An anti-inflammatory repair composition containing multiple B vitamins and a cornflower extract and uses thereof

Abstract

The application provides an anti-inflammatory repair composition containing multiple B vitamins and cornflower extract and an application thereof, and comprises panthenol, nicotinamide, folic acid, cobalamin and cornflower extract. The application uses panthenol, nicotinamide, folic acid, cobalamin and cornflower extract in combination, and each component is mutually matched and promoted, and a synergistic effect is obtained in up-regulating the expression of two key factors FGF2 and CAT, so that the skin repair effect is promoted. In addition, the composition provided by the application can also significantly reduce the expression of inflammatory factors such as IL-8, and has a stronger effect of inhibiting inflammation.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of daily chemical technology, and particularly relates to an anti-inflammatory repair composition containing multiple B vitamins and cornflower extract and application thereof. BACKGROUND

[0002] Skin, as the largest organ of the human body, has a total area of 1.5-2 square meters, not only bears the physical barrier function of resisting external stimuli, but also plays a key role in moisturizing, immune defense, etc. In modern life, the skin is continuously exposed to multiple harmful environmental factors such as ultraviolet radiation, air pollution, electronic device blue light, etc. At the same time, the accelerated pace of life leads to irregular work and rest, increased stress, and frequent use of irritating cosmetics, which further exacerbates skin health problems. With age, the sebaceous glands secrete less, the synthesis of hyaluronic acid decreases, and the lipid bilayer of the stratum corneum is disordered. UVA and UV in ultraviolet light can induce collagen fiber degradation and damage to keratinocyte DNA, respectively. In addition, improper skin care such as excessive cleansing, frequent exfoliation, and disease factors such as atopic dermatitis and rosacea can exacerbate barrier dysfunction, leading to dryness, desquamation, redness, and sensitivity.

[0003] Therefore, it is of great practical significance and broad application prospects to develop an anti-inflammatory repair composition to meet market demand and improve skin health. SUMMARY

[0004] Therefore, the technical problem to be solved by the present application is to provide an anti-inflammatory repair composition containing multiple B vitamins and cornflower extract, a product and application thereof. The composition provided in the present application has a synergistic effect in up-regulating the expression of two key factors FGF2 and CAT, thereby playing a role in promoting skin repair.

[0005] The present application provides an anti-inflammatory repair composition containing multiple B vitamins and cornflower extract, comprising: panthenol, nicotinamide, folic acid, cobalamin and cornflower extract.

[0006] The mass ratio of the panthenol, nicotinamide, folic acid, cobalamin and cornflower extract is 3-7:1-5:0.0005-0.1:0.00005-0.01:0.01-0.5:0.001-0.3.

[0007] Panthenol, also known as pantothenyl alcohol, pro-Vitamin B5, can be converted into pantothenic acid as a coenzyme A precursor to participate in energy metabolism, and provide energy for the repair activities of damaged skin. After penetrating into the stratum corneum, panthenol can stimulate the synthesis of hyaluronic acid, significantly improve skin hydration, and relieve dryness. In the early stage of inflammation, panthenol can quickly inhibit the inflammatory signaling pathway, reduce the release of inflammatory factors such as prostaglandin E2, relieve skin redness, tingling symptoms, accelerate the synthesis of lipids such as ceramide and cholesterol in the skin barrier, stabilize the "brick wall structure", and enhance the ability of the skin to resist external stimuli.

[0008] Nicotinamide, also known as niacinamide, is an amide compound of nicotinic acid (VB3). By inhibiting keratinocyte protease, it reduces the decomposition of lipids such as ceramide from the source, and directly strengthens the lipid structure of the skin barrier. Nicotinamide has the ability to block the NF-κB signaling pathway, and can efficiently inhibit the release of inflammatory factors such as IL-6 and TNF-α, and reduce inflammation. In addition, nicotinamide can also regulate melanosome transport, inhibit tyrosinase activity, reduce post-inflammatory pigmentation, and improve uneven skin tone. It can not only fight inflammation, but also whiten and repair the skin barrier.

[0009] Folic acid belongs to vitamin B9 (VB9). As a key coenzyme for DNA synthesis, it can accelerate keratinocyte mitosis and increase cell proliferation rate after damage from ultraviolet light and other factors, and rapidly replace damaged and aged cells. The metabolic product of folic acid, tetrahydrofolate, has strong antioxidant properties, can neutralize free radicals, reduce oxidative stress damage to skin cell DNA, proteins and lipids, prevent inflammation chain reactions, and help the skin maintain normal physiological functions and structural integrity in an inflammatory state.

[0010] Cobalamin, also known as cyanocobalamin, is vitamin B12 (VB12). Cobalamin participates in the methylation reaction of methionine cycle to precisely regulate the expression of cell differentiation-related genes, guide the normal maturation of keratinocytes, and is particularly important in the repair of skin nerve endings. Damaged skin often has nerve ending dysfunction, leading to sensitivity and burning sensation. Cobalamin can improve nerve conduction and relieve discomfort. In combination with other ingredients, it can reduce the abnormal sensations caused by inflammation from the nerve level and promote overall skin repair.

[0011] Centaurea cyanus flower extract is involved in a variety of cellular functions, and as an FGF receptor cofactor, it can be involved in calcium homeostasis regulation and phosphate, vitamin D metabolism, and maintain normal cell function. Centaurea cyanus flower extract can inhibit the insulin signaling pathway, activate cell health and longevity gene transcription, block Wnt signaling pathway and ligand binding, prevent cell cycle arrest and reduce inflammatory mediator expression, delay aging, promote collagen type I and elastin synthesis, reduce skin redness, improve luster, effectively inhibit aging symptoms, and has great potential in skin repair and anti-inflammatory fields. In some specific implementations, the main active ingredient of the Centaurea cyanus flower extract is an indole alkaloid, and the content is not less than 0.1%. In some specific implementations, the indole alkaloid is coumaroylmethoxyltryptamine, which is obtained by combining 5-methoxytryptamine and coumaric acid through an amide bond, rich in active ingredients such as flavonoids and anthocyanins, and the common CAS number is 581-35-5 (cyanidin). The source of the Centaurea cyanus flower extract is not specially limited in the present application and can be purchased on the market, such as Guangzhou Yunli Biological Technology Co., Ltd., Guangzhou Gallop Biological Technology Co., Ltd., Sollice Biotech of France, and Merck of Germany.

[0012] The present application uses panthenol, nicotinamide, folic acid, cobalamin and Centaurea cyanus flower extract in combination, and each component cooperates and promotes each other, and a synergistic effect is obtained in up-regulating the expression of two key factors FGF2 and CAT, thereby playing a role in promoting skin repair. In addition, the composition provided by the present application can also significantly reduce the expression of inflammatory factors such as IL-8, and has a stronger effect of inhibiting inflammation.

[0013] In some specific implementations, the mass ratio of the panthenol, nicotinamide, folic acid, cobalamin and Centaurea cyanus flower extract is 4-6:2-4:0.001-0.05:0.0001-0.005:0.01-0.2.

[0014] In some specific implementations, the mass ratio of the panthenol, nicotinamide, folic acid, cobalamin and Centaurea cyanus flower extract is 4.5-5.5:2.5-3.5:0.005-0.015:0.0005-0.0015:0.05-0.15.

[0015] In some specific implementations, the mass ratio of the panthenol, nicotinamide, folic acid, cobalamin and Centaurea cyanus flower extract is 5:3:0.01:0.001:0.1.

[0016] The present application also provides the use of the above-mentioned composition in the preparation of anti-inflammatory and / or repair products.

[0017] The present application also provides the use of the above-mentioned composition in the preparation of a product for reducing the expression of inflammatory factors, such as IL-8.

[0018] The present application also provides the use of the above-mentioned composition in the preparation of a product for up-regulating the expression of FGF2 and / or CAT genes.

[0019] The present application also provides a product comprising the above-mentioned composition.

[0020] In some embodiments of the present application, the above-mentioned product includes, but is not limited to, personal care products, pharmaceutical products, etc. Those skilled in the art can understand that, in addition to the above-mentioned composition, other excipients, such as pharmaceutical excipients or personal care product excipients, are also included.

[0021] In some embodiments of the present application, the personal care product includes a basic care product and / or a color cosmetic product.

[0022] In some embodiments of the present application, the color cosmetic product includes, but is not limited to:

[0023] (1) Foundation:

[0024] Liquid / foundation cream: used for uniform skin color, covering blemishes;

[0025] BB cream / CC cream: light and thin foundation product, with skin care and modification functions;

[0026] Concealer: for covering local areas such as acne, dark circles, etc.

[0027] Powder / face powder: setting makeup, reducing facial shine.

[0028] (2) Eye color cosmetics:

[0029] Eye shadow: adding color and depth to the eyes;

[0030] Eyeliner pen / liquid / gel: outlining the eye line to make the eyes look more lively;

[0031] Mascara: lengthening and thickening eyelashes to increase the depth of eye expression;

[0032] Eyebrow pencil / eyebrow powder / eyebrow gel: filling the gaps in the eyebrows and shaping the ideal eyebrow shape.

[0033] (3) Cheek color cosmetics:

[0034] Blush: adding natural blood color to the cheeks to improve complexion;

[0035] Contour cake / stick: using shadow techniques to make the face profile more three-dimensional;

[0036] (4) Lip makeup:

[0037] Lipstick / lip gloss / lip color: change or emphasize the color of the lips;

[0038] Lip liner: draw a clear lip shape boundary to prevent lipstick from spilling.

[0039] (5) Multi-functional makeup:

[0040] Highlighter stick / liquid / powder: brighten the high points of the face (such as the nose bridge, cheekbones), and create a sense of gloss.

[0041] In addition, there are also products specially designed for special occasions, such as waterproof and sweat-resistant eye liner, long-lasting and non-fading lipstick, etc.

[0042] In some embodiments of the present application, the basic care products include but are not limited to:

[0043] Face wash / cleansing foam: gently remove facial dirt, oil and cosmetic residues;

[0044] Makeup remover oil / water / cream: specially used to thoroughly remove makeup, especially waterproof makeup;

[0045] Toner: used after cleansing, can further clean the skin surface residues, at the same time, supply water to the skin, restore the skin pH balance, and lay a good foundation for the absorption of subsequent skin care products;

[0046] Serum: contains high concentration of active ingredients, provides deep nourishment and repair for specific skin problems (such as anti-aging, moisturizing, whitening, etc.);

[0047] Eye cream: specially designed for sensitive areas around the eyes, with the function of lightening fine lines, dark circles and tightening the skin around the eyes, usually with light and easy-to-absorb texture;

[0048] Day / night emulsion or cream: with the functions of protection, repair and nourishment, promotion of cell regeneration, etc., can provide necessary nourishment to the skin and lock in the supplied water;

[0049] Sunscreen: used for UV protection, prevention of photoaging, etc.

[0050] Face mask: provides additional nourishment and care to the skin, such as hydration, pore cleaning or skin lightening, etc.

[0051] In some specific implementations, the personal care products include 0.1wt%-10wt% of the composition of the above technical solution, preferably include 0.5wt%-9.5wt% of the composition of the above technical solution, more preferably include 1wt%-9wt% of the composition of the above technical solution.

[0052] The panthenol, nicotinamide, folic acid, cobalamin and cornflower extract are used in combination, and each component promotes each other and has a synergistic effect on up-regulating the expression of two key factors FGF2 and CAT, thereby playing a role in promoting skin repair. In addition, the composition provided by the application can also significantly reduce the expression of inflammatory factors such as IL-8, and has a stronger effect of inhibiting inflammation. BRIEF DESCRIPTION OF DRAWINGS

[0053] Figure 1 A columnar graph of the results of testing the effect of the sample on the expression of inflammatory factors of keratinocytes;

[0054] Figure 2 The results of testing the effect of the sample on the expression of FGF2;

[0055] Figure 3 The results of testing the effect of the sample on the expression of CAT. DETAILED DESCRIPTION

[0056] The application provides an anti-inflammatory repair composition containing a plurality of B vitamins and cornflower extract, a product and an application thereof, and those skilled in the art can refer to the content herein, appropriately improve process parameters, and realize the application. The method and application of the application have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application herein without departing from the content, spirit and scope of the application, to realize and apply the application technology.

[0057] The terms "comprising", "having", or "including", including the use of their grammatical synonyms, should generally be understood as open-ended and non-limiting, for example, not excluding other non-recited elements or steps, unless otherwise specifically stated or understood from the context.

[0058] It should be understood that the order of steps or the order of performing certain actions is not important as long as the application is still operable. In addition, two or more steps or actions can be performed simultaneously.

[0059] The use of any and all examples, or exemplary language herein, such as "for example" or "including", is intended merely to better illustrate the application and does not pose a limitation on the scope of the application unless claim language is used. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the application.

[0060] Also, the numerical ranges and parameters given herein are by way of approximation. While the exact values and ranges are always preferred, close approximation are acceptable. It must be understood that all numbers noted herein are "about" the value in some amount. Often, but not always, this manner will be expressly connoted where, explicitly or implicitly, presented in the context of the specification.

[0061] The present application provides an anti-inflammatory repair composition containing multiple B vitamins and cornflower extract, comprising panthenol, nicotinamide, folic acid, cobalamin and cornflower extract;

[0062] The mass ratio of the panthenol, nicotinamide, folic acid, cobalamin and cornflower extract is 3-7: 1-5: 0.0005-0.1: 0.00005-0.01: 0.001-0.3.

[0063] The present application uses panthenol, nicotinamide, folic acid, cobalamin and cornflower extract in combination, and each component cooperates and promotes each other, and a synergistic effect is obtained in up-regulating the expression of two key factors FGF2 and CAT, thereby playing a role in promoting skin repair. In addition, the composition provided by the present application can also significantly reduce the expression of inflammatory factors such as IL-8, and has a stronger effect of inhibiting inflammation.

[0064] The present application will be further described below in combination with examples.

[0065] In the following examples, the raw materials are purchased from the market, wherein the cornflower extract is purchased from Guangzhou Gala Biological Technology Co., Ltd.

[0066] Example 1, Comparative Examples 1-2

[0067] The composition is prepared according to the formula shown in Table 1:

[0068] Table 1 Formula provided by the present application examples and comparative examples

[0069]

[0070] Test Example 1 Inhibition of inflammatory factor interleukin 8 (IL-8)

[0071] Test method:

[0072] 1. Cell inoculation

[0073] Immortalized keratinocytes are selected, after recovering the cells, when the plating rate reaches about 60%, the cells are inoculated into 12-well plates, and incubated in a CO2 incubator (37°C, 5% CO2) overnight.

[0074] 2. Test groups

[0075] The test was performed according to the grouping and conditions shown in Table 2, and blank control group (BC), negative control group (NC) and sample group (Example 1, Comparative Example 1 and Comparative Example 2) were set up, and 3 repeated holes were set up for each group.

[0076] Table 2 Test grouping and conditions

[0077]

[0078] 3. Drug administration

[0079] According to Table 2, the culture medium was replaced with a culture medium containing the sample to be tested, and incubated for 24 hours. All experimental conditions were performed in n=3. At the end of the incubation, the cells were collected and immediately frozen at 80°C.

[0080] 4. q-PCR analysis:

[0081] The total RNA of the cells collected under each experimental condition was extracted using TriPure Isolation Reagent, reversed to cDNA, and the expression of the target gene was detected by RT-qPCR method, and each primer is shown in Table 3. Each example and untreated group has 3 repeated samples.

[0082] Table 3 Target gene primers

[0083]

[0084] 5. Statistical analysis of results

[0085] GraphPad Prism was used for plotting, and the results were expressed as Mean±SD. One-way ANOVA statistical analysis was used for comparison between groups. P<0.05 was considered to have significant difference, and P<0.01 was considered to have extremely significant difference.

[0086] The results are shown in Table 4 and Figure 1 Table 4 is the result of the effect of the test sample on the expression of the inflammatory factor of keratinocyte, Figure 1 is the column chart of the effect of the test sample on the expression of the inflammatory factor of keratinocyte, Figure 1 When one-way ANOVA method was used for statistical analysis, the significance was represented by *, P-value<0.05 was represented by *, and P-value<0.01 was represented by **.

[0087] Table 4 Effect of test sample on expression of inflammatory factor of keratinocyte

[0088]

[0089] By Figure 1 and Table 4:

[0090] Compared with the BC group, the expression of inflammatory factors in the NC group was significantly up-regulated, indicating that the test stimulation condition was effective.

[0091] Compared with the NC group, the expression of IL-8 in Comparative Example 1, Comparative Example 2 and Example 1 was significantly reduced, indicating that the examples and comparative examples could effectively inhibit inflammation.

[0092] In addition, compared with Comparative Example 1, Comparative Example 2 and Example 1, the expression of inflammatory factors was significantly reduced, indicating that Example 1 could further reduce the expression of inflammatory factors on the basis of Comparative Example 1, and had a stronger effect of inhibiting inflammation.

[0093] According to the inhibition rate calculation formula, the inhibition rates of the expression of corresponding genes of all comparative examples and examples were calculated, as shown in Table 4. According to the King's method synergy calculation formula q = E A+B / (E A +E B -E A ×E B ), Example 1 compared with Comparative Examples 1 & 2, calculated q = 1.22 > 1.15. Therefore, we believe that the vitamin B combination composition has a synergistic effect with the cornflower extract.

[0094] Test Example 2 Repair and Anti-oxidation Related Pathway

[0095] Test method:

[0096] 1. Cell inoculation

[0097] Fibroblasts were selected, and after the cells were recovered, when the plating rate reached about 60%, the cells were inoculated into 12-well plates and incubated in a CO2 incubator (37°C, 5% CO2) overnight.

[0098] 2. Test grouping

[0099] According to Table 5, the experiment was carried out according to the grouping and conditions, and blank control group (BC) and sample group (Example 1, Comparative Example 1 and Comparative Example 2) were set.

[0100] Table 5 Test grouping and dosing concentration

[0101]

[0102] 3. Dosing

[0103] The medium was replaced with medium containing the sample to be tested, and incubated for 24 hours. All experimental conditions were performed in n = 1. At the end of the incubation, the cells were collected and immediately frozen dry at -80°C.

[0104] 4. RNA-seq analysis:

[0105] Total RNA was extracted from the cells collected under each experimental condition using TriPure Isolation Reagent, using the MJzol Animal RNA Isolation Kit (Majorivd) and according to the standard operating procedure provided by the manufacturer. The purified total RNA was subjected to mRNA isolation, fragmentation, first-strand cDNA synthesis, second-strand cDNA synthesis, end repair, 3' end A addition, linker ligation, and enrichment steps to complete the construction of the mRNA sequencing library. Sequencing was performed according to the effective concentration of the library and the data output requirements. The sequencing platform used was the Illumina NovaSeq6000, and the sequencing mode was PE150 (Pair-end 150 bp), i.e., double-end sequencing of 150 bp per end. NGS (Next Generation Sequencing) technology was used to sequence the cDNA using a high-throughput sequencer (such as the Illumina NovaSeq6000).

[0106] 5. Data processing

[0107] First, Seqtk was used to filter the initial data, and the spliced mapping algorithm of Hisat2 (version: 2.0.4) was used to perform Genome mapping on the preprocessed reads. Stringtie (version: 1.3.0) was used to count the number of Fragments for each gene after Hisat2 alignment, then the TMM (trimmed mean of M values) method was used for normalization, and finally the perl script was used to calculate the FPKM value of each gene.

[0108] The calculation formula is as follows:

[0109]

[0110] Where:

[0111] total exon fragments: the number of fragments mapped to the gene exon (fragment: a pair of reads);

[0112] Exon length: total length of exons of a gene

[0113] Mapped reads: total number of reads mapped to the reference genome

[0114] The edgeR was applied for the differential gene analysis between samples, and the multiple hypothesis test correction was performed after P-value, and the threshold of P-value was determined by controlling FDR (False Discovery Rate), and the corrected P-value was q-value. At the same time, we calculated the differential expression fold according to FPKM value, that is, Fold-change.

[0115] The differential gene screening conditions are as follows:

[0116] 1) Q-value <= 0.05 (when the total number of differential genes in all comparisons is less than 50, it will be automatically adjusted to P-value <= 0.05);

[0117] 2) | log2FC | >=1;

[0118] The results are shown in Figure 2 , Figure 3 , Table 6 and Table 7, Figure 2 , which are the results of the effect of the test sample on the expression of FGF2, Figure 3 , which is the effect of the test sample on the expression of CAT, Table 6 is the effect of the test sample on the gene expression profile of fibroblasts, and Table 7 is the relative promotion rate of the test sample on the gene expression of fibroblasts.

[0119] Table 6 Effect of test sample on gene expression profile of fibroblasts

[0120]

[0121] Table 7 Relative promotion rate of test sample on gene expression of fibroblasts

[0122]

[0123] BC is the blank control group, Comparative Example 1 is a mixture obtained by dissolving panthenol, nicotinamide, folic acid and cobalamin in an aqueous solution at a ratio of 5:3:0.01:0.001, Comparative Example 2 is a mixture obtained by dissolving cornflower extract in an aqueous solution, and Example 1 is a mixture obtained by dissolving panthenol, nicotinamide, folic acid, cobalamin and cornflower extract in an aqueous solution at a ratio of 5:3:0.01:0.001:0.1.

[0124] From Figure 2 , Figure 3 , Table 6 and Table 7, it can be seen that:

[0125] In the two indicators of FGF2 and CAT, Example 1 significantly up-regulated the gene expression of fibroblasts.

[0126] Fibroblast growth factor 2 (FGF2) is a key regulator of skin repair, secreted by keratinocytes, fibroblasts and endothelial cells. It is essential for skin repair and is involved in all stages of the repair process: cell migration, cell proliferation and differentiation, neovascularization, regulation of synthesis and deposition of various extracellular matrix components, and regulation of collagenase activity.

[0127] Catalase (CAT) is a key enzyme in the skin antioxidant defense system, expressed by epidermal keratinocytes and dermal fibroblasts. It achieves triple protection by efficiently decomposing hydrogen peroxide: first, it scavenges free radicals, blocking ROS-induced oxidative damage to DNA, lipids and proteins; second, it regulates inflammation by inhibiting the H2O2-activated NF-κB pathway, reducing the inflammatory storm; and finally, it maintains cell function: protecting repair cells (such as fibroblasts) from apoptosis.

[0128] The composition provided by the present application can promote skin repair by up-regulating two key factors, FGF2 and CAT, and is superior to B vitamin composition and cornflower extract in regulating these two factors.

[0129] According to the promotion rate calculation formula, the promotion rates of the corresponding gene expressions of all comparative examples and examples are calculated as shown in Table 7. According to the Jin's method synergy calculation formula q = E A+B / (E A +E B -E A ×E B ), the q of Example 1 is greater than or equal to 1.15 compared with Comparative Examples 1 and 2, therefore, the vitamin B composition and the cornflower extract composition have synergistic effect.

[0130] The above is only a preferred embodiment of the present application, it should be pointed out that, for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. An anti-inflammatory repair composition containing multiple B vitamins and cornflower flower extract, characterized in that: Composed of panthenol, niacinamide, folic acid, cobalamin, and cornflower flower extract; The mass ratio of panthenol, niacinamide, folic acid, cobalamin and cornflower flower extract is 3-7:1-5:0.0005-0.1:0.00005-0.01:0.001-0.

3.

2. The anti-inflammatory repair composition according to claim 1, characterized in that The mass ratio of panthenol, niacinamide, folic acid, cobalamin and cornflower flower extract is 4-6:2-4:0.001-0.05:0.0001-0.005:0.01-0.

2.

3. The anti-inflammatory repair composition according to claim 2, characterized in that The mass ratio of panthenol, niacinamide, folic acid, cobalamin and cornflower flower extract is 4.5-5.5:2.5-3.5:0.005-0.015:0.0005-0.0015:0.05-0.

15.

4. The anti-inflammatory repair composition according to claim 3, characterized in that The mass ratio of the panthenol, niacinamide, folic acid, cobalamin and cornflower flower extract is 5:3:0.01:0.001:0.

1.

5. Use of the anti-inflammatory and repairing composition according to any one of claims 1 to 4 in the preparation of anti-inflammatory and / or repairing products.

6. Anti-inflammatory and repairing products, characterized by: include: The anti-inflammatory repair composition according to any one of claims 1 to 4.

7. The anti-inflammatory and repairing product according to claim 6, characterized in that: Including personal care products.

8. The anti-inflammatory and repairing product according to claim 6, characterized in that: The content of the anti-inflammatory repair composition is 0.1 wt% to 10 wt%.

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