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Abstract Background The ginseng endophyte Paenibacillus polymyxa Pp-7250 (Pp-7250) has multifaceted roles such as preventing ginseng diseases, promoting growth, increasing ginsenoside accumulation, and degrading pesticide residues, however, these effects still have room for improvements. Composite fungicides are an effective means to improve the biocontrol effect of fungicides, but the effect of Pp-7250 in combination with its symbiotic bacteria on ginseng needs to be further investigated, and its mechanism of action has not been elucidated. In this study, a series of experiments was conducted to elucidate the effect of Paenibacillus polymyxa and Bacillus cereus co-bacterial agent on the yield and quality of understory ginseng, and to investigate their mechanism of action. Results The results indicated that P. polymyxa and B. cereus co-bacterial agent (PB) treatment improved ginseng yield, ginsenoside accumulation, disease prevention, and pesticide degradation. The mechanism is that PB treatment increased the abundance of beneficial microorganisms, including Rhodanobacter, Pseudolabrys, Gemmatimonas, Bacillus, Paenibacillus, Cortinarius, Russula, Paecilomyces, and Trechispora, and decreased the abundance of pathogenic microorganisms, including Ellin6067, Acidibacter, Fusarium, Tetracladium, Alternaria, and Ilyonectria in ginseng rhizosphere soil. PB co-bacterial agents enhanced the function of microbial metabolic pathways, biosynthesis of secondary metabolites, biosynthesis of antibiotics, biosynthesis of amino acids, carbon fixation pathways in prokaryotes, DNA replication, and terpenoid backbone biosynthesis, and decreased the function of microbial plant pathogens and animal pathogens. Conclusion The combination of P. polymyxa and B. cereus may be a potential biocontrol agent to promote the resistance of ginseng to disease and improve the yield, quality, and pesticide degradation.

Abstract Multi-generational asexual reproduction of Gastrodia elata Bl. will cause seedling species degeneration. Sexual reproduction of Gastrodia elata Bl. seed is an effective method to solve the problem of degeneration. The development of Gastrodia elata Bl. seeds cannot be separated from the germination fungus. However, there are few strains of germination fungus in production, and there is also the problem of species degradation in application for many years. It is very important for the sexual reproduction of Gastrodia elata Bl. to isolate more new strains of excellent germination fungus from the origin. This study used the Gastrodia elata Bl. f. glauca S. chow seeds germination vegetative propagation corms capture method to isolate its symbiotic germination fungus, and comprehensively identified the species of germination fungus by colony morphology, ITS, sporocarps regeneration and germination function, and compared the growth characteristics and germination ability with other germination fungus (Mycena purpureofusca, Mycena dendrobii and Mycena osmundicola). The germination fungus was isolated from the vegetative propagation corms of Gastrodia elata Bl. f. glauca S. chow seeds and named GYGL-1. After comprehensive identification, GYGL-1 was Mycetinis scorodonius. Compared with other germination fungus, GYGL-1 has fast germination speed, vigorous growth, and high germination ability for Gastrodia elata Bl. f. glauca S. chow seeds. Innovated the isolation method of Gastrodia elata Bl. seeds germination fungus, obtained the regenerated sporocarps of the germination fungus, and discovered that Mycetinis scorodonius has a new function of germinating Gastrodia elata Bl. f. glauca S. chow seeds, enriching the resource library of Gastrodia elata Bl. germination fungus.

IntroductionAsexual reproduction of Gastrodia elata Bl. f. glauca S. chow (GeB) produces degeneration with increasing number of GeB. Therefore, we analyzed the microorganisms of GeB seedlings and surrounding soil by Illumina Miseq high-throughput sequencing technology.MethodsIn this study, Illumina Miseq high-throughput sequencing technology was applied to analyze the types and quantities of GeB seedlings and surrounding soil microorganisms in the first to third generations of asexual reproduction, isolated and identified the dominant strains of GeB in the first to third generations and screened the antagonistic bacteria of its pathogenic fungi, and evaluated the effects of beneficial bacteria on the production performance of seedlings planted with GeB.ResultsWith an increase in the number of asexual reproductive generations, the number of pathogenic fungi and bacteria in GeB seedlings and the surrounding soil increased, and the number of beneficial fungi and bacteria decreased. Pseudomonas sp., Agrobacterium rhizomes, and Herbaspirillum hiltneri were isolated and identified in the first generation, and Trichoderma harzianum, Penicillium viridiatum, Fusarium oxysporum, and Novosphingobium sp. Were isolated and identified in the third generation. Antagonistic strains of the three pathogenic bacterial strains were screened. In conclusion, beneficial bacteria significantly improved the production performance of asexual reproductive seedlings planted with GeB.DiscussionIn conclusion, our findings suggested that the microorganisms of GeB seedlings and the surrounding soil change as the number of generations of GeB reproduction increases, disrupts the microecological balance of surrounding soil and endophytic microbiomes.This study provides a theoretical basis for the degradation of asexual reproduction in GeB.

Skin cancer is a malignant tumour that can spread in the skin and is a serious threat to human health. Excessive ultraviolet radiation is one of the most common factors that leads to skin cancer. The purpose of this study was to evaluate the anti-skin tumour and anti-UVB damage effects of Armillaria mellea (Vahl) P. Kumm transformed Gastrodia elata Bl products (ATGP). Gastrodia elata Blume f.glauca S. Chows from Changbai Mountain and 4 Armillaria mellea (Vahl) P. Kumm strains were used as experimental materials. First, the optimum fungal strains of Armillaria mellea used to transform gastrodin were screened. Then, the MTT assay and scratch test proved that ATGP had anti-B16 melanoma cell activity in vitro. Furthermore, the anti-UVB damage effect of ATGP was proven by a UVB damage mouse skin model and HaCat photoaging model, and its mechanism was preliminarily discussed.

Abstract Background Gastrodia elata Bl. f. glauca, a perennial herb of G.elata Bl. in Orchidaceae, is one of the most valuable traditional Chinese medicines. G. elata Bl. is a chlorophyll-free myco-heterotrophic plant, which must rely on the symbiotic growth of Armillaria, but not all Armillaria strains can play the symbiotic role. Additionally, Armillaria is easy to degenerate after multiple generations, and the compatibility between the strains from other areas and G. elata Bl. f. glauca in Changbai Mountain is unstable. Therefore, it is incredibly significant to isolate, identify and screen the symbiotic Armillaria suitable for the growth of G. elata Bl. f. glauca in Changbai Mountain, and to explore the mechanism by which Armillaria improves the production performance of G. elata Bl. f. glauca. Results Firstly, G. elata Bl. f. glauca tubers, and rhizomorphs and fruiting bodies of Armillaria were used for the isolation and identification of Armillaria. Five Armillaria isolates were obtained in our laboratory and named: JMG, JMA, JMB, JMC and JMD. Secondly, Armillaria was selected based on the yield and the effective component content of G. elata Bl. f. glauca. It was concluded that the yield and quality of G. elata Bl. f. glauca co-planted with JMG is the highest. Finally, the mechanism of its high quality and yield was explored by investigating the effects of different Armillaria strains on the soil, its nutrition element contents and the soil microbial diversity around G. elata Bl. f. glauca in Changbai Mountain. Conclusions Compared with commercial strains, JMG significantly increased the content of Na, Al, Si, Mn, Fe, Zn, Rb and the absorption of C, Na, Mg, Ca, Cr, Cu, Zn and Rb in G. elata Bl. f. glauca; it improved the composition, diversity and metabolic functions of soil microbial communities around G. elata Bl. f. glauca at phylum, class and genus levels; it markedly increased the relative abundance of bacteria such as Chthoniobacter and Armillaria in the dominant populations, and enhanced such functions as Cell motility, amino acid metabolism and Lipid metabolism; it dramatically decreased the relative abundance of Bryobacter and other fungi in the dominant populations, and reduced such functions as microbial energy metabolism, translation and carbohydrate metabolism. This is the main reason why excellent Armillaria strains promote the high quality and yield of G. elata Bl. f. glauca in Changbai Mountain.

Gastrodia elata Bl. f. glauca is an important traditional Chinese medicinal plant. The yield and quality of Gastrodia elata Bl. have significantly decreased due to multigenerational asexual reproduction. Therefore, it is necessary to have sexual reproduction of Gastrodia elata Bl. to supplement the market supply. Seeds of G. elata Bl. have no endosperm, and their sexual reproduction depends on the nutrients provided by the embryo cells infected by Mycena fungi to complete seed germination. However, Mycena fungi are small and have many species, and not all Mycena fungi can promote the germination of G. elata Bl. seeds. Therefore, it is of great significance to isolate and identify suitable germination fungi and explore the mechanism for improving the production performance and yield, and quality of G. elata Bl. Six closely related Mycena isolates, JFGL-01, JFGL-02, JFGL-03, JFGL-04, JFGL-05, and JFGL-06, were isolated from the leaves and protocorms of G. elata Bl. f. glauca and were identified as Mycena purpureofusca. The mycelial state and number of germinating protocorms were used as indicators to preferentially select Mycena fungi, and it was concluded that JFGL-06 had the best mycelial state and ability to germinate G. elata Bl. seeds. Finally, a mechanism to increase the yield of G. elata Bl. was explored by comparing the changes in nutrient elements and microbial diversity in the soil around G. elata Bl. with different strains. JFGL-06 proved to be an excellent Mycena fungal strain suitable for G. elata Bl. f. glauca. Compared with the commercial strain, JFGL-06 significantly increased the C, N, Na, Mg, S, Cl, K, Ca, and Fe contents of the soil surrounding the protocorms of G. elata Bl. f. glauca. JFGL-06 improved the composition, diversity, and metabolic function of the surrounding soil microbial community of G. elata Bl. f. glauca protocorms at the phylum, class, and genus levels, significantly increased the relative abundance of bacteria such as Acidobacteria and fungi such as Trichoderma among the dominant groups, and increased the abundance of functional genes in metabolic pathways such as nucleotide metabolism and energy metabolism. There was a significant reduction in the relative abundance of bacteria, such as Actinomycetes, and fungi, such as Fusarium, in the dominant flora, and a reduced abundance of functional genes, such as amino acid metabolism and xenobiotic biodegradation and metabolism. This is the main reason why the JFGL-06 strain promoted high-quality and high-yield G. elata Bl. f. glauca in Changbai Mountain.

Abstract Background Several recent studies have found that Osteocalcin (OCN), a multifunctional protein secreted exclusively by osteoblasts, is beneficial to glucose metabolism and type 2 diabetes mellitus (T2DM). However, the effects of OCN on islets function especially islet ɑ cells function in patients with type 2 diabetes mellitus characterized by a bi-hormonal disease are still unclear. The purpose of this cross-sectional study was to investigate the relationship between serum OCN and the secretion of islet β cells and ɑ cells in Chinese patients with type 2 diabetes mellitus. Methods 204 patients with T2DM were enrolled. Blood glucose (FBG, PBG0.5h, PBG1h, PBG2h, PBG3h), insulin (FINS, INS0.5h, INS1h, INS2h, INS3h), C-peptide (FCP, CP0.5h, CP1h, CP2h, CP3h), and glucagon (GLA0, GLA0.5 h, GLA1h, GLA2h, GLA3h) levels were measured on 0 h, 0.5 h, 1 h, 2 h, and 3 h after a 100 g standard bread meal load. Early postprandial secretion function of islet β cells was calculated as Δcp0.5h = CP0.5-FCP. The patients were divided into low, medium and high groups (T1, T2 and T3) according to tertiles of OCN. Comparison of parameters among three groups was studied. Correlation analysis confirmed the relationship between OCN and pancreatic secretion. Multiple regression analysis showed independent contributors to pancreatic secretion. Main results FBG, and PBG2h were the lowest while Δcp0.5h was the highest in the highest tertile group (respectively, p

Abstract Background Osteocalcin, a protein secreted mainly by mature osteoblasts, has been shown to be involved in glucose metabolism through various pathways. However, few studies has explored the association between osteocalcin and Time in range (TIR). Continuous glucose monitoring (CGM) -derived metrics, such as TIR and other indexes have been gradually and widely used in clinical practice to assess glucose fluctuations. The main purpose of this study was to investigate the correlation between osteocalcin and indexes from CGM in patients with type 2 diabetes mellitus (T2DM). Method The total number of 376 patients with T2D were enrolled, all of them performed three consecutive days of monitoring. They were divided into four groups on account of the quartile of osteocalcin. Time in range, Time below range (TBR), Time above range(TAR) and measures of glycemic variability (GV) were assessed for analysing. After a 100 g standard steamed bread meal, blood glucose (Glu0h Glu0.5 h, Glu1h, Glu2h, GLu3h), C-peptide (Cp0h, Cp0.5 h, Cp1h, Cp2h, Cp3h), serum insulin (INS0h, INS0.5 h, INS1h, INS2h, INS3h) concentrations at different time points were obtained. HOMA-IS, HOMA-βwas calculated to evaluate insulin sensitivity and insulin secreting of the participants. Results Patients with higher osteocalcin level had higher TIR (P 0.05). Multiple stepwise regression showed that osteocalcin was an independent contributor to TIR, TAR and HOMA-IS. Conclusion Circulating osteocalcin is positively correlated with TIR and negatively correlated with MODD, ADDR, and MAGE. Osteocalcin may have a beneficial impact on glucose homeostasis in T2DM patients.

Abstract Background Ghrelin may ameliorate cancer cachexia (CC) by preventing anorexia, muscle, and fat loss. However, the mechanisms mediating these effects are not fully understood. This study characterizes the pathways involved in muscle mass and strength loss in the Lewis lung carcinoma (LLC)‐induced cachexia model, and the effects of ghrelin in mice with or without its only known receptor: the growth hormone secretagogue receptor‐1a ((GHSR‐1a), Ghsr+/+ and Ghsr−/−). Methods Five to 7‐month‐old male C57BL/6J Ghsr+/+ and Ghsr−/− mice were inoculated with 1 × 106 heat‐killed (HK) or live LLC cells (tumour implantation, TI). When tumours were palpable (7 days after TI), tumour‐bearing mice were injected with vehicle (T + V) or ghrelin twice/day for 14 days (T + G, 0.8 mg/kg), while HK‐treated mice were given vehicle (HK + V). Body weight and grip strength were evaluated before TI and at termination (21 days after TI). Hindlimb muscles were collected for analysis. Results Less pronounced body weight (BW) loss (87.70 ± 0.98% vs. 83.92 ± 1.23%, percentage of baseline BW in tumour‐bearing Ghsr+/+ vs. Ghsr−/−, P = 0.008), and lower upregulation of ubiquitin‐proteasome system (UPS, MuRF1/Trim63, 5.71 ± 1.53‐fold vs. 9.22 ± 1.94‐fold‐change from Ghsr+/+ HK + V in tumour‐bearing Ghsr+/+ vs. Ghsr‐/‐, P = 0.036) and autophagy markers (Becn1, Atg5, Atg7, tumour‐bearing Ghsr+/+

Abstract Background Gastrodia elata Bl. f. glauca S. Chow is a medicinal plant. G. elata f. glauca is unavoidably infected by pathogens in their growth process. In previous work, we have successfully isolated and identified Penicillium oxalicum from fungal diseased tubers of G. elata f. glauca. As a widespread epidemic, this fungal disease seriously affected the yield and quality of G. elata f. glauca. We speculate that the healthy G. elata F. glauca might carry resistance genes, which can resist against fungal disease. In this study, healthy and fungal diseased mature tubers of G. elata f. glauca from Changbai Mountain area were used as experimental materials to help us find potential resistance genes against the fungal disease. Results A total of 7540 differentially expressed Unigenes (DEGs) were identified (FDR 2). The current study screened 10 potential resistance genes. They were attached to transcription factors (TFs) in plant hormone signal transduction pathway and plant pathogen interaction pathway, including WRKY22, GH3, TIFY/JAZ, ERF1, WRKY33, TGA. In addition, four of these genes were closely related to jasmonic acid signaling pathway. Conclusions The immune response mechanism of fungal disease in G. elata f. glauca is a complex biological process, involving plant hormones such as ethylene, jasmonic acid, salicylic acid and disease-resistant transcription factors such as WRKY, TGA.

Abstract Trace elements are essential nutrients for the growth of microorganisms and play an important role in their proliferation. Hence, the purpose of this paper is to explore the optimal C and N sources for large-scale culture of Paenibacillus polymyxa, and to screen trace elements that can promote their proliferation and improve the activity. First, the concentration of Paenibacillus polymyxa Pp-7250, the number of spores were used as evaluation index. It was found that the four trace elements Cu2+, Fe2+, Mn2+, and Zn2+ could promote the proliferation of Paenibacillus polymyxa at their optimal concentrations. Next, when using wheat starch as carbon source and soybean meal as nitrogen source, it was most suitable for large-scale culture. Finally, field experiments were carried out, and it was discovered that the combination of four trace elements plus the wheat soybean meal group could significantly improve the disease prevention, growth promotion ability of Pp-7250 and its colonization in ginseng. Moreover, the ability of Pp-7250 to transform ginseng roots and leaf saponins were also significantly improved. The group also affected the rhizosphere bacterial community of ginseng and the number showed a significant promotion or inhibition.

Objective. The objective of this study is to investigate the relationship between time in range (TIR), a new metric of continuous glucose monitoring (CGM) and cardiovascular autonomic neuropathy (CAN) in individuals with type 2 diabetes mellitus (T2DM). Methods. A total of 349 individuals with T2DM were enrolled in this study. Evaluating by the standard cardiac autonomic reflex tests (CARTs), there were 228 diabetic individuals without cardiovascular autonomic neuropathy (without confirmed CAN) including absent CAN (n=83 cases) and early CAN (n=145 cases) and 121 diabetic individuals complicated with cardiovascular autonomic neuropathy (CAN) including definite CAN (n=109 cases) and severe CAN (n=12 cases). All patients underwent 3-day CGM. TIR is defined as the time percent during a 24-hour period when the glucose is in the range of 3.9-10 mmol/L. The Spearman analysis was used to analyze the correlation between TIR and CART parameters, total CAN score. The logistic regression was applied to analyze the relationship between TIR and CAN by adjusting for the age, duration of diabetes, sex, lipid situation, serum creatinine, body mass index, blood pressure, HbA1c (%), and other glycemic variability (GV) metrics. Results. The total presence of CAN was 34.67% (definite CAN 31.23% and severe CAN 3.44%). Patients with more severe CAN had lower TIR (P

Objective. Type 2 diabetes (T2DM) is a global epidemic and increases mortality due to its vascular complications. Chemerin has been found to exert a major role in glucose and lipid metabolism. The aim of this study was to explore the correlation between plasma chemerin levels and microangiopathy in patients with T2DM. Methods. A total of 598 T2DM patients were classified into two groups: with and without microvascular complications. Anthropometric parameters and blood pressure were taken. The amounts of glycosylated hemoglobin, glucose, lipid profiles, creatinine, and chemerin concentrations in the blood were determined. The presence and severity of nephropathy, retinopathy, and neuropathy were also evaluated by specific tests. Results. Plasma levels of chemerin in diabetic subjects with microvascular complications were markedly elevated compared to those without. The number of microvascular complications increased with high plasma chemerin levels. Patients with high chemerin levels had an increased incidence of nephropathy and retinopathy. Furthermore, the chemerin plasma concentrations increased with the progression of diabetic nephropathy with highest values in macroalbuminuria groups. In contrast, no significant difference was observed in plasma chemerin levels between subjects with and without peripheral neuropathy. Pearson correlation analysis showed that plasma chemerin levels were positively related to duration of diabetes, serum creatinine, and 24-hour urine albumin excretion, even after multiple adjustments. Using logistic regression analysis, plasma chemerin concentrations were independently associated with the presence of nephropathy and retinopathy, not neuropathy. Conclusion. This study elucidated a positive correlation between increased chemerin levels and the development of some subtypes of diabetic microangiopathy.

Background: Bovine viral diarrhea virus (BVDV) infections are endemic in cattle populations worldwide and cause major economic losses. Thus, an effective vaccine is needed against the transmission of BVDV. The glycoprotein Erns is one of the envelope proteins of this virus and shows BVDV-related immunogenicity. Here, we report the use of Panax ginseng as an alternative production platform for the expression of glycoprotein Erns via Agrobacterium-mediated transformation. Result: Polymerase chain reaction (PCR) and reverse transcription (RT)-PCR analyses showed that pBI121-Erns was stably integrated into the chromosome of transformants. ELISA assay and Western blot analysis confirmed the antigenicity of plant-derived Erns glycoprotein. Immunogenicity was evaluated subcutaneously in deer using a soluble protein extract of dried transgenic ginseng hairy roots. Specific humoral and cell-mediated immune responses against BVDV were detected following immunization. Conclusion: These results demonstrated that the Erns glycoprotein could be expressed in ginseng hairy roots and that plant-derived glycoprotein E retained its antigenicity and immunogenicity. [ABSTRACT FROM AUTHOR]

The bovine viral diarrhea virus (BVDV), a single-stranded RNA virus, can cause fatal diarrhea syndrome, respiratory problems, and reproductive disorders in herds. Over the past few years, it has become clear that the BVDV infection rates are increasing and it is likely that an effective vaccine for BVDV will be needed. In this study, transgenic Astragalus was used as an alternative productive platform for the expression of glycoprotein E0. The immunogenicity of glycoprotein E0 expressed in transgenic Astragalus was detected in deer. The presence of pBI121-E0 was confirmed by polymerase chain reaction (PCR), transcription was verified by reverse transcription- (RT-) PCR, and recombinant protein expression was confirmed by ELISA and Western blot analyses. Deer that were immunized subcutaneously with the transgenic plant vaccine developed specific humoral and cell-mediated immune responses against BVDV. This study provides a new method for a protein with weak immunogenicity to be used as part of a transgenic plant vaccine. [ABSTRACT FROM AUTHOR]