Object To study the feasibility of suspension culture of protocorm in Dendrobium candidum Wall ex Lindl and effect of inoculum and medium volume on the growth of protocorm Methods Effect of four basic media MS, 1/2 MS, 67 V, and B 5, inoculum and medium volume on the growth of protocorm were studied by completely random experimental design and orthogonal test design Results The growth of D candidum protocorms in liquid medium was markedly better than that in solid medium (P0 05), B 5 was much better than 1/2 MS (P

Pseudolaricis Cortex and its adulterants were identified using DNA barcoding technique in this study. The sequences of ITS2 of Pseudolaricis Cortex and its five adulterants including 37 samples were analyzed. Se-quences were assembled using CodonCode Aligner 3.5.7. K2P distances were calculated and NJ tree was per-formed applying MEGA5.0. The ITS2 secondary structure was predicted using the ITS2 database and website. The results showed that the haplotypes of ITS2 regions of Pseudolaricis Cortex were the same as that of the original plant leaves. The inter-specific K2P distances of ITS2 were obviously higher than the intra-specific ones. Sam-ples of Pseudolaricis Cortex were clustered into a single clade in the NJ tree. The secondary structure of ITS2 of Pseudolaricis Cortex was significantly different from its adulterants. Therefore, ITS2 could powerfully discriminate Pseudolaricis Cortex and its adulterants.

Objective:To investigate the characteristics of polypoidal lesions and abnormal branching vascular network (BVN) in patients with polypoidal choroidal vasculopathy (PCV) by optical coherence tomography angiography (OCTA).Methods:A case series study was conducted.Untreated 45 eyes of 42 patients with initial diagnosis of PCV from May 2016 to April 2017 in Peking Union Medical College Hospital were enrolled.There were 26 males (27 eyes) and 16 females (18 eyes) with the average age of (68.59±8.52) years.All patients underwent fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA) and OCTA.The OCTA image features and detection rate of polypoidal lesion and BVN were analyzed.The study protocol was approved by the Ethics Committee of Peking Union Medical College Hospital (No.S-K631). Written informed consent was obtained from each patient prior to any examination.Results:ICGA images showed obvious polyps with higper fluorescence in all PCV patients, and BVN structure was observed in the ICGA images of 38 eyes.Ring, circle, cluster or node like hyper-reflective structures were detected in polypoidal lesions in the en face OCTA images of 35 eyes, and the detection rate was 77.8% (35/45). The BVN structure was detected in the OCTA images of all the 38 eyes, with the 100% (38/38) detection rate.Conclusion:OCTA can detect polypoidal lesions and BVN of PCV non-invasively, which can locate the lesion in combination with en face image and B-scan image.

Salvia miltorrhiza Bge. is a perennial deciduous flowering plant. Its medicinal root and rhizomes part is widely used in the treatment of various diseases. In this study, bioinformatics analysis was performed to identify 4832 genome SSR loci with length longer than or equal to 40 bp from the draft genome assembly of S. miltorrhiza. The re-sults showed that the dinucleotide repeat motifs and trinucleotide repeat motifs constitute the main types of genome SSR loci, accounting for 37.3% and 61.3% respectively. SSR types enriched with A/T bases showed significantly higher abundance than other types, including AT/TA AAT/ATT, ATA/TAT, TAA/TTA, accounting for 30.5%, 21.6%, 17.1%, 20.4% of the total number of SSR loci, respectively. 1079 primer pairs were designed for these genome SSR loci. These primers can be used for genomic diversity analysis, genetic map construction, genetic marker screening. These data could lay the foundation for population genetics and genomics research of S. miltorrhiza.

Cytochrome P450 (CYP450) is a key element in the Ganoderma triterpenoid biosynthetic pathway. The catalytic reaction process for CYP450 requires NADPH / NADH for electron transfer. After searching the genome dataset of Ganoderma lucidum, the unique sequence encoding CYP450 and NADPH were discovered, separately. The open reading frames of GLCYP450 and GLNADPH were cloned separately using RT-PCR strategy from G lucidum. The appropriate restriction enzyme cutting sites were introduced at the 5' and 3' ends of gene sequence. The genes of GLCYP450 and GLNADPH were recombined into the yeast expression vector pESC-URA, leading to the formation of the yeast expression plasmid pESC-GLNADPH-GLCYP450. This study provides a foundation for researching Ganoderma triterpene biosynthesis using the approach of synthetic biology.

Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.

Objective To compare the fast blood flow density (FBFD) of intermediate choroid between endogenous Cushing syndrome (ECS) patients and healthy control subjects.Methods Thirteen eyes of 7 eligible ECS patients (ECS group) and 13 eyes of 7 gender,age,axial length matched healthy volunteers (control group) were enrolled in this study.For each subject,macular radial scan with swept source optical coherence tomography (SS-OCT) was performed and subfoveal choroidal thickness (SCT) was measured.Then 3.0 mm× 3.0 mm macular scan with SS-OCT angiography was performed,and selected blood flow image at intermediate choroid level or 1/2 SCT beneath Bruch membrane.The grayscale images were then binarized for the analysis of FBFD.Results The SCT in ECS group was (394.7±77.7) μm,which was significantly thicker than (332.1 ± 68.1) μm in control group (t=2.923,P=0.008).The FBFD of intermediate choroid in ECS group were (76.35± 14.46)％,which were significantly greater than (63.57± 13.42)％ in control group (t=2.775,P=0.01).Conclusion ECS patients had increased FBFD at intermediate choroid level compared with healthy controls.

Lack of biocompatibility and bioactivity is a big problem for the synthetic materials that have been generated for neural tissue engineering. To get around the problem and generate better scaffold for neural tissue repair, we intended to generate nano-fibers by self-assembly of polypeptide IKVAV. Bioactive IKVAV Peptide-Amphiphile (IKVAV-PA) was first synthesized and purified, the property of which was analyzed and determined by high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Then, by addition of hydrogen chloride (HCl), self-assembly of IK-VAV-PA was induced in vitro and nano-fibers formed as shown by transmission electron microscopy (TEM). The effect of IKVAV nanofibers on adherence of PC12 cells was assayed in cell culture and the results showed that the rates of adherence of PC12 increased significantly when the density of IKVAV was within a certain range (0.58 microg/cm2 to 15.6 microg/cm2). However, its effect on the rates of adherence did not significantly alter with time, whether after 1 hour or 3 hours of culture. In general, we showed that IKVAV-PA can successfully self-assemble to form nanofiber, and promote rapid and stable adherence of PC12 cells, and the effect of the self-assembled IKVAV to promote PC12 cells adherence is dosage-dependent within a certain range of densities.