SPA, a Stigma-style-transmitting tract Physical microenvironment Assay for investigating mechano-signaling in pollen tubes.

Accurate sensing and responding to physical microenvironment are crucial for cell function and survival, but the underlying molecular mechanisms remain elusive. Pollen tube (PT) provides a perfect single-cell model for studying mechanobiology since it's naturally subjected to complex mechanical instructions from the pistil during invasive growth. Recent reports have revealed discrepant PT behaviors between in vivo and flat, two-dimensional in vitro cultures. Here, we established the Stigma-style-transmitting tract (TT) Physical microenvironment Assay (SPA) to recapitulate pressure changes in the pistil. This biomimetic assay has enabled us to swiftly identify highly redundant genes, GEF8/9/11/12/13, as new regulators for maintaining PTs integrity during style-to-TT emergence. In contrast to normal growth on solid medium, SPA successfully phenocopied gef8/9/11/12/13 PT in vivo growth-arrest deficiency. Our results suggest the existence of distinct signaling pathways regulating in vivo and in vitro PT integrity maintenance, underscoring the necessity of faithfully mimicking the physical microenvironment for studying plant cell biology.

Use of High-Throughput Sequencing to Identify Fungal Communities on the Surface of Citri Reticulatae Pericarpium During the 3-Year Aging Process.

Citri Reticulatae Pericarpium (CRP) is a natural product that is used widely in food and is an ingredient in traditional Chinese medicine. CRP improves gradually with aging; this process typically takes 3 years or more. During the aging process, CRP can be colonized with fungi and mildew. Molds and mildew may result in an increased flavonoid content; however, this has been observed only in response to fungi of the genera Penicillium and Aspergillus. As fungal colonization may alter the quality and properties of CRP, it is critical to have an understanding of the fungal communities detected on the surface of CRP during the aging process. We used a high-throughput sequencing (HiSeq) platform to sequence internal transcribed spacers (ITS) region to identify the contaminants associated with CRP during the 3-year aging process. We also evaluated the distribution of the dominant fungi of the genera Aspergillus and Penicillium over time. At the phylum level, we identified Ascomycota (36.26%) and Basidiomycota (18.98%), along with smaller populations of Mucoromycota, Glomeromycota, and Mortierellomycota. At the genus level, the fungi detected include Wallemia (12.40%), Cystofilobasidium (4.62%), Zasmidium (4.52%), Cladosporium (3.72%), Hanseniaspora (3.55%), Fusarium (3.49%), Kurtzmaniella (2.03%), Candida (1.74%), Passalora (1.47%), Ceramothyrium (1.33%), Mucor (1.07%), and Aspergillus (1.03%). Fungi of the genus Penicillium were detected primarily during the first year of storage. By contrast, fungi of the genus Aspergillus were not detected during the early stages (fresh peel-8 months), but appeared gradually at later stages of the aging process. Taken together, our results indicate that HiSeq is an effective method to study the changes in fungal communities that develop on the CRP surface over time. These findings provide a basis for further research into the correlation between dominant fungi and the mechanisms underlying the successful aging of CRP.

Effect of exercise and butyrate supplementation on microbiota composition and lipid metabolism.

The composition and activity of the gut microbiota depend on the host genome, nutrition, and lifestyle. Exercise and sodium butyrate (NaB) exert metabolic benefits in both mice and humans. However, the underlying mechanisms have not been fully elucidated. This study aimed to examine the effect of exercise training and dietary supplementation of butyrate on the composition of gut microbiota and whether the altered gut microbiota can stimulate differential production of short-chain fatty acids (SCFAs), which promote the expression of SESN2 and CRTC2 to improve metabolic health and protect against obesity. C57BL/6J mice were used to study the effect of exercise and high-fat diet (HFD) with or without NaB on gut microbiota. Bacterial communities were assayed in fecal samples using pyrosequencing of 16S rRNA gene amplicons. Western blot was performed using relevant antibodies to detect the protein expressions in liver and HepG2 cell extracts. Exercise and butyrate administration significantly reversed metabolic dysfunctions induced by HFD (P < 0.05). The number of Firmicutes and the proportion of Firmicutes to Bacteroidetes order were predominant in all HFD groups (P = 0.001). Exercise and butyrate supplementation significantly inhibited the relative abundance of lipopolysaccharide-producing phyla (P = 0.001). SESN2 and CRTC2 expression in the liver of mice were significantly increased after exercise (P < 0.05) and/or supplementation of butyrate (P < 0.05). Exercise enhances butyrate-producing fecal bacteria and increases butyrate production and consequently improves lipid metabolism through the butyrate-SESN2/CRTC2 pathway. Excess butyrate may reduce the proportion of probiotics and reverse the metabolic effects.

The Increase of Flavonoids in Pericarpium Citri Reticulatae (PCR) Induced by Fungi Promotes the Increase of Antioxidant Activity.

There are thousands of traditional Chinese medicines in China, and they all have to be stored for a period of time for the reason of market price or other factors. But some traditional Chinese medicines especially need to be stored longer, and the clinical efficacy will be better. The cause and mechanism of this phenomenon have attracted much attention in recent years. In this study, we analyze the reason of "the longer storage period the better" of Pericarpium Citri Reticulatae (PCR); method of microscopic and molecular identification was used to identify the fungi separated and purified from PCR. The HPLC and UV spectrophotometry methods were used to determine the contents of flavonoids in PCR. The isolated fungi were inoculated into sterile samples to screen the fungi closely related to the change of flavonoids. The results indicate that the strain of Aspergillus niger could obviously promote the contents of flavonoids, and it could also increase the antioxidant effect of PCR. In conclusion, this study explains the reason of "the longer storage period the better" of PCR from the perspective of microbe, proving the beneficial effect of microorganism on the surface of PCR.

Effects of Governor Vessel electroacupuncture on the systematic expressions of NTFs in spinal cord transected rats.

This study evaluated the effects Governor Vessel electroacupuncture (GVEA) on the systematic regulation of neurotrophic factors (NTFs) in the spinal segments caudal (CSS) to the site of transection in rats subjected to spinal cord transection (SCT). Using RT-PCR, we amazingly found the gene expressions of NGF, IGF-1, FGF-2, CNTF, PDGF, TGF-ß1, TrkA, TrkB and TrkC were downregulated following GVEA treatment. However, the number of GAP-43 and Synaptophysin profiles in the CSS in the GVEA rats showed a significant increase, compared with non-EA animals, although both the 5-HT and corticospinal fibers have no statistical differences in the CSS. Simultaneously, there was significant recovery in hindlimb locomotor and sensory functions after GVEA treatment. Therefore, these findings challenge the past view that GVEA promotes functional restoration, which is linking to the up-regulation of NTFs in rats subjected to SCT. The present findings may give some novel indication on the mechanism of acupuncture for the treatment of SCI.

[Soil nutrient accumulation and its affecting factors during vegetation succession in karst peak-cluster depressions of South China].

Taking the typical karst peak-cluster depressions in Huanjiang County of northwest Guangxi as the objects, and by using the method of replacing time with space, an analysis was made on the dynamic changes of top soil (0-15 cm) nutrients and their dominant controlling factors during the process of vegetation succession. With the positive succession of vegetation (herb-shrub-secondary forest-primary forest), the soil organic carbon (SOC), total nitrogen (TN), and total phosphorus (TP) contents increased significantly, with the soil SOC, TN, and TP increased from 29.1 g x kg(-1), 2.48 g x kg(-1), and 0.72 g x kg(-1) in herb community to 73.9 g x kg(-1), 8.10 g x kg(-1), and 1.6 g x kg(-1) in primary forest, respectively, which indicated that the positive succession of vegetation was helpful to the soil nutrient accumulation. The soil cation exchange capacity (CEC) had close relationships with the soil SOC and TN, being the primary controlling factor for the accumulation of the soil C and N. The litter P content, C/P ratio, and N/P ratio were the major factors controlling the P accumulation in the topsoil. The litters higher P content and N/P ratio and smaller C/P ratio were helpful for the P accumulation. Topographic indices (slope, aspect, and rock exposure ratio) had little effects on the soil nutrients.

Electro-acupuncture treatment improves neurological function associated with downregulation of PDGF and inhibition of astrogliosis in rats with spinal cord transection.

The aim of the current study was to determine the effects of electro-acupuncture (EA) on the improvement of locomotor function in injured spinal cord and underlying mechanism. Forty-five female Sprague-Dawley rats (180~200 g) were randomly divided into three groups, sham operation control group (sham), spinal cord transection group (SCT) and EA group. The Basso, Beattie, and Bresnahan (BBB) Locomotor Rating Scale was used to evaluate functional recovery of rats in hindlimbs at 1, 3, 5 weeks after injury and EA therapy. The gene and protein expression of glial fibrillary acidic protein (GFAP) and platelet derived growth factor (PDGF) were measured by reverse transcription-polymerase chain reaction and Western blot analysis, respectively, and the number of GFAP-positive cells was counted, also. Compared with SCT group, the locomotor function in hindlimbs of rats was improved after 1, 3, 5 weeks following EA therapy. EA treatment not only decreased effectively the number of GFAP immunostaining and GFAP expression, but also downregulated the PDGF expression both gene and protein, in addition decreased the number of PDGF immunostaining in injured spinal cord of rats with transection. It therefore concluded that EA therapy can significantly promote the recovery of locomotor function, and this may be linked to the inhibition of astrogliosis, together with the downregulation of PDGF.

[Determination of geniposide, crocin and crocetin in different processing products of fructus gardeniae by HPLC-ELSD].

OBJECTIVE: To develop a HPLC- ELSD method for determination the contents of geniposide, crocin and crocetin in different processing products of Fructus Gardeniae. METHODS: The separation was performed in the HyperClone ODS C18 column (250 mm x 4. 6 mm, 5 microm) with linear gradient elution using methanol-water and 0.05% phosphoric acid in water, the flowing rate was 0.8 mL/min, the column temperature was 30 degrees C, and the ELSD parameter was as follow: 70 degrees C as atomization temperature and 2.0 L/min as the gas flowing rate. RESULTS: The contents of geniposide and crocin in raw, yellowish, carbocoal and scorched Fructus Gardeniae decreased with the deepening of processing degree. However, the content of crocetin in carbocoal and scorched Fructus Gardeniae increased comparing with the raw one. CONCLUSION: This is a simple and credible quality control method, and can be used for the quality control and comprehensive evaluation for different processed products of Fructus Gardeniae.