The saffron phenylpropane synthesis pathway and Fusarium oxysporum cell wall-degrading enzymes play key roles in their early interactions. Saffron (Crocus sativus) is a highly important crop with diverse medicinal properties. F. oxysporum is a widely-distributed soil-borne fungus, causing the serious saffron rot disease. Currently, there is no effective management strategy to control this disease because of no resistant cultivars and limited information about the resistance and pathogenic mechanisms. In this study, we first characterized the infection process and physiological responses of saffron infected by F. oxysporum. The molecular mechanism of these infection interactions was revealed by dual RNA-seq analysis. On the 3rd day of infection, the hyphae completely entered, colonized and spread in the corm cells; while on the 6th day of infection, hyphae had appeared in the xylem cells, blocking these vessels. Transcriptome results indicate that within the host, phenylpropanoid metabolism, plant hormone signal transduction and plant pathogen interaction pathways were activated during infection. These pathways were conducive to the enhancement of cell wall, the occurrence of hypersensitivity, and the accumulation of various antibacterial proteins and phytoantitoxins. Meanwhile, in the fungus, many up-regulated genes were related to F. oxysporum cell wall degrading enzymes, toxin synthesis and pathogenicity gene, showing its strong pathogenicity. This study provides new ideas for the control of saffron corm rot, and also provides a theoretical basis for mining the key functional genes.

Cypripedium palangshanense is a rare and endangered protected plant unique to China. It is an extremely small population of wild distributed in a narrow area. To study its genetic diversity CDDP molecular marker technology was used to evaluate the genetic diversity of 92 materials from 6 experimental sites in 2 wild populations of C. palangshanense in Wolong and Wanglang Nature Reserves, and further explore the relationship between genetic diversity and environmental factors. The results of the genetic analysis showed that a total of 131 locus were detected at the species level with 12 primers, the percentage of polymorphic loci (PPL) was 100%, the observed allele (Na) was 2, the effective allele (Ne) was 1.5026, the Nei′s genetic diversity index (H) was 0.3141, and the Shannon diversity index (I) was 0.4856; At the population level, the genetic diversity of Wolong population was higher than that of Wanglang population, and the variation range of Na was 1.4504 ~ 1.9160; the variation range of Ne was 1.2446 ~ 1.4336; H was between 0.1464 and 0.2679; I was between 0.2231 and 0.4153; the variation range of PPL was 45.04% ~ 91.60%. The genetic differentiation coefficient (Gst) of C. palangshanense was 02863. The genetic structure and AMOVA analysis showed that there was genetic differentiation among the populations of C. palangshanense. Mantel test showed that there was a significant correlation between genetic distance and geographical distance among Cypripedium palang shanense populations in Balang mountain (R2 = 0.3830, P<0.05). According to UPGMA cluster analysis, Wolong population and Wanglang population are clustered into one branch. The correlation analysis between genetic diversity and environmental factors showed that there was a significant positive correlation among observed allele (Na), the number of polymorphic loci (Np), percentage of polymorphic loci (PPL) and available potassium content (AK); The number of effective alleles (Ne), Nei′s genetic diversity index (H) and Shannon information index (I) were significantly positively correlated with altitude (Alt); There was no significant correlation between remaining environmental factors and genetic diversity index. The results showed that CDDP molecular markers technology was suitable for the study of the genetic diversity of C. palangshanense, and was highly polymorphic.

巴郎山杓兰(Cypripedium palangshtanense)是中国特有的珍稀濒危保护植物,属于狭域分布的极小种群野生植物.为研究其遗传多样性,采用CDDP分子标记技术对卧龙和王朗自然保护区巴郎山杓兰的2个野生居群6个试验点的92份材料进行了遗传多样性评价,并进一步探讨了遗传多样性与环境因子之间的相关性.遗传分析结果表明,筛选的12条引物,在物种水平上,共检测到131个位点,多态位点条带百分率(PPL)为100％,观测等位基因(Na)为 2.0000,有效等位基因(Ne)为1.5026,Nei's遗传多样性指数(H)为0.3141,Shannon多样性指数(I)为0.4856,巴郎山杓兰表现出较高的遗传多样性;在居群水平上,卧龙居群的遗传多样性高于王朗居群,其中,Na的变化范围为1.4504～1.9160;Ne的变化范围为1.2446～1.4336;H在0.1464～0.2679之间;I在0.2231～0.4153之间;PPL的变化范围为45.04％～91.60％.巴郎山杓兰的遗传分化系数(Gst)为0.2863,遗传结构和AMOVA分析均表明巴郎山杓兰居群间出现了遗传分化.Mantel检验表明巴郎山杓兰居群之间的遗传距离和地理距离之间有显著的相关性(R2=0.3830,P＜0.05).UPGMA聚类分析表明卧龙居群和王朗居群各聚为一支.遗传多样性与环境因子的相关性分析表明,观测等位基因(Na)、多态位点数(Np)、多态位点百分率(PPL)与速效钾含量(AK)呈显著正相关;有效等位基因数(Ne)、Nei's遗传多样性指数(H)和Shannon信息指数(I)与海拔(Alt)呈显著正相关;其余环境因子与遗传多样性指数之间没有显著相关性.本研究结果表明CDDP分子标记适用于巴郎山杓兰的遗传多样性研究,并且表现出较高的多态性.

Probiotics participate in the regulation of various micro-ecosystems of the host and have the potential to treat a variety of diseases related to microecological imbalance. In recent years, some studies have found that probiotics have a good application prospect in the prevention and treatment of oral diseases, especially periodontal diseases, however, the potential of probiotics in the biomedical field has not been fully developed. This paper is to review the mechanism, effect, and application prospect of probiotics in the prevention and treatment of oral diseases, especially periodontal diseases and peri-implant diseases.

Antioxidant and hepatoprotective activities in vitro of saffron petals were examined in this study for better utilizing saffron (Crocus sativus L.) biowaste. Using the DPPH and ABTS radical scavenging method, we compared the antioxidant activity and the content of total flavonoid extracts from petals (TFESP), stamens (TFESS), and both saffron petals and stamens (TFEMS). The results showed that the antioxidant capacity and the flavonoid content of TFESP were higher than those of TFESS and TFEMS. Then, the hepatoprotective activity of TFESP was determined, and the silymarin was used as a positive control. The main components of TFESP were analysed by ultrahigh performance liquid chromatography (UPLC) photodiode array (PDA)/mass spectrometry (MS) and nuclear magnetic resonance (NMR). The result showed that (1) TFESP could release oxidative liver injury induced by tert-butyl hydroperoxide (t-BHP). (2) TFESP could reduce the accumulation of reactive oxygen species (ROS); enhance the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH); and then improve the total antioxidant capacity (T-AOC) in BRL-3A cells. (3) TFESP could enhance the expression of B-cell lymphoma-2 (BCL-2) and decrease the expression of caspase-3 and caspase-9; increase the expression of Kelch-like ECH-associated protein-1 (Keap-1), nuclear factor, erythroid 2-related factor 2 (Nrf2), superoxide dismutase, and heme oxygenase 1 (HO-1); and downregulate inducible nitric oxide synthase (INOS), interleukin-6 (IL-6), and nuclear factor kappa B-9 (NF-κB-9). (4) The main hepatoprotective component of TFESP was identified as kaempferol-3-o-sophoroside. The mechanism may be that kaempferol-3-o-sophoroside can protect t-BHP-induced cell injury by regulating the expression of antioxidant, antiapoptotic, and anti-inflammatory genes. Thus, saffron petals are a potential hepatoprotective resource worthy of development.

Gingival crevicular fluid (GCF) is a physiological fluid and an inflammatory serum exudate derived from the gingival plexus of blood vessels and mixed with host tissues and subgingival plaque flows. In addition to proteins, GCF contains a diverse population of cells, including desquamated epithelial cells, cytokines, electrolytes, and bacteria from adjacent plaques. Recently, matrix metalloproteinases(MMPs), which are endopeptidases that are active against extracellular macromolecules, in GCF have been revealed as potential utility biomarkers for the diagnosis and follow-up of oral and systemic diseases, thereby facilitating the early evaluation of malignancy risk and the monitoring of disease progression and treatment response. Tissue inhibitors of metalloproteinases (TIMPs) are specific inhibitors of matrixins that participate in the regulation of local activities of MMPs in tissues. This review provides an overview of the latest findings on the diagnostic and prognostic values of MMPs and TIMPs in GCF of oral and systemic diseases, including periodontal disease, pulpitis, peri-implantitis and cardiovascular disease as well as the extraction, detection and analytical methods for GCF.

In order to explore the allelopathy of leaf litter of Larix kaempferi on native plants, the effects of different concentrations (0.01, 0.05, 0.1 g/mL) of water extracts of Larix kaempferi litters (LkLWE) on seed germination and seedling growth of native plants were studied using petri dish filter paper method and pot soil culture method with 5 native plants in Wolong. The results showed that LkLWE had an effect on the seed germination and seedling growth of 5 native plants.1) The intensity of the comprehensive effect of the allelopathy of LkLWE on seed germination was in order of M.sativa > P.vulgaris > M.officinalis > P.depressa > E.dahuricus. The LkLWE mainly inhibited the seed germination of M.sativa, M.officinalis and P.vulgaris, and the inhibition degree was positively correlated with the concentration of the extract; the seed germination of E.dahuricus and P.dipressa showed a phenomenon of low promotion and high inhibition. 2) The intensity of allelopathic comprehensive effect of LkLWE on the growth of seedlings was in the order of M.sativa > M.officinalis >P.vulgaris > E.dahuricus > P.depressa. Among them, the extract mainly inhibited the seedlings growth of M.sativa and M.officinalis, and was positively correlated with the concentration. The LkLWE has a lo-promoting and high-inhibiting effect on the seedlings growth of E. dahuricus and P. vulgaris, and has always been a positive effect on the growth of P.depressa seedlings. The results suggest some management measures for artificial replanting under Larix kaempferi forest and increasing plant diversity.

In order to promote the reuse of Larix kaempferi waste resources， this study used the litter needles of Larix kaempferi as raw materials to optimize the extraction process of total flavonoids by ultrasonic-assisted extraction method, and studied the antioxidant activity of flavonoids. By three-factor three-level response surface analysis, the best extraction process for total flavonoids from Larix kaempferi litter needles was: the ethanol concentration is 60%, the liquid-to-material ratio is 33:1 mL/g, the extraction temperature is 80 ℃, the ultrasonic power is 270 W, the ultrasonic frequency is 40 kHz, and the extraction time is 58 min. Studies on the antioxidant activity of different extraction phases in vitro showed that the antioxidant activity was n-butanol phase>water residue phase>ethyl acetate phase>petroleum ether phase from large to small. The semi-inhibitory rate of n-butanol on DPPH free radical was 7.438 μg/mL, which is slightly weaker than that of vitamin C (7.259 μg/mL); the IC50 value of ABTS radical scavenging was 6.425 μg/mL, showing a higher scavenging capacity than vitamin C. These results showed that the extraction process was stable and feasible, with a high yield of total flavonoids and good antioxidant activity. The research results show that Larix kaempferi waste has the potential to develop into plant-derived natural antioxidants.