This study investigated the choroplast genome sequence of wild Atractylodes lancea from Yuexi in Anhui province by high-throughput sequencing, followed by characterization of the genome structure, which laid a foundation for the species identification, analysis of genetic diversity, and resource conservation of A. lancea. To be specific, the total genomic DNA was extracted from the leaves of A. lancea with the improved CTAB method. The chloroplast genome of A. lancea was sequenced by the high-throughput sequencing technology, followed by assembling by metaSPAdes and annotation by CPGAVAS2. Bioiformatics methods were employed for the analysis of simple sequence repeats(SSRs), inverted repeat(IR) border, codon bias, and phylogeny. The results showed that the whole chloroplast genome of A. lancea was 153 178 bp, with an 84 226 bp large single copy(LSC) and a 18 658 bp small single copy(SSC) separated by a pair of IRs(25 147 bp). The genome had the GC content of 37.7% and 124 genes: 87 protein-coding genes, 8 rRNA genes, and 29 tRNA genes. It had 26 287 codons and encoded 20 amino acids. Phylogenetic analysis showed that Atractylodes species clustered into one clade and that A. lancea had close genetic relationship with A. koreana. This study established a method for sequencing the chloroplast genome of A. lancea and enriched the genetic resources of Compositae. The findings are expected to lay a foundation for species identification, analysis of genetic diversity, and resource conservation of A. lancea.
Phylogeny ; Atractylodes/genetics* ; Genome, Chloroplast ; Whole Genome Sequencing ; Microsatellite Repeats ; Lamiales

Objective:To explore the application value of metagenomics sequencing in the etiological diagnosis of bacterial meningitis(BM) in children and improve the diagnosis and treatment effect of BM in children.Methods:All BM cases were collected from Jiangxi Provincial Children′s Hospital from February 1, 2018 to January 31, 2020.Meanwhile, such biological samples as blood and cerebrospinal fluid were collected for traditional etiological testing and metagenomics sequencing.The results of traditional etiological testing were regarded as the gold standard, and the specificity and sensitivity of metagenomics sequencing in the diagnosis of BM in children were confirmed.Results:A total of 45 cases were collected in this study, including 31 males and 14 females, with the age between (74.74 ± 58.67) months.Twenty-six cases were identified by metagenomics sequencing, with the positive rate being 57.78%.Among them, there were 8 cases of Streptococcus pneumoniae, 2 cases of Escherichia coli, 2 cases of Neisseria meningitidis, 2 cases of Staphylococcus, 2 cases of Salmonella, 2 cases of Mycobacterium multiplex, 1 case of Streptococcus intermedius, 1 case of Streptococcus pyogenes, 1 case of Streptococcus paris, 1 case of Streptococcus salivarius, 1 case of Haemophilus influenzae, 1 case of Pseudomonas aeruginosa, 1 case of Acinetobacter baumannii, and 1 case of Aspergillus.The traditional etiological positive rate was 17.78%, and the metagenomic next-generation sequencing (mNGS) positive rate was 57.78% ( P=0.014, kappa=0.273). As per the comparison results, the sensitivity, specificity, positive predictive value, negative predictive value, Youden index and misdiagnosis rate were 100.00%, 51.35%, 30.76%, 100.00%, 51.36%, 48.64% and 0, respectively. Conclusions:Metagenomics examination had high sensitivity, which can improve the etiological diagnosis rate of acute BM in children, especially in case of high clinical suspicion of infection.Therefore, metagenomics examination should be selected as early as possible when the etiology cannot be determined by traditional approaches.

Objective:To explore the clinical characteristics of pediatric glucose transporter type 1 deficiency syndrome (GLUT1 DS), evaluate the efficacy and safety of ketogenic diet therapy (KDT).Methods:Clinical data of 19 children with GLUT1 DS admitted to Children′s Hospital of Fudan University, Tianjin Children′s Hospital, Shenzhen Children′s Hospital, Children′s Hospital of Nanjing Medical University and Jiangxi Provincial Children′s Hospital between 2015 and 2019 were collected retrospectively. The first onset symptom, main clinical manifestations, cerebrospinal fluid features and genetic testing results of patients were summarized, the efficacy and safety of ketogenic diet treatment were analyzed. Results:Among the 19 cases, 13 were males and 6 females. The age of onset was 11.0 (1.5-45.0) months,the age of diagnosis was 54.0 (2.8-132.0) months. Epilepsy was the first onset symptom of 13 cases. Different forms of tonic-clonic seizures were the most common types of epilepsy (7 cases with generalized tonic-clonic seizures, 5 cases with focal tonic or clonic seizures, 4 cases with generalized tonic seizures). Antiepileptic drugs were effective in 4 cases. Paroxysmal motor dysfunction was present in 12 cases and ataxia was the most common one. All patients had different degrees of psychomotor retardation. Among 17 patients received cerebrospinal fluid examination, cerebrospinal fluid (CSF) glucose level was lower than 2.2 mmol/L and CSF glucose/glycemic index was<0.45 in 16 cases, only 1 case presented normal CSF glucose level (2.3 mmol/L) and normal CSF glucose/glycemic index(0.47). SLC2A1 gene mutations were found in 16 patients, missense, frameshift and nonsense mutations were the common types with 5 cases, 5 cases and 3 cases respectively. All 19 patients were treated with ketogenic diet, which was effective in 18 cases in seizure control, 11 cases in dyskinesia improvement and 18 cases in cognitive function improvement. No serious side effects were reported in any stage of KDT.Conclusions:The diagnosis of GLUT1 DS is often late. It is necessary to improve the early recognition of the disease and perform CSF glucose detection and genetic testing as early as possible. The KDT is an effective and safe treatment for GLUT1 DS, but a small number of patients have not response to diet therapy.

To achieve a comprehensive understanding of heavy metals and harmful elements residues in Niuhuang Qingwei Pills,49 samples from 18 manufactures were collected from 31 provinces in China.Risk assessment and control preparations were applied innovatively in evaluation of exogenous pollution in traditional Chinese Medicine.Determination methods for Pb,Cd,As,Hg and Cu were established by inductively coupled plasma mass spectrometry(ICP-MS).Based on the procedures including hazard identification,hazard characterization,exposure assessment and risk characterization,risk assessment was performed and residual limits for Pb,Cd,As,Hg and Cu in the drug were formulated.The results showed that the hazardous quotients(HQ) of the elements were decreased in the following order:Pb>As>Cu>Hg>Cd,and the total hazardous index(HI) of heavy metals and harmful elements in Niuhuang Qingwei Pills was above 1,implying health risk of the drug.Under the proposed limits,5 elements in the control preparation as well as Cd and Cu in the samples were within the limits range,but the excess rates of Pb,As and Hg in the samples were 12%,12% and 14%,respectively.For the first time,basic steps for risk assessment of Chinese patent medicine were established,which provided model and reference for risk assessment and limit formulation of other drugs.
China ; Drug Contamination ; Drugs, Chinese Herbal ; analysis ; standards ; Metals, Heavy ; analysis ; Risk Assessment

OBJECTIVETo investigate the immunological mechanism of prednisone in the treatment of infantile spasm (IS) by evaluating the immune function of IS children before and after treatment.

METHODSThirty children with IS were enrolled as IS group. Thirty healthy infants who underwent physical examination were enrolled as healthy control group. Fasting venous blood was collected for both groups before and after prednisone treatment. Chemiluminescence was used to measure serum levels of interleukin-1B (IL-1B), interleukin-2R (IL-2R), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α). Immunoturbidimetric assay was used to measure serum levels of immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin G (IgG). Flow cytometry was used to measure the percentages of T lymphocyte subsets (CD3, CD4, and CD8). The clinical outcome and electroencephalographic findings were evaluated for all IS children after prednisone treatment.

RESULTSThe IS group had significantly higher serum levels of IL-2R, IL-8, and TNF-α than the healthy control group before treatment (P<0.05). The mean number of daily ictal clusters was positively correlated with the levels of IL-2R, IL-8, and TNF-α in IS children, the mean number of total daily seizures was positively correlated with IL-8 level, and any two indices out of IL-2R, IL-8, and TNF-α were positively correlated with each other (P<0.05). Among the 30 IS children treated with prednisone, 19 achieved seizure control; electroencephalography showed that 18 children achieved complete remission of hyperarrhythmia. After treatment, the IS group had significant reductions in the numbers of daily ictal clusters and total daily seizures, significant improvement in developmental quotient (P<0.05), and significant reductions in serum levels of IL-2R, L-8, and TNF-α, the percentage of CD4T lymphocytes, and CD4/CD8ratio (P<0.05), as well as a significant increase in the percentage of CD8T lymphocytes (P<0.05).

CONCLUSIONSIS children have immune dysfunction. Prednisone can control seizures in IS children, possibly by regulating and improving immune dysfunction.

CD4-CD8 Ratio ; Cytokines ; blood ; Electroencephalography ; Female ; Humans ; Infant ; Male ; Prednisone ; therapeutic use ; Spasms, Infantile ; drug therapy ; immunology

Objective By studying the changes of the seizures of infantile spasm(IS)、EEG and HPA axis function before and after the treatment of prednisone,to explore the efficacy of prednisone in treating infan-tile spasm,the role of HPA axis in the pathogenesis of IS,and elucidate the HPA axis mechanism of prednisone in controling seizure.Methods A total of 30 patients with IS (IS group) and 30 cases of healthy infants and young children (control group) were recruited.Number of seizures、EEG、HPA axis function was detected be-fore and after the treatment of prednisone in patients with infantile spasm.Serum cortisol,ACTH were deter-mined by the chemiluminescence analysis,serum CRH was measured by enzyme-linked immunosorbent assay. Results serum CRH levels of IS group was significantly higher than normal control group(P<0.05).Serum cortisol,ACTH in IS group were no evidently different compared with control group (P>0.05).The average number of daily ictal clusters and the average daily total seizure number positively correlated with CRH respec-tively.After the application of the prednisolone,seizure of 19 cases of the IS were controlled,11 cases were not controlled,18 cases of hyperarrhythmia were completely remited and 12 cases of hyperarrhythmia were not com-pletely remited.The average number of daily ictal clusters and The average daily total seizure number after treat-ment were significantly lower than before treatment(P<0.05);DQ after treatment was higher than DQ before treatment(P<0.05);The pathogenesis was the main influencing factor of the prednisone treatment effect,the length of the disease,the worse the treatment(P<0.05).CRH、cortisol、ACTH after treatment were significantly lower than before treatment(P<0.05).Conclusion Prednisone can effectively control the onset of infantile spasms,and early treatment is better.IS patient has HPA axis dysfunction,and prednisone can regulate HPA axis dysfunction to control spasm.

Objective To analyze the potential risk factors of infantile spasms (IS) relapse through following up the respondents with IS after different treatment protocols.Methods Sixty-nine cases were collected in the Department of Pediatric Neurology,Jiangxi Children's Hospital from May 2011 to September 2013,who had complete cessation of spasms for at least 28 days or more after the different treatment protocols.The follow-up was performed on these patients until spasms seizure relapse or at least 1 year for those without recurrence.According to the literature review,8 possible risk factors of IS recurrence (gender,age of onset,course of diseases,etiology,high irregular types of electroencephalogram,development quotient,onset time,treatment protocols) were selected,and then Logistic multiple regression was used to analyze the relationship of various potential risk factors with the relapse of spasms.Results (1) The recurrence rate at 6 months and 12 months were 40.6％ (28/69 cases)and 43.5 ％ (30/69 cases),respectively.(2) Among the various potential factors,the age at onset and the time to response were closely related to the IS recurrence.Namely,the non-classic onset(early-onset and late-onset) of IS were more likely to relapse than the classic onset[66.7％ (14/21 cases) vs 33.3％ (16/48 cases),x2 =6.605,P =0.010];the responders beyond 1 week were more likely to relapse than those within 1 week[63.6％ (14/22 cases) vs 34.0％ (16/47 cases),x2 =5.341,P =0.021].There were significant differences between the 2 groups (all P ＜ 0.05).(3) Logistic multiple regression analysis demonstrated that age at onset (Wald =3.603) was most closely related to the relapse of spasms.Conclusions (1) The relapse rate of IS in children was high,and the majority of them relapsed within 6 months.So a long-term,rational and effective clinical management solution should be explored.(2) The age at onset and the time to response are very important risk factors of the IS recurrence,and the former was more significant.So,early diagnosis and early treatment are more likely to improve the efficacy of IS,and reduce the risks of recurrence and improve the prognosis.

Objective To discuss the key points of endovascular therapy for complex subclavian artery occlusive diseases. Methods During the period from January 2012 to December 2013, a total of 92 patients with complex subclavian artery occlusive disease were admitted to Xuanwu Hospital of Capital Medical University, Beijing, China. The clinical data were retrospectively analyzed. The features of the lesions, the success rate of endovascular therapy, the use of combined approaches, the relief of symptoms after treatment, etc. were evaluated. Results The complex subclavian artery occlusive diseases could be divided into three types. Type Ⅰ: long segment of the left subclavian artery was occluded; type Ⅱ: ostial stenosis or occlusion of the right subclavian artery; and type Ⅲ: subclavian artery stenosis or occlusion was associated with the ostial disorder of the vertebral artery, or the opening of vertebral artery was affected by the subclavian artery stenosis or occlusion. The technical success rate was 82.6%. Combination use of femoral artery and brachial artery approach was employed in 27.2% of patients, which had improved the technical success rate. After the treatment the symptom improvement rate was 81.6%. Conclusion Upper limb artery approach can improve the re-canalization rate of left subclavian artery with long segment occlusion, and can ensure the accurate positioning of stent at the site of right subclavian artery opening. During the procedure of endovascular intervention for subclavian artery occlusion disease, attention should be paid to the protection of the vertebral artery.

OBJECTlVE To establish Tg(-6.3CYP3A65∶EGFP) transgenic zebrafish for quick, intuitive detection of heavy metals ( copper, cadmium and zinc) , dioxin-like PCBs ( PCB126) and other environmental pollutants. METHODS Tol2 transposon system was used to generate transgenic zebrafish lines Tg(-6.3CYP3A65∶EGFP) in which CYP3A65 promoter regualated labeled fluorescence. The effect of heavy mentals ( copper, cadmium and zinc ) and PCB126 on the relative amounts of CYP3A65 gene expression was determined by observing the change in fluorescence intensity. RESULTS The relative gene expression of CYP3A65 was significantly increased after 96 h exposure to copper 0.1 and 0.2μmol·L-1 , cadmium 0.35 and 0.7μmol·L-1 , zinc 1.5 and 3μmol·L-1 , and PCB126 2-32μmol·L-1 , respectively ( P<0.01) , but decreased after 96 h exposure to copper 0. 9 μmol·L-1 , cadmium 2. 7 and 5.4 μmol·L-1 , and zinc 24μmol·L-1 , respectively( P<0.01) . CYP3A65 gene expression was significantly increased after 168 h exposure to copper 0.1 and 0.2 μmol·L-1 , cadmium 0.35 and 0.7 μmol·L-1 , zinc 1.5 and 3 μmol·L-1, and PCB126 2-32 μmol·L-1, respectively(P<0.01), but decreased after 168 h exposure to copper 0.9 μmol·L-1, cadmium 2.7 and 5.4 μmol·L-1, and zinc 12 and 24 μmol·L-1( P<0.05) , in a concentration-dependent manner. CONCLUSlON The results suggest that zebrafish CYP3A65 gene expression and the CYP3A65 labeled fluorescence lines can be another candidate biomarker for detecting environmental pollutants.

Objective To evaluate the clinical efficacy and safety of different doses of prednisone combined with topiramate(TPM) in the treatment of infantile spasms(IS),in order to provide a new choice of the therapy of IS.Methods Fifty-six cases were collected in the Department of Neurology of Jiangxi Children's Hospital from May.2011 to Dec.2012.They were randomly divided into 2 groups:control group and trial group.The patients in control group took prednisone tablet of 1 mg/kg,2 times a day for 2 weeks;and the patients in trial group took prednisone tablet of 10 mg/ d,4 times a day for 2 weeks.In addition,TPM was used in both groups by initial dose 1 mg/(kg · d) or 2 times a day,and then was gradually increased to 3-5 mg/(kg · d) within 2 weeks.For those children in whom the spasms seizure completely ceased after 2 weeks,prednisone was then reduced by degrees to be discontinued for a 7-weeks course(extending to 4 weeks with the initial doses if spasms continued after 2 weeks).All patients underwent the assessment of spasms seizure and a 3-12 h video-electroencephalogram monitoring including wake and sleep states,which were performed before treatment,after 2 weeks and the end of the courses (7 or 9 weeks after treatment),respectively.Meanwhile,the side effects of the drugs during the treatment were recorded.The developmental quotient (DQ) tests of children with complete cessation of spasms more than 6 months were performed before treatment and after 6 months.All patients had been followed up for 2-18 months.Results 1.After 2 weeks of the therapy,the rate of cessation of spasms were 75.00％ (21/28 cases) and 28.57％ (8/28 cases) in the trial group and the control group,respectively,there was significant difference (x2 =12.087,P =0.001).And in the same term,the rate of complete resolution of hypsrrhythmia were 60.71％ (17/28 cases) and 21.43 ％ (6/28 cases),respectively,there was significant difference (x2 =8.928,P =0.003).At the end of treatment,the rate of cessation of spasms were 67.86％ (19/28 cases) and 35.71％ (10/28 cases) in the trial group and control group,respectively,there was significant difference (x2 =5.793,P =0.016).And in the mean time,the rate of complete resolution of hypsrrhythmia were 57.14％ (16/28 cases) and 14.29％ (4/28 cases),respectively,there was significant difference (x2 =11.200,P =0.001).2.Weight gain and increased appetite were the most frequent side effects.The incidence of side effects were 82.14％ (23/28 cases) and 67.86％ (19/28 cases) in the trial group and control group,respectively,there was no significant difference between the 2 groups (x2 =1.524,P =0.217).No death occurred in this clinical trail and no one discontinued the treatment protocol as result of the adverse events.3.The recurrence rate in the trial group and the control group were 31.82％ and 72.73％,respectively.And there was significant difference between the 2 groups (x2 =4.950,P =0.026).In the trial group,there were 9 cases with cessation of spasms more than 6 months,whose average business development values had no significant difference before and after treatment (t =2.271,P =0.053).Conclusion The efficacy of large-dose prednisone combined with TPM for IS was significantly better than that of conventional dose prednisone combined with TPM.