Uso de canabidiol como terapia adjuvante em paciente com síndrome de Zellweger: relato de caso / Cannabidiol as adjuvant therapy in a patient with Zellweger syndrome: a case report

RESUMO: Também denominada síndrome cerebrohepatorenal, a síndrome de Zellweger é uma doença autossômica recessiva rara, pertencente ao espectro de erros inatos do metabolismo que afetam os peroxissomos. São causados principalmente por mutações em qualquer um dos 14 genes PEX diferentes que codificam para proteínas envolvidas na montagem do peroxissoma, sendo a mais comum do PEX1. O quadro clínico geralmente é observado no período neonatal e primeira infância, incluindo alterações faciais, hipotonia profunda e ausência de reflexos neonatais, além de disfagia, disfunção hepática e convulsões. O diagnóstico é feito a partir da clínica e testes bioquímicos e confirmados pela visualização da mutação em um dos 14 genes PEX. Como não há tratamento específico, é feito tratamento sintomático. Nosso paciente masculino de 1 ano e 9 meses apresentou a hipotonia congênita como sintoma marcante, além de crises convulsivas recorrentes logo após o nascimento. Evoluiu com necessidade de gastrostomia e estagnação de marcos neuromotores. O diagnóstico foi confirmado aos seis meses, através da dosagem de ácidos graxos de cadeia longa. Crises convulsivas evoluíram de maneira refratária a diversos anticonvulsivantes e com elevada frequência diária, por isso iniciamos canabidiol (CBD-RSHO GOLD) por via enteral que reduziu significantemente as crises. Não há tratamento definitivo para esta enfermidade, sendo importante tratamento sintomático das crises convulsivas e terapias de reabilitação, nesse caso, o uso de (CBD- RSHO GOLD) provocou uma redução de 92% na frequência de crises diárias do paciente. No entanto, não é possível concluir, ainda, melhoras em outros sinais e sintomas. (AU)

ABSTRACT: Also referred to as "brain-liver-kidney" syndrome, the Zellweger syndrome is a rare autosomal recessive disorder, belonging to the spectrum of inborn errors of metabolism that affect peroxisomes. They are caused mainly by mutations in any of the 14 different PEX genes that code for proteins involved in the assembly of peroxisome, being the most common of PEX1. The clinic is usually observed in the neonatal and early childhood period, including facial changes, deep hypotonia, and absence of neonatal reflexes in childhood, in addition to dysphagia, hepatic dysfunction, and seizures. The diagnosis is made from clinical and biochemical tests and confirmed by the visualization of the mutation in one of the 14 PEX genes. Since there is no specific treatment, symptomatic treatment is done. Our 1-year and 9-month-old male patient presented congenital hypotonia as a striking symptom in addition to recurrent seizures shortly after birth. It evolved with the need for gastrostomy and stagnation of neuromotor frames. The diagnosis was confirmed at six months by the measurement of long-chain fatty acids. Convulsive seizures evolved in a manner that was refractory to several anticonvulsants and with a high daily frequency, so we initiated cannabidiol (CBD-RSHO GOLD) by an enteral route that significantly reduced the seizures. Since there is no avail-able treatment for seizures, in this case, the use of CBD-RSHO GOLD reduced by 92% the daily seizure frequency. However, it is not possible to conclude further improvements in other signs and symptoms. (AU)

Abnormal expression of PEX10 gene may be related to epilepsy associated with 1p36 copy number variations / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of PEX10 gene and 1p36 copy number variations in 1p36 region with concurrent epilepsy through analyzing 3 cases.</p><p><b>METHODS</b>The karyotypes of 3 patients were determined by high resolution chromosome banding, multiplex ligation dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH) combined with single nucleotide polymorphism array (SNP) technology. Real-time PCR was carried out to determine the mRNA levels of PEX10 gene in peripheral blood of the patients.</p><p><b>RESULTS</b>No abnormality was found upon high resolution karyotyping. MLPA analysis showed that all of the 3 patients had a copy number variation of subtelomeric region in the short arm of chromosome 1, which was confirmed by FISH and SNP chip analyses. Case 1 and case 2 both had an epilepsy phenotype, and their copy number variations have encompassed the PEX10 gene. On the other hand, case 3 has absent epilepsy, and its PEX10 gene copy number was normal. Family investigation confirmed that the chromosome abnormalities in all of the 3 cases were of de novo type. Compared with healthy controls, real-time PCR showed that mRNA of the PEX10 gene was increased in case 1 but decreased in case 2.</p><p><b>CONCLUSION</b>The abnormal expression of PEX10 gene resulting from copy number variations of 1p36 region may be associated with the epilepsy phenotype.</p>

Screening the hepatitis B virus PreS1 associated protein by the yeast two-hybrid system / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen the hepatitis B virus PreS1 associated protein from normal human liver cDNA library by the yeast two-hybrid system and explore the role of PreS1 protein in the infection of hepatitis B virus (HBV).</p><p><b>METHODS</b>PCR was preformed to amplify the PreS1 gene containing EcoRI and PstI from HBV positive serum, and the production was inserted into plasmid pAS2-1 after digesting with the former two restricted endonuclease, then the bait vector pAS2-1-PreS1 was verified by auto-sequencing assay. The PreS1-BD fusion protein expressed in the yeast cells was confirmed by western blot, after pAS2-1-PreS1 was transfected into the yeast cell AH109. Yeast cells co-transfected with pAS2-1-PreS1 and the normal human liver cDNA library grew in selective SC/-trp-leu-his-ade2 medium, and the second screening was performed with LacZ report gene. Furthermore, segregation analysis and mating experiment were done to eliminate the false positive, then the true positive clones were submitted for PCR and sequencing. The results were submitted to the BLAST notebook of World Wide Wed Site NCBI to seek homologous sequence.</p><p><b>RESULTS</b>Bait vector pAS2-1-PreS1 included the anticipated fragment of PreS1 gene. Western blot showed that pAS2-1-PreS1 could correctly express PreS1-BD fusion protein in the yeast cells. After yeast cells co-transfected with pAS2-1-PreS1 and the normal human liver cDNA library, 97 colonies grew in the selective SC/-trp-leu-his-ade2 medium, only one clone was positive and showed high homology with Homo sapiens nascent-polypeptide-associated complex alpha polypeptide.</p><p><b>CONCLUSION</b>Bait vector pAS2-1-PreS1 is successfully constructed, and nascent-polypeptide-associated complex alpha polypeptide protein expressed in hepatocyte can interact with PreS1 by the yeast two-hybrid system.</p>