the detection and transmission risk assessment of PERV in xenotransplantation test were reviewed, aiming to provide reference for alleviating severe shortage of donor organs and driving the advancement of xenotransplantation technologies.

Evaluación de una prueba de reacción en cadena de la polimerasa múltiplex anidada para la detección de HTLV-1

El Perú es un área endémica al virus linfotrópico T humano tipo 1 (HTLV-1) y para su confirmación diagnóstica se usa pruebas serológicas que pueden dar resultados no concluyentes. Objetivos: evaluar una prueba de PCR múltiplex anidada para diagnosticar el HTLV-1. Métodos: la validación de la PCR se realizó con primers dirigidos a las regiones Pol y LTR del HTLV-1. Se empleó el gen ß-globina como control endógeno interno y el límite de detección se evaluó con células MT2. Los parámetros de precisión diagnóstica se evaluaron frente a 95 muestras sanguíneas de Referencia. Resultados: la prueba evaluada obtuvo un límite de detección de 0,5 ng/µL de ADN sensibilidad diagnóstica=97,1%, especificidad diagnóstica y analítica=100%, vpn=97,2%, vpp, repetibilidad y reproducibilidad=100%; Kappa, Índice Youden=0,97. Conclusiones: la prueba evaluada presenta un alto rendimiento diagnóstico y debido a su bajo costo se recomienda su implementación en el algoritmo del diagnóstico de HTLV-1 en Perú.

Peru is an endemic area for human T-lymphotropic virus type 1 (HTLV-1) and for its diagnostic confirmation serological tests are used, which can give inconclusive results. Objectives: to evaluate a nested multiplex PCR test to diagnose HTLV-1. Methods: PCR validation was performed with primers targeting the Pol and LTR regions of HTLV-1. The ß-globin gene was used as an internal endogenous control and the detection limit was evaluated with MT2 cells. Diagnostic accuracy parameters were evaluated against 95 Reference blood samples. Results: the evaluated test obtained a detection limit of 0.5 ng/µL of DNA; diagnostic sensitivity=97.1%, diagnostic and analytical specificity=100%, vpn=97.2%, vpp, repeatability and reproducibility=100%; Kappa, Youden Index=0.97. Conclusions: the evaluated test has a high diagnostic performance and due to its low cost, its implementation in the HTLV-1 diagnosis algorithm in Peru is recommended.

Virus de la leucemia e inmunodeficiencia felina: un estudio retrospectivo en clínicas veterinarias particulares en Bogotá y Chía (Colombia), 2015-2019 / Leukemia viruses and feline immunodeficiency: a retrospective study in private veterinary clinics in Bogotá and Chía (Colombia), 2015-2019

Resumen Los virus de inmunodeficiencia y leucemia felina representan un problema de gran envergadura para los felinos domésticos debido a la multiplicidad de sintomatologías que manifiestan. El objetivo del presente estudio fue establecer, retrospectivamente, la prevalencia en la presentación de ViLeF y VIF en pacientes de seis clínicas de pequeños animales en Bogotá y Chía, en relación con factores como su edad, raza y género. Se realizó un estudio transversal y retrospectivo, mediante la recopilación de datos de 1.014 historias clínicas de pacientes felinos que ingresaron a seis clínicas de la ciudad de Bogotá y Chía, para determinar la prevalencia de VIF y ViLeF y la asociación de estas con factores como edad, género y raza, entre 2015 y 2019, a través de la prueba OR. La detección de los virus se realizó mediante una prueba rápida basada en inmunocromatografía. La mayor prevalencia para cada enfermedad por año fue: 12,3% para VIF en 2012 y 18% para ViLeF en 2019. Los machos presentaron mayores seroprevalencias para ambas enfermedades durante la mayoría los años evaluados. Factores como raza (criolla: VIF: 1,85; ViLeF: 2,01), género (macho: VIF: 1,53 OR; ViLeF: 1,64) y edad (> 7 años: VIF: 3,82; ViLeF: 3,21) se relacionaron positivamente con la presentación de ambas enfermedades en la población felina evaluada.

Abstract Immunodeficiency virus and feline leukemia virus represent major problems for domestic felines due to the multiplicity of symptoms they manifest. The objective of the present study was to establish, retrospectively, the prevalence in the presentation of FeLV and FIV in patients from six small animal clinics in Bogota and Chia, related to factors such as age, race, and gender. A cross-sectional and retrospective study was carried out, collecting data from 1.014 clinical records of feline patients who were admitted to six clinics in the city of Bogota and Chia, to determine the prevalence of FIV and FeLV and their association with factors such as age, gender, and race, between 2015 and 2019 through the OR test. The detection of the viruses was carried out through a rapid test based on immunochromatography. The highest prevalence for each disease per year was 12,3% for FIV in 2012 and 18% for FeLV in 2019. Males presented higher seroprevalences for both diseases during most of the years evaluated. Factors such as race (Creole: FIV: 1,85; FeLV: 2,01), gender (male: FIV: 1.53 OR, FeLV: 1,64), and age (> 7 years: FIV: 3.82; FeLV: 3.21) were positively related to the presentation of both diseases in the feline population evaluated.

Autoimmune manifestations in pediatric patients with human type I T - cell lymphotropic virus (HTLV-1) infection / Manifestaciones autoinmunes en pacientes pediátricos con infección por virus linfotrópico humano de células T tipo I (HTLV-1)

ABSTRACT HTLV-1 is a retrovirus that has an impact on human health due to its T-lymphocyte tropism. It occurs worldwide, but is more prevalent in tropical regions. Although most infected individuals will remain asymptomatic, the infection may manifest with complications such as uveitis, myelopathy, and leukemia, among others. The diagnosis is a chieved by the detection of anti-HTLV antibodies and a confirmatory test (Western Blot or proviral load). Although there is no specific treatment, medical treatments are aimed towards the management of secondary diseases. Three cases are described of pediatric patients diagnosed with HTLV-1 infection and associated autoimmune manifestations.

RESUMEN El virus linfotrópico humano de células T tipo I (HTLV-1) es un retrovirus que causa impacto en la salud del ser humano debido al tropismo para infectar a linfocitos T. Está distribuido mundialmente, pero es más prevalente en regiones tropicales. La mayoría de las personas afectadas permanecen asintomáticas, sin embargo, al manifestarse puede causar complicaciones como uveítis, mielopatía y leucemia, entre otras. Su diagnóstico se hace mediante la determinación de anticuerpos anti-HTLV y prueba confirmatoria (Western Blot o carga proviral). No tiene tratamiento específico, las medidas están dirigidas a la prevención y el manejo de las afecciones secundarias. Se describen tres pacientes en edad pediátrica con diagnóstico de infección por HTLV-1 y manifestaciones autoinmunes.

Detecção do vírus da imunodeficiência felina em gatos domésticos (Felis catus) pela técnica de reação em cadeia da polimerase / Detection of feline immunodeficiency virus in domestic cats (Felis catus) by polymerase chain reaction technique

O objetivo deste trabalho foi detectar a presença de DNA do Vírus da Imunodeficiência Felina (FIV) em gatos domesticos (Feliz catus) assintomáticos. Foi realizada a tecnica de reação em cadeia da polimerase (PCR) em 50 animais. Para tal, foram coletadas amostras de sangue, por venopunção da jugular, de forma asséptica para armazenamento de 1-2 mL de sangue total. Os animais que participaram do estudo fizeram parte do projeto de castração "Vida digna" da Universidade Federal Rural da Amazônia. E a escolha dos animais foi realizada de maneira aleatória, sem distinção por sexo ou idade, resultando em 29 foram fêmeas e 21 machos. Para o diagnóstico, foi realizada a extração do DNA, em seguida as amostras foram testadas em duas reações de PCR utilizando- se dois conjuntos de primers do Gene gag de FIV. Achou-se uma prevalência de 2% (1/50), confirmando assim a presença do vírus na cidade de Belém. Assim, evidenciando a importância de testar os felinos mesmo sendo assintomáticos. Desta forma, faz-se necessário a realização de trabalhos futuros que amplie o número amostral dos animais testados para assim elucidar o perfil epidemiológico da doença na região de Belém do Pará, considerando a relevância clínica desta infecção e a correta conduta médica veterinária para evitar novas infecções.

The objective of this work was to detect the presence of Feline Immunodeficiency Virus (FIV) proviral DNA in asymptomatic domestic cats (Feliz catus). The polymerase chain reaction technique was performed from 50 animals. For this, blood samples were collected by jugular venipuncture, aseptically for storage of 1-2 mL of whole blood. The animals that participated in the study were part of the castration project "Vida digna" at the Universidade Federal Rural da Amazônia. And the choice of animals was performed randomly, without distinction by sex or age, resulting in 29 females and 21 males. For diagnosis, DNA extraction was performed, then the samples were tested in two PCR reactions using two sets of FIV gag gene primers. A prevalence of 2% (1/50) was observed, thus confirming the presence of the virus in the city of Belém. Thus, highlighting the importance of testing the felines even if they are asymptomatic. Therefore, it is necessary to carry out future work that expands the sample number of animals tested in order to elucidate the epidemiological profile of the disease in the region of Belém do Pará, considering the clinical relevance of this infection and the correct veterinary medical conduct to avoid new infections.

Caracterización clínico-epidemiológica de pacientes con VIH/sida en el municipio Manuel Tames, Guantánamo

RESUMEN Introducción: En el municipio Manuel Tames, de la provincia de Guantánamo, no se han realizado estudios que caractericen los pacientes con infección por VIH/sida. Objetivo: Caracterizar clínico-epidemiológicamente a los pacientes con VIH/sida, pertenecientes al municipio Manuel Tames, Guantánamo. Método: Se realizó un estudio descriptivo, transversal, retrospectivo que abarca el periodo comprendido entre 1988-2020. La población estudiada fue de 46 pacientes con el diagnóstico de VIH/sida. Se estudiaron las siguientes variables: edad, sexo, estado civil, año de diagnóstico, clasificación clínica, fuente de pesquisa, esquema de tratamiento, enfermedades concomitantes. Resultados: Los pacientes más afectados se encontraron entre las edades de 25 a 34 años (n=18; 31,9 %), en el sexo masculino (n=28; 60,9 %). El mayor número de los pacientes se encontró soltero (n=37; 80,4 %). La fuente de pesquisa más usual fue la categoría de captado (n=11; 23,9 %). El mayor número de pacientes se halló en la categoría A (n=33; 71,7 %), lo que respalda la cifra de que 32 pacientes (69,6 %) no tuvieran ninguna enfermedad asociada a la infección por el virus. El mayor porcentaje de los pacientes utilizó el esquema terapéutico de Dulutegravir + Trubada (n=26; 56,5 %). Conclusiones: Se puede concluir que la infección por VIH/sida es una enfermedad que se observa en las edades medias de la vida, con un aumento del número de infecciones en tanto pasan los años. Cuando los pacientes mantienen un esquema de tratamiento se observan menos enfermedades concomitantes y se mantienen en las categorías clínicas que menos complicaciones presentan.

ABSTRACT Introduction: No studies have been conducted to characterize patients with HIV/AIDS infection in Manuel Tames municipality, Guantánamo. Objective: To characterize clinically and epidemiologically the HIV/AIDS infected patients in Manuel Tames municipality, Guantanamo. Method: A descriptive, cross-sectional, retrospective study was carried out, covering the period 1988-2020. The studied population consisted of 46 patients with a diagnosis of HIV/AIDS. Variables studied were as follow: age, sex, marital status, year of the diagnosis report, clinical classification, source of information, scheme of treatment, concomitant diseases. Results: Patients 25 to 35 years old were found as the most affected (n=18; 31.9%), with a predominance of males (n=28; 60.9%). Most of patients were found single (n=37; 80.4%). The most common source of information was the capitated category (n=11; 23.9%). Most patients were found in category A (n=33; 71.7%), which supports the figure that 32 patients (69.6%) had no disease associated with virus infection. The highest percentage of patients used the Dulutegravir plus Trubada treatment scheme (n=26; 56.5%). Conclusions: As an end point of the outcome it can be said that HIV/AIDS infection is a disease seen in the middle ages of life, with an increasing number of infections as the years go by. When patients maintain a correct treatment schedule, fewer concomitant diseases are observed and they remain in the clinical categories with the fewest complications.

RESUMO Introdução: No município de Manuel Tames, província de Guantánamo, não foram realizados estudos que caracterizassem pacientes com infecção por HIV/AIDS. Objetivo: Caracterizar clínico-epidemiologicamente os pacientes com HIV/AIDS, pertencentes ao município de Manuel Tames, Guantánamo. Método: Foi realizado um estudo descritivo, transversal e retrospectivo abrangendo o período de 1988-2020. A população estudada foi de 46 pacientes com diagnóstico de HIV/AIDS. Foram estudadas as seguintes variáveis: idade, sexo, estado civil, ano do diagnóstico, classificação clínica, fonte de investigação, esquema de tratamento, doenças concomitantes. Resultados: Os pacientes mais acometidos estavam na faixa etária de 25 a 34 anos (n=18; 31,9%), no sexo masculino (n=28; 60,9%). O maior número de pacientes era solteiro (n=37; 80,4%). A fonte de pesquisa mais comum foi a categoria recrutada (n=11; 23,9%). O maior número de pacientes foi encontrado na categoria A (n=33, 71,7%), corroborando a cifra de que 32 pacientes (69,6%) não apresentavam nenhuma doença associada à infecção viral. O maior percentual de pacientes usava o esquema Dulutegravir + Trubada (n=26; 56,5%). Conclusões: Pode-se concluir que a infecção pelo HIV/AIDS é uma doença que se observa na idade média da vida, com aumento do número de infecções com o passar dos anos. Quando os pacientes mantêm um esquema de tratamento, menos doenças concomitantes são observadas e eles permanecem nas categorias clínicas que apresentam menos complicações.

Aplasia medular asociado con virus linfotrópico humano de células T tipo 1 (HTLV-1): Reporte de un caso / Medullary aplasia associated with Human T-Cell Lymphotropic Virus Type 1 (HTLV-1): A Case Report

Antecedentes: La aplasia medular es una enfermedad caracterizada por pancitopenia con médula ósea hipocelular. La mayor parte de estas son adquiridas pudiendo ser idiopáticas o secundarias al daño de la médula ósea producido por agentes físicos, químicos o infecciosos. El HTLV-1 es un retrovirus intracelular que se transmite por la lactancia materna prolongada, las relaciones sexuales y las transfusiones sanguíneas. Descripción del caso: Reportamos el caso de una mujer de 28 años que presentó petequias, gingivorragia y palidez generalizada. En sus exámenes se encontró pancitopenia severa, HTLV-1 positivo y en el aspirado de médula ósea, hipocelularidad marcada. Se administró antibióticos, antifúngicos, antivirales y agentes inmunosupresores con una evolución estacionaria. Conclusiones: La infección por HTLV-1 es frecuente en Perú; sin embargo, su asociación con aplasia medular es poco descrita. El trasplante de células madre hematopoyéticas es una alternativa de tratamiento en caso de no responder a la terapia inmunosupresora.

Background:Medullary aplasia is a disease characterized by pancytopenia with hypocellular bone marrow. Most of these are acquired and can be idiopathic or secondary to bone marrow damage produced by physical, chemical or infectious agents. HTLV-1 is an intracellular retrovirus that is transmitted by prolonged breastfeeding, sexual intercourse, and blood transfusions. Case description: We report the case of a 28-year-old woman who presented petechiae,gingivorrhagia,andgeneralizedpaleness.Inthelaboratory examination, severe pancytopenia was found, HTLV-1 positive, and in the bone marrow aspirate, marked hypocellularity. She received antibiotics, antifungals, antivirals, and immunosuppressive agents with a steady course. Conclusions:HTLV-1 infection is common in Peru; however, its association with medullary aplasia is poorly described. Hematopoietic stem-cell transplantation is an alternative treatment in case of failure to respond to immunosuppressive therapy.

Mielopatía asociada con infección por HTLV-1: paraparesia espástica tropical / Myelopathy associated with HTLV-1 infection: tropical spastic paraparesia

RESUMEN La mielopatía asociada con infección por HTLV-1 o paraparesia espástica tropical (MAH/PET) es una enfermedad crónica degenerativa del sistema nervioso central que afecta principalmente a la médula espinal. Está asociada a la infección por el virus linfotrópico humano tipo 1 (HTLV-1) que pertenece a la familia Retroviridae. América Latina, Asia y África Subsahariana son zonas endémicas de infección por HTLV-1 y MAH/ PET, entre el 2 % y el 17 %, la mayoría asintomáticos. Colombia tiene una alta prevalencia y en especial en su costa pacífica. En las mujeres la infección por HTLV-1 es más prevalente que en los hombres sin que sea clara la causa; se transmite por transfusiones sanguíneas (principalmente de sangre completa), lactancia materna o relaciones sexuales, que es la forma de transmisón más común en pacientes con MAH/PET. La MAH/PET se presenta principalmente en adultos, su periodo de incubación hasta el desarrollo de síntomas varía de dos años hasta décadas. Las principales manifestaciones clínicas de esta enfermedad son la paraparesia y espastici-dad lentamente progresiva que afecta la marcha, asociada con síntomas de esfínteres, como vejiga neurogénica o estreñimiento. Para su diagnóstico se requiere resonancia magnética (RM) de cerebro y médula espinal y la prueba de Elisa, las más usada, con confirmación mediante western blot (WB). En la actualidad no existe un tratamiento específico para la MAH/PET. En el futuro, el uso de biomarcadores ayudará a la detección temprana de la enfermedad e incluso como posible diana terapéutica.

SUMMARY Myelopathy associated with HTLV-1 infection or tropical spastic paraparesis (MAH / PET) is a chronic degenerative disease of the Central Nervous System that mainly affects the spinal cord. It is associated with infection by human lymphotropic virus type 1 (HTLV-1) that belongs to the Retroviridae family. Latin America, Asia and Sub-Saharan Africa are endemic areas of infection by HTLV-1 and MAH / PET between 2 % and 17 %, most asymptomatic. Colombia has a high prevalence and especially on its Pacific coast. In women, HTLV-1 infection is more prevalent than in men without the cause being clear, it is transmitted by blood transfusions (mainly whole blood), breastfeeding and / or sexual intercourse, which is the most common form of transmission in patients with MAH / PET. MAH / PET occurs mainly in adults, its incubation period until the development of symptoms varies from 2 years to decades. The main clinical manifestations of this disease are slowly progressive paraparesis and spasticity affecting gait, associated with sphincter symptoms, such as neurogenic bladder or constipation. For its diagnosis, magnetic resonance imaging (MRI) of the brain, spinal cord and the most used ELISA test are required with confirmation by western blot (WB). Currently there is no specific treatment for HAM / PET. In the future, the use of biomarkers will help early detection of the disease and even as a possible therapeutic target.

Clinical profile of human T-lymphotropic virus type I: a forgotten infection in pediatrics / Perfil clínico del virus linfotrópico humano tipo I: una infección olvidada en pediatría

Summary The Human T-lymphotropic virus type 1 (HTLV-1), a retrovirus with oncogenic properties, affects around ten to twenty million people worldwide. The most common disorders associated with HTLV-1 infection are T-cell leukemia/lymphoma (ALT) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Studies have reported other clinical manifestations in HTLV-1 seropositive patients, including inflammatory disorders, co-infections with opportunistic agents, and pulmonary diseases. Objective: Here, we aim to describe a cohort of juvenile patients with confirmed HTLV-1 infection that showed clinical manifestations other than neurological symptoms. Methodology and patients: Retrospective analysis of clinical data describing background and clinical findings of 12 juvenile patients with confirmed HTLV-1 infection, attended during January 2018 to February 2020 in a pediatric referral hospital in Cali, Colombia. Results: 11 out 12 patients were from Colombia´s Pacific coast, 10 suffered from significant nutritional deficiencies. Six exhibited dermatological findings, 3 compatible with infective dermatitis. None of the cases exhibited clinical or laboratory findings suggesting ALT or HAM/TPS. Eight patients had structural lung disease assessed by chest Computed Tomography (CT) scans; 4 of them tested positive for galactomannan antigen in bronchoalveolar fluid suggesting pulmonary aspergillosis, and 2 others exhibited a positive PCR testing for tuberculosis. Three patients were diagnosed with autoimmune disorders; 1 patient with Crohn´s Diseases, 1 case of autoimmune thrombocytopenic purpura, and a patient with Vogt-Koyanagi-Harada syndrome (non-granulomatous uveitis). Conclusions: There is a broad range of clinical manifestations in pediatric HTLV-1 patients, and the clinician should consider structural pulmonary disease, opportunistic co-infections and autoimmune disorders in the diagnostic algorithm.

Resumen El Virus Linfotrópico de células T humanas tipo 1 (HTLV-1), retrovirus con propiedades oncogénicas, afecta alrededor de 10-20 millones de personas mundialmente. Las manifestaciónes más comúnmente asociadas a HTLV-1 incluyen leucemia/linfoma de células T (ALT) y mielopatía asociada a HTLV-1/ paraparesia espástica tropical (HAM/TSP). Estudios han reportado otras manifestaciones clínicas en pacientes positivos para HTLV-1, incluyendo enfermedades inflamatorias, coinfecciones con gérmenes oportunistas y enfermedad pulmonar. Objetivo: es describir clínicamente una cohorte de pacientes pediátricos con infección por HTLV-1 confirmada que presentan manifestaciones clínicas diferentes a síntomas neurológicos. Metogolodía y pacientes: Análisis retrospectivo de historia clínica describiendo procedencia y hallazgos clínicos en 12 pacientes con infección por HTLV-1 confirmada, atendidos durante el periodo de Enero de 2018 a Febrero de 2020 en un hospital pediátrico de referencia en Cali, Colombia. Resultados: Once de 12 pacientes procedían de la costa Pacífica Colombiana, 10 con deficiencias nutricionales significativas. Seis mostraron compromiso dermatológico, 3 compatibles con dermatitis infectiva. Ningún paciente presentó hallazgos clínicos o paraclínicos sugestivos de ALT o HAM/TPS. Ocho pacientes presentaron enfermedad pulmonar estructural evidenciada por TAC de tórax; 4 de ellos con antígeno galactomanan positivo en lavado broncoalveolar, sugiriendo aspergilosis pulmonar, y otros 2 resultaron con PCR positiva para tuberculosis. Tres pacientes presentaron enfermedades autoinmunes concomitantes: uno con Enfermedad de Crohn, uno con Púrpura Trombocitopénica Autoinmune, y un paciente con Síndrome de Vogt-Koyanagi-Harada. Conclusiones: Existe un amplio rango de manifestaciones clínicas en pacientes pediátricos con HTLV-1, considerando enfermedad pulmonar estructural, coinfecciones oportunistas y enfermedades autoinmunes dentro del algoritmo diagnóstico.

Preliminary report of preclinical trial of multi-genome engineering pig-to-macaque heart, liver and kidney transplantation / 器官移植

Objective To investigate the application prospect of the most extensive genome engineering pig internationally in preclinical xenotransplantation. Methods Porcine endogenous retrovirus (PERV) knockout combined with 3 major heterologous antigen gene knockouts and 9 humanized genes for inhibition of complement activation, regulation of coagulation disorders, anti-inflammatory and anti-phagocytosis were transferred into a pig (PERV-KO/3-KO/9-TG) as a donor, and the heart, liver and kidney were obtained and transplanted to 3 Rhesus macaque recipients respectively to establish a preclinical research model of pig-to-Rhesus macaque xenotransplantation. The functional status of xenografts after blood flow reconstruction was observed and the survival of recipients was summarized. The hemodynamics of xenografts were monitored. The change of hematological indexes of each recipient was compared. The histopathological manifestation of xenografts was observed. Results After the blood flow was reconstructed, all xenografts showed ruddy color, soft texture and good perfusion. The transplant heart, liver and kidney showed full arterial and venous blood flow and good perfusion at 1 d after operation. The postoperative survival time of heart, liver, and kidney transplant recipients was 7, 26, and 1 d, respectively. The levels of creatine kinase, creatine kinase isoenzyme, and lactate dehydrogenase increased in heart transplant recipient at 1 d after operation, and gradually recovered to near normal levels at 6 d after operation. All indexes increased sharply at 7 d after operation. The level of aspartate aminotransferase increased in liver transplant recipients at 2 d after operation, and the alanine aminotransferase basically returned to normal at 10 d after operation, but the total bilirubin continued to increase. Both aspartate aminotransferase and alanine aminotransferase increased at 12 d after operation, and reached a peak at 15 d after operation. The kidney transplant recipient developed mild proteinuria at 1 d after operation, and died of sudden severe arrhythmia. Histopathology showed that the tissue structure of cardiac and renal xenografts was close to normal, and liver xenografts presented with patchy necrosis, the liver tissue structure was disordered, accompanied by inflammatory damage, interstitial hemorrhage and thrombotic microangiopathy. Conclusions PERV-KO/3-KO/9-TG pig shows advantages in overcoming hyperacute rejection, mitigating humoral rejection and coagulation dysregulation. However, whether it can be used as potential donor for clinical xenotransplantation needs further evaluation.

Expression of HHLA2 in hepatocellular carcinoma tissues and its clinical significance / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of human endogenous retrovirus subfamily H long terminal repeat associating protein 2 (HHLA2) in hepatocellular carcinoma (HCC) tissues and its correlation with the clinicopathological characteristics and prognosis of patients with HCC. Methods: Based on TCGA database, the correlation between HHLA2 mRNA expression and B7 family genes in human HCC tissues was analyzed. HHLA2 expression in 90 pairs of HCC tissues and their adjacent tissues was detected by tissue microarry and immunohistochemical staining. Wilcoxon rank sum test was used to compare the difference of HHLA2 expression between HCC tissues and its adjacent tissues. The chi-square test was used to analyze the relationship between HHLA2 expression in human HCC tissues and clinicopathological features of the patients. Kaplan-Meier survival analysis was performed to analyze the correlation between HHLA2 expression and patients’ overall survival (OS), and the Cox model was used to evaluate the prognostic value of different indices. Results: The expression level of HHLA2 mRNA in HCC tissues was correlated with B7 family CD274, C10orf54, PDCD1LG2, ICOSLG and CD276. The expression level of HHLA2 in HCC tissues was significantly correlated with tumor size (χ2=4.531, P<0.05). The OS of HCC patients with high HHLA2 expression was significantly shorter than that of the patients with lower HHLA2 expression (HR=1.878, 95%CI: 1.066-3.309, P<0.05). The COX model showed that tumor size (HR=2.493, 95%CI: 1.310-4.742, P<0.01) could be used as an independent risk factor for the prognostic prediction of the patients. Conclusion: HHLA2 is significantly correlated with the prognosis of HCC patients, and can be used as a potential target for HCC immunotherapy.

Clinical implication of xenotropic and polytropic retrovirus receptor 1 in papillary thyroid carcinoma / 浙江大学学报·医学版

To investigate the expression of xenotropic and polytropic retrovirus receptor 1 （） in papillary thyroid cancer （PTC） and its clinical implication. The HPA and UALCAN databases were used to explore the expression of XPR1 in PTC and normal tissues. The cBioPortal database was used to obtain the clinical data of PTC patients and gene expression profile. The correlation of expression with gender，age，sub-types，T stage，N stage，M stage and clinical stage of patients were analyzed. Cox regression was conducted to analysis the factors affecting the prognosis of PTC patients. The mutation of was assessed through cBioPortal database. GO and KEGG analyses were used to explore the related biological pathway of involved in PTC. HPA database analysis showed that XPR1 was highly expressed in PTC tissue compared with normal tissues. UALCAN analysis displayed that expression was significantly higher in PTC tissue compared with normal tissues （0.05）. Cox regression analysis showed that was an independent prognostic factor of PTC patients （=2.894，<0.05）. The cBioPortal database indicated that the mutation appeared in 6% PTC patients; the mutation type mainly was missense and the mutation point was located at the E615K. Enrichment analysis indicated that might affect the PTC progression through involvement in metabolic pathway. is highly expressed in PTC tissues，which is associated with the prognosis of patients. Metabolic pathway associated with might play an important role in PTC progression，indicating that might be a novel biomarker for diagnosis and treatment of PTC.

Avaliação de dois testes sorológicos comerciais para diagnóstico das infecções pelo FIV e pelo FeLV / Evaluation of two point-of-care tests to diagnosis of FIV and FeLV infections

FIV e FeLV são retrovírus associados principalmente com neoplasias. Dois testes rápidos são disponibilizados no Brasil para o diagnóstico dessas infecções: um kit de imunocromatografia de fluxo bidirecional (SNAP® Combo IDEXX) e um kit de imunocromatografia de fluxo lateral unidirecional (ALERE/BIONOTE Anigen Rapid). O objetivo deste estudo foi comparar o teste SNAP® com o teste ALERE. Amostras de sangue de 178 gatos foram testadas utilizando-se ambos os kits. A reação em cadeia de polimerase em tempo real (qPCR) foi empregada como método confirmatório para todos os resultados. O teste SNAP® apresentou sensibilidade e especificidade de 100% para FIV; a sensibilidade e a especificidade do teste ALERE foram de 96,15% e 98,68%, respectivamente. A sensibilidade e a especificidade para o FeLV foram de 93,02% e 96,30% para o teste SNAP® e de 90,70% e 97,78% para o teste ALERE. Ainda em relação ao FeLV, três amostras com resultado positivo na qPCR obtiveram resultado falso-negativo em ambos os testes. Não houve diferença estatisticamente significante entre os métodos. Considerando a qPCR como padrão-ouro, o teste SNAP® apresentou maior sensibilidade e especificidade para o FIV, e o teste ALERE apresentou maior especificidade para o FeLV. Os resultados mostraram uma boa correlação entre os testes.(AU)

FIV and FeLV are Retrovirus associated mainly with feline neoplasms. Two point-of-care tests are commercially available in Brazil for diagnosis of these infections: a bidirectional flow immunochromatography kit (IDEXX SNAP ® Combo) and a lateral unidirectional flow immunochromatography kit (ALERE/BIONOTE Anigen Rapid). The aim of this study was to compare SNAP ® and ALERE tests. Blood samples obtained from 178 cats were evaluated using both tests. Quantitative real-time polymerase chain reaction (qPCR) was used as confirmatory test for all samples. The sensitivity and specificity of SNAP ® test was 100% for FIV, and for ALERE test was 96.15% and 98.68%, respectively. The sensitivity and specificity for FeLV was 93.02% and 96.30% for SNAP ® test and 90.70% and 97.78% for ALERE test. Three samples with a qPCR positive result for FeLV obtained a false negative result in both SNAP ® and ALERE tests. There was no statistically significant difference between the two methods. Considering qPCR as gold standard method, the SNAP® test showed higher sensitivity and specificity for FIV, and the ALERE test presented higher specificity for FeLV. The results showed good agreement among the tests.(AU)

Analysis on biological functions of restriction factor SAMHD1 of primates / 吉林大学学报(医学版)

Objective: To discuss the antiviral effect, the inhibitory effect on LINE-1 retrotransposon activity and the redection of interferon production signal pathway of restriction factor SAMHD1 of the primates, and to provide the basis for further study of the SAMHD1 of the primates. Methods: The U937 cells stably expressing the SAMHD1 of primates were established; the U937-control cells established with pLVX -puro were used as negative control group, and the U937-SAMHD1 cells stably expressing the SAMHD1 protein of the different primates were used as experimental groups; the cells were treated with PMA to induce cell differentiation. The virus infection rates of HIV-1 in the cells in various groups were determined by flow cytometry. The HEK293T cells transfected with the expression plasmid of SAMDH1 were used as control group, and the cells co-transfected with the SAMHD1 and HIV-2/SIV Vpx expression plasmids were used as experimental groups. The cells were obtained 48 h after transfection, and the expression levels of SAMHD1 protein were determined by Western blotting method. The intracellular location of SAMHD1 protein was determined by immunofluorescence. The HEK293T cells transfected with LINE-1-GFP report plasmid were used as control group, and the cells co-transfected with LINE-1-GFP and SAMHD1 expression plasmids were used as experimental groups. The rates of GFP positive cells (activity of SAMHD1 to LINE-1 transposon) were determined by flow cytometry. The HEK293T cells transfected with IFN- Luc report plasmid were used as control group, and the cells co-transfected with IFN-Luc and pSAMHDl expression plasmids were used as experimental groups. The expression levels of luciferase in HEK293T cells were determined by chemiluminescence instrument. Results: Compared with negative control group, the virus infection rates of HIV-1 in experimental groups with stable expression of SAMHD1 in the primates were significantly decreased (P<0. 01). Compared with control group, the expression levels of SAMHD1 protein of the primates in experimental groups were decreased (P<0. 05 or P<0. 01). The immunofluorescence results showed that the SAMHD1 protein of the primates was localized in the nucleus. Compared with control group, the rates of GFP positive cells (activity of SAMHD1 to LINE-1 transposon) in experimental groups were significantly decreased (P< 0.05 or P<0.01). Compared with control group, the expression levels of luciferase in the HEK293T cells in experimental groups were significantly decreased (P<0. 05). Conclusion: The SAMHD1 protein of the different primates can resist the HIV-1 infection, inhibit the LINE-1 retrotransposon and antagonize the IFN production by natural immune system.

Cystic fibrosis gene therapy / 国际儿科学杂志

Cystic fibrosis(CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane conductance regulator(CFTR) gene that encodes a cAMP-regulated anion channel.Although CF is a multi-organ system disease, most people with CF die of progressive lung disease that begins early in childhood and is characterized by chronic bacterial infection and inflammation.Nearly 90% of people with CF have at least one copy of the ΔF508 mutation, but there are hundreds of CFTR mutations that result in a range of disease severities.A CFTR gene replacement approach would be efficacious regardless of the disease-causing mutation.After the discovery of the CFTR gene in 1989, the in vitro proof-of-concept for gene therapy for CF was quickly established in 1990.In 1993, the first of many gene therapy clinical trials attempted to rescue the CF defect in airway epithelia.Despite the initial enthusiasm, there is still no FDA-approved gene therapy for CF.Here we discuss the history of CF gene therapy, from the discovery of the CFTR gene to current state-of-the-art gene delivery vector designs.While implementation of CF gene therapy has proven more challenging than initially envisioned; thanks to continued innovation, it may yet become a reality.

Regulation of porcine endogenous retrovirus by dual LTR1+2 (Long Terminal Region) miRNA in primary porcine kidney cells

Porcine endogenous retroviruses (PERVs) integrate into germline DNA as proviral genome that enables vertical transmission from parents to their offspring. The provirus usually survives as part of the host genome rather than as an infectious agent, but may become pathogenic if it crosses species barriers. Therefore, replication-competent PERV should be controlled through selective breeding or knockout technologies. Two microRNAs (miRNAs), dual LTR1 and LTR2, were selected to inhibit the expression of PERV in primary porcine kidney cells. The inhibition efficiency of the miRNAs was compared based on their inhibition of different PERV regions, specifically long terminal repeats (LTRs), gag, pol, and env. Gene expression was quantified using real-time polymerase chain reaction and the C-type reverse transcriptase (RT) activity was determined. The messenger RNA (mRNA) expression of the PERV LTR and env regions was determined in HeLa cells co-cultured with primary porcine kidney cells. The mRNA expression of the LTR, gag, pol, and env regions of PERV was dramatically inhibited by dual miRNA from 24 to 144 h after transfection, with the highest inhibition observed for the LTR and pol regions at 120 h. Additionally, the RT activity of PERV in the co-culture experiment of porcine and human cells was reduced by 84.4% at the sixth passage. The dual LTR 1+2 miRNA efficiently silences PERV in primary porcine kidney cells.

Cystic fibrosis gene therapy / 国际儿科学杂志

Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene that encodes a cAMP-regulated anion channel.Although CF is a multi-organ system disease,most people with CF die of progressive lung disease that begins early in childhood and is characterized by chronic bacterial infection and inflammation.Nearly 90％ of people with CF have at least one copy of the △F508 mutation,but there are hundreds of CFTR mutations that result in a range of disease severities.A CFTR gene replacement approach would be efficacious regardless of the diseasecausing mutation.After the discovery of the CFTR gene in 1989,the in vitro proof-of-concept for gene therapy for CF was quickly established in 1990.In 1993,the first of many gene therapy clinical trials attempted to rescue the CF defect in airway epithelia.Despite the initial enthusiasm,there is still no FDA-approved gene therapy for CF.Here we discuss the history of CF gene therapy,from the discovery of the CFTR gene to current state-of-the-art gene delivery vector designs.While implementation of CF gene therapy has proven more challenging than initially envisioned;thanks to continued innovation,it may yet become a reality.

Hemotropic mycoplasmas in naturally infected cats in Northeastern Brazil / Micoplasmas hemotróficos em gatos naturalmente infectados no Nordeste do Brasil

Abstract This study aimed to determine the prevalence, factors associated, laboratory findings (with and without coinfection by retroviruses) among naturally infected cats by hemoplasmas in northeastern Brazil. For convenience, 200 domesticated and healthy cats were selected. Blood samples were taken to perform complete blood counts, serum biochemical, immunochromatography tests and nPCR for FIV and FeLV, and PCR for hemoplasma recognition. An interview was conducted to determine the factors associated with hemoplasmas. A total of 71/200 (35.5%) cats were positive for at least one hemoplasma species. Isolated infections were observed in 12,5% for 'Candidatus Mycoplasma haemominutum', 12% for Mycoplasma haemofelis and 3% for 'Candidatus Mycoplasma turicensis'. Regarding copositivity, 2% of the animals were positive for M. haemofelis and 'Candidatus Mycoplasma haemominutum', 1.5% for M. haemofelis and 'Candidatus Mycoplasma turicensis', and 4.5% for ' Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis'. No clinical and laboratory changes were observed in the animals that were concomitantly positive for retroviruses and hemoplasmas. Periurban region cats were more likely to be infected by M. haemofelis, while contact with other cats and infection by ' Candidatus Mycoplasma turicensis' were associated with 'Candidatus Mycoplasma haemominutum'. This study indicates that infection by hemoplasmas is a common find in cats from northeastern Brazil.

Resumo Objetivou-se com este estudo determinar a prevalência, fatores associados, achados laboratoriais (com e sem coinfecção com retrovírus) em gatos naturalmente infectados por hemoplasmas no Nordeste do Brasil. Selecionou-se, por conveniência, 200 gatos domiciliados, hígidos, sendo colhidas amostras de sangue para realização do hemograma, bioquímica sérica, imunocromatografia e nested-PCR para FIV e FeLV, e PCR para identificação dos hemoplasmas. Uma entrevista foi realizada para determinação dos fatores associados aos hemoplasmas. A frequência de positividade foi de 35,5% (71/200). Infecções isoladas foram observadas em 12,5% dos animais para 'Candidatus Mycoplasma haemominutum', 12% para Mycoplasma haemofelis e 3% para 'Candidatus Mycoplasma turicensis'. Quanto a co-positividades, 2% dos animais foram positivos para M. haemofelis e 'Candidatus Mycoplasma haemominutum', 1,5% foram positivos para M. haemofelis e 'Candidatus Mycoplasma turicensis', e 4,5% foram positivos para 'Candidatus Mycoplasma haemominutum' e 'Candidatus Mycoplasma turicensis'. Não foram observadas alterações clínicas ou laboratoriais nos animais positivos para retrovírus e hemoplasmas, concomitantemente. A região periurbana foi identificada como fator de risco associado a M. haemofelis. Enquanto o contato com outros gatos e a infecção por 'Candidatus Mycoplasma turicensis' foi associado à 'Candidatus Mycoplasma haemominutum'. Este estudo indica que a presença dos agentes da micoplasmose hemotrópica felina é comum no Nordeste brasileiro.

Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus

Abstract Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp ® plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD450 nm) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a κ value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle.

Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus

Abstract Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp ® plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD450 nm) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle.