Identification and expression analysis of R2R3-MYB gene family in Andrographis paniculata / 中国中药杂志

The plant growth, development, and secondary metabolism are regulated by R2 R3-MYB transcription factors. This study identified the R2 R3-MYB genes in the genome of Andrographis paniculata and analyzed the chromosomal localization, gene structure, and conserved domains, phylogenetic relationship, and promoter cis-acting elements of these R2 R3-MYB genes. Moreover, the gene expression profiles of R2 R3-MYB genes under abiotic stress and hormone treatments were generated by RNA-seq and validated by qRT-PCR. The results showed that A. paniculata contained 73 R2 R3-MYB genes on 21 chromosomes. These members belonged to 34 subfamilies, 19 of which could be classified into the known subfamilies in Arabidopsis thaliana. The 73 R2 R3-MYB members included 36 acidic proteins and 37 basic proteins, with the lengths of 148-887 aa. The domains, motifs, and gene structures of R2 R3-MYBs in A. paniculata were conserved. The promoter regions of these genes contains a variety of cis-acting elements related to the responses to environmental factors and plant hormones including light, ABA, MeJA, and drought. Based on the similarity of functions of R2 R3-MYBs in the same subfamily and the transcription profiles, ApMYB13/21/35/67/73(S22) may regulate drought stress through ABA pathway; ApMYB20(S11) and ApMYB55(S2) may play a role in the response of A. paniculata to high temperature and UV-C stress; ApMYB5(S7) and ApMYB33(S20) may affect the accumulation of andrographolide by regulating the expression of key enzymes in the MEP pathway. This study provides theoretical reference for further research on the functions of R2 R3-MYB genes in A. paniculata and breeding of A. paniculata varieties with high andrographolide content.

Screening of Reference Genes of Andrographis paniculata Under MeJA and Abiotic Stresses by Real-time Fluorescence-based Quantitative PCR / 中国实验方剂学杂志

ObjectiveTo screen the appropriate reference genes for real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）analysis of the Andrographis paniculata under methyl jasmonate（MeJA）and various abiotic stresses. MethodThe actin 1（ACT1），actin 2（ACT2），elongation factor（EF-1α），glyceraldehyde-3-phosphate dehydrogenase（GAPDH），tubulin（TUB），polyubiquitin（UBQ）， and 18S rRNA（18S）gene were selected as candidate reference genes based on the RNA-seq data of high temperature，drought， UV， and MeJA. The expression of seven candidate reference genes in the A. paniculata leaves was assessed by Real-time PCR，and the stability was analyzed by geNorm，NormFinder，BestKeeper， and Refinder. ResultThe results of stability evaluated by geNorm，NormFinder， and BestKeeper were not the same due to different indicators. As analyzed by Refinder， for the stability of the expression， the genes were ranked as UBQ>18S>EF-1α>ACT2>ACT1>GAPDH>TUB under high temperature stress， ACT1>UBQ>EF-1α>18S>ACT2>GAPDH>TUB under drought stress， EF-1α>TUB>ACT2>UBQ>18S>GAPDH>ACT1 under UV stress， and ACT1>EF-1α>UBQ>ACT2>18S>TUB>GAPDH under MeJA stress. Among them，18S gene was not suitable as an internal reference gene duo to its high expressive abundance. This study also verified the relative expression level of andrographolide synthesis-related gene hydroxy-methylglutaryl-CoA synthase （HMGS） in the four stresses on the basis of transcriptome data，and found that the Real-time PCR results of appropriate internal reference genes were accurate and reliable. ConclusionUBQ-ACT1-UBQ，EF-1α-TUB，and ACT1-EF-1α were the suitable combinations under stresses of high temperature，drought，UV， and MeJA. This study is expected to provide references for the research on function regulation and expression of genes in A. paniculata under high temperature，drought，UV， and MeJA stresses.

Directing construction of CRISPR/Cas9 vector of SmPAL1 in Salvia miltiorrhiza by target efficiency detection in vitro / 中国中药杂志

To construct CRISPR/Cas9 vectors for the editing of SmPAL1 in the phenylpropane metabolic pathway of Salvia miltiorrhiza, CIRSPR/Cas9 target sites of SmPAL1 were designed by online software. Its target efficiencies were detected in vitro by enzyme digestion and sequences with highly efficiency were constructed into CRISPR/Cas9 vectors. Three possible CRISPR target sequences (SmPAL1-g1, SmPAL1-g2, SmPAL1-g3) were designed and the enzyme digestion efficiencies were 53.3%, 76.6% and 10.0%. SmPAL1-g1 and SmPAL1-g2 were constructed into vector VK005-03 named as VK005-03-g1 and VK005-03-g2. The results of sequencing showed that the two CRISPR/Cas target sequences were all constructed into VK005-03. Here we first laid the foundation for the study of SmPAL1 and provided an effective strategy for the screening of sgRNA.

Management of an impacted maxillary central incisor with dilacerated root

Dilaceration is a comparatively uncommon dental deformity generally characterized by an angulation between crown and root, and consequently causing non-eruption of the tooth. Dilaceration generally occurs following trauma to the apices of deciduous dentition, which lies close to the permanent tooth buds. As a result, surgical extraction used to be the first choice when making the treatment plan for a case with severely dilacerated teeth. This case report presents the orthodontic alignment of a permanent maxillary right central incisor in an 8-year-old boy who had an impacted inverted maxillary central incisor, with distoangular root dilaceration through the crown. Following surgical exposure with the closed-eruption technique and appropriate orthodontic traction, the tooth was successfully aligned into the dental arch and the root was radiologically shown to be straightened and relatively well developed. The impacted dilacerated incisor diagnosed in the early mixed dentition should be treated with the aid of orthodontic traction

DSA diagnosis of nontumorous arterioportal shunt in cirrhosis / 介入放射学杂志

Objective To assess the diagnostic value of DSA for nontumorous arterioportal shunt in cirrhosis.Methods One hundred seventy cirrhotic patients without HCC under going transhepatic artery self bone marrow stem cell transplantation were taken liver arterial DSA together with 165 CT data.Arterioportal shunt(APS)appearance phase,persisting time and characteristics of liver parenchyma staining caused by APS were analyzed dynamically.Small HCC was differentiated from nontumorous APS.TACE procedure was done when HCC lesion was confirmed by DSA findings.Results 31.7%(54/170)DSA presented the sign of APS. Among the 142 cases with no evidence of APS on CT examinations,but APS was seen in DSA of 31 cases (21.8%).All patients with CT findings of liver abnormal enhancement of APS,23 cases including 16 cases as nontermorous and 7 cases whose CT arterial phase showing local persistant enhancement without declination in portal venous phase which were recognized to be HCC later by DSA characteristics with only one having tumorous APS.Conclusions DSA has a comparative high visualization rate of nontumorous APS in liver cirrhosis and benefits the differentiation with malignant lesions guiding the interventional procedure.

Subintimal recanalization for peripheral artery total occlusion / 中华放射学杂志

Objective To assess the technique and clinical outcome of subintimal recanolization (SIR)for peripheral artery total occlusion.Methods Eighteen patients with 20 arterial occlusions underwent SIR.The occlusive lesions were located in the lilac-femoral artery(14 lesions),the infrapopliteal arteries(4 lesions)and the left subclavian artery(2 lesions),respectively.Basic procedure of SIR was to create a subintimal path in the wall of the occluded artery and reentry the true lumen by using 0.035 inch diameter TERUMO guidwire and angiographic catheter.Subsequently,balloon angioplasty was performed in all recanalized lesions,and stent was placed in initial parts or whole recanalized path except one lesion in the lilac-femoral artery,and the lesions in the infrapopliteal arteries.Results Seventeen occlusive lesions in 15 patients were successfully recanalized by means of SIR,and the corresponding symptoms were disappeared or improved obviously.Technical failure occurred in 3 patients due to inability to reenter the distal true lumen or balloon angioplasty was not satisfactory.No complications of artery rupture,thrombosis or embolization were found.Conclusion SIR is an effective and safe therapy for peripheral artery total occlusion.This technique provides a feasible alternative to conventional percutaneous transluminal angioplasty especially in the complicated cases.