Cloning and expression analysis of CtbHLH transcription factor gene from Carthamus tinctorius and construction of plant expression vector / 中国中药杂志

To clone bHLH( basic helix-loop-helix) gene from Carthamus tinctorius,analyze the expression level in different plant tissues and construct the plant expression vector. The bHLH1 gene was cloned by RT-PCR techniques,and the protein characteristics were analyzed by bioinformatics,and phylogenetic tree was constructed. The expression of bHLH1 gene in different tissues and the roots after inoculated by Fusarium oxysporum were analyzed using real time-PCR,and the plant expression vector p BASTA-bHLH1 was constructed. The obtained ORF sequence of bHLH1 gene was 897 bp,encoded a protein of 298 amino acids. Sequence alignment and phylogenetic tree analyses showed that C. tinctorius bHLH1 had a certain homology with other species of amino acids,and was the most similar to the amino acid sequence of tobacco. Real-time PCR results showed significant differences,CtbHLH1 gene in red flower petals in different tissues and different flowering period had remarkable difference in expression level,its high amount expressed in petals,flowers third day after blossom expressed the highest quantity,at the end of the flowering the expression quantity is low. In addition,it is expressed in the root,and the expression in the stem and leaves is extremely low. The bHLH1 gene of C. tinctorius is successfully cloned,and the expression is analyzed. The plant expression vector p BASTA-bHLH is constructed.

Predictive Value of Fetal Umbilical Artery Doppler on Adverse Pregnancy Outcomes in Women with Systemic Lupus Erythematosus / 中山大学学报(医学科学版)

[Objective]To explore the adverse pregnancy outcomes(APO)and examine the predictive value of umbil-ical artery Doppler in systemic lupus erythematosus(SLE)pregnancies.[Methods]Data of 273 pregnancies from 2010 to 2016 were analyzed retrospectively. Pulsatility index(PI),resistance index(RI),and systolic/diastolic ratio(S/D)of the umbilical artery flow velocity data were monitored by Doppler ultrasound.[Results]One or more APO occurred in 61.9% of patients with SLE,among which pregnancy loss occurred in 60 cases;preterm birth in 56 cases;intrauterine growth retardation occurred in 20 cases;and fetal distress occurred in 21 cases.Twelve of pregnancies resulted in neona-tal Lupus. In total,118 patients underwent fetal umbilical artery Doppler during 28~34 gestational weeks. Doppler PI, RI,and S/D were significantly higher in the APO groups than in the patients without APO.[Conclusion]Pregnancies in lupus still had an increased risk of APO.Umbilical artery Doppler was useful in predicting APO in lupus pregnancies.

Molecular properties of grass carp reovirus in southern China and establishment of a duplex PCR detection method / 病毒学报

A strain of grass carp reovirus was isolated from sick grass carp with symptoms of haemorrhage in Guangdong province in 2009. The strain was tentatively named as GCRV-GD108 because it was isolated from grass carp and possessed 11 segments of dsRNA. Complete genome sequence analysis showed that significant differences existed between GCRV-GD108 and GCRV, as well as other known species of aquareovirus. In this study, molecular characteristics of diseased grass carp collected from different farms in Guangdong, Fujian and Hunan provinces were assayed. Based on the sequences of the 11 segments of GCRV-GD108, PCR primers corresponding to each of the segments were designed and synthesized. Total RNA of the diseased fish was extracted and used as templates of RT-PCR reaction. Specific amplification bands were obtained from all of the samples whereas no band was produced from GCRV standard strain. While using the primers specific to GCRV produced specific band in GCRV standard strain rather than in these collected samples. Sequencing of the amplification products showed that these samples displayed high similarities with each other (95.2%-99.4%), and they also shared high sequence similarities with that of GCRV-GD108 (95.0%-99.8%), suggesting that these samples shared similar molecular characteristics with those of GCRV-GD108, and were quite different from GCRV as well as the known species of genus aquareovirus. The results indicated that there are different molecular types of reovirus existed in the pond-cultured grass carp in China, and GCRV-GD108 is a representative strain in southern China, therefore great attention should be paid in order to control the disease efficiently, especially in vaccine preparation. Two pairs of primers were chosen to establish a duplex PCR assay method by combining each pair of the primers specific to GCRV-GD108 with the GCRV primer pair respectively. The duplex PCR assay method will enable the identification of GCRV-GD108 or GCRV by only a single PCR reaction.