Cloning and Expression Analysis of PaAACT Gene from Phytolacca americana / 中国实验方剂学杂志

Objective: To clone the key enzyme gene involved in the biosynthesis of esculentoside A(EsA),acetoacetyl-CoA transferase(AACT) gene was cloned from Phytolacca americana for bioinformatics analysis and prokaryotic expression. Method: Total RNA was extracted from the root of P. americana, and then cDNA was synthesized through the reverse transcription. Based on analysis of the transcriptome data of P. americana, the specific primers of PaAACT gene were designed,and the cDNA sequence of PaAACT gene was amplified by polymerase chain reaction(PCR) method. Prokaryotic induction,expression and purification of the target protein were induced through the construction of the prokaryotic expression vector pET-32a-PaAACT. Result: The open reading frame (ORF) of PaAACT gene was 1 254 bp,and encoded 417 amino acid residues. Bioinformatic analysis showed that the molecular formula of PaAACT protein was C1 914H3 120N538O576S17,inferring that its molecular weight was 43.43 kDa,the theoretical isoelectric point was 8.90,and the instability index of PaAACT protein was 32.27,which was a stable protein. According to bioinformatics analysis,PaAACT protein was a member of the thiolase family and contained one conserved site and one active site of the thiolase family at the C-terminal. PaAACT protein may be located in the cytoplasm,without a signal peptide or transmembrane domain. The phylogenetic analysis indicated that PaAACT protein showed the highest homology with AACT protein from polygonaceae plants (such as Beta vulgaris). The recombinant PaAACT protein was successfully expressed in Escherichia coli BL21(DE3) strain through IPTG induction, and the purified target protein was obtained by Ni2+ affinity chromatography. Conclusion: In this study,the PaAACT gene was cloned from P. americana,which lays a foundation for further determination of enzyme activity assay of PaAACT and preparation of antibody,and provides the theoretical basis for studying its role in the biosynthesis pathway of EsA.

Transcriptome analysis reveals candidate genes involved in esculentoside A biosynthesis in Phytolacca americana / 药学学报

In order to study the biosynthesis pathway of esculentoside A, the Illumina HiSeq 4000 highthroughput sequencing method was used to analyze the transcriptome of Phytolacca americana seedlings. The 9.60 Gb clean data were obtained after the transcriptome of P. americana assembled by Trinity software. The total 63 957 unigenes were obtained after assembly and the average length was 988.82 bp, among them 24 517 unigenes (38.33%) were annotated in the public databases Nr, Swiss-Prot, COG, KOG, Pfam, GO and KEGG. According to the assignment of KEGG pathway, 53 unigenes were involved in terpenoid backbone biosynthesis and 8 unigenes involved in triterpenoid biosynthesis. Additionally, there were 417 unigenes assigned to other secondary metabolic pathways in P. americana. The post-modification enzyme genes involved in the esculentoside A biosynthesis were also analyzed in the transcriptome of P. americana. The results indicated that 130 unigenes may have the function of CYP450 which was involved in oxidation/hydroxylation modification of P. americana secondary metabolites. Furthermore, 46 unigenes had the function of glycosyltransferase UGT. The transcriptome data of P. americana laid a foundation for studying the biosynthesis pathway of esculentoside A and other secondary metabolites, and also provided theoretical basis for formation of medicinal materials quality.

Research progress on chemical constituents, pharmacological effects, and clinical applications of Phytolaccae Radix / 中草药

Phytolaccae Radix, including Phytolacca acinosa and P. Americana, is recorded by Chinese Pharmacopoeia for its important medicinal property. The most characteristic chemical constituents of Phytolaccae Radix are triterpenoid saponins, and there are also flavone, phenolic acid, sterol, and polysaccharides in it. Phytolaccae Radix has various pharmacological activities, such as diuretic, antibacterial, antiviral, anti-inflammation, anticancer effects, and so on, and it is usually used to treat hepatitis B, psoriasis, irritability, and so on. In this paper, we summarize the chemical composition, pharmacyological action, and clinical curative effect of Phytolaccae Radix to prompt the development and utilization of them.

Four Cases of Phytolacca Esculenta Van Houtte Intoxication due to Misidentification as a Ginseng

Poisoning by Phytolacca esculenta commonly occurs by misidentification as other edible plants. The root of Phytolacca esculenta is similar to other roots, such as kudzu, balloon flower, codonopsis lanceolata, and ginseng. The author experienced four cases of Phytolacca esculenta intoxication due to misidentification as a ginseng. We report on these cases with a review of the literature.

Clinical Features of Pokeweed Intoxication Experienced in a University Hospital

PURPOSE: Although Pokeweed intoxication is relatively less severe, thereis little data onthe clinical presentation ofPokeweed intoxication in Korea. This study examined the clinical aspectsto providebasic data for evaluating Pokeweed intoxication. METHODS: A retrospective study by a chart review was performedon 19 patients who ingested Pokeweed and presented to anacademic emergency department with an annual census of 40,000 between March 2012 and May 2012. RESULTS: Nineteen patients were identified. All patients wereintoxicated unintentionally. The most common symptoms were vomiting with diarrhea and abdominal pain. The onset time varied, but occurs 30 minutes to 5 hours post ingestion of Pokeweed. All patients were discharged without fatal complications. CONCLUSION: Compared to previous reports, mostpokeweed poisoning patients complain of gastrointestinal symptoms. Supportive care is the mainstay of the management of pokeweed intoxication. All symptoms were resolved over a 24 to 48 hour period.

Exploración del efecto de altas diluciones sucusionadas (medicamento homeopático) de phytolacca americana sobre linfocitos humanos. Fase III / Exploration of the effect of high dilutions succussed (homeopathic medicine) of phytolacca americana on human lymphocytes. Phase III

Phytolacca americana es una planta que preparada bajo el método homeopático se ha usado para el manejo de enfermedades que cursan con adenopatías, sabemos que in Vitro exhibe un efecto sobre la proliferación de células mononucleares de sangre periférica, principalmente linfocitos y sobre la producción de diversas citoquinas. El objetivo de este trabajo fue la exploración del efecto de Phytolacca americana sobre la proliferación de células mononucleares de sangre periférica y sobre la producción de TNFα, en diluciones consecutivas entre la 12 CH y la 18 CH. Para la evaluación de la viabilidad celular se usó el método MTT (Bromuro de 3(4,5 dimetil-2-tiazoil)-2,5- difeniltetrazólico) y para la cuantificación de la producción de TNFα el método ELISA, se analizaron muestras de sangre periférica de 10 individuos sanos. Se encontró de manera general aumento máximo de la proliferación celular 48 horas posterior a la exposición a Phytolacca, y un aumento en la producción de TNF α para la dilución 14 CH. Concluimos que Phytolacca americana produce un aumento de la proliferación de células mononucleares de sangre periférica y que en las diluciones 13, 14 y 18 CH se estimula la producción de TNFα, sin embargo, para el resto de diluciones, el efecto varía de manera individual.

Two Cases of Phytolacca Americana Intoxication with Confusion and Abdominal Cramping

Phytolacca americana poisoning is a benign plant intoxication that causes gastrointestinal symptoms, including abdominal cramps, vomiting, diarrhea, and gastrointestinal bleeding. Other signs and symptoms include diaphoresis, salivation, visual disturbance, and seizures or mental changes. We report two cases of patients who experienced confusion and abdominal pain, vomiting, and hematemesis after oral ingestion of pokeweed. A 60-year-old female with confusion and a 67-year-old female with abdominal pain, vomiting, and diarrhea were admitted to the emergency department after pokeweed poisoning. After supportive treatment of hydration and gastrointestinal medication, the two patients showed full recovery within 24 h and were discharged from the hospital.