Asperosaponin VI Protects Against Bone Loss Due to Hindlimb Unloading in Skeletally Growing Mice Through Regulating Microbial Dysbiosis Altering the 5-HT Pathway.

Osteoporosis is a complex multifactorial disease that can lead to an increased risk of fracture. However, selective and effective osteoporosis drugs are still lacking. We showed that Asperosaponin VI (AVI) has the implications to be further developed as an alternative supplement for the prevention and treatment of bone loss. AVI has been found to have beneficial effects on metabolic diseases such as bone loss, obesity, and atherosclerosis. Our study was designed to determine the effect and mechanism of action of AVI against bone loss through regulating microbial dysbiosis. A hindlimb unloading mouse model was established to determine the effect of AVI on bone microarchitecture, gut microbiota, and serum metabolites. Eighteen female C57BL/6 J mice were divided into three groups: control, hindlimb unloading with vehicle (HLU), and hindlimb unloading treated with AVI (HLU-AVI, 200 mg/kg/day). AVI was administrated orally for 4 weeks. The results demonstrated that AVI improved the bone microstructure by reversing the decrease in bone volume fraction and trabecular number, and the increase in trabecular separation and structure model index of cancellous bone in hindlimb suspension mice. The results of 16sRNA gene sequencing suggested that the therapeutic effect of AVI on bone loss may be achieved through it regulating the gut microbiota, especially certain specific microorganisms. Combined with the analysis of ELISA, immunohistochemistry, and serum metabolome results, it could be speculated that AVI played an important role in adjusting the balance of bone metabolism by influencing specific flora such as Clostridium and its metabolites to regulate the 5-hydroxytryptophan pathway. The study explored the novel mechanism of AVI against osteoporosis, and has implications for the further development of AVI as an alternative supplement for the prevention and treatment of bone loss.

First Report of Cucumber mosaic virus in Agastache rugosa in China.

Agastache rugosa (family Lamiaceae) is one of the most common herbs of traditional Chinese medicine in China, and the species increasingly gains popularity on the international market. In June 2012, typical mosaic symptoms were observed on many A. rugosa plants in a field in Shanxi Province. The incidence of this disease reached more than 60% in a 2.6-ha field. Seven symptomatic plants were tested by DAS-ELISA using monoclonal antibodies specific for Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), and Tomato mosaic virus (ToMV); all antisera used in DAS-ELISA were generated and validated in our laboratory. CMV was found in all seven samples tested, but not TMV and ToMV. Double-stranded RNAs (dsRNA) extracted from infected leaves were used as templates in the subsequent two-step RT-PCR reaction (1). In order to further confirm the presence of CMV, a pair of specific primers (forward: 5'-ACGTCGACCATGGACAAATC-3', and reverse: 5'-TACCCGGGTCAGACTGGTAGCACC-3') based on the coat protein gene sequence of CMV were used to amplify PCR products of the expected size (657 bp) from ELISA-positive samples (2). These PCR products were cloned into pUCm-T Vector (Sangon Biotech, Shanghai) and sequenced. Five independent clones have been sequenced to obtain the consensus sequence. This consensus sequence (GenBank Accession No. JQ403529) was compared with other CMV sequences available in GenBank using DNAMAN. The partial CMV coat protein sequence showed the highest 97.9% nucleotide identity with a subgroup IB CMV isolate from China (DQ459481). To our knowledge, this is the first report of the natural occurrence of CMV on A. rugosa. References: (1) M. Krajacic et al. J. Chromatogr. A. 1144:111, 2007. (2) F. Li et al. J. Zhejiang Univ. 26:261, 2000.

Energy, amino acid, and phosphorus digestibility of phytase transgenic corn for growing pigs.

Three experiments were conducted to evaluate energy, AA, and P digestibility in a phytase transgenic corn (PTC) containing a phytase gene (phyA2) isolated from Aspergillus niger compared with a nontransgenic near-isoline conventional corn (CC) grown in the same environmental conditions for growing pigs. Experiment 1 was an energy balance experiment conducted to measure DE and ME in PTC and CC. Eighteen growing barrows (initial BW 25.8±0.3 kg) from 9 litters were allotted by BW and litter to 1 of 2 dietary treatments with 9 pigs per treatment in a randomized complete block design. Pigs were individually placed in metabolism cages and fed diets based on the 2 corns. The DE and ME in PTC (3,967 and 3,941 kcal/kg of DM, respectively) were greater (P<0.05) than those in CC (3,917 and 3,848 kcal/kg of DM, respectively). Experiment 2 was conducted to measure apparent ileal digestibility (AID) and standardized ileal digestibility (SID) values of CP and AA in the 2 corns. Eighteen growing barrows (initial BW 41.8±0.7 kg) were equipped with a T-cannula in the distal ileum. Pigs were placed in metabolism cages in a completely randomized design with 3 dietary treatments of 6 pigs each. An N-free diet was used to estimate basal endogenous losses of CP and AA. The AID and SID values for CP and all AA did not differ between the 2 corns. Experiment 3 was conducted to measure apparent total tract digestibility (ATTD) and standardized total tract digestibility (STTD) values of P in the 2 corns. Eighteen growing pigs (initial BW 30.5±0.5 kg) from 6 litters were placed in metabolism cages in a randomized complete block design with 3 dietary treatments of 6 pigs each based on BW and litter. Two diets were based on the 2 corns, and a P-free diet was used to measure endogenous P losses. The ATTD and STTD values of P were greater (P<0.05) in the PTC diet (71.4% and 76.9%, respectively) than those in the CC diet (27.6% and 33.4%, respectively). Pigs fed the PTC diet had a greater (P<0.05) P retention (70.7%) than those fed the CC diet (27.1%). It was concluded that PTC had a greater digestibility of energy and P than CC for growing pigs. As a consequence, if PTC replaces CC in a pig diet, the DE and ME in the diet will increase, and less inorganic P will need to be supplemented to the diet, and thus P excretion in manure will be decreased.

The beneficial effect of Radix Dipsaci total saponins on bone metabolism in vitro and in vivo and the possible mechanisms of action.

UNLABELLED: The purpose of this study is to investigate the anti-osteoporotic effects of Radix Dipsaci total saponins (RTS). We showed that RTS was able to improve bone properties by either an increase of osteoblastic activity or a decrease in osteoclastic activity. INTRODUCTION: Radix Dipsaci has long been used as an anti-osteoporotic drug. The present study investigates the anti-osteoporotic effects of RTS. METHODS: Three-month-old female rats were randomly assigned into a sham-operated group (sham) and five ovariectomy (OVX) subgroups, namely, OVX with vehicle (OVX), OVX with 17ß-ethinylestradiol (E(2)), and OVX with graded doses of RTS (50, 100, or 200 mg/kg/d). RTS and E(2) were administered orally, daily from 1 week after OVX treatment for 4 months. Bone mass, turnover, and strength were evaluated by dual-energy X-ray absorptiometry, biochemical markers, and the three-point bending test. The trabecular bone microarchitecture was assessed by microCT. In vitro experiments were performed to determine the potential molecular mechanisms of the anti-osteoporotic effect of RTS. RESULTS: RTS prevented the loss of bone mass induced by OVX. The preventive effect on bone loss was primarily indicated by decreasing levels of bone turnover markers and confirmed by the changes in urinary calcium and phosphorus excretion. The treatment also enhanced the biomechanical strength of bone and prevented the deterioration of trabecular bone microarchitecture. RTS induced MC3T3-E1 and primary osteoblastic cell maturation and differentiation and increased bone formation by increasing BMP-2 synthesis. In addition, RTS inhibited osteoclastogenesis through an increase in osteoprotegrin and a decrease in NF-kB ligand expression in vitro. CONCLUSIONS: RTS treatment can effectively suppress the loss of bone mass induced by OVX and in vitro evidence suggests this could be through actions on both osteoblasts and osteoclasts.

First Report of Tobacco mosaic virus in Abutilon theophrasti in China.

Velvetleaf (Abutilon theophrasti Medic), a tall, fecund, self-fertile annual plant in the family Malvaceae, is widely grown in China as a fiber crop and for its medicinal properties. In July of 2008, we observed diffuse chlorotic and necrotic spots on the oldest leaves of velvetleaf plants in the field in Shanxi Province. Sap extracts from six symptomatic plants were tested by direct antigen coated ELISA using polyclonal antibodies specific to Potato virus Y (PVY) and monoclonal antibodies specific to Tomato mosaic virus (ToMV), Cucumber mosaic virus (CMV), and Tobacco mosaic virus (TMV). Five of the six samples were negative for ToMV, CMV, and PVY but positive for TMV. Double-stranded RNA extracted from leaves of the five velvetleaf plants was used as template for reverse transcription (RT)-PCR as described by Krajacica et al. (1) and Li et al. (2) with some modifications. One-step RT-PCR was performed using a Quant One Step RT-PCR Kit (TIANGEN BIOTECH CO., LTD., Beijing, China) with sense (5'-CTGTTTAGCCGGTTTGGT-3'), and antisense (5'-TCCCTTTACGGACATCAC-3') primers (3) designed to specifically amplify a fragment of the movement protein coding region of TMV. The expected 470-bp fragments were amplified from dsRNA from these five plants and the amplicon from each plant was cloned and sequenced (GenBank Accession No. FJ873800). Comparisons of a consensus sequence derived from the five amplicons with the nucleotide sequences available in the NCBI database using BLAST showed 99% identity with TMV from South Korea (GenBank Accession No. AB354955) and Spain (Accession No. AJ308692) and 98% with TMV from China (Accession No. AF165190). The serological, RT-PCR results and sequence data revealed that these velvetleaf plants were infected by TMV. On the basis of the serological analysis and genome sequence comparisons, this isolate was tentatively designated as TMV-Velvetleaf. To our knowledge, this is the first report of TMV in Abutilon theophrasti Medic in China. References: (1) M. Krajacic et al. J. Chromatogr A. 1144:111, 2007. (2) H. Li et al. Agric. Sci. Chin. 6:86, 2007. (3) Y. B. Niu et al. Chin. Biotechnol. 29:76, 2009.