Acceleration of melanocyte senescence by the proinflammatory cytokines IFNγ and TNFα impairs the repigmentation response of vitiligo patients to narrowband ultraviolet B (NBUVB) phototherapy.

Vitiligo is a chronic autoimmune disease characterized by the T helper 1 (Th1) cytokine-driven immune destruction of melanocytes (MCs). Although narrowband ultraviolet B (NBUVB) phototherapy has been proven to be an effective therapeutic option, the repigmentation response to that phototherapy varies greatly in different vitiligo patients. Here, we demonstrate that there is an increase of NBUVB-induced cellular senescence in vitiligo MCs exposed to Th1 cytokine interferon γ (IFNγ) and/or tumor necrosis factor α (TNFα) in lesional vitiligo skin from poor responders who had undergone NBUVB phototherapy. Supplementation with exogenous recombinant human stem cell factor (rhSCF) in the culture medium as well as the lentiviral vector-mediated overexpression of cKIT could prevent the MCs from the IFNγ/TNFα-accelerated cellular senescence. Mechanistic studies indicated that the reduced ratio of membrane-bound KIT (mKIT) to the soluble form of KIT (sKIT) is directly related to the cellular senescence of vitiligo MCs following exposure to IFNγ and TNFα. Furthermore, the matrix metalloprotease 9 (MMP9) inhibitor GM6001 attenuates the production of sKIT via the suppression of cKIT ectodomain shedding. Altogether, our study indicates that the presence of Th1 cytokines IFNγ and/or TNFα in the epidermal milieu might impair the repigmentation response of vitiligo patients to NBUVB phototherapy.

[Effects of tillage with mulching on potato yield and the characteristics of soil water and temperature in arid area of southern Ningxia]. / 宁南旱区耕作覆盖对马铃薯产量及土壤水热特征的影响.

To clarify the effects of tillage with mulching on potato yield and soil water and heat characteristics, we conducted a field experiment for two consecutive years in arid region of southern Ningxia. The results showed that tillage depths and mulching materials had significant impacts on soil water storage at 0-100 cm layer during the potato sowing period. The interactive effects of tillage depths and mulching materials were not significant. In 2019, the highest soil water storage was obtained in the subsoiling 30 cm with plastic film mulching, while soil water storage under the subsoiling 40 cm with straw mulch was the highest in 2020. Subsoiling 30 cm with plastic film mul-ching and subsoiling 40 cm with straw mulch significantly increased soil water storage by 16.9% and 33.4% compared with the plowing 15 cm with no mulch (CK), respectively. Tillage depths and mulching materials significantly affected soil water storage in the key growth period of potato. Among the tillage systems, the straw mulching plots and plastic film mulching plots had the strongest effect of soil water conservation. Irrespective of the mulching materials, soil water storage was significantly improved in the subsoiling 30-40 cm plots. Mulching materials and the interaction between tillage depths and mulching materials significantly affected soil effective accumulated temperature at 0-25 cm soil layer after sowing to budding. Among the tillage systems, the plastic film mulching plots significantly increased the average soil effective accumulated temperature by 9.3%, whereas the straw mulching plots significantly reduced the temperature by 18.7%, in comparison with no mulching plots. The highest soil effective accumulated temperature during the whole growth period was obtained in the subsoiling 30 cm with plastic film mulching and subsoiling 40 cm with plastic film mulching treatments in 2019 and 2020. The highest potato tuber yield and economic benefit in 2019 were found in the subsoiling 30 cm with straw mulching treatment, respectively, being 84.6% and 107.9% higher than CK. In 2020, the improvement effect of subsoiling 40 cm with straw mulch on potato tuber yield and economic benefit was the strongest, respectively, which were significantly increased by 81.7% and 105.7%, compared with CK. Tillage depths and mulching materials had significant interactive effects on the water and heat use efficiency of crop. The higher water use efficiency was obtained in the subsoiling 30-40 cm with straw mulch treatments, whereas the accumulated temperature use efficiency was increased significantly under different tillage depths with straw mulching treatments compared with CK. Soil water and effective accumulated temperature during the tuber formation stage were the main factors affecting potato total yield, with stronger effect of soil water than that of soil effective accumulated temperature. Therefore, the treatments of subsoiling 30-40 cm with straw mulch could improve soil moisture and heat condition, and realize potato yield and income increase and efficient use of water and heat resources, which have application and popularization value in dryland potato cultivation of southern Ningxia.

Efficacy of neoadjuvant hyperthermic intraperitoneal chemotherapy in advanced high-grade serous ovarian cancer (the NHIPEC trial): study protocol for a randomised controlled trial.

BACKGROUND: Neoadjuvant chemotherapy (NACT) is an important treatment option for patients with ovarian cancer. Although intravenous NACT can improve optimal resection rates and decrease surgical morbidity and mortality, these advantages do not translate into a survival benefit. Ovarian carcinoma is mainly confined to the peritoneal cavity, which makes it a potential target for hyperthermic intraperitoneal chemotherapy (HIPEC). Our previous study showed that HIPEC could be used in the neoadjuvant setting, which was named neoadjuvant HIPEC (NHIPEC). Since hyperthermia is an excellent chemosensitiser, we hypothesised that the combination of NHIPEC and intravenous NACT could show superior efficacy to intravenous NACT alone. METHODS: This study is a single-centre, open-label, randomised (1:1 allocation ratio) phase 2 trial. A total of 80 patients will be randomly assigned into an experimental group (NHIPEC+intravenous NACT) or a control group (intravenous NACT). Patients in the experimental group will receive NHIPEC following laparoscopic evaluation, and four tubes will be placed via the laparoscopic ports, which will be used to administer NHIPEC. Then, perfusion with docetaxel (60-75 mg/m2) will be performed (43°C for 60 min, Day 0) followed by cisplatin (75 mg/m2, Day 1) infusion (43°C for 60 min) 24 hours later. After NHIPEC, two cycles of intravenous NACT will be given. Patients in the control group will receive three cycles of intravenous NACT. The primary endpoint is the proportion of patients who achieve a Chemotherapy Response Score (CRS) of 3 according to the CRS system. The secondary endpoints include progression-free survival, overall survival and the rates of complete resection and NHIPEC-related adverse events. ETHICS APPROVAL AND DISSEMINATION: This study was approved by the Ethics Committee of Sun Yat-sen Memorial Hospital (approval number: 2020-ky-050). Results will be submitted to peer-reviewed journals and presented at national and international conferences. TRIAL REGISTRATION NUMBER: ChiCTR2000038173.

[Promoting tanshinone synthesis of Salvia miltiorrhiza root by a seed endophytic fungus, Phoma herbarum D603].

The interaction of endophytes and host plant is an effective mean to regulate the growth and secondary metabolism of medicinal plants. Here we want to elucidate the effects and mechanism of Phoma herbarum D603 on the root development and tanshinone synthesis in root of Salvia miltiorrhiza by endophyte-plant coculture system. The mycelium of P. herbarum D603 was colonized in the root tissue space, and formed a stable symbiotic relationship with host plant. The in vitro activities analysis showed that the concentration of IAA produced by D603 can reach(6.45±0.23) µg·mL~(-1), and this strain had some abilities of phosphorus solubilization and siderophore production activities. The coculture experiment showed that strain D603 can significantly promote the synthesis and accumulation of tanshinones in the root of S. miltiorrhiza, in which after 8 weeks of treatment with D603, the content of tanshinone Ⅱ_A in the roots reached up to(1.42±0.59) mg·g~(-1). By the qRT-PCR analysis results, we found that D603 could improve the expression levels of some key genes(DXR, DXS, GGPP, HMGR, CPS) of tanshinone biosynthesis pathway in host plant S. miltiorrhiza, but the promoting effect mainly occurred in the early stage of the interaction, and the enzyme activity level decreased in varying degrees of the later stage. In summary, seed-associated endophyte P. herbarum D603 can promote the growth and root development of S. miltiorrhiza by producing hormones, promoting nutrient absorption and siderophore production, and promote the synthesis and accumulation of tanshinones by regulating the expression level of key genes in the synthetic pathway in S. miltiorrhiza.

Role of the p53­TRPM1/miR­211­MMP9 axis in UVB­induced human melanocyte migration and its potential in repigmentation.

Clinical studies have proven that ultraviolet B (UVB) based phototherapy can induce perifollicular and marginal repigmentation patterns in the skin of vitiligo patients. It is, however, difficult to conceive how melanocytes can easily exit from their tightly interconnected epidermal microenvironment to re­enter a different location in the skin to establish a new network with neighboring keratinocytes. While it is known that matrix metalloprotease 9 (MMP9) is involved in the degradation of the extracellular matrix in physiological or pathological processes, little is known about whether MMP9 affects melanocyte migration in vitiligo repigmentation. To investigate the effects of the p53­ transient receptor potential cation channel subfamily M member 1 (TRPM1)/microRNA (miR/miRNA)­211­MMP9 axis to regulate melanocyte migration following exposure to UVB, the expression profile of MMP9 in cultured human melanocytes transfected with or without the miR­211­mimic and p53­GFP lentiviral vector, respectively were determined. Quantitative polymerase chain reaction and western blotting were used to examine p53, TRPM1 and MMP9 mRNA and protein levels in UVB­exposed and unexposed cells. The capacity of melanocytes to migrate on collagen IV substrate was estimated using a Transwell migration assay. Interestingly, the upregulation of p53 and MMP9 at the mRNA and protein levels was evident in melanocytes treated with single or repeat exposures to UVB, whereas levels of TRPM1 and miR­211 were significantly suppressed in UVB­exposed melanocytes compared with the UVB­unexposed control cells. These results indicate that the p53­TRPM1/miR­211­MMP9 axis is significantly activated in melanocytes exposed to UVB. Notably, the ability of melanocyte migration was altered by the overexpression of p53 using a lentiviral vector and by the upregulation of miR­211 using an miRNA mimic. That altered migration could be neutralized by co­treatment with GM6001 (a broad­spectrum MMP inhibitor). Overall, these results show that the MMP9­mediated migration of melanocytes is regulated by a novel mechanism driven by the p53­TRPM1/miR­211­MMP9 axis. Activation of the p53­TRPM1/miR­211­MMP9 axis potentially represents an attractive therapeutic target to improve repigmentation outcomes in vitiligo patients.

Core Microbiome of Medicinal Plant Salvia miltiorrhiza Seed: A Rich Reservoir of Beneficial Microbes for Secondary Metabolism?

Seed microbiome includes special endophytic or epiphytic microbial taxa associated with seeds, which affects seed germination, plant growth, and health. Here, we analyzed the core microbiome of 21 Salvia miltiorrhiza seeds from seven different geographic origins using 16S rDNA and ITS amplicon sequencing, followed by bioinformatics analysis. The whole bacterial microbiome was classified into 17 microbial phyla and 39 classes. Gammaproteobacteria (67.6%), Alphaproteobacteria (15.6%), Betaproteobacteria (2.6%), Sphingobacteria (5.0%), Bacilli (4.6%), and Actinobacteria (2.9%) belonged to the core bacterial microbiome. Dothideomycetes comprised 94% of core fungal microbiome in S. miltiorrhiza seeds, and another two dominant classes were Leotiomycetes (3.0%) and Tremellomycetes (2.0%). We found that terpenoid backbone biosynthesis, degradation of limonene, pinene, and geraniol, and prenyltransferases, were overrepresented in the core bacterial microbiome using phylogenetic examination of communities by reconstruction of unobserved states (PICRUSt) software. We also found that the bacterial genera Pantoea, Pseudomonas, and Sphingomonas were enriched core taxa and overlapped among S. miltiorrhiza, maize, bean, and rice, while a fungal genus, Alternaria, was shared within S. miltiorrhiza, bean, and Brassicaceae families. These findings highlight that seed-associated microbiomeis an important component of plant microbiomes, which may be a gene reservoir for secondary metabolism in medicinal plants.

Baicalin increases hair follicle development by increasing canonical Wnt/ß­catenin signaling and activating dermal papillar cells in mice.

Baicalin is a traditional Chinese herbal medicine commonly used for hair loss, the precise molecular mechanism of which is unknown. In the present study, the mechanism of baicalin was investigated via the topical application of baicalin to reconstituted hair follicles on mice dorsa and evaluating the effect on canonical Wnt/ß­catenin signaling in the hair follicles and the activity of dermal papillar cells. The results indicate that baicalin stimulates the expression of Wnt3a, Wnt5a, frizzled 7 and disheveled 2 whilst inhibiting the Axin/casein kinase 1α/adenomatous polyposis coli/glycogen synthase kinase 3ß degradation complex, leading to accumulation of ß­catenin and activation of Wnt/ß­catenin signaling. In addition, baicalin was observed to increase the alkaline phosphatase levels in dermal papillar cells, a process which was dependent on Wnt pathway activation. Given its non­toxicity and ease of topical application, baicalin represents a promising treatment for alopecia and other forms of hair loss. Further studies of baicalin using human hair follicle transplants are warranted in preparation for future clinical use.

New dihydro-ß-agarofuran sesquiterpenes from Parnassia wightiana wall: isolation, identification and cytotoxicity against cancer cells.

Five new (4-8) and three known (1-3) dihydro-ß-agarofuran sesquiterpene polyesters were isolated from the whole plants of Parnassia wightiana. The structures of all compounds were elucidated through spectroscopic analysis including 2D-NMR and HR-MS. The absolute configuration of these compounds was established by X-ray diffraction analysis, comparison of NOESY spectra and biogenetic means. The cytotoxities of compounds 2-8 were evaluated in vitro against HL-60, SMMC-7721, A549, MCF-7 and SW480 cell lines. Compounds 5-7 exhibited the highest activities with IC50 values of 11.8-30.1 µM in most cases. The SAR revealed that the introduction of hydroxyl group was able to significantly improve the activities of the compounds for most of the cell lines.

In vitro antifungal activity of sanguinarine and chelerythrine derivatives against phytopathogenic fungi.

In order to understand the antifungal activity of some derivatives of sanguinarine (S) and chelerythrine (C) and their structure-activity relationships, sixteen derivatives of S and C were prepared and evaluated for in vitro antifungal activity against seven phytopathogenic fungi by the mycelial growth rate method. The results showed that S, C and their 6-alkoxy dihydro derivatives S1-S4, C1-C4 and 6-cyanodihydro derivatives S5, C5 showed significant antifungal activity at 100 µg/mL against all the tested fungi. For most tested fungi, the median effective concentrations of S, S1, C and C1 were in a range of 14-50 µg/mL. The structure-activity relationship showed that the C=N+ moiety was the determinant for the antifungal activity of S and C. S1-S5 and C1-C5 could be considered as the precursors of S and C, respectively. Thus, the present results strongly suggested that S and C or their derivatives S1-S5 and C1-C5 should be considered as good lead compounds or model molecules to develop new anti-phytopathogenic fungal agents. can't login to work station for 2hrs--took 2 hrs vacation

[Ecological adaptability of leaf epidermis of erosion-resistant plants in hilly-gully area of Loess Plateau, Northwest China].

By the temporary slide method of leaf epidermis, an observation was made on the morphological characteristics of the leaf epidermis of six erosion-resistant plant species in different soil erosion environments (gully, inter-gully, and inter-gully artificial Robinia pseudoacacia forest land) in hilly-gully area of Loess Plateau. Compared with those in the gully, the stomata aperture, stomata density, stomata index, stomata apparatus length/width plasticity, stomata apparatus area plasticity, epidermal hair density, and epidermal cell density of the leaf upper and lower epidermis of the plants in the inter-gully were 93.8% and 90.4%, 66.8% and 76.6%, 17.9% and 9.8%, 36.4% and 47.1%, 42.3% and 43.9%, 199.4% and 98.2%, and 46.5% and 50.1% higher, respectively; while in the inter-gully artificial R. pseudoacacia forest land, the same morphological indices of the leaf upper and lower epidermis of the plants were 66.7% and 106.7%, 20.5% and 45.8%, 11.9% and 11.9%, 37.9% and 41.3%, 19.8% and 21.2%, 113.1% and 52.2%, and 10.8% and 28.1% higher than those in the gully, respectively. The epidermal hair length and epidermal cell area of the leaf upper and lower epidermis of the plants in the inter-gully were 58.8% and 29.7%, and 40.3% and 37.0% lower than those in the gully, and the same morphological indices of the leaf upper and lower epidermis of the plants in the intergully artificial R. pseudoacacia forest land were respectively 25.0% and 23.6%, and 22.2% and 19.2% lower than those in the gully, respectively. The results suggested that the erosion-resistant plants in the study area were able to adapt to various soil erosion environments by increasing their leaf stomata aperture, stomata density, stomata index, stomata apparatus length/width plasticity, stomata apparatus area plasticity, epidermal hair density, and epidermal cell density, and by reducing their epidermal hair length and epidermal cell area.

[Study on antibacterial active components from Viola yedoensis].

OBJECTIVE: Study on the antibacterial activity of Viola yedoensis and the antibacterial active compounds. METHOD: The chemical compositions were isolated by means of solvent extraction, column chromatography on silica gel, sephadex LH-20 and crystallization. The antibacterial activities were tested by Neo-Sensitab disk-diffusion method, nephelometric analysis and plating method. RESULT: One new compound (4) along with three known compounds were isolated from this plant for the first time and were identified as aesculetin (1), 6,7-dimethoxycoumarin (2), scopoletin (3) and 5-methoxy-7-hydroxymethylcoumarin (4), respectively. All the compounds showed antibacterial and antibactericidal activities at varying degree on Streptococcus Aureas, S. agalactiae, S. uberis, S. dysgalactiae, E. coli and Salmonella, of which 1 was most active with 0.031- 0.313 g x L(-1) of minimal inhibitory concentrations (MIC) and 0.313 - 0.625 g x L(-1) of minimal bactericidal concentrations (MBC). CONCLUSION: Viola yedoensis has a broad spectrum of antibacterial activity on animal pathogenic bacteria, and coumarins may be the main antibacterial activity ingredients.

Two new bufadienolides from the rhizomes of Helleborus thibetanus Franch.

Two new bufadienolides, named tigencaoside A(1) and tigencaoside B(2), were isolated from the rhizomes of Helleborus thibetanus Franch., along with two known bufadienolides, hellebrigenin (3) and 5beta,14beta-dihydroxy-19-oxo-3beta-[(alpha-L-rhamnopyranosyl)oxy]bufa-20,22-dienolide (4). Their structures were elucidated on the basis of extensive spectroscopic analysis. Two new compounds were evaluated for their cytotoxic activities against four strains of cultured tumor cells.

Further studies on the chemical constituents of Chinese folk medicine Gentiana apiata N.E. Br.

One new secoiridoid glycoside with conjugated diene, named 3-epi-swertiajaposide C (1), was isolated from the whole plants of Gentiana apiata N.E. Br., together with 11 known compounds, 7-deoxyloganic acid (2), isoorientin (3), gentiopicroside (4), silybin B (5), swertiamarin (6), asystasioside A (7), 6'-O-beta-D-glucopyranosylgentiopicroside (8), umbelliferone (9), oleanolic acid (10), kaempferol (11), and beta-sitosterol (12). The structure of the new compound (1) was elucidated on the basis of spectroscopic evidence including UV, IR, MS, NMR, HMBC, HMQC, and NOESY. Compounds 1, 2, 5, 6, 8, 9, and 11 were found in this plant for the first time. Moreover, silybin B (5) was isolated from the other plants besides Silybum marianum (L.) Gaertn for the first time by the present study.