Specific DNA barcodes screening, germplasm resource identification, and genetic diversity analysis of Platycodon grandiflorum / 药学学报

Platycodonis Radix is the dry root of Platycodon grandiflorum of Campanulaceae, which has a variety of pharmacological effects and is a commonly used bulk Chinese medicine. In this study, the chloroplast genome sequences of six P. grandiflorum from different producing areas has been sequenced with Illumina HiSeq X Ten platform. The specific DNA barcodes were screened, and the germplasm resources and genetic diversity were analyzed according to the specific barcodes. The total length of the chloroplast genome of 6 P. grandiflorum samples was 172 260-172 275 bp, and all chloroplast genomes showed a typical circular tetrad structure and encoded 141 genes. The comparative genomics analysis and results of amplification efficiency demonstrated that trnG-UCC and ndhG_ndhF were the potential specific DNA barcodes for identification the germplasm resources of P. grandiflorum. A total of 305 P. grandiflorum samples were collected from 15 production areas in 9 provinces, for which the fragments of trnG-UCC and ndhG_ndhF were amplificated and the sequences were analyzed. The results showed that trnG-UCC and ndhG_ndhF have 5 and 11 mutation sites, respectively, and 5 and 7 haplotypes were identified, respectively. The combined analysis of the two sequences formed 13 haplotypes (named Hap1-Hap13), and Hap4 is the main genotype, followed by Hap1. The unique haplotypes possessed by the three producing areas can be used as DNA molecular tags in this area to distinguish from the germplasm resources of P. grandiflorum from other areas. The haplotype diversity, nucleotide diversity and genetic distance were 0.94, 4.79×10-3 and 0.000 0-0.020 3, respectively, suggesting that the genetic diversity was abundant and intraspecific kinship was relatively close. This study laid a foundation for the identification of P. grandiflorum, the protection and utilization of germplasm resources, and molecular breeding.

Screening of specific DNA barcode, identification of germplasm resources, and analysis of genetic diversity of Atractylodes chinenesis / 药学学报

italic>Atractylodes chinensis has important medicinal and economic values. In this study, the chloroplast genome sequences of four A. chinensis samples from different producing areas were sequenced using the Illumina platform. The specific DNA barcodes were screened and the germplasm resources of A. chinensis samples from different producing areas and the genetic diversity of the population were analyzed basing on the specific barcodes. The whole chloroplast genomes of the four A. chinensis samples had a typical cyclic tetrad structure, with 112 genes annotated. The comparative genomics results indicated that ccsA and trnC-GCA_petN were potential specific DNA barcodes for intraspecific identification of A. chinensis. Polymerase chain reaction (PCR) analysis of ccsA and trnC-GCA_petN was performed on 256 samples from 14 areas in 9 provinces, and the amplification efficiency was 100%. Sequence analysis showed that ccsA and trnC-GCA_petN had 11 and 22 variant positions, which could identify 16 and 22 haplotypes, respectively. The combined sequence analysis identified 39 haplotypes, named Hap1-Hap39, of which the most abundant and widely distributed genotype was Hap9. Haplotype diversity (Hd) = 0.896 and nucleotide diversity (Pi) = 0.002 22 indicated high genetic diversity at the species level in A. chinensis. The genetic distances of the haplotypes were 0.000 00-0.004 88, indicating that there were small genetic differences among the haplotypes. The results of phylogenetic tree analysis showed that 39 haplotypes had very close genetic relationship, and formed two obvious branches with other groups of the same genus except Atractylodes macrocephala. This study plays an important role in the identification of the origin of A. chinensis and the protection and breeding of germplasm resources.

Specific DNA barcodes, germplasm resources, and genetic diversity of Eleutherococcus senticosus / 中国中药杂志

Eleutherococcus senticosus is one of the Dao-di herbs in northeast China. In this study, the chloroplast genomes of three E. senticosus samples from different genuine producing areas were sequenced and then used for the screening of specific DNA barcodes. The germplasm resources and genetic diversity of E. senticosus were analyzed basing on the specific DNA barcodes. The chloroplast genomes of E. senticosus from different genuine producing areas showed the total length of 156 779-156 781 bp and a typical tetrad structure. Each of the chloroplast genomes carried 132 genes, including 87 protein-coding genes, 37 tRNAs, and 8 rRNAs. The chloroplast genomes were relatively conserved. Sequence analysis of the three chloroplast genomes indicated that atpI, ndhA, ycf1, atpB-rbcL, ndhF-rpl32, petA-psbJ, psbM-psbD, and rps16-psbK can be used as specific DNA barcodes of E. senticosus. In this study, we selected atpI and atpB-rbcL which were 700-800 bp and easy to be amplified for the identification of 184 E. senticosus samples from 13 genuine producing areas. The results demonstrated that 9 and 10 genotypes were identified based on atpI and atpB-rbcL sequences, respectively. Furthermore, the two barcodes identified 23 genotypes which were named H1-H23. The haplotype with the highest proportion and widest distribution was H10, followed by H2. The haplotype diversity and nucleotide diversity were 0.94 and 1.82×10~(-3), respectively, suggesting the high genetic diversity of E. senticosus. The results of the median-joining network analysis showed that the 23 genotypes could be classified into 4 categories. H2 was the oldest haplotype, and it served as the center of the network characterized by starlike radiation, which suggested that population expansion of E. senticosus occurred in the genuine producing areas. This study lays a foundation for the research on the genetic quality and chloroplast genetic engineering of E. senticosus and further research on the genetic mechanism of its population, providing new ideas for studying the genetic evolution of E. senticosus.

Cloning and subcellular location of the PnMYB1R1 gene from Panax notoginseng and expression analysis with different stresses / 药学学报

MYB transcription factors, one of the largest transcription factor families in plants, play an important role in signal transduction, plant growth and plant resistance. In this study a full-length cDNA of the PnMYB1R1 gene was cloned from Panax notoginseng. Sequence analysis, prokaryotic expression and purification, subcellular location, transcriptional activity analysis, tissue-specific analysis and expression analysis under different abiotic stresses was performed. The open reading frame (ORF) of PnMYB1R gene was 738 bp, encoding a protein of 245 amino acids with a predicted molecular mass (MW) of 27.0 kD. The sequence analysis and polygenetic analysis indicated that the PnMYB1R1 protein contains a conserved R3 domain, belonging to TRF-like protein in 1R-MYB-type transcription factors. The recombinant PnMYB1R1 protein was expressed in Escherichia coli BL21(DE3) cells using the prokaryotic expression vector pET28a-PnMYB1R1 and was purified. Subcellular localization analysis showed that PnMYB1R1 was localized in the nucleus. Transcriptional activity analysis indicated that the PnMYB1R1 transcription factor has transcriptional activation activity. Expression analysis indicated that PnMYB1R1 was primarily expressed in roots, followed by stems and leaves, and then rootlets. The expression level of PnMYB1R1 in root, stems, leaves and rootlets was influenced by salt, low temperature and drought treatment, while the abundance of PnMYB1R1 was significantly induced by salt stress in these tissues. These results provide valuable insights into the role of 1R-MYB transcription factors in plant defense.

Association of vitamin D level with asthma control and pulmonary function in asthmatic children aged 4-12 years / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the association of vitamin D level with asthma control and pulmonary function in children with asthma.</p><p><b>METHODS</b>A total of 150 children with asthma were enrolled as observation group, and 55 healthy children were enrolled as control group. According to the level of asthma control, the children were divided into good control group, partial control group, and non-control group. Chemiluminescence microparticle immunoassay was used to measure the serum level of 25-hydroxyvitamin D [25(OH)D] for all groups. According to the level of 25(OH)D, the asthmatic children were divided into normal vitamin D group, vitamin D insufficiency group, and vitamin D deficiency group. Pulmonary function was measured for all asthmatic children.</p><p><b>RESULTS</b>The observation group had a significantly lower serum level of 25(OH)D than the control group (25± 7 ng/mL vs 29± 4 ng/mL; P<0.05). The normal vitamin D group had the highest asthma control rate, followed by the vitamin D insufficiency group and the vitamin D deficiency group (P<0.05). There was no significant difference in pulmonary function among the three groups (P>0.05).</p><p><b>CONCLUSIONS</b>Asthmatic children have a lower serum level of 25(OH)D than healthy children. The serum level of 25(OH)D is associated with the level of asthma control and has no association with pulmonary function.</p>

Application of flexible bronchoscopy in children with respiratory diseases / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the significance of flexible bronchoscopy in children with respiratory diseases.</p><p><b>METHODS</b>A retrospective analysis was performed for the clinical data of 80 children who were hospitalized due to respiratory diseases (including severe pneumonia, Mycoplasma pneumoniae pneumonia with atelectasis/lung consolidation/local emphysema, protracted pneumonia, coughing and wheezing of unknown cause, chronic cough of unknown cause, and laryngeal stridor) and who underwent flexible bronchoscopy/alveolar lavage.</p><p><b>RESULTS</b>Bronchoscopy found that all the 80 children had endobronchial inflammation, among whom 28 children had severe airway obstruction by secretion. Twenty-four children had congenital airway dysplasia besides endobronchial inflammation, and three children had bronchial foreign bodies. In the children with coughing and wheezing of unknown cause and laryngeal stridor, some had congenital airway dysplasia or bronchial foreign bodies. Among the 27 children with Mycoplasma pneumoniae pneumonia, 26 had severe airway obstruction/embolization by secretion; 25 children (93%) underwent chest imaging again at 2 weeks after alveolar lavage, and the results showed complete or partial lung recruitment. Among the 80 children who underwent bronchoscopy, 3 had severe hypoxemia during surgery, 1 had epistaxis, 1 had minor bleeding during alveolar lavage, 3 had transient bronchospasm, and 5 had postoperative fever; these children were all improved after symptomatic treatment.</p><p><b>CONCLUSIONS</b>Flexible bronchoscopy is safe and reliable in children with respiratory diseases. Early alveolar lavage under a flexible bronchoscope is recommended for children with severe/refractory Mycoplasma pneumoniae pneumonia to improve prognosis. Flexible bronchoscopy is recommended for children with recurrent coughing and wheezing and persistent laryngeal stridor, in order to directly observe the throat and airway under an endoscope.</p>

Observation on therapeutic effect of acupuncture on restless legs syndrome / 中国针灸

<p><b>OBJECTIVE</b>To compare therapeutic effects of acupuncture combined with TDP radiation and western medicine on restless legs syndrome.</p><p><b>METHODS</b>One hundred and fifty-eight cases of restless legs syndrome were randomly divided into an observation group and a control group, 79 cases in each group. The treatment group were treated with acupuncture at Shenshu (BL 23), Dachangshu (BL 25), Guanyuanshu (BL 26), Zhibian (BL 54), Taixi (KI 3), etc., and TDP radiation. The control group were treated with L-Dopa. Clinical observation was made for 30 days.</p><p><b>RESULTS</b>In the observation group, 42 cases were cured, 30 cases were effective and 7 cases were ineffective with a total effective rate of 91.1%; in the control group no case was cured, 24 cases were effective and 55 cases were ineffective with a total effective rate of 30.4%, with a significant difference between the two groups in the total effective rate.</p><p><b>CONCLUSION</b>Acupuncture combined with TDP radiation has a better therapeutic effect on restless legs syndrome.</p>

Estimation on the level of birth and death rates of population in the three gorges area by means of capture-mark-recapture method / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluate quality of surveillance and emendate rates of birth and death of population of the Three Gorges area.</p><p><b>METHODS</b>Data on the two samples collected were designed based on principle of capture-recapture method. An investigation of missing report of birth and death was conducted in 7061 families selected through stratified random sampling method. We collected and registered the data of birth and death in every family investigated and checked with correlative records reported in disease surveillance system of the Three Gorges area. The missing report rates and the 95% confidence intervals of birth rate and death rate were calculated.</p><p><b>RESULTS</b>The underreporting rates of birth and death were 13.91% and 15.60% and death of infant was 33.33%. The emended birth rate was 8.92 per thousandth and the 95% confidence interval of birth rate was 8.38 per thousandth-9.45 per thousandth. The emended report rate of death was 6.88 per thousandth and the collectivity 95% confidence interval was 6.37%-7.38 per thousandth.</p><p><b>CONCLUSION</b>Results showed that the quality of birth and death in the disease surveillance reporting system of Three Gorges area was competent to the quality level of the standard set for national disease surveillance system. The birth and death rates of population in the Three Gorges area were under 10.00 per thousandth.</p>

Alteration of the content of minor elements in apoptotic cells induced by anti-cancer drugs with synchronous radiation / 浙江大学学报·医学版

OBJECTIVE: To explore the change of content of minor elements and its biological implication in apoptotic processes. METHODS: The content of minor elements of apoptotic cells from breast cancer induced by anti-cancer drugs was quantitatively analysed with synchronous radiation X-ray fluorescence. RESULTS: Seven kinds of minor elements including Ti, Cr, Fe, Cu, Zn, Ga and Ge as well as five major elements P, S, Ca, Cl and K were detected in the apoptotic and control cells. The content of elements Zn and P in apoptotic cells after 48 h treatment with taxol and colchicine was significantly increased higher than that in the control cells P<0.05). The content of element Fe in 48 h treatment with taxol was significantly decreased lower than that in the control cells P<0.05). The same trend of change of elements was observed during the process of apoptotic cells death induced with VP-16 or cycloheximide. CONCLUSION: This results suggest that the elements Zn, Fe and P should be involved in apoptosis induced by anti-cancer drugs.