Análise epidemiológica dos casos de Aids no Sudeste brasileiro de 2010 a 2019 / Epidemiological analysis of Aids cases in Southeastern Brazil from 2010 to 2019 / Análisis epidemiológico de casos de Sida en el sudeste de Brasil de 2010 a 2019

Resumo Introdução: A epidemia de HIV e Aids representa um dos maiores problemas de saúde pública atualmente. No Brasil, estima-se que apenas 83 % das pessoas vivendo com HIV conhecem seu diagnóstico e somente 46 % possuem carga viral indetectável. Isso evidencia a necessidade de estudos voltados à compreensão do HIV e da Aids no Brasil, para a melhora das políticas públicas. Assim, o objetivo deste estudo é realizar um levantamento de dados sobre o perfil epidemiológico de pacientes portadores de Aids no Sudeste brasileiro. Métodos: Trata-se de um estudo epidemiológico descritivo, de caráter quantitativo. Os dados foram obtidos no SINAN. A análise restringiu-se ao período de janeiro de 2010 a junho de 2019. Resultados: No período analisado, o ano de 2017 apresentou a maior prevalência da doença (11,97 %). Além disso, observa-se uma tendência de crescimento no número de casos entre 2010 e 2014 e uma tendência decrescente nos últimos anos. O Rio de Janeiro foi o estado com maior número de novos casos (21,90 %) da Região Sudeste. Observou-se que os grupos com maior incidência de casos foram: sexo masculino (71,1 %), jovens e adultos (79,37 %), cor branca (46,67 %), ensino médio completo (19,71 %) e heterossexuais (46,94 %). Contudo, foi observado um aumento relativo da incidência entre homens homossexuais e entre os pardos. Além disso, a principal forma de transmissão foi via sexual (78,09 %). Conclusão: O estudo concluiu que são necessárias mais pesquisas acerca do tema, para acompanhar a epidemiologia da Aids e orientar medidas governamentais eficientes.

Abstract Introduction: The HIV and Aids epidemic represents one of the biggest public health problems today. In Brazil, it is estimated that only 83 % of people living with HIV know their diagnosis and only 46 % have an undetectable viral load. This highlights the need for studies aimed at understanding HIV and Aids in Brazil, to improve public policies. Thus, this study aims to conduct a survey of data on the epidemiological profile of Aids patients in Southeast Brazil. Methods: It is a descriptive, quantitative epidemiological study. The data was extracted from SINAN. The analysis was restricted to the period from January 2010 to June 2019. Results: In the analyzed period, 2017 had the highest prevalence of the disease (11,97 %). In addition, there was an increase in the number of cases between 2010 and 2014 and a drop over the last few years. Rio de Janeiro was the state with the highest number of new cases (21,90 %) in the Southeast Region. Results: The groups with the highest incidence of cases were: man (71,1 %), young and adults (79,37 %), white (46,67 %), high school graduates (19,71 %) and heterosexuals (46,94 %). However, a relative increase in incidence was observed among homosexual men and among browns. Also, sexual transmission was the most prevalent (78,09 %). Conclusion: The study concluded that more research on the topic is needed, to monitor the epidemiology of Aids and to guide efficient government measures.

Resumen Introducción: La epidemia de VIH y Sida representa un gran problema de salud pública actualmente. En Brasil, se estima que solo el 83 % de las personas que viven con el VIH conocen su diagnóstico y solo el 46 % tiene una carga viral indetectable. Esto resalta la necesidad de estudios que comprendan el VIH y el Sida en Brasil, para mejorar las políticas públicas. El objetivo de este estudio es realizar una encuesta de datos sobre el perfil epidemiológico de los pacientes con Sida en el sudeste de Brasil. Métodos: Es un estudio epidemiológico descriptivo y cuantitativo. Los datos se obtuvieron del SINAN. El análisis se restringió al período de enero de 2010 a junio de 2019. Resultados: En el período analizado, el año 2017 presentó la mayor prevalencia de la enfermedad (11,97 %). Existe una tendencia creciente en el número de casos entre 2010 y 2014 y una tendencia decreciente en los últimos años. Rio de Janeiro fue el estado con mayor número de casos nuevos (21,90 %) en la Región Sudeste. Se observó que los grupos con mayor incidencia de casos fueron: hombres (71,1 %), jóvenes y adultos (79,37 %), blancos (46,67 %), escuela secundaria completa (19,71 %) y heterosexuales (46,94 %). Sin embargo, se observó un aumento relativo en la incidencia entre hombres homosexuales y entre pardos. Además, la principal forma de transmisión fue la sexual (78,09 %). Conclusión: Se necesita más investigación sobre el tema, para monitorear la epidemiología del Sida y orientar medidas gubernamentales eficientes.

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.

Self-reported HIV/HAART-associated Lipodystrophy and Modifiable Risk Factors for Cardiovascular Disease

Abstract Background Patient self-report is the most common diagnostic tool in the literature to detect HIV/HAART-associated lipodystrophy. However, data on the association of cardiovascular risk factors with HIV/HAART-associated lipodystrophy assessed by self-report are still missing. Objectives To determine the prevalence of self-reported HIV/HAART-associated lipodystrophy and to identify independent associations between traditional modifiable cardiovascular risk factors and self-reported lipodystrophy. Methods We conducted a retrospective observational study at an outpatient infectious disease clinic in the Central-West of Brazil to identify the association between traditional modifiable cardiovascular risk factors and self-reported lipodystrophy. Sedentary lifestyle, smoking status, family history of cardiovascular disease, hypertension, diabetes, dyslipidemia, increased waist circumference and overweight were the cardiovascular risk factors assessed. Self-reported HIV/HART-associated lipodystrophy was categorized as: mild (noticeable by patients' close inspection), moderate (easily noticeable by patient and physician) or severe (readily noticeable by a casual observer). Prevalence ratio (PR) and 95% confidence interval (CI95%) were calculated. Multivariate Poisson's regression was used to analyze factors associated to HIV/HAART-associated lipodystrophy assessed by self-report considering a significance level of 5%. Results A total of 183 patients were included, with a mean age of 39.3±10.9 years. Most of the sample were male (77.6%), non-white (50.8%) and single (53.0%). The overall prevalence of HIV/HAART-associated lipodystrophy was 52.5% (95% CI 44.96 - 59.88). Severe lipodystrophy was observed in more than half patients (55.2%). No traditional modifiable cardiovascular risk factor was independently associated with lipodystrophy. Female sex (PR 1.49; 95% CI 1.15 - 1.95; p =0.003), time of HIV infection diagnosis of 1-3 years (PR 1.83; 95% CI 1.09 - 3.08; p =0.002) and a positive family history of CVD (PR 1.62; 95% CI 1.11 - 2.36; p <0.001) were independently associated with lipodystrophy. Conclusion HIV/HAART-associated lipodystrophy assessed by patient self-report was not associated with traditional modifiable cardiovascular risk factors. Int J Cardiovasc Sci. 2020; [online].ahead print, PP.0-0

Seroprevalencia de los virus linfotrópicos de células T humanas de tipos I y II en donantes del Banco de Sangre del Hospital Pablo Tobón Uribe, entre 2014 y 2015 / HTLV-I/II seroprevalence in blood donors of Hospital Pablo Tobón Uribe Blood Bank during the period 2014-2015

Resumen Introducción . El virus linfotrópico humano de células T (HTLV) es un retrovirus del cual se conocen varios tipos, entre ellos el HTLV-I y el HTLV-II, los cuales son de importancia clínica por ser los causantes de diferentes enfermedades, como la leucemia y el linfoma de células T del adulto, la paraparesia espástica tropical y la mielopatía asociada al HTLV. Objetivo . Obtener la prevalencia de las reacciones presuntiva y confirmatoria de los virus HTLV-I y HTLV-II en los donantes del Banco de Sangre del Hospital Pablo Tobón Uribe de Medellín, entre el 2014 y el 2015. Materiales y métodos . La información se obtuvo de la base de datos del Banco de Sangre del Hospital Pablo Tobón Uribe. Se analizaron la edad, el sexo y el lugar de procedencia y de residencia de los donantes, así como la reacción en la prueba de tamización (ELISA) y en la prueba confirmatoria (inmunoblot). Resultados . La población de donantes estudiados incluyó a 6.275 hombres y 8.148 mujeres, para un total de 14.423 donantes reclutados entre el 1° de marzo de 2014 y el 30 de junio de 2015. De ellos, 25 resultaron positivos para HTLV-I o HTLV-II en la prueba de tamización (ELISA). En la prueba confirmatoria (inmunoblot), nueve (36 %) pacientes fueron positivos para el HTLV-I o HTLV-II , y de ellos ocho (32 %) lo fueron para el HTLV-I y uno (4 %) para el HTLV-II; la seroprevalencia global fue de 0,06 % (IC95% 0,10-0,25). Conclusiones . Los hallazgos del estudio concordaron con los de estudios similares en áreas no endémicas del país y con los de los estudios consultados a nivel internacional.

Abstract Introduction: The human-T cell lymphotropic virus is a retrovirus with various types known so far. HTLV-I and HTLV-II are of clinically importance as they cause different diseases such as adult T-cell leukemia/lymphoma, tropical spastic paraparesis, and human T-lymphotropic virus type I-associated myelopathy (HAM). Objective: To estimate the prevalence of presumptive and confirmatory reactivity to HTLV-I/II in blood donors of Hospital Pablo Tobón Uribe Blood Bank between 2014 and 2015. Materials and methods: The information was obtained from the Hospital Pablo Tobón Uribe Blood Bank database. We analyzed age, sex, place of origin, and place of residence of donors, and the reactivity using the screening test (ELISA) as well as the confirmatory test (immunoblot). Results: The donor population studied included 6,275 men and 8,148 women, for a total of 14,423 donors recruited between March 1, 2014, and June 30, 2015. Of all tested donors, 25 were positive for HTLV-I/II by the screening test (ELISA). After performing the confirmatory test (immunoblot), only nine patients were positive for HTLV-I/II (36%), of whom eight were reactive to HTLV-I (32%) and one to HTLV-II (4%), for a global seroprevalence of 0.06% (CI 95%: 0.10-0.25). Conclusions: Our findings were consistent with those found in similar studies in non-endemic areas of the country and with those from studies at international level reported in the literature.

Analysis of swine leukocyte antigen class I gene profiles and porcine endogenous retrovirus viremia level in a transgenic porcine herd inbred for xenotransplantation research

Molecular characterization of swine leukocyte antigen (SLA) genes is important for elucidating the immune responses between swine-donor and human-recipient in xenotransplantation. Examination of associations between alleles of SLA class I genes, type of pig genetic modification, porcine endogenous retrovirus (PERV) viral titer, and PERV subtypes may shed light on the nature of xenograft acceptance or rejection and the safety of xenotransplantation. No significant difference in PERV gag RNA level between transgenic and non-transgenic pigs was noted; likewise, the type of applied transgene had no impact on PERV viremia. SLA-1 gene profile type may correspond with PERV level in blood and thereby influence infectiveness. Screening of pigs should provide selection of animals with low PERV expression and exclusion of specimens with PERV-C in the genome due to possible recombination between A and C subtypes, which may lead to autoinfection. Presence of PERV-C integrated in the genome was detected in 31.25% of specimens, but statistically significant increased viremia in specimens with PERV-C was not observed. There is a need for multidirectional molecular characterization (SLA typing, viremia estimation, and PERV subtype screening) of animals intended for xenotransplantation research in the interest of xeno-recipient safety.

Factors associated with pain in individuals infected by human T-cell lymphotropic virus type 1 (HTLV-1)

Abstract Introduction Despite the high prevalence of chronic pain in individuals infected with HTLV-1, predictive and protective factors for its development are still unclear. Objective To identify factors associated with chronic pain in individuals with HTLV-1. Methods This cross-sectional study was conducted in a reference center for treatment of patients infected with HTLV-1 in Salvador, Bahia, Brazil. The study included individuals infected with HTLV-1, over 18 years, and excluded those with difficulty to respond the pain protocol. Data on sociodemographic, health behavior, and clinical characteristics were collected in a standardized way. The prevalence ratio (PR) of pain is described, as well as the factors independently associated with the presence of pain, which were assessed by multiple logistic regression. Results A total of 142 individuals were included in the study, mostly female (62.7%), aged 20–64 years (73.2%), married (61.3%), with less than eight years of education (54.2%), and with a steady income (79.6%). Multivariate analysis showed that being symptomatic for HTLV-1 – sensory manifestations, erectile dysfunction, overactive bladder, and/or HAM/TSP (PR = 1.21, 95% CI: 1.05 to 1.38), self-medication (PR = 1.29, 95% CI: 1.08–1.53), physiotherapy (PR = 1.15, 95% CI: 1.02–1.28), and depression (PR = 1.14, 95% CI: 1.01–1.29) were associated with an increased likelihood of presenting pain. On the other hand, physical activity (PR = 0.79, 95% CI: 0.67–0.93) and religious practice (PR = 0.83, 95% CI: 0.72–0.95) were associated with a decreased likelihood of having pain. Conclusion The use of self-medication, physiotherapy and the presence of depression are independently associated with neurological symptoms in HTLV-1 infected patients. Religious practice and physical activity are both protective for the development of pain.

Enfoque bioético y social de la infección por el Virus de Inmunodeficiencia Humana / Bioethics and social approach of the Acquired Immunodeficiency Syndrome

Las personas portadoras o enfermas del virus de inmunodeficiencia humana han experimentado consecuencias devastadoras en el plano personal, familiar y social desde la aparición de los primeros casos. El objetivo del presente trabajo es analizar el cumplimiento de los principios bioéticos, así como la repercusión derivada de las implicaciones éticas del actuar profesional con estos pacientes, lo cual demuestra la necesidad de vencer los tabúes y la discriminación que han sido objeto desde los inicios de la epidemia.

Persons who are carrier of the Acquired Immunodeficiency Syndrome or persons who are sick have experienced destructive consequences in the personal, familiar and social spheres since the appearance of the first cases. The objective of this work is to analyze the fulfilment of the bioethical principles, as well as the repercussion derived from the ethics dilemma of the professional behaviour with these patients, what demonstrate the necessity of overcoming taboos and the discrimination they have suffered from since the beginnings of the disease.

LIM mineralization protein expression in LIM mineralization protein-1 gene-transfected bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:In vitro experiments suggest that LIM mineralization protein-1 (LMP-1) gene can increase the expression of LMP-1 protein in osteoporotic bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the LMP-1 expression in osteoporotic bone marrow mesenchymal stem cells transfected with RV-LMP-1-GFP in vitro. METHODS:Twelve SD female rats were selected and subjected to bilateral ovariectomy for establishment of osteoporosis models. After 2 months of feeding, bilateral femurs, tibiae, and humeri of rats were taken to isolate and culture bone marrow mesenchymal stem cells. Passage 3 cells were taken and randomly divided into ovariectomized group and LMP-1 transfection group. Another six rats only underwent removal of the same amount of fat tissue around the ovary, and passage 3 cells which were harvested as those in the former two groups served as sham group. RT-PCR and western blot assay were performed to determine the expression of LMP-1 protein and mRNA. RESULTS AND CONCLUSION:Under an inverted fluorescence microscope, transfected bone marrow mesenchymal stem cells from osteoporotic rats showed green fluorescent expression. The three groups al could express LMP-1 at the protein and mRNA levels. The expression levels of LMP-1 protein and mRNA in the LMP-1 transfection group were significantly higher than those in the ovariectomized group and sham group (P0.05). The successful expression of the RV-LMP-1-GFP gene in osteoporotic SD rat bone marrow mesenchymal stem cells was realized at the protein and mRNA levels;moreover, the expression of LMP-1 was increased dramatical y. These findings indicate that LMP-1 gene is successful y transferred into osteoporotic SD rat bone marrow mesenchymal stem cells, and significantly elevates the expression of LMP-1 mRNA and protein.

Expression of recombinant retroviral vector carrying nerve growth factor gene in neural stem cells / 中国组织工程研究

BACKGROUND:Nerve growth factor (NGF) belongs to a biological macromolecule that is difficult to pass through the blood-brain barrier. However, a retroviral vector carrying exogenous gene can be stably inserted and integrated into the host cellgenome, which is suitable for gene therapy. OBJECTIVE:To study the gene expression of recombinant retroviral vector carrying rat NGF gene in neural stem cels. METHODS: The rat NGF gene was inserted into a retroviral vector pLEGFP-N1, which was transferred into packaging cels PT67 by Lipofectamine 2000. The positive clones in virus supernatant were colected by G418 selection and used to infect neural stem cels. After that, the NGF expression was tested by enzyme linked immunosorbent assay and the biological competence by PC12 cels, and then morphological change of neural stem cels and celltyping were examined by fluorescent microscope. RESULTS AND CONCLUSION:The neural stem cels could express extrinsic source NGF protein after the recombinant plasmid was infected into neural stem cels. The PC12 cels increased in the experimental group and stretched out long neurites. And the NGF protein could maintain the neural stem cellsurvival and stimulate their differentiation. These findings suggest that the neural stem cels carrying extrinsic source NGF gene could express NGF successfuly, and the NGF protein induced the survival and differentiation of neural stem cels.

Paraparesia espástica tropical y anestesia: reporte de caso y revisión temática / Tropical Spastic Paraparesis and Anesthesia: Case Report and Topic Review

Introducción: La paraparesia espástica tropical es una infección endémica en Colombia, causada por el retrovirus HTLV-1. Se caracteriza por una mielopatía de lenta instauración que compromete principalmente los miembros inferiores. Las complicaciones como escaras por decúbito prolongado, retención urinaria por disfunción esfinteriana, fracturas, etc. hacen de estos pacientes candidatos quirúrgicos potenciales. Objetivo: Reporte de caso y revisión temática de la fisiopatología, la epidemiología, la clínica y el tratamiento, y de los aspectos anestésicos básicos de la enfermedad. Metodología: Reporte de caso y revisión temática. Se incluyeron en la búsqueda ensayos clínicos, metaanálisis, guías para la práctica, ensayos controlados aleatorizados, revisiones, reportes de casos, artículos clásicos, estudios comparativos, conferencias de consenso, clases magistrales y libros de texto, de artículos publicados sobre paraparesia espástica tropical/mielopatía asociada al HTLV-1 (PET/MAH) e implicaciones anestésicas. Se incluyeron las publicaciones cuyo tema central fuese etiología, fisiopatología, epidemiología, manifestaciones clínicas, tratamiento y repercusiones anestésicas de PET/MAH. Se realizó una búsqueda en PubMed, MdConsult, EBSCOhost, OvidSP y Scielo de artículos en inglés y español. Se utilizaron los términos MeSH: paraparesis, tropical spastic, anesthesia y los términos DeCS: paraparesia espástica tropical, anestesia. Se estudiaron de forma independiente los títulos y resúmenes de los artículos identificados en las bases de datos. Resultados: Se describe el caso de un adulto masculino quien fue llevado a cirugía para reconexión uretral, después de presentar una de las complicaciones características de la paraparesia espástica tropical. La búsqueda arrojó 1.829 estudios. Veinte escritos cumplieron con los criterios de inclusión. Se hace una presentación de implicaciones anestésicas y de la enfermedad.

Introduction: Tropical spastic paraparesis is an endemic infection in Colombia caused by the HTLV-1 retrovirus. It is characterized by a slow and progressive myelopathy that initially targets lower limbs. Complications such as eschars due to a prolonged decubitus, urinary retention to sphincter dysfunction, fractures, etc. make these patients potential surgery candidates. Objective: To report a case and to review the physiopathology, epidemiology, clinical manifestations, treatment and basic anesthetic considerations of this disease. Methods: Case report and topic review. The research included clinical trials, meta-analysis, practice guides, randomized controlled assays, revisions, case reports, classic articles, comparative studies, consensus conferences, magisterial classes and textbooks regarding published articles on tropicalsSpastic paraparesis/HTLV-1 (TSP/HAM) Associated myelopathy and anesthetic implications. Publications focused on etiology, physiopatology, epidemiology, clinical manifestations, treatment and anesthetic repercussions of TSP/ HAM were included in this article. Research was carried out through PubMed, MdConsult, EBSCOhost, OvidSP, and Scielo, of articles in English and Spanish. The MeSH terms used were: paraparesis, tropical spastic, anesthesia and the DeCS terms were: paraparesia espástica tropical, anestesia. Titles and abstracts of articles identified in the database were studied independently. Results: We describe the case of a male adult patient who was admitted to surgery for urethral reconnection after presenting a classic complication of tropical spastic paraparesis. Research on the topic yielded 1829 studies. A total 20 writings met the inclusion criteria. We present implications regarding anesthesia and the disease.

Paraparesia espástica tropical/mielopatia associada ao HTLV-1: relato de dois casos com manifestações dermatológicas em familiares de doador de sangue HTLV-1 positivo assintomático no Hemocentro do Amazonas - Hemoam / Tropical spastic paraparesis/myelopathy associated with HTLV-1: report of two related cases with dermatological involvement

Dois casos de paraparesia espástica tropical / mielopatia associada ao HTLV-1 (HAM/TSP) foram diagnosticados no Hemocentro do Amazonas - Hemoam, em familiares de doador de sangue soropositivo para o HTLV-1 assintomático. Aqui descrevemos a investigação familiar, as características clínicas dos casos e as manifestações dermatológicas associadas.

Herein we report on two cases of tropical spastic paraparesis / myelopathy associated with HTLV-1 (HAM/TSP) diagnosed in relatives of a blood donor found positive for HTLV-1 at serologic screening. The donor himself was asymptomatic. Family studies, the clinical characteristics of the cases and the associated dermatologic manifestations are reported.

Epidermal growth factor receptor mutation type III promotes the migration of glioma cells by increasing FAK phosphorylation / 肿瘤

Objective: To investigate the mechanism underlying the promoting effects of epidermal growth factor receptor mutation type III (EGFRv III) on the migration of glioma cells. Methods: Human glioma cell line U87EGFRv III was established by retroviral infection of the parental glioma cells. The effect of EGFRv III on the migration of the cells was detected by gel-drop assay and Transwell migration assay. The phosphorylation of FAK was detected by Western blotting. The FAK Y397F mutant was transfected into U87EGFRv III cells and the change in cell migration was observed. Results: EGFRv III promoted the migration of human glioma U87 cells and increased the FAK phosphorylation level at Tyr397. Transfection with FAK Y397F mutant reduced FAK phosphorylation level in glioma cells U87EGFRvH[ and induced the decrease in the migration abilities of the cells. Conclusion: EGFRv III promoted the migration of glioma cells by up-regulating FAK phosphorylation levels.

Construction of mIL-12 recombinant retrovirus vector and establishment of viral package cell line / 吉林大学学报(医学版)

Objective To construct mIL-12 recombinant retrovirus vector and evaluate the effect of mIL-12 expression on glioma in rats.Methods mIL-12 DNA was cloned into vector pLEGFP using standard procedures to develop recombinant plasmid pLEGFP-mIL 12,then it was transferred into PA317 cells.Results Products of enzyme digestion and PCR of recombinant plasmid pLEGFP-mIL12 were analysed using electrophoresis and a(2 290 bp) fragment of DNA as same as mIL-12 gene in size was found,transfected NIH3T3 expressed GFP-mIL12 fusion protein.Conclusion Transfected PA317 cells can produce retrovirus which can infect NIH3T3 cells,and the expression of mIL-12 gene can be detected in NIH3T3 cells.

Effect of human factor Ⅸ intron 1 inserted in forward orientation into retroviral vectors on expression of factor Ⅸ in muscle cells / 第二军医大学学报

Objective:To investigate the effects of human factor Ⅸ (FⅨ) intron 1 inserted in forward orientation into retroviral vector on the expression of FⅨ in muscle cells. Methods: Two FⅨ mini-genes, FⅨm1 and FⅨm2, were inserted in forward orientation into retroviral vector backbone, LⅨSN, constructing 2 vectors named LⅨm1SN and LⅨm2SN, containing shortened intron of 1.4 and 0.3 kb, respectively. Retrovirus was made by transfecting PA317 packaging cells. Transient and stable expression of FⅨ was tested in SCID mouse muscle cells (SCID-MB). Results: The vector titers and expression levels of LⅨm1SN and LⅨm2SN in SCID-MB were 1 to 2 folds higher than that of LⅨSN. Some SCID-MB colonies transfected with LⅨm2SN contained intact FⅨm2 inton elements, suggesting that splicing of virus mRNA may not be complete in packaging cells. Conclusion: FⅨ intron 1 inserted in forward orientation in retroviral vectors can increase retroviral titer and the expression level of FⅨ in muscle cells.

Construction of two retroviral vectors for interleukin-1 receptor antagonist and interleukin-10 and in vitro transfection of rabbit synoviocytes / 中华风湿病学杂志

Objectives To construct two retroviral vectors, one containing human interleukin-1 recep-tor antagonist (hIL-1Ra) gene and the other containing human interleukin-10 (hIL-10) gene and to transfect rabbit synoviocytes in vitro and detect the expression level of target genes. Methods RNA from human peripheral blood mononuclear cells were extracted and target genes were amplified by RT-PCR. The target genes were cloned into retroviral vector pLXSN, which was then transducted into GP2-293 cells to produce recombinant retrovirus. Rabbit synoviocytes were transfected and the expression of target genes was detected by RT-PCR, immunohistochemistry and western-blot. Results The retroviral vector containing hIL-1Ra gene or hIL-10 gene was constructed successfully. The hIL-1Ra gene and hIL-10 gene were transduced respectively into rabbit synoviocytes in vitro. The mRNA level of both genes reached peak in 5 days. In positive cell clones, the protein level of hIL-1Ra reached peak within 30 days and maintained at least 60 days; the protein level of hIL-10 maintained at least 40 days. Conclusion The hIL-1Ra gene and hIL-10 gene can be transduced successfully into rabbit synoviocytes by recombinant retrovirus.

Construction of pancreatic duodenal homeobox-1 recombinant retroviral vector and its stable expression in liver progenitor cells / 第二军医大学学报

Objective:To construct a recombinant retroviral vector expressing pancreatic duodenal homeobox-1(PDX-1) gene and express the PDX-1 expression cassette in liver progenitor cells(LEPCs),so as to study the conversion of hepatic cells to pancreatic-like cells.Methods: The full length of PDX-1 gene was amplified by PCR and subcloned into pMSCVpuro vector.The pMSCV PDX-1 puro was then introduced into Phoenix package cells and the cell culture supernatant was used for Ping-Pong infection of another packaging cell line PT67 to obtain stable virus-producing cell line.Results: The pMSCV PDX-1 puro vector was successfully constructed and transfected into PT67 cells.The viral supernatants of PT67 cells efficiently infected LEPCs in vitro and the infected LEPCs stably expressed PDX-1 gene.Conclusion: The pMSCV PDX-1 puro vector and PDX-1 expressing LEPCs have been successfully constructed,laying a basis for studying the transdifferentiation of liver progenitor cells to pancreas lineage cells.

Effects of anti-leukemia by antisense c-myc by retrovirus vector / 中国病理生理杂志

AIM: To study the role of c-myc oncogene in L6565 leukemia oncogenesis and the effects of therapy by inhibition of its expression with antisense c-myc. METHODS: A recombinant retroviral vector containing antisense c-myc of the murine (pGNCas)was constructed and then transfected into PA317 cells by the method of calcium phosphate precipitation. L6565 clone cells were infected with retrovirus particles. Stable integretion of antisense c-myc was shown by PCR. The change of the malignance and phenotype of L6565as were detected by the examination of the growth, morphology, cells cycle, agar assay and expression of c-myc. RESULTS: The shape of most L6565as cells became spherical. The growth of L6565as was inhibited compared to control cells. The analysis of cells cycle: L6565as cells were arrest in G 0/G 1 phase, decreased in S phase. The ability of L6565as cells to form colony in soft agarose was significantly suppressed. c-myc in L6565as cells was lowly expressed. CONCLUSION: (1)c-myc plays a critical role in L6565 leukemia oncogenesis; (2)Inhibition of expression of c-myc makes partly reversion of malignant phenotype of L6565 murine leukemia clone cells.

Killing effects on human hilar cholangiocarinoma cells with CD and HSV-tk double suicide genes coexpression / 中国现代普通外科进展

Objective:To investigate the different killing effects on human hilar cholangiocarinoma cells FRH with cytosine deaminase(CD) and herpes simplex virus thymidine kinase(HSV tk)double suicide genes coexpression compared with single gene mediated by retrovirus.To find a more efficient and low toxicity suicide gene therapy for hilar cholangiocarinoma.Methods:CD and HSV tk double suicide genes were transfected into PA317 cells using lipofectamine.The positive clones were picked out and cultured after G418 selected.The viral supernatant was collected.The FRH cells were infected with the virus containing the double suicide genes.After G418 selection,RT PCR was resorted to demonstrated the successful transcription of CD and HSV tk genes.The FRH/CD+tk and FRH cells in culture were respectively treated with 5 Fc and /or GCV.The cytoxicity efficacy was evaluated by microculture tetrajolium test (MTT) method.Results:The virus containing double suicide gene was produced in PA317 cells.Double suicide genes were stably expressed in RFH cells after being infected with the virus.The killing effect of combination 5 Fc with GCV on FRH/CD+tk cells is more effective than that of using 5 Fc or GCV alone.Conclusion:The CD+TK/5 Fc+GCV co expression system is more effective for killing effects on FRH cells than that of CD/5 Fc or tk/GCV system alone.

Construction and identification of recombinant retroviral vector with controllable human preproenkephalin gene / 中华麻醉学杂志

Objective To construct a recombinant retroviral vector with human preproenkephalin gene regulated by tetracycline. Methods Human preproenkephalin gene was amplificated by polymerase chain reaction (PCR) and was cloned into retrovirus tetracycline responsive plasmid. Then this recombinant plasmid and regulatory plasmid pRev Tet-On were transferred into packaging cell PT67 respectively. The transfected PT67 / Tet-On and PT 67 / TREhPPE cells were selected by the corresponding antibiotics and identified by RT-PCR. The selected cells were enriched and virus liter was assayed using NIH3T3. Results The restriction endonuclease digestion, PCR analysis and DNA sequencing confirmed that the recombinant RevTRE/hPPE vector was constructed successfully. The virus liter of PT 67/TREhPPE was 3.2 ? 103 CFU?ml-1 and the virus titer of PT67/Tet-On was 2.6?105 CFU?ml-1.Conclusion A recombinant retroviral vector with human preproenkephalin gene regulated by trtracycline and stable virus producing lines have been successfully constructed, providing a good basis for further research on regulated cell therapy of chronic pain.