Integrative analysis of bulk and single-cell RNA-seq reveals the molecular characterization of the immune microenvironment and oxidative stress signature in melanoma.

Background: The immune microenvironment and oxidative stress of melanoma show significant heterogeneity, which affects tumor growth, invasion and treatment response. Single-cell and bulk RNA-seq data were used to explore the heterogeneity of the immune microenvironment and oxidative stress of melanoma. Methods: The R package Seurat facilitated the analysis of the single-cell dataset, while Harmony, another R package, was employed for batch effect correction. Cell types were classified using Uniform Manifold Approximation and Projection (UMAP). The Secreted Signaling algorithm from CellChatDB.human was applied to elucidate cell-to-cell communication patterns within the single-cell data. Consensus clustering analysis for the skin cutaneous melanoma (SKCM) samples was executed with the R package ConsensusClusterPlus. To quantify immune infiltrating cells, we utilized CIBERSORT, ESTIMATE, and TIMERxCell algorithms provided by the R package Immuno-Oncology Biological Research (IOBR). Single nucleotide variant (SNV) analysis was conducted using Maftools, an R package specifically designed for this purpose. Subsequently, the expression levels of PXDN and PAPSS2 genes were assessed in melanoma tissues compared to adjacent normal tissues. Furthermore, in vitro experiments were conducted to evaluate the proliferation and reactive oxygen species expression in melanoma cells following transfection with siRNA targeting PXDN and PAPSS2. Results: Malignant tumor cell populations were reclassified based on a comprehensive single-cell dataset analysis, which yielded six distinct tumor subsets. The specific marker genes identified for these subgroups were then used to interrogate the Cancer Genome Atlas Skin Cutaneous Melanoma (TCGA-SKCM) cohort, derived from bulk RNA sequencing data, resulting in the delineation of two immune molecular subtypes. Notably, patients within the cluster2 (C2) subtype exhibited a significantly more favorable prognosis compared to those in the cluster1 (C1) subtype. An alignment of immune characteristics was observed between the C2 subtype and unique immune functional tumor cell subsets. Genes differentially expressed across these subtypes were subsequently leveraged to construct a predictive risk model. In vitro investigations further revealed elevated expression levels of PXDN and PAPSS2 in melanoma tissue samples. Functional assays indicated that modulation of PXDN and PAPSS2 expression could influence the production of reactive oxygen species (ROS) and the proliferative capacity of melanoma cells. Conclusion: The constructed six-gene signature can be used as an immune response and an oxidative stress marker to guide the clinical diagnosis and treatment of melanoma.

Recombinant hirudin attenuates pulmonary hypertension and thrombosis in acute pulmonary embolism rat model.

Background: Acute pulmonary embolism (APE) is classified as a subset of diseases that are characterized by lung obstruction due to various types of emboli. Current clinical APE treatment using anticoagulants is frequently accompanied by high risk of bleeding complications. Recombinant hirudin (R-hirudin) has been found to have antithrombotic properties. However, the specific impact of R-hirudin on APE remains unknown. Methods: Sprague-Dawley (SD) rats were randomly assigned to five groups, with thrombi injections to establish APE models. Control and APE group rats were subcutaneously injected with equal amounts of dimethyl sulfoxide (DMSO). The APE+R-hirudin low-dose, middle-dose, and high-dose groups received subcutaneous injections of hirudin at doses of 0.25 mg/kg, 0.5 mg/kg, and 1.0 mg/kg, respectively. Each group was subdivided into time points of 2 h, 6 h, 1 d, and 4 d, with five animals per point. Subsequently, all rats were euthanized, and serum and lung tissues were collected. Following the assessment of right ventricular pressure (RVP) and mean pulmonary artery pressure (mPAP), blood gas analysis, enzyme-linked immunosorbnent assay (ELISA), pulmonary artery vascular testing, hematoxylin-eosin (HE) staining, Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining, immunohistochemistry, and Western blot experiments were conducted. Results: R-hirudin treatment caused a significant reduction of mPAP, RVP, and Malondialdehyde (MDA) content, as well as H2O2 and myeloperoxidase (MPO) activity, while increasing pressure of oxygen (PaO2) and Superoxide Dismutase (SOD) activity. R-hirudin also decreased wall area ratio and wall thickness to diameter ratio in APE rat pulmonary arteries. Serum levels of endothelin-1 (ET-1) and thromboxaneB2 (TXB2) decreased, while prostaglandin (6-K-PGF1α) and NO levels increased. Moreover, R-hirudin ameliorated histopathological injuries and reduced apoptotic cells and Matrix metalloproteinase-9 (MMP9), vascular cell adhesion molecule-1 (VCAM-1), p-Extracellular signal-regulated kinase (ERK)1/2/ERK1/2, and p-P65/P65 expression in lung tissues. Conclusion: R-hirudin attenuated pulmonary hypertension and thrombosis in APE rats, suggesting its potential as a novel treatment strategy for APE.

Corrigendum: The fabrication of a gellan gum-based hydrogel loaded with magnesium ions for the synergistic promotion of skin wound healing.

[This corrects the article DOI: 10.3389/fbioe.2021.709679.].

Clinical and histopathological characteristics, diagnosis and treatment, and comorbidities of Bowen's disease: a retrospective study.

Background: Bowen's disease (BD) is a slow-growing precancerous skin condition, often concurrent with other diseases, with a high misdiagnosis rate. Previous studies show that patients with BD in different populations have differentiated characteristics. Materials and methods: A retrospective study was conducted in a tertiary hospital in Shenzhen, China. Data about demographic information, diagnosis and treatment, clinical and pathological characteristics, and comorbidities of 50 patients with BD were collected and analyzed. Results: Clinical data of onset age and disease course of 43 patients with BD were available, the average onset age of male and female patients are 55.1 (standard deviation (SD) = 15.29) and 58.2 (SD = 15.59) years old, respectively; the average disease course of male and female patients are 25.3 (SD = 28.63) and 33.9 (SD = 49.65) months, respectively. The onset age (p = 0.52) and disease course (p = 0.49) between male and female patients are not significantly different. Interestingly, there is a negative correlation between onset age and disease course (r = -0.245, p = 0.11). The correct rate of clinical diagnosis is relatively low (54.00%); Some patients with BD are misdiagnosed as Bowenoid papulosis (10.00%), actinic keratosis (8.00%), basal cell carcinoma (8.00%), seborrheic keratosis (6.00%), and pigmented naevus (4.00%). Trunk and limbs are the most common distribution sites of BD lesions, and 94.00% patients with BD are treated with surgical resection; 66.00% patients with BD had comorbidities, including skin diseases (48.48%), cardiovascular diseases (39.39%), gastrointestinal diseases (30.30%), respiratory diseases (27.27%), and tumors (18.18%). The most commonly observed histopathological characteristics of BD are squamous-cell hyperplasia (86.00%), disordered maturation with atypical keratinocytes (74.00%), atypical mitoses (60.00%), hyperkeratosis with hypokeratosis (48.00%), dermal inflammatory cell infiltration (36.00%), and koilocytosis (22.00%). Conclusion: BD often occurs in middle-aged and elderly people and is easily misdiagnosed. The onset age and disease course of patients with BD are not significantly different between males and females, whereas there is a negative correlation between the onset age and disease course. BD is more likely to occur in trunk and limbs in the Chinese population, and most patients with BD are concurrent with comorbidities.

Cosmetic therapies for Chinese patients with acne taking concomitant or recent intake of oral isotretinoin: A retrospective study.

BACKGROUND: The current standard recommendation is to initiate the cosmetic therapies after discontinuing taking oral isotretinoin for at least 6 months. However, this recommendation has been questioned in several recent publications, and it is difficult to operate in clinical practice as early initiation of effective treatment is desirable for patients with acne sequelae. OBJECTIVE: The purpose of this study is to evaluate the efficacy and safety of chemical peeling and light/laser or radiofrequency treatments combined with oral isotretinoin for patients with acne vulgaris and acne scars. METHOD: A retrospective study of 511 patients on/or recently administered with isotretinoin treated with glycolic acid, intense pulsed light, nonablative fractional laser, fractional radiofrequency, and ablative carbon dioxide laser. A total of 1352 interventions were performed. The medical follow-up lasted for at least 1 year. The efficacy and safety of different procedures were evaluated. RESULTS: A total of 511 patients, who were treated with isotretinoin orally or stopped for <6 months, received 477 sessions of glycolic acid chemical peeling treatment, 588 sessions of intense pulsed light treatment, 61 sessions of nonablative fractional laser treatment, 101 sessions of fractional radiofrequency treatment, and 125 sessions of ablative fractional carbon dioxide laser treatment. No hypertrophic scars and keloids were found, and the incidence of serious adverse reactions such as scarring, erythema, blisters, and postinflammatory hyperpigmentation did not increase. CONCLUSIONS: It is safe to perform skin procedures in patients with acne and acne scars during or after discontinuation of isotretinoin for <6 months. Invasive treatments such as ablative fractional carbon dioxide laser treatment can be performed, as appropriate, by an experienced physician. The guideline of avoiding chemical and physical procedures in such patients taking oral isotretinoin should to revised.

National situation, trends, and predictions of disease burden of atopic dermatitis in Chinese children and adolescents.

Background: Atopic dermatitis (AD) is an important global health problem affecting children and adolescents and detailed national information of disease burden in China is lacking. We aimed to evaluate the national disease burden of AD in Chinese children and adolescent, to provide the temporal trends over the past 30 years and to predict the burden for the next 10 years. Methods: The data of AD in China, including incidence, prevalence, and DALY, and population data were obtained from the Global Burden of Disease Study 2019 (GBD study 2019), which were estimated using the DisMod-MR 2.1. We analyzed the three measures by age and sex; the age groups were <5 years, 5-9 years, 10-14 years, and 15-19 years. The joinpoint regression analyses was conducted to assess the temporal trends from 1990 to 2019. The Bayesian age-period cohort (BAPC) model was used to predict measures from 2020 to 2030. Results: In 2019, the highest incidence case and rate were observed in <5 years group; for prevalence and disability adjusted life year (DALY), the groups of <5 years and 5-9 years showed similar higher levels and the groups of 10-14 years and 15-19 years had similar relatively lower levels. Overall, the male-to-female ratios were >1 in <5 years group and <1 in 10-14 and 15-19 age groups. The trend analyses found an overall trend of decrease in cases of the three measures; in recent about 3 years, slight increase trends were shown in cases and rates of the three measures in <5 years group. The prediction analyses found a slight decreasing trend for cases of these measures and a slight increasing trend for rates of these measures in the <5 years group in the next 10 years; the 5-9 years group was predicted to increase slightly in rates of the three measures. Conclusion: In conclusion, the groups of <5 years and 5-9 years are two important populations that need targeted measures to reduce disease burden of AD in China. Regarding sex disparity, we should pay more attention to males in <5 years group and to females in 10-19 years group.

Screening DNA aptamers that control the DNA cleavage, homology-directed repair, and transcriptional regulation of the CRISPR-(d)Cas9 system.

Accurate genome editing based on various molecular tools has always been the focus of gene-editing research and the primary goal for therapeutic application. The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system is a well-established gene-editing method that is preferred due to its simplicity and high efficiency. In this study, a group of single-stranded DNA aptamers with high affinity and high specificity for the Cas9 protein were obtained by the systematic evolution of ligands through the exponential enrichment method. Their binding affinity and possible binding domains to the Cas9 protein were analyzed. In addition, we demonstrated the effectiveness of aptamers in regulating dCas9-modulated gene transcription, in terms of both transcriptional activation and repression. Additionally, the aptamers successfully reduced the off-target effect and improved the efficiency of gene homologous recombination repair mediated by CRISPR-Cas9. The findings suggest a potential method to better control precise gene editing and enrich the diversity of modulating tools for the CRISPR-Cas9 system.

Roles of AIM2 Gene and AIM2 Inflammasome in the Pathogenesis and Treatment of Psoriasis.

Psoriasis is an immune-mediated chronic inflammatory skin disease caused by a combination of environmental incentives, polygenic genetic control, and immune regulation. The inflammation-related gene absent in melanoma 2 (AIM2) was identified as a susceptibility gene for psoriasis. AIM2 inflammasome formed from the combination of AIM2, PYD-linked apoptosis-associated speck-like protein (ASC) and Caspase-1 promotes the maturation and release of inflammatory cytokines such as IL-1ß and IL-18, and triggers an inflammatory response. Studies showed the genetic and epigenetic associations between AIM2 gene and psoriasis. AIM2 gene has an essential role in the occurrence and development of psoriasis, and the inhibitors of AIM2 inflammasome will be new therapeutic targets for psoriasis. In this review, we summarized the roles of the AIM2 gene and AIM2 inflammasome in pathogenesis and treatment of psoriasis, hopefully providing a better understanding and new insight into the roles of AIM2 gene and AIM2 inflammasome in psoriasis.

Acitretin enhances the cytotoxic effect of 5-aminolevulinic acid photodynamic therapy on squamous cell carcinoma cells.

The efficacy of 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) on invasive cutaneous squamous cell carcinoma (cSCC) remains to be improved due to the limited penetration of this treatment. Previous study showed that acitretin and ALA-PDT had synergistic effect on cSCC, but whether acitretin can enhance the cytotoxic effect of ALA-PDT on cSCC is unclear. OBJECTIVE: To investigate whether acitretin can enhance the cytotoxic effect of ALA-PDT on SCL-1 cells, as well as the possible mechanism involved. METHODS: Inverted microscopy, trypan blue exclusion assay, and flow cytometry were used to studied the morphology, viability and apoptosis of SCL-1 cells treated with acitretin, ALA-PDT and acitretin followed by ALA-PDT treatment, respectively. Confocal microscopy was applied to detect the ROS formation of SCL-1 cells treated with acitretin of four different concentrations. The ROS formation of SCL-I cells treated with acitretin of four different concentrations followed by ALA-PDT treatment was detected using confocal microscopy and flow cytometry. RESULTS: SCL-1 cells exhibited a significant morphological alteration when treated with acitretin followed by ALA-PDT. The combination of acitretin and ALA-PDT induced a higher cell death rate and apoptosis than that with acitretin or ALA-PDT treatment alone. ROS could be induced when incubated with acitretin at a concentration of 6.4 × 10-4mg /mL or above. However, a higher level of ROS formation was observed when SCL-1 cells were treated with acitretin followed by ALA-PDT than that with ALA-PDT or acitretin alone. CONCLUSION: Acitretin can enhance the cytotoxic effect of ALA-PDT on SCL-1 cells, possibly via the ROS pathway.

Successful treatment of oral condyloma acuminata with photodynamic therapy: Two case reports.

Oral condyloma acuminatum (OCA) is a less prevalent subtype of condyloma acuminatum and is challenging to treat. We reported two cases of OCA successfully treated with 5-aminolevulinic acid photodynamic therapy (ALA-PDT). After three treatments spaced one week apart, both cases were completely cured without significant adverse reactions and were followed up for one year with no recurrence.

COVID-19 online teaching intervention and learning performance of college foreign language students.

This quasi-natural experimental study examined an online teaching intervention implemented in response to COVID-19 in China in 2020. It applied the difference-in-difference model to examine the impact and path of the intervention on students' learning performance of a college foreign language (LPCFL). Based on data from records of withdrawing and changing courses, classroom learning, and teaching evaluations; a questionnaire survey of teachers and students; and relevant school documents during the last seven terms, the results indicated that the online teaching intervention could significantly improve students' LPCFL. This finding remained robust after adopting a placebo test approach to mitigate possible endogeneity issues. Additionally, this study also conducted a group test through sub-sample regression based on students' discipline characteristics and intervention organization methods. The results showed that the students who participated in the intervention significantly improved in the three disciplines: humanities was most significantly affected, science and engineering were least significantly affected, and economics and management were in the middle. A range effect was observed for organizational methods. The two downward transmission methods by college teaching management terms had significant positive effects, whereas the other two methods of downward transmission by college student management had significant negative effects. An analysis of the action mechanism indicated that the online teaching intervention mostly improved LPCFL through two channels: students' learning input and learning support. Overall, these findings not only help expand the research framework on macro environmental intervention policy and micro-learning behavior but also have implications for the in-depth understanding of the real learning effect of online learning interventions for college students and their design in the post-COVID-19 era.

Long noncoding RNA ALOX12-AS1 inhibits cervical cancer cells proliferation via targeting miR-3171.

Cervical cancer is a common female malignancy worldwide, and the molecular mechanism of cervical tumorigenesis remains poorly understood. A large piece of evidence have demonstrated the important roles of long noncoding RNAs (lncRNAs) in tumorigenesis, cancer progression and drug resistance. In this study, we comprehensively analyzed the lncRNAs expression pattern in cervical cancer using RNA sequencing and microarray data from the cancer genome atlas, gene expression omnibus and Genotype Tissue Expression. Moreover, we assessed the correlation between lncRNA expression levels and cervical cancer patient's survival. We uncovered hundreds of lncRNAs that are upregulated or downregulated in cervical cancer tissues. Among these aberrantly lncRNAs, some are significantly associated with cervical patients' poorer prognosis, such as ALOX12-AS1 and LINC00173. ALOX12-AS1 expression is downregulated in cervical cancer, and over-expression of ALOX12-AS1 could inhibit cervical cancer cells proliferation in vitro. Further, mechanistically investigation revealed that ALOX12-AS1 could interact with AGO2 and sponge miR-3171, thereby antagonizing its' repression of tumor suppressor phosphatase and tensin homolog expression in cervical cancer cell. Taken together, this study provides lncRNA candidates in cervical cancer and highlights the critical role of ALOX12-AS1 in cervical cancer.

PUM1 modulates trophoblast cell proliferation and migration through LRP6.

Preeclampsia is a severe pregnancy complication characterized by hypertension and may cause maternal morbidity and mortality. A better understanding of the essential genes involved in preeclampsia pathophysiology is urgently needed. This study investigated the function and molecular mechanisms of pumilio RNA binding family member 1 (PUM1) in extravillous trophoblast cells (EVTs). The interaction between protein and mRNA was verified by RNA pull-down assays, RNA immunoprecipitation assays, and luciferase reporter assays. The mRNA and protein levels of the genes involved were determined by RT-qPCR and western blot assays, respectively. Our results demonstrated that PUM1 could bind to the 3'-untranslated region of low-density lipoprotein receptor-related protein 6 (LRP6) mRNA, resulting in reduced expression of LRP6 mRNA and protein. Repression of PUM1 resulted in enhanced colony formation, cell proliferation, migration, and invasion of EVTs. The PUM1-depletion-mediated promotion effects on EVTs could be abrogated by LRP6 knockdown. PUM1 regulates the growth and mobility of EVTs by modulating LRP6 expression. Developing strategies to balance PUM1 and LRP6 levels may be beneficial for the management of preeclampsia patients.

Cosmc transfection decreases malignant behavior of Tn+ cells and enhances sensitivity to apoptosis when induced by Apo2L/TRAIL via alteration of O-glycan structure.

Cosmc mutations may cause abnormal O-glycosylation and result in Tn antigen expression. In the current study, it was discovered that proliferation and migration of Tn+ cells (Jurkat T and LS174T-Tn+ cells) with mutant Cosmc decreased after transfected Cosmc, and their sensitivity to apoptosis induced by Apo2L/TRAIL increased. Core 1-, 2-, and 3-derived O-glycans were absent in Tn+ cells. After Cosmc transfection, normal extended core 1-derived O-glycans appeared and were accompanied by increased T-synthase activity. Core 2-derived O-glycans appeared in transfected LS174T-Tn+ cells, and their structural types and levels were lower than those in LS174T-Tn- cells. Core 3-derived O-glycans were present only in LS174T-Tn- cells. The activity of C3GnT in LS174T-Tn+ cells was lower than that in LS174T-Tn- cells, and it was absent in Jurkat T cells. Cosmc transfection did not alter C3GnT activity or core 3-derived O-glycans in Jurkat T and LS174T-Tn+ cells. The results demonstrated that the composition and structure of O-glycans were different among various Tn+ cells, which not only affected cell malignant behavior but also modulated sensitivity to apoptotic stimuli. Thus, Cosmc transfection may effectively decrease the malignant behavior of Tn+ tumor cells and enhance their sensitivity to apoptosis when induced by Apo2L/TRAIL through modification of O-glycans.

The Fabrication of a Gellan Gum-Based Hydrogel Loaded With Magnesium Ions for the Synergistic Promotion of Skin Wound Healing.

To accelerate serious skin burn wound healing in a convenient manner, an interpenetrating network of hydrogel consisting of gellan gum and polyacrylamide was synthesized by chemical crosslinking and Mg2+ ion immersion techniques. The prepared Mg2+@PAM/GG hydrogel was characterized by morphology, water vapor loss, swelling ratio, rheological properties, tensile mechanical, biocompatibility, and flow cytometry study. The results show that Mg2+@PAM/GG hydrogel's mechanical strength could be enhanced by the dual network structure and physical crosslinking agent Mg2+ ions. In addition, the tension strength of Mg2+@PAM/GG hydrogel is obviously increased from 86 to 392 kPa, the elongation at break increased from 84 to 231%, and crosslinking density N increased from 4.3 to 7.2 mol/m3 compared with pure GG hydrogel. The cumulative release curve of Mg2+ ions shows that the multiple release mechanism of Mg2+ ions belong to non-Fick's diffusion. Meanwhile, in vitro experiments show that Mg2+@PAM/GG double network hydrogel has favorable proliferation and an NF-κB pathway inhibition property for fibroblast cells. Finally, the healing effect of the Mg2+@PAM/GG was evaluated in a rat full-thickness burn model. The animal study demonstrates that Mg2+@PAM/GG could accelerate the healing efficiency in case of the sustained-released Mg2+ ions in wound beds. Considering this excellent performance, this convenient prepared hydrogel has great potential as a commercial application for skin full-thickness burn healing materials.

Silencing of AFAP1-AS1 lncRNA impairs cell proliferation and migration by epigenetically promoting DUSP5 expression in pre-eclampsia.

As a unique and common obstetric complication of pregnant women, pre-eclampsia (PE) has been the first leading cause of maternal and perinatal morbidity and mortality in the world. Mounting studies have demonstrated that an abnormality of long noncoding RNA (lncRNA) expression was related to the pathological process of PE. Here, we showed that lncRNA AFAP1-AS1 was markedly downregulated in pre-eclamptic placentas. We further investigated the mechanism underlying the regulatory role of AFAP1-AS1 in PE using human trophoblast cells. In vitro functional assays revealed that AFAP1-AS1 knockdown inhibited trophoblast proliferation, migration, and invasion. Moreover, AFAP1-AS1 interacts with EZH2 and inhibits DUSP5 expression through modulating H3K27m3 in the DUSP5 promoter of trophoblast cells, thus being involved in PE pathogenesis. Overall, these findings suggest that AFAP1-AS1 could potentially become a prognostic biomarker as well as a new therapeutic target for PE.

Upregulated long noncoding RNAs LINC02163 and FEZF1-AS1 exert oncogenic roles in colorectal cancer.

A growing number of evidence has revealed that aberrantly expressed long noncoding RNAs (lncRNAs) are involved in the development of a variety of malignancies, including colorectal cancer (CRC). However, the clinical relevance of most lncRNAs and their potential biological functions in CRC remains poorly understood. The aim of this study was to identify the key lncRNAs related to patient prognosis as well as their biological function and underlying mechanism in CRC. Therefore, five independent datasets containing CRC and normal tissue RNA sequencing, microarray data and the corresponding clinical data from The Cancer Genome Atlas and Gene Expression Omnibus were screened. Hundreds of significantly differentially expressed lncRNAs in CRC were determined, and Kaplan-Meier analyses revealed that some of these lncRNAs were related to the overall survival and progression-free survival of patients with CRC, such as RP11-108K3.2, FOXD3-AS1, H19 and AP001469.9. Among these dysregulated lncRNAs, LINC02163 and FEZF1-AS1 were significantly upregulated in CRC tissues, suggesting that they may have oncogenic roles in CRC. Furthermore, loss of function assays revealed that downregulation of LINC02163 and FEZF1-AS1 impaired CRC cell proliferation. In addition, RNA Immunoprecipitation and Chromatin Immunoprecipitation assays determined that FEZF1-AS1 regulates CRC cell growth via interacting with LSD1 and repressing KLF2 expression. Collectively, hundreds of dysregulated lncRNAs and their associated biological roles identified in this study may provide potentially useful biomarkers and therapeutic targets for CRC.

5-aminolevulinic acid photodynamic therapy inhibits invasion and metastasis of SCL-1 cells probably via MTSS1 and p63 gene related pathways.

OBJECTIVE: To investigate the effect of 5-aminolevulinic acid (ALA) mediated photodynamic therapy (PDT) on the invasion and metastasis in cutaneous squamous cell carcinoma (cSCC) cell line(SCL-1) and to study whether the effect was via the MTSS1 gene and p63 gene related pathways. METHODS: SCL-1 cells were cultured and submitted to ALA-PDT treatment (ALA-PDT group), ALA treatment alone (ALA group), LED illumination alone (LED group) and remains untreated (control group). Scratch test, Transwell migration chamber assay and Matrigel cell invasion assay were used to detect the ability of migration and invasion of SCL-1 cells after treatment. The mRNA levels and protein expressions of tumor metastasis suppressor gene (MTSS1) and p63 gene were further detected by using quantitative real-time PCR and flow cytometry assay respectively after treatment. RESULTS: The migration and invasion abilities of SCL-1 cells after treatment were significantly reduced in the ALA-PDT groups than that in ALA group, LED group and control group (P＜0.05). Both the mRNA and protein expression levels of MTSS1 gene were up-regulated, while the mRNA and protein expression levels of p63 gene were down-regulated after ALA-PDT treatment. CONCLUSION: ALA-PDT suppressed the migration and invasion of human cSCC cell line, probably via the MTSS1 gene and p63 gene related pathways. This study put forward a possible mechanism of invasion in SCL-1 cell, also providing a potential target for the therapy of cSCC.

lncRNA GHET1 has effects in development of pre-eclampsia.

To explain long noncoding RNA (lncRNA) gastric carcinoma high expressed transcript 1 (GHET1) affects the mechanism in development of pre-eclampsia. The pathological changes of normal, mild, and severe pre-eclampsia were evaluated by hematoxylin and eosin staining and measured the lncRNA GHET1 expression in different tissues by reverse-transcription polymerase chain reaction. In the cell experiment, the BeWo cells were randomly divided into three groups: normal control (NC) group, model group, and lncRNA group. The JEG3 cells of the model and lncRNA groups were cultured in the hypoxia condition. The JEG3 cells invasion and migration abilities were measured by Tanswell and wound-healing assays. The relative protein expressions of different groups were evaluated by Western blot (WB) assay. Compared with normal puerperal, the lncRNA GHET1 gene expression of pre-eclampsia was significantly downregulated (P < 0.05, respectively). In the cell experiment, the invasion cell number and wound-healing rate of the model group were significantly suppressed compared with the NC group (P < 0.05, respectively). However, the invasion cell number and wound-healing rate of lncRNA group were enhanced by lncRNA GHET1 overexpression (P < 0.05, respectively). In WB assay, the E-cadherin, fibronectin, and vimentin proteins expression showed significant differences between the model and lncRNA groups (P < 0.05, respectively). lncRNA GHET1 overexpression had restored cell invasion and migration abilities reduced by hypoxia in pre-eclampsia.

Resveratrol promotes trophoblast invasion in pre-eclampsia by inducing epithelial-mesenchymal transition.

Impairment spiral arteries remodelling was considered to be the underlying cause of pathogenesis of pre-eclampsia (PE). Resveratrol (RE) was reported that it could modulate cellar phenotype to ameliorate diverse human diseases. However, the biological function of RE in PE remains poorly understood. In this report, we investigated the effect of RE on trophoblast phenotype both in vivo and in vitro. We conducted MTT and transwell assays to explore cell proliferation and invasion events in HTR-8/SVneo. In mice model, the clinical characteristics of PE were established through the injection of NG-nitro-l-arginine methyl ester (L-NAME). Furthermore, related experiments were performed to detect cellar phenotype-associated signalling pathway, including epithelial-mesenchymal transition (EMT) and Wnt/ß-catenin. Cell assays indicated that RE could increase trophoblasts migration and invasion. In addition, hypertension and proteinuria were markedly ameliorated by RE compared with the controls in PE mice model. Moreover, treatment by RE in trophoblasts or in PE model, we found that RE activated EMT progress through the regulation of E-cadherin, ß-catenin, N-cadherin, vimentin expression, and further altered the WNT-related gene expression, including WNT1, WNT3 and WNT5B. Our findings demonstrated that RE might stimulate the invasive capability of human trophoblasts by promoting EMT and mediating the Wnt/ß-catenin pathway in PE.