N6-methyladenosine reader YTHDF3 regulates melanoma metastasis via its 'executor'LOXL3.

BACKGROUND: A number of studies have demonstrated that N6-methyladenosine (m6A) plays a vital role in the pathological process of various tumours. Recently, it was found that m6A writers or erasers affect the tumourigenesis of melanoma. However, the relationship between m6A readers such as YTH domain family (YTHDF) proteins and melanoma was still elusive. METHODS: RT-qPCR, Western blot and immunohistochemistry were conducted to measure the expression level of YTH N6-methyladenosine RNA binding protein 3 (YTHDF3) and lysyl oxidase-like 3 (LOXL3) in melanoma tissues and cells. The effects of YTHDF3 and LOXL3 on melanoma were verified in vitro and in vivo. Multi-omics analysis including RNA-seq, MeRIP-seq, RIP-seq and mass spectrometry analyses was performed to identify the target. The interaction between YTHDF3 and LOXL3 was verified by RT-PCR, Western blot, MeRIP-qPCR, RIP-qPCR and CRISPR-Cas13b-based epitranscriptome engineering. RESULTS: In this study, we found that m6A reader YTHDF3 could affect the metastasis of melanoma both in vitro and in vivo. The downstream targets of YTHDF3, such as LOXL3, phosphodiesterase 3A (PDE3A) and chromodomain helicase DNA-binding protein 7 (CHD7) were identified by means of RNA-seq, MeRIP-seq, RIP-seq and mass spectrometry analyses. Besides, RT-qPCR, Western blot, RIP-qPCR and MeRIP-qPCR were performed for subsequent validation. Among various targets of YTHDF3, LOXL3 was found to be the optimal target of YTHDF3. With the application of CRISPR-Cas13b-based epitranscriptome engineering, we further confirmed that the transcript of LOXL3 was captured and regulated by YTHDF3 via m6A binding sites. YTHDF3 augmented the protein expression of LOXL3 without affecting its mRNA level via the enrichment of eukaryotic translation initiation factor 3 subunit A (eIF3A) on the transcript of LOXL3. LOXL3 downregulation inhibited the metastatic ability of melanoma cells, and overexpression of LOXL3 ameliorated the inhibition of melanoma metastasis caused by YTHDF3 downregulation. CONCLUSIONS: The YTHDF3-LOXL3 axis could serve as a promising target to be interfered with to inhibit the metastasis of melanoma.

Etanercept treatment of Stevens-Johnson syndrome and toxic epidermal necrolysis.

BACKGROUND: Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) is a severe cutaneous adverse reaction to drugs with considerable morbidity and mortality. Immunomodulators for SJS/TEN including systemic corticosteroids and intravenous immunoglobulin (IVIG) have been widely used in clinical practice. Emerging evidence suggested the therapeutic effects of tumor necrosis factor-α antagonists on SJS/TEN. OBJECTIVE: To compare the efficacy and safety of IVIG and systemic steroids in conjunction with or without etanercept, a tumor necrosis factor-α inhibitor, for patients with SJS/TEN. METHODS: We undertook a retrospective review of 41 patients with SJS/TEN admitted to our institution from 2015 to February 2021. A total of 25 patients with integrated data were involved in this study, of which 14 patients were treated with IVIG and corticosteroids and 11 were in addition given etanercept. The clinical characteristics, duration of hospitalization, exposure time to high-dose steroids, and the total amount of systemic steroids were analyzed. RESULTS: In comparison to conventional therapy, conjunction with etanercept reduced the duration of hospitalization (13.5 vs 19.0 days; P = .01), the exposure time of high-dose steroids (7.1 vs 14.9 days; P = .01), and the overall amount of systemic steroid (925 mg vs 1412.5 mg; P = .03) in patients with SJS/TEN. No pronounced adverse effects were observed within 6 months of follow-up after the treatment. CONCLUSION: The add-in of etanercept at the time of initiating conventional therapy could be a superior option to accelerate disease recovery and reduce the high dose and total amount of systemic steroids without pronounced adverse events in patients with SJS/TEN.

[Regeneration characteristics of three natural Juniperus forests in the Three-River Headwater Region of Qinghai Province, China]. / 青海三江源地区三种天然圆柏林更新特征.

The aims of this study were to clarify the regeneration characteristics and dominant factors affecting the regeneration of three natural Juniperus forests in the Three-River Headwater Region of Qinghai Province, and thus to provide a reference for the protection and management of natural forests. We evaluated the natural regeneration levels of Juniperus forests, and the effects of stand factors and soil factors on natural regeneration. The results showed that three natural Juniperus forests were poorly regenerated, with insufficient regeneration potential. The average regeneration density of J. tibetica forest, J. przewalskii forest and J. convallium forest was 332, 279 and 202 ind·hm-2, respectively. The height range of regenerate individuals was concentrated in 1-3 m. Only a few seedlings (12 ind·hm-2) were found under the J. tibetica forest, and no seedlings were found under the J. convallium and J. przewalskii forests. The regeneration density of J. tibetica forest was significantly positively correlated with stand density, soil organic matter and available phosphorus, and negatively correlated with shrub coverage. The regeneration density of J. convallium forest was significantly negatively correlated with herb coverage, human disturbance degree, woodland slope and soil total nitrogen, and positively correlated with soil water content. The regeneration density of J. przewalskii forest was significantly positively correlated with stand density, soil available potassium and available phosphorus, but negatively correlated with herb coverage. Results of multiple regression analysis showed that the regeneration of J. tibetica forest was mainly affected by understory shrub coverage and soil available phosphorus, that of J. convallium forest was mainly affected by understory herb coverage, soil total nitrogen and human disturbance, and that of J. przewalskii forest was mainly affected by understory herb coverage and soil available potassium. It was necessary to strengthen forest enclosure, management and protection, rationally regu-late the coverage of understory vegetation, increase soil fertility and improve biotope in the forest, which would promote the protection and natural regeneration of natural Juniperus forests in the Three-River Headwater Region.

[Criteria of "arrival of qi" of acupuncture from "reaction of spirit produces curative effect": inheriting SHI Xue-min's thoughts of treating spirit].

From "arrival of qi produces curative effect", it is considered that arrival of qi, qi reaching and acupuncture sensation cannot contain the effect attribute, location attribute and performance attribute of acupuncture at the same time, so it is necessary to find a more suitable standard to measure the arrival of qi and clinical curative effect. Inspired by "the arrival of qi should be reaction of spirit, and reaction of spirit produces curative effect", the authors propose to use the term "reaction of spirit" to summarize the numerous external manifestations of arrival of qi, and to coordinate the attributes of arrival of qi, qi reaching and acupuncture sensation. Academician SHI Xue-min guides the clinical treatment with "reaction of spirit produces curative effect", attaches importance to the spirit treatment to improve the clinical curative effect, and evaluates whether acupuncture is successful or not with "reaction of spirit".

Iatrogenic Arsenism Characterized by Palmoplantar Hyperkeratosis and Diffused Skin Cancers for Over Decades.

ABSTRACT: Chronic arsenism usually occurs after a long-term unawareness of arsenic exposure from environment, occupation, food, and water. We here reported 3 cases with diffused arsenic keratosis and skin cancers derived from long-term arsenic medication ingestion. In these cases, hyperkeratotic skin lesions were initially found on palms and soles, slowly progressed to every part of the skin and lasted maximally for over 30 years. Skin cancers were diagnosed and removed intermittently within decades, but with no malignancies in other organs. Oral retinoids combing with topical 5- fluorouracil and photodynamic treatment yielded a desirable outcome.