A putative new emaravirus isolated from Ailanthus altissima (Mill.) Swingle with severe crinkle symptoms in China.

A putative new emaravirus, named "ailanthus crinkle leaf-associated emaravirus" (ACrLaV), was detected in Ailanthus altissima with severe crinkle symptoms by RNA-Seq and RT-PCR. Four viral segments associated with ACrLaV were identified and fully sequenced, except for a few nucleotides at the genomic termini. The RNA-dependent RNA polymerase (RNA1), glycoprotein (RNA2), nucleocapsid protein (RNA3), and movement protein (RNA4), showed 26.5%-57%, 17%-49.9%, 14.4%-40.4%, and 14.1%-65.9% amino acid sequence identity, respectively, to those of known emaraviruses. All four ACrLaV genomic RNA segments are most closely related to those of common oak ringspot-associated virus from Germany, as supported by sequence comparisons and phylogenetic analysis. ACrLaV is considered a distinct member of the genus Emaravirus, and this is the first report of an emaravirus in A. altissima.

Complete genome sequence of a putative new and distinct caulimovirus from Metaplexis japonica (Thunb.) Makino in China.

In 2019, a new disease characterized by yellow mottling on leaves was observed on Metaplexis japonica (Thunb.) Makino plants in Liaoning, China. Using RNA sequencing and PCR with overlapping primers, the complete nucleotide sequence of the circular double-stranded DNA genome of a new virus similar to caulimoviruses in diseased samples was determined. The genome comprised 7733 base pairs and was predicted to encode six proteins. It had the closest relationship to a caulimovirus, strawberry vein banding virus, based on sequence comparisons and phylogenetic analysis. Species-level sequence divergences from caulimoviruses were also observed in the conserved genomic regions. A complete episomal viral genome sequence in symptomatic M. japonica plants was identified, and the name "metaplexis yellow mottle-associated virus" is proposed for this virus.

First report of tomato spotted wilt virus (TSWV) isolated from nasturtium (Tropaeolum majus L.) with a serious leaf mosaic disease in China.

Tomato spotted wilt virus (TSWV, Genus Orthotospovirus, Family Tospoviridae) is a thrips-transmitted negative-stranded RNA virus with a large host range. Major economic losses caused by TSWV have been recorded in various crops such as tomato, pepper and lettuce (Pappu et al., 2009; Adams et al., 2017). Nasturtium (Tropaeolum majus L.), a common flowering plant species native to Peru, is generally cultivated as a garden and greenhouse ornamental plant in China. It is also known for its medical, edible and cosmetic values (Jakubczyk et al., 2018). In June 2019, a serious leaf mosaic disease (Figure S1) was observed in ~77% of T. majus plants in Beiling Park in Shenyang City, Liaoning, China. To reveal the possible viral agent associated with the disease, leaf tissue was collected from 9 symptomatic and 2 asymptomatic plants, followed by total RNA extraction from each of the samples using RNASimple Total RNA Kit (Tiangen, Beijing, China). The RNA from one representative symptomatic sample (Figure S1, d) was used to construct an rRNA-depleted library using the Ribo-Zero™ rRNA Removal Kit (Plant Leaf) (Illumina, CA, USA). The library was subjected to RNA-Seq using a BGISEQ platform (Shenzhen Huada Gene Science and Technology Service Co., Ltd., Shenzhen, China). A total of 109,279,540 quality-filtered reads were obtained using the CLC Genomics Workbench 9.5 software (Qiagen, Valencia, CA, USA). Clean reads were assembled into 88,091 contigs ranging in length from 200 to 19,695 bp using Velvet (Zerbino and Birney 2008). Among the contigs, three sized at 8801, 4617 and 2909 bp were found to share a sequence identity of 98.74-99.35% with the RNA segments (L, M and S) of TSWV. No sequences of other viruses, viroids included, were detected. Ten primer pairs were designed based on the TSWV contigs to obtain the full genome sequence of the virus (Table S1). Five, three, and two amplicons were obtained for the TSWV RNA segments L, M and S, respectively, from the same RNA sample used in the RNA-Seq process. These amplicons were then cloned into the pMD18T vector (TaKaRa, Dalian, China) and Sanger sequenced. The resulting sequences were assembled and analyzed using the DNAMAN version 8.0 (LynnonBiosoft, Quebec, Canada) and DNAStar version 6.0 software (DNAStar Inc, Madison, WI, USA) (Table S1). The RNA segments were determined to be 8,914 nt (L, accession no. MT241883), 4,791 nt (M, MT241884) and 2,922 nt (S, MT241885) in length. They shared a sequence identity of >99% with the "LL-N.05" (segment L, KP008128) and "SPAIN-1" (segment S, AY744479) isolates from tomatoes in Spain and the "Beijing" isolate (segment M, MH717046) from chrysanthemum in China. These results, together with phylogenetic analysis, suggest that the isolate (designated as isolate LN-HJL) was likely resulted from genome reassortment between other isolates. No reliable recombination events were detected in the RNA segments of LN-HJL by the RDP4 program (Martin et al., 2015). RT-PCR assay with primer pair S-1F/1R on all samples led to positive detection of TSWV only in the symptomatic samples (Figure S1, g), indicating that the virus was likely responsible for the disease symptoms observed in T. majus. To the best of our knowledge, this is the first report of the TSWV infection in T. majus in China, and the second in the world since the first report in the USA (Ie, 1964). TSWV-infected T. majus plants not only lose their aesthetic and economic values (Figure S1, f), but also may act as reservoirs for the spread of TSWV to other important crops. T. majus should therefore be monitored regularly for the virus and managed accordingly in China.

Full genome sequence of a novel potexvirus from Euonymus bungeanus Maxim based on RNA-Seq analysis.

In July 2018, a yellow mottle leaf disease was observed on the leaves of Euonymus bungeanus Maxim plants in Liaoning Province of China. Typical flexuous filaments (diameter, 13 nm; length, ca. 500 nm) were observed in extracts on the symptomatic leaves. Transcriptome sequencing indicated the presence of a potexvirus in the infected samples. The complete viral genome, determined to be 6,784 nucleotides in length, excluding the poly(A) tail, contains five open reading frames and is most closely related to that of euonymus yellow vein associated virus (EuYVAV, MF078061) (41.0%). Based on the coat protein gene, this virus shares the highest sequence similarity with cymbidium mosaic virus (CyMV, EF125179), with 43.9% nucleotide and 38.8% amino acid sequence identity, respectively. Phylogenetic analysis indicated that the virus clustered with potexviruses and is most closely related to EuYVaV. This virus is a distinct member of the genus Potexvirus, for which the name "euonymus yellow mottle associated virus" (EuYMaV) is proposed.

The complete nucleotide sequence and genome organization of a novel virus of the order Tymovirales isolated from Prunus davidiana (Carr.) Franch. in Liaoning, China.

In September 2017, a yellow spot leaf disease was noted on the leaves of Prunus davidiana (Carr.) Franch. plants in Liaoning, China, and spherical virions (approx. 30 nm in diameter) were later observed in preparations of symptomatic leaves. Subsequent deep sequencing of small RNA revealed the presence of a virus in these symptomatic leaves The complete genome of this viral isolate consists of 6,072 nucleotides, excluding the poly(A) tail. The virus showed the closest genetic relationship to grapevine-associated tymo-like virus, reported in Colmar, France (GaTLV, MH383239), which is the sole member of the newly proposed genus "Gratylivirus" within the order Tymovirales, which is currently unassigned to a particular family. The virus clustered closely with GaTLV in a phylogenetic tree constructed based on complete genomic sequences. On the basis of the nucleotide and amino acid sequences of the replicase and coat protein genes, this virus shares the highest (although still relatively low) sequence similarity with those of GaTLV (41.6%-60.8% identity), indicating that the virus is a distinct member of the order Tymovirales, for which the name "prunus yellow spot-associated virus" (PYSaV) is proposed. To our knowledge, this is the first report of a virus naturally infecting P. davidiana.

[Effect of retinoic acid on alkaline phosphatase expression in rat adipose-derived stem cells].

OBJECTIVE: To investigate whether adipose-derived stem cells (ADSCs) induced by retinoic acid (RA) in vitro express primordial germ cell marker alkaline phosphatase (ALP) and vasa. METHODS: ADSCs were isolated from adult female SD rats and cultured in vitro. The third passage of ADSCs was identified by adipogenic differentiation, osteogenic differentiation and cell surface marker detection. The ADSCs were treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA for 7 or 14 days, and the cellular expression of ALP was detected. vasa mRNA expression in ADSCs treated with 1×10(-5) mol/L RA for 7 days was detected using RT-PCR. RESULTS: The OD value of ADSCs treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA was 0.59∓0.04, 0.27∓0.07, and 0.15∓0.03 after a 7-day treatment, and was 0.42∓0.02, 0.34∓0.01, and 0.19∓0.02 after a 14-day treatment, respectively, demonstrating significantly enhanced ALP expression in RA-treated ADSCs compared with that in the control cells (0.07∓0.01 and 0.07∓0.01 at 7 and 14 days, respectively, P<0.01). The ADSCs showed a negative vasa mRNA expression after 1×10(-5) mol/L RA treatment for 7 days. CONCLUSION: RA-induced ADSCs express ALP, a marker of primordial germ cells, but does not express the primordial germ cell marker vasa.

[Effect of allogenic adipose-derived stem cell grafting on serum biochemical indicators and sporting ability of endurance trained rats].

OBJECTIVE: To observe the effect of transplantation of allogeneic adipose-derived stem cells (ADSCs) on serum biochemical indicators and sporting ability in highly endurance-trained rats from a fasciological perspective. METHODS: The ADSCs were cultured in vitro. Forty male Wistar rats were randomized into 4 equal groups, namely the blank control group, overtraining (model) group, transplantation without training group and overtraining plus transplantation group. The rats in the two overtraining groups were subjected to exhaustive swimming for 1 week, and in the two transplantation groups, cultured allogeneic ADSCs (2×10(6)/ml) were injected via the tail vein. The exhaustion time in swimming and the serum levels of BUN, LDH, BLa, and Hb of the rats were recorded after the treatments. RESULTS: The rats in the model group showed significantly increased serum BUN, LDH and BLa levels and decreased Hb level with a extended exhaustion time as compared with those in the blank control group (P<0.01). The BUN, LDH and BLa was significantly lower, Hb level higher and the exhaustion time significantly longer in the overtraining plus transplantation group than those in the model group (P<0.01). CONCLUSIONS: ADSCs can effectively prolong the exhaustion time of rats during exhaustive swimming and enhance their sporting ability.

Correlations between fasciology and yin yang doctrine.

The aim of this study is to explore the correlations between fasciology and yin yang doctrine. Professor Yuan developed fasciology by three-dimensional reconstruction of connective tissue (fascia) in the trunk and limbs of the human body and tracing back to tissue origins in light of biological evolution and developmental biology. Fasciology states that the human body can be divided into two systems: the supporting-storing system and the functional system. This article elaborates on the roles of the two systems and their mutual relationship. The two systems are used to analyze the yin, the yang, and their relationship. The two systems are promoted but also restricted in different contexts. The supporting-storing system is formed by undifferentiated connective tissue and provides undifferentiated cells and nutrients for differentiated cells of the functional system. Thus, the supporting-storing system could be classified as quiet, similar to yin. The functional system continuously maintains the various functional activities of the human body. Thus, the functional system could be classified as active, similar to yang. In interpreting the yin yang doctrine from the point of view of fasciology, yin can be compared with the supporting-storing system and yang can be compared with the functional system.

[Effect of allogenic adipose-derived stem cell transplantation on bone mass in rats with glucocorticoid-induced osteoporosis].

OBJECTIVE: To investigate the effects of systemic transplantation of allogenic adipose-derived stem cells (ADSCs), the main undifferentiated cells in the supporting-storing system based on the fasciology hypothesis, on the bone mineral density (BMD) and histomorphometry in rats with glucocorticoid-induced osteoporosis (GIOP) rats, and explore a new therapeutic approach for osteoporosis. METHODS: Forty female adult Wistar rats were randomized equally into blank control group (A), model group (B), control treatment group (C) and treatment group (D). In groups B, C, and D, osteoporosis was induced by injection of prednisolone (8 mg/kg) via the tail vein 3 times a week for 12 consecutive weeks. After successful establishment of osteoporosis, allogenic ADSCs (3×10(6)/ml) were transplanted via the tail vein. The BMD at the L3-L5 levels and of the right femurs were detected, and the histomorphometry of the right tibias was analyzed in all the rats. RESULTS: After prednisolone injection, the BMD of L3-L5 vertebrae and the right femurs, the percent trabecular area, trabecular thickness and trabecular number of the right tibias were all obviously lowered while the trabecular separation and osteoclast number increased in group B as compared to those in group A (P<0.05). Four weeks after ADSC transplantation, all these indices were significantly improved in group D, showing significant differences from those in group B (P<0.05) and also group C. CONCLUSION: Transplantation of allogenic ADSCs can restore the BMD and bone histomorphometric properties of rats with GIOP, and may serve as a potential treatment for GIOP. These results also provide partial experimental evidence supporting the hypothesis of fasciaology.