In vitro study of cold atmospheric plasma-activated liquids inhibits malignant melanoma by affecting macrophage polarization through the ROS/JAK2/STAT1 pathway.

Melanoma is a prevalent malignant skin tumor known for its high invasive ability and a high rate of metastasis, making clinical treatment exceptionally challenging. Tumor-associated macrophages (TAMs) are the most abundant immune cells in the tumor microenvironment and play a crucial role in tumor survival and development. Cold atmospheric plasma (CAP) is an emerging tool for tumor treatment that has garnered attention from scholars due to its interaction with non-tumor cells in the tumor microenvironment. Here, we used the macrophage lines THP-1 and RAW264.7, as well as the melanoma cell lines A375 and MV3, as research subjects to investigate the effect of plasma-activated liquid (PAL) on macrophage differentiation and its inhibitory effect on melanoma cell proliferation. We confirmed that the killing effect of PAL on melanoma cells was selective. Using flow cytometry and PCR, we discovered that PAL can influence macrophage differentiation. Through in vitro cell coculture, we demonstrated that PAL-treated macrophages can significantly impede tumor cell development and progression, and the effect is more potent than that of PAL directly targeting tumor cells. Furthermore, we have proposed the hypothesis that PAL promotes the differentiation of macrophages into the M1 type through the ROS/JAK2/STAT1 pathway. To test the hypothesis, we employed catalase and fludarabine to block different sites of the pathway. The results were then validated through Western Blot, qPCR and ELISA. This study illustrates that PAL therapy is an effective tumor immunotherapy and expands the scope of tumor immunotherapy. Furthermore, these findings establish a theoretical foundation for potential clinical applications of PAL.

A multicenter, randomized, double-blinded, placebo-controlled, phase â ¢ study evaluating the efficacy and safety of Xeligekimab (GR1501) in patients with moderate-to-severe plaque psoriasis.

BACKGROUND: Xeligekimab is a fully human monoclonal antibody that selectively neutralizes IL-17A and had shown potential efficacy in preliminary trials. OBJECTIVE: To evaluate the efficacy and safety of Xeligekimab in Chinese patients with moderate-to-severe psoriasis. METHODS: A total of 420 Chinese patients were randomized to 200 mg Xeligekimab every 2 weeks (n = 281) or placebo (n = 139) for the first 12 weeks, followed by extending the treatment schedule to GR1501 every 4 weeks for further 40 weeks. Efficacy was assessed by evaluating the Physician's Global Assessment (PGA) 0/1 and Psoriasis Area and Severity Index (PASI) 75/90/100 improvement. The safety profile was also evaluated. RESULTS: At week 12, The PASI 75/90/100 were achieved in 90.7%/74.4%/30.2%% patients in GR1501 group compared with 8.6%/1.4%/0% patients in placebo group, respectively. The PGA 0/1 were achieved in 74.4% patients of GR1501 group and 3.6% patients in placebo group, respectively. The PASI 75 and PGA 0/1 maintained until week 52. No unexpected adverse events were observed. CONCLUSION: Xeligekimab showed high efficacy and is well tolerated in Chinese patients with moderate-to-severe plaque psoriasis.

Toxicity study of rats treated by plasma-activated solution.

Background: Cold atmospheric plasma (CAP) is an effective treatment for various skin diseases. Plasma-activated solution (PAS) is an indirect method of CAP treatment that produces biological effects similar to those of direct treatment with plasma devices. The anticancer and bacteriostatic effects of PAS have been demonstrated in vitro experiments; however, on the basis of the lack of toxicological studies on PAS, its effects on living mammals when administered by subcutaneous injection is poorly known. Purpose: The purpose of this study was to evaluate the effects of PAS on local skin tissue cells, blood system, heart, liver, lungs, kidneys and other vital organs of the rat when injected subcutaneously. Methods: PAS was prepared by CAP irradiation of phosphate-buffered saline (PBS). PBS and different PBS groups (CAP irradiation for 1, 3, or 5 min) were injected subcutaneously once every 48 h. The rats were euthanized immediately after 10 cycles of therapy. Results: No adverse effects were observed during the entire period of the experiment. Histopathological examination of organs and tissues revealed no structural changes. Moreover, no obvious structural changes were observed in skin tissue. DNA damage and cancerous proliferative changes were not detected in skin tissue treated with PAS. Subsequently, RNA sequencing and western blotting were performed. The results showed that PAS increased the expression of growth factors like transforming growth factor beta (TGF-ß) and vascular endothelial growth factor A (VEGFA). These results might be directly linked to the role of PAS in stimulating TGF-ß receptor signaling pathway and angiogenesis. Conclusion: The results showed that multiple subcutaneous injections of PAS did not show significant toxic side effects on local skin tissues and some vital organs in rats, providing a scientific basis to support the future treatment of skin diseases with PAS.

Basic research and clinical exploration of cold atmospheric plasma for skin wounds.

Skin wounds, such as burns, diabetic foot ulcers, pressure sores, and wounds formed after laser or surgical treatment, comprise a very high proportion of dermatological disorders. Wounds are treated in a variety of ways; however, some wounds are greatly resistant, resulting in delayed healing and an urgent need to introduce new alternatives. Our previous studies have shown that cold atmospheric plasma (CAP) has antibacterial activity and promotes cell proliferation, differentiation, and migration in vitro. To further advance the role of CAP in wound healing, we evaluated the safety and efficacy of CAP in vitro by irradiation of common refractory bacteria on the skin, irradiation of normal skin of rats and observing reactions, treatment of scald wounds in rats, and treating clinically common acute wounds. Our findings revealed that CAP can eliminate refractory skin bacteria in vitro; CAP positively affected wound healing in a rat scalding wound model; and direct CAP irradiation of low intensity and short duration did not lead to skin erythema or edema. CAP promises to be a new, economical, and safe means of wound treatment.

Plasma activated Ezhangfeng Cuji as innovative antifungal agent and its inactivation mechanism.

Candida albicans is a highly drug-resistant fungus for which new treatments are urgently needed due to the lack of clinically effective options. In this study, we evaluated the antifungal activity and mechanism of plasma-activated Ezhangfeng Cuji (PAEC) against Candida albicans and compared it with physiological saline (PS), plasma-activated physiological saline (PAPS) and Ezhangfeng Cuji (EC). After dielectric barrier discharge (DBD) plasma treatment with EC for 20 min followed by a 10 min immersion of Candida albicans, the fungus was reduced by approximately 3 orders of magnitude. High performance liquid chromatography (HPLC) results showed an increase of 41.18% and 129.88% in the concentration of oxymatrine and rhein, respectively, after plasma-treated EC. The concentrations of reactive species (RS), such as H2O2, [Formula: see text], and O3, were found to be higher and the pH value was getting lower in PS after plasma treatment. Detailed analysis of intracellular material leakage, reactive oxygen species (ROS), apoptosis for Candida albicans and observation by transmission electron microscopy (TEM) and scanning electron microscopy (SEM) demonstrated that PAPS, EC and PAEC disrupt the morphological structure of Candida albicans to varying degrees.Additionally, specific analyses on Candida albicans virulence factors, such as adhesion to tissue surfaces, cell surface hydrophobicity (CSH), the transition of yeast-phase cells to mycelium-phase cells, and the secretion of hydrolytic enzymes for Candida albicans were conducted and found to be inhibited after PAPS/EC/PAEC treatment. In our investigation, the inhibitory effects on Candida albicans were ranked from strong to weak as follows: PAEC, EC, PAPS, and PS.

Application of improved high-risk sign board for older people inpatients: a retrospective study.

AIM: To observe the effect of the implementation of improved high-risk sign boards for older people inpatients. METHOD: The older people inpatients of the Department of Geriatrics at our hospital were selected as the research subjects and divided into two groups. The control group used the single-strip high-risk sign, and the intervention group used the improved double-layer card slot, push-pull integrated high-risk sign board (national patent). The sign-related nurse operation time, patient/attendant satisfaction, and high-risk-related adverse events were observed and compared between the two groups. RESULTS: After the adoption of the improved high-risk sign board, the nurse operation time was reduced from 94.3 ± 16.2 s to 53.9 ± 12.5 s, and patient/attendant satisfaction increased from 6.65 ± 0.38 points to 9.30 ± 0.52 points (P < 0.001). The incidence of high-risk-related adverse events decreased from 6.08 to 1.86%, but the difference was not statistically significant (χ2 = 3.675, P = 0.055). The implementation of the improved high-risk sign board can increase nursing efficiency and enhance the awareness of risk prevention in high-risk patients among nurses, older people inpatients, and attendants. CONCLUSION: The application of double-layer card slot and push-pull comprehensive high-risk identification card to older people inpatients can alert nurses, patients, and nursing staff more prominently, which can improve patient satisfaction, reduce installation time and reduce the incidence of adverse events to a certain extent.

Research on risk factors of ischemic cerebrovascular disease in postmenopausal women based on the social-ecological model.

OBJECTIVE: Based on the social-ecological model, this study aimed to comprehensively explore factors affecting the risk of ischemic cerebrovascular disease (ICVD) in postmenopausal women to provide theoretical bases for further prevention and intervention for postmenopausal women. METHODS: Postmenopausal women who underwent medical examinations in one health-checkup agency in Tianjin from May 2015 to October 2015 were enrolled in this study. The ICVD 10-year Risk Assesment Form developed by the research team of the National "Tenth Five-Year Plan" research project was used to assess the factors affecting the risk of ICVD. Based on the social-ecological model, multiple types of scales, including physical activities, depression, Type D personality, social supports, and environment score, were used to comprehensively explore the factors associated with ICVD in postmenopausal women. RESULTS: 300 valid questionnaires were obtained, with an effective rate of 92.0%. The subjects aged 44-74 years, with the average age of 62.06 ± 7.09 years. Among them, 58.67% of the subjects only obtained high-school diploma, 32.67% obtained college or university diploma, 90.33% were retirees, 95.33% were married, 92.33% experienced the natural menopause, 93.33% lived in urban or suburban areas, and 1.00% had a history of breast cancer. Multivariate logistic regression analysis suggested that monthly income (¥), parity, exposure to second-hand or third-hand smoke, easy access to healthy food, physical activities, depression, Type D personality, social support and environmental factors were associated with the risk of ICVD in postmenopausal women (P < 0.05). Among them, easy access to healthy food (OR = 0.242), social support (OR = 0.861) and environmental factors (OR = 0.866) were protective factors from ICVD. OR < 1 indicates that the exposure factor is negatively correlated with the disease, and the exposure factor has a protective effect on preventing the occurrence of the disease. Parity (OR = 3.795), exposure to second-hand or third-hand smoke (OR = 2.886), depression (OR = 1.193), and Type D personality (OR = 1.148) were risk factors of ICVD. OR > 1 means that the exposure factor is positively correlated with the disease, and the exposure factor increases the risk of disease occurrence. CONCLUSIONS: For postmenopausal women, in the future, in addition to prevention and management of the conventional risks, the conditions of their mentality and social support should be paid attention to, at the same time, and if they can, try to choose a good community environment to live in, which could better reduce the incidence and mortality of ICVD in postmenopausal women.

Membrane Attack Complex C5b-9 Promotes Renal Tubular Epithelial Cell Pyroptosis in Trichloroethylene-Sensitized Mice.

Trichloroethylene (TCE), a commonly used organic solvent, is known to cause trichloroethylene hypersensitivity syndrome (THS), also called occupational medicamentosa-like dermatitis due to TCE (OMDT) in China. OMDT patients presented with severe inflammatory kidney damage, and we have previously shown that the renal damage is related to the terminal complement complex C5b-9. Here, we sought to determine whether C5b-9 participated in TCE-induced immune kidney injury by promoting pyroptosis, a new form of programed cell death linked to inflammatory response, with underlying molecular mechanisms involving the NLRP3 inflammasome. A BALB/c mouse-based model of OMDT was established by dermal TCE sensitization in the presence or absence of C5b-9 inhibitor (sCD59-Cys, 25µg/mouse) and NLRP3 antagonist (MCC950, 10 mg/kg). Kidney histopathology, renal function, expression of inflammatory mediators and the pyroptosis executive protein gasdermin D (GSDMD), and the activation of pyroptosis canonical NLRP3/caspase-1 pathway were examined in the mouse model. Renal tubular damage was observed in TCE-sensitized mice. GSDMD was mainly expressed on renal tubular epithelial cells (RTECs). The caspase-1-dependent canonical pathway of pyroptosis was activated in TCE-induced renal damage. Pharmacological inhibition of C5b-9 could restrain the caspase-1-dependent canonical pathway and rescued the renal tubular damage. Taken together, our results demonstrated that complement C5b-9 plays a central role in TCE-induced immune kidney damage, and the underlying mechanisms involve NLRP3-mediated pyroptosis.

Evaluation of Susceptibility Genes/Loci Associated with Male Androgenetic Alopecia (MAGA) for Female-Pattern Hair Loss in a Chinese Han Population and a Brief Literature Review.

BACKGROUND Female-pattern hair loss (FPHL) is a common disorder affecting women, and FPHL can cause psychological dysfunction and affect the social activities of patients. The disease-causing mechanisms are believed to be similar to those of male androgenetic alopecia (MAGA). Although genome-wide association studies (GWAS) have confirmed susceptibility genes/loci for MAGA, the associations between these genetic loci and FPHL are largely unknown. We investigated the associations between susceptibility loci for MAGA and FPHL in a Chinese Han population; a literature review of susceptibility loci associated with MAGA for FPHL was also performed. MATERIAL AND METHODS Twenty-two previously reported sites were analyzed with the Sequenom iPlex platform, and the genotype statistical analysis consisted of a trend test and conservative accounting. The samples comprised 82 patients diagnosed with FPHL by dermatoscopy and 381 healthy controls from the Chinese Han population. RESULTS No significantly associated variants were found in this FPHL study. The examined 22 tag SNPs in MAGA may not be associated with FPHL. The results of the current study in a Chinese Han population support the previous negative association obtained for a European population. CONCLUSIONS This was the first study exploring whether identified MAGA-associated loci confer susceptibility to FPHL in a Chinese Han population, and dermatoscopy was used to improve the diagnostic accuracy. However, there was no evidence of a relationship between susceptibility genes for MAGA and FPHL, and the results indicated that FPHL and MAGA are etiologically separate entities. Therefore, a systematic GWAS approach to FPHL may be required to clarify associated pathophysiological uncertainties.

Dermoscopic Features Summarization and Comparison of Four Types of Cutaneous Vascular Anomalies.

Objective: Dermoscopic features of cutaneous vascular anomalies have been reported, but the described features currently known are limited and not well-understood. The aim of this study is to comprehensively summarize and compare the dermoscopic features of the four different types of cutaneous vascular anomalies [infantile hemangiomas (IH), cherry angioma (CA), angiokeratomas (AK), and pyogenic granuloma (PG)] in the Chinese Han population. Materials and Methods: Dermoscopic features of 31 IH, 172 CA, 31 AK, and 45 PG were collected based on the contact non-polarized mode of dermoscopy at 20-fold magnification. Dermoscopic features including background, lacunae, vessel morphology and distribution were collected and summarized. Additionally, we compared these features by age stage, gender, and anatomical locations in CA. Results: The dermoscopic features of IH included the red lacunae, red/red-blue/red-white backgrounds, and vessel morphology such as linear curved vessels, serpiginous vessels, coiled vessels. For CA, the lacunae appeared reddish brown to reddish blue or only red. In terms of vascular morphology, serpentine vessels, coiled vessels, looped vessels, and curved vessels could be seen in the lesions. A few lesions were black or presented with a superficial white veil. There were statistical differences in red background (P = 0.021), unspecific vessel distribution (P = 0.030), black area (P = 0.029), and white surface (P = 0.042) among different age groups. Red-brown lacunae (P = 0.039), red-blue (P = 0.013), red-white background (P = 0.015), black area (P = 0.016), and white surface (P = 0.046) were of statistical difference in terms of the locations of lesions. Lacunae were also observed in AK, which presented with red, dark purple, dark blue, black. Global dermoscopic patterns that were characterized by a homogeneous area were obvious in all PG lesions, among which 30 (66.7%) were red-white and 15 (33.3%) were red. As for local features, "white rail" lines were detected in 19 (42.2%) lesions and white collarette was seen in 34 (75.6%) lesions. Conclusions: Dermoscopy is an applicable diagnostic tool for the diagnosis of cutaneous vascular anomalies. It is necessary to take into account the age stage and lesion location when we diagnose CA using dermoscopy.

In vitro antimicrobial effect and mechanism of action of plasma-activated liquid on planktonic Neisseria gonorrhoeae.

Neisseria gonorrhoeae (Ng) is highly resistant to treatment, and there is an urgent need for new treatments to alleviate gonococcal resistance caused by antibiotic monotherapy. The antimicrobial effect and mechanism of plasma-activated liquid (PAL) on Ng were evaluated in this study. Upon PAL treatment, extensively analyses on cell culturability, metabolic capacity, intracellular reactive oxygen species (ROS),membrane integrity and nucleic acids for Ng were carried out and significant antimicrobial effects observed.PAL exerted antibacterial effect on Ng and induced bacterial death (6.71-log) following immersion for 30 min and treatment for 120 s. However, bacterial viability test revealed that after immersion in the same PAL, 10.17% of bacteria retained their metabolic capacity. This indicates that bacteria enter a physiological viable but non-culturable state to protect themselves from environmental stress. Confocal fluorescence microscopy and transmission electron microscopy demonstrated that PAL exerts bactericidal effect on Ng and disrupts its morphological structure. PAL may upregulate inflammatory factors and genes to modulate the resistance of Ng and affect the immune status of the host during infection.

Erratum: MIST1 regulates SNAI1 and acts through the PTEN/AKT signaling axis to promote anoikis resistance in human melanoma cells.

[This corrects the article DOI: 10.3892/etm.2018.6225.].

Identification of key genes and pathways for melanoma in the TRIM family.

Certain members of the TRIM family have been shown to have abnormal expression and prognostic value in cancer. However, in the development and progression of melanoma, the role of different TRIM family members remains unknown. To address this issue, this study used the Oncomine, UCSC, Human Protein Atlas, DAVID, and GEPIA databases to study the role of TRIMs in the prognosis of melanoma. Differential expression of TRIM2, TRIM7, TRIM8, TRIM18 (MID1), TRIM19 (PML), TRIM27, and TRIM29 may play an important role in the development of melanoma. The expression TRIM7 and TRIM29 appeared to be helpful in the identification of primary tumors and metastases. Survival analysis suggested that the expression of TRIM27 significantly affected the overall survival and disease-free survival of melanoma, and its expression was confirmed by qRT-PCR. Our results indicated that the expression level of TRIM27 might be a prognostic marker of melanoma.

Inhibition of basal cell carcinoma cells by cold atmospheric plasma­activated solution and differential gene expression analysis.

Basal cell carcinoma is a common skin tumor. Cold atmospheric plasma (CAP) has attracted increasing attention for its antitumor effects. The aim of the present study was to investigate the effects and related mechanisms of two CAP­activated solutions on the TE354T basal cell carcinoma and HaCat keratinocyte cell lines. Plasma­activated solution (PAS) was prepared by CAP irradiation of DMEM and PBS. TE354T cells were treated with PAS in vitro and the effect on cell viability was evaluated by an MTT assay. The apoptosis rate was detected by Annexin V/propidium iodide staining. Furthermore, western blotting and RNA­sequencing were performed. The present results demonstrated that PAS induced apoptotic signaling in basal cell carcinoma cells, and that this effect was associated with the activation of the MAPK signaling pathway. Therefore, the present study demonstrated that PAS may serve as a novel treatment for basal cell carcinoma.

Cold atmospheric plasma­activated Ringer's solution inhibits the proliferation of osteosarcoma cells through the mitochondrial apoptosis pathway.

The present study aimed to investigate the effects of cold atmospheric plasma (CAP)­activated Ringer's solution on osteosarcoma cell lines MG63 and U2OS, and to identify the molecular mechanism underlying these effects. CAP­activated Ringer's solution was used to treat osteosarcoma cell lines MG63 and U2OS for 30 min. Cell viability was measured using the MTT method. The apoptosis rate was detected using Annexin­V and propidium iodide. The expression levels of cytochrome c, caspase­3 and polyADP ribose polymerase (PARP) in MG63 cells were analyzed via western blotting. The change in mitochondrial membrane potential was detected via the JC­1 dye method and verified by the level of reactive oxygen species (ROS). CAP­activated Ringer's solution inhibited the proliferation of MG63 and U2OS cells in a dose­ and time­dependent manner. Furthermore, CAP­activated Ringer's solution induced the apoptosis of MG63 cells, increased the intracellular ROS level, decreased the mitochondrial membrane potential level, and induced the release of cytochrome c. CAP­activated Ringer's solution inhibits osteosarcoma cell proliferation through intracellular ROS­mediated mitochondrial apoptosis.

Clinical Observation and Follow-Up of Subungual Splinter Hemorrhages of Toenails in Chinese Adults.

BACKGROUND: Subungual splinter hemorrhage (SSH) is a common nail disorder and is difficult to differentiate from other nail diseases because of their similar characteristics. The epidemiological study of SSH is lacking and it is unclear whether SSH needs treatment or not. AIMS: The aims of this study were to observe the clinical characteristics of SSH of toenails in Chinese adults and to provide a reference for the diagnosis and prognosis of patients with SSH. MATERIALS AND METHODS: We collected and followed up 63 SSH patients who were diagnosed through dermoscopic examination. The clinical and dermoscopic characteristics, disease course, and recovery results were recorded. Data sets were analyzed through the use of SPSS 16.0. Literature concerning SSH were reviewed and compared with the results in this study. RESULTS: Sixty-three SSH patients were included with an age range of 17-58 years. Nearly 93.7% of SSH patients occurred in the right, left, or right and left first toenails; 66.7% of SSH patients occurred only in one toenail; there were 60.3% of SSH patients with a clear stimulus causing it and 31.8% of SSH patients with five types of comorbidities. The common characteristics of dermoscopy showed a reddish brown or brown stain with a clear boundary visible under the nail in which the color gradually faded outward from the center, with no blue or white structure. After following up for 24 weeks, the disappearance of hemorrhage was observed in 95.2% of SSH patients. For the recovered SSH patients, data analysis showed no significant difference in the course of SSH in males and females (P = 0.645); the statistical analysis showed that there was no significant correlation (r 2 = -0.002) between age and course in recovered SSH patients (P = 0.986). CONCLUSIONS: It is unnecessary to do any special treatment during the 1st year. Measures ought to be taken for SSH patients if the course of the disease exceeds more than 1 year.

Bactericidal efficacy of cold atmospheric plasma treatment against multidrug-resistant Pseudomonas aeruginosa.

Aim: The global of spread multidrug-resistant Pseudomonas aeruginosa has become a public health threat. Cold atmospheric plasma (CAP) is reported to have bactericidal efficacy; however, its effects on clinical super multidrug-resistant P. aeruginosa are unclear. The aim of this study was to investigate the bactericidal efficacy of CAP on a strain of super multidrug-resistant P. aeruginosa. Materials & methods: The effects of CAP treatments were evaluated using assays for the detection of growth, viability, metabolism, virulence factors and reactive oxygen species (ROS) levels. Results: Both CAP treatments dose-dependently inhibited cell viability and metabolic activity, and decreased the expression of several virulence factors. CAP treatment induced a significant increase in intracellular ROS levels, and ROS scavengers inhibited this effect. Conclusion: CAP treatment is a promising option for the clinical inhibition of multidrug-resistant P. aeruginosa, and the development of CAP technologies might be the key to solving the long-standing problem of drug-resistant bacteria.

MiRNA-27a decreases ultraviolet B irradiation-induced cell damage.

MiRNAs were involved in the various biological process through mediating the posttranscriptional gene silencing. The abnormal expression of miRNAs is also involved in various disorders. Our previous study showed that miRNA-27a (miR-27a) was upregulated after ultraviolet B (UVB) irradiation. However, the function of miR-27a in UVB-induced cell damage is still unclear. In this study, we used the miR-27a overexpression and knockdown lentivirus to transfect UVB irradiated HaCaT cell line and observed the influence of miR-27a on UVB irradiated damages in cells. We found that miR-27a removed cyclobutane pyrimidine dimers (CPDs) and decreased the cell apoptosis after UVB radiation. Further studies showed that miR-27a directly decreased the expression and luciferase activity of target genes transactive response DNA-binding protein (TARDBP) and apoptotic protease activating factor-1 (APAF-1). In conclusion, miR-27a can inhibit CPDs, reduce the cell apoptosis and down-regulate its target genes TARDBP and APAF-1 induced by UVB irradiation in HaCaT cells. It is indicated that miR-27a may serve as a target for UVB irradiation protection.