Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.

Objective:To investigate the pathogenic characteristics of viral encephalitis or meningitis in children living in Shijiazhuang city and surrounding areas, and to study the relationship between pathogenic and clinical findings.Methods:A total of 132 cerebrospinal fluid specimens were randomly collected from hospitalized children diagnosed with viral encephalitis or meningitis (January 2018 to December 2018) in the Department of Neurology of Hebei Children′s Hospital in Shijiazhuang city and surrounding areas. The nucleic acids of four viruses in cerebrospinal fluid were detected by real-time quantitative PCR. SPSS 21.0 software was used for statistical analysis.Results:Among the 132 cases, 78 were boys and 54 were girls, with a sex ratio of 1.44∶1. However, in the gender composition of children in each age group, there was no significant difference (χ 2=3.901, P=0.272). Of the 132 children, 121 had signs of fever, 109 had symptoms of headache, 92 had symptoms of vomiting, 17 had abnormal electroencephalogram(EEG), and 15 had abnormal head magnetic resonance imaging(MRI). 132 cerebrospinal fluid specimens were tested for pathogenic pathogens, and 80 of them were successfully detected. There was 1 case of human herpesvirus type I(HHV-I), 2 cases of varicella-zoster virus (VZV) and 77 cases of enterovirus(EV). The age group of 1~3 years′s EV detection rate was 66.67%, it is the highest, but overall, the EV detection rate, there was no significant difference among the four age groups (χ 2=3.147, P=0.369). The detection rate of EV in summer and autumn were 65.52% and 70.83%, respectively, which were significantly higher than those in spring and winter (33.33% and 0.00%), and there was a significant difference (χ 2=22.504, P=0.000). There was no significant difference in the positive rates of fever, headache and vomiting between EV-positive and non-EV-positive children ( P>0.05). There was no significant difference in the incidence of abnormal EEG and abnormal head MRI between EV-positive and non-EV-positive children ( P>0.05). Conclusions:In 2018, EV was the main pathogen of viral encephalitis and meningitis in children in Shijiazhuang city and surrounding areas, and EV detection rate was high in summer and autumn.

Objective: To investigate the pathogenic characteristics of viral encephalitis in children living in Hebei province. Methods: We randomly collected cerebrospinal fluid specimens from a total of 399 children diagnosed with viral encephalitis in Hebei Children′s Hospital from May to December 2017. Real-time fluorescence quantitative PCR and Sanger sequencing were used to detect viral nucleic acids in cerebrospinal fluid by an automatic laboratory station. Statistical analysis was performed on the experimental data using SPSS 21.0 software and the clinical data were analyzed. Comparison of infection rates of EV encephalitis in different months, using line × column chi-square test. The MRI and EEG positive rates of different viral encephalitis and viral encephalitis patients not infected with the virus were analyzed by Fisher′s exact probability test. The positive rate of infection with different viruses and non-virus agents was analyzed by Fisher′s exact probability test. Results: The result showed that 80 of 399 samples were positive, and the positive rate was 20.05%. It included 22 cases of enterovirus, 4 cases of influenza A virus, 3 cases of mumps virus, 2 cases of herpes simplex virus type 1, 1 case of herpes simplex virus type 2, 4 cases of EB virus, 7 cases of cytomegalovirus, 7 cases of herpes zoster virus, 8 cases of adenovirus, 14 cases of human herpesvirus type 6. Eight cases had combined viral infection. Eight cases had concurrent infections: 3 cases had enterovirus and herpesvirus type 6 concurrent infection, 1 case had enterovirus and Japanese encephalitis virus concurrent infection and 1 case had herpes simplex virus type 2 and adenovirus, 1 case had influenza A virus herpesvirus type 6, 1 case had mumps virus and herpesvirus type 6, 1 case had mumps virus and herpesvirus type 6, 1 case had herpes simplex virus type 1 and herpes zoster virus concurrent infections. Children with EV viral encephalitis in Hebei Province were highly prevalent in May and June (P=0.016). HHV6 virus encephalitis was more susceptible to infection than non-HHV6 virus (P=0.016); The rate of MRI positive findings in patients with different viral encephalitis was not statistically significant (P>0.05). The result of EEG of different viral encephalitis were P>0.05, which was not statistically significant. Conclusions EV was the most common pathogen of children with viral encephalitis in Hebei province. Encephalitis caused by influenza A virus cannot be ignored in clinical practice.