High-resolution Tandem Mass Spectrometry for Studying Chemical Constituents of Gynura bicolor DC.

The separation and analysis of the desired chemical components are important subjects for the fundamental research of traditional Chinese medicine (TCM). Ultra-high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) has gradually become a leading technology for the identification of TCM ingredients. Gynura bicolor DC. (BFH), a perennial stemless herb used for medicine and food in China has medicinal effects such as clearing heat, moistening the lung, relieving cough, dispersing stasis, and relieving swelling. Polyphenols and flavonoids contain numerous isomers, which hinder the identification of the complex compounds in BFH. This paper presents a systematic protocol for studying chemical constituents of BFH based on solvent extraction and integrated data via UPLC-Q-TOF-MS. The method described here includes systematic protocols for sample pretreatment, MS calibration, MS acquisition, data processing, and analysis of results. Sample pretreatment includes collection, cleaning, drying, crushing, and extraction. MS calibration consists of multipoint and single-point correction. Data processing includes data importing, method establishment, analysis processing, and result presentation. Representative results of the typical fragmentation pattern of phenolic acids, esters, and glycosides in Gynura bicolor DC. (BFH) are presented in this paper. In addition, organic solvent selection, extraction, data integration, collision energy selection, and method improvement are discussed in detail. This universal protocol can be widely used to identify complex compounds in TCM.

Self-stigmatization of high-school students seeking professional psychological help: the chain-mediating effect of perceived social support and optimism.

Background: Research has shown that stigmatization of professional psychological help-seeking is an important factor influencing attitudes toward seeking professional psychological help (ATSPPH). However, how perceived social support (PSS) and optimism have a mediating role is not clear. Objective: Examine the associations between ATSPPH, self-stigmatization of seeking help, PSS, and optimism in a cohort of Chinese high-school students (HSSs). Methods: An offline survey was conducted in three high schools in Chongqing (China) from 20 February to 20 May 2023. Participants were HSSs recruited through their teachers. A total of 2,159 HSSs completed a survey on demographic information as well as the Self-Stigmatization of Seeking Help (SSOSH) score, ATSPPH, Perceived Social Support Scale (PASS), and Life Orientation Test (LOT). Mediation analyses were conducted using the "Process" macro in SPSS 26.0 to estimate the direct and indirect effects of self-stigmatization of seeking psychological help on ATSPPH. Results: Self-stigmatization of seeking psychological help was significantly and negatively related to ATSPPH among HSSs. Self-stigmatization of psychological help-seeking influenced ATSPPH through three pathways: (a) separate mediating effect of PSS (effect = -0.029); (b) separate mediating effect of optimism (effect = -0.069); (c) chain-mediating effect of PSS and optimism (effect = -0.017). These data suggested that self-stigmatization of psychological help-seeking could influence ATSPPH directly and indirectly through PSS and optimism. Conclusion: PSS and optimism mediated the relationship between self-stigmatization of seeking help and ATSPPH. Improving the ability of HSSs to perceive social support and cultivating optimism could help improve the self-stigmatization of help-seeking and promote a positive attitude toward professional help-seeking.

Highly catalytic and stable Au@AuPt nanoparticles for visual and quantitative detection of E. coli O157:H7.

A visual and quantitative ELISA-like method for Escherichia coli O157:H7 is developed based on highly catalytic and stable Au@AuPt nanoparticles. The proposed enhanced ELISA method can visually detect 100 CFU mL-1 O157:H7 with high specificity and without the need for strict low-temperature reagent storage, thereby increasing the utility. Moreover, it is applicable to spiked tap water and milk tea samples without additional treatments.

Assessment of the predictive ability of the Johns Hopkins Fall Risk Assessment Tool (Chinese Version) in inpatient settings.

AIMS: This study was to assess the predictive ability of the Johns Hopkins Fall Risk Assessment Tool (Chinese Version) in inpatient settings. DESIGN: A case-control study. METHODS: This study was conducted in a tertiary hospital based on 2019 data. With a case-control design in a 1:2 ratio, the predictive ability of the Johns Hopkins Fall Risk Assessment Tool (Chinese Version) was determined by ROC curve. The best cut point was identified based on sensitivity, specificity, positive predict value and negative predict value. Conditional logistical regression analysis was conducted to test the predictive ability of each indicator. RESULTS: The study included 309 patients, with 103 in the case group and 206 in the control groups. Generally, the predictive ability was acceptable with the area under ROC curve value at 0.73 (95% CI: 0.67-0.79). Positive predict value and negative predict value performed best at the cut point of 13. Sensitivity at cut point 6 was much higher than that at cut point 13, though specificity was lower. Except for age, all indicators in the Johns Hopkins Fall Risk Assessment Tool (Chinese Version) demonstrated significant predictive ability as to occurrence of fall. CONCLUSION: The Johns Hopkins Fall Risk Assessment Tool (Chinese Version) is a reliable assessment instrument in the inpatient settings. IMPACT: This is the first study that evaluated the predictive ability of the Johns Hopkins Fall Risk Assessment Tool (Chinese version) in the inpatient settings, and proved that the instrument is reliable for assessing inpatient fall risks. Further studies could be carried out to assess the predict ability of Johns Hopkins Fall Risk Assessment Tool (Chinese version) among specific populations.

miR-489-3p Inhibits Prostate Cancer Progression by Targeting DLX1.

PURPOSE: Prostate cancer (PCa) is the third most common cancer in men and the second leading cause of cancer-related death in men. DLX1 belongs to the DLX homeobox family and exhibits antitumor activity in many kinds of tumors. MicroRNAs (miRNAs) play important roles in the progression of cancer. However, whether miRNAs affect the development of PCa by targeting DLX1 has not been determined. In this study, we aimed to investigate the role of miR-489-3p in the regulation of DLX1 expression and PCa progression and to provide a potential therapeutic target for PCa treatment. METHODS AND MATERIALS: The Cancer Genome Atlas database was used to analyze the divergent expression of DLX1 in carcinomas and adjacent normal tissues. The expression level of DLX1 in malignant and normal prostate cells was also measured using RT-qPCR and Western blotting. A dual-luciferase reporter assay was performed to determine whether miR-489-3p directly targets DLX1. We transfected 22Rv1 and DU145 cells with miR-489-3p mimics to overexpress miR-489-3p and then evaluated its effect on cellular function. MTT, EdU, colony formation and cell cycle assays were used to evaluate cell growth. JC-1 and ROS assays with flow cytometry were performed to indirectly analyze apoptosis. Transwell assays were conducted to investigate metastasis. RESULTS: The expression level of DLX1 was upregulated in both PCa tissues and cell lines. MiR-489-3p directly targeted DLX1 and downregulated its expression. Overexpression of miR-489-3p significantly suppressed cell growth. MiR-489-3p induced apoptosis through mitochondrial function impairment. Overexpression of miR-489-3p also inhibited cell migration and invasion. DLX1 overexpression reversed the above effects induced by miR-489-3p. CONCLUSION: We identified the involvement of the miR-489-3p/DLX1 pathway in PCa for the first time. In this pathway, miR-489-3p acts as a tumor suppressor by negatively regulating the expression of DLX1. MiR-489-3p may be a potential therapeutic target for PCa treatment.

Ethanol extract of Gynura bicolor (GB) protects against UVB-induced photodamage of skin by inhibiting P53-mediated Bcl-2/BAX/Caspase-3 apoptosis pathway.

To investigate the protective effect of ethanol extract of Gynura bicolor (GB) against UVB-induced photodamage of skin and the possible mechanisms. DPPH (1,1-diphenyl-2-pico radical) test was used to detect the antioxidant capacity of ethanol extract of Gynura bicolor (GB). The protective effects of GB against UVB irritation were detected both in Hacat cells and photodamage rat models. UVB irradiation could inhibit viability and induce apoptosis of Hacat cells in a dose-dependent manner. The pretreatment of Hacat cells by GB could obviously reverse the effects in a dose-dependent manner. The mRNA and protein expressions of p53, Bax, caspase-3 were increased, while anti-apoptotic protein Bcl-2 was decreased and this effect could be reversed by GB pretreatment in a dose-dependent manner. In vivo, the application of GB could alleviate the skin damage of SD rats and improve the superficial inflammation of the dermis as well as inhibit the expressions of P53 and Caspase-3 induced by UVB irradiation. Ethanol extract of Gynura bicolor could protect the photodamage of human Hacat keratinocytes and SD rats against UVB irradiation by inhibiting P53-mediated Bcl-2/ BAX/Casaspe-3 apoptosis pathway.

When Al-Doped Cobalt Sulfide Nanosheets Meet Nickel Nanotube Arrays: A Highly Efficient and Stable Cathode for Asymmetric Supercapacitors.

Although cobalt sulfide is a promising electrode material for supercapacitors, its wide application is limited by relative poor electrochemical performance, low electrical conductivity, and inefficient nanostructure. Here, we demonstrated that the electrochemical activity of cobalt sulfide could be significantly improved by Al doping. We designed and fabricated hierarchical core-branch Al-doped cobalt sulfide nanosheets anchored on Ni nanotube arrays combined with carbon cloth (denoted as CC/H-Ni@Al-Co-S) as an excellent self-standing cathode for asymmetric supercapacitors (ASCs). The combination of structural and compositional advantages endows the CC/H-Ni@Al-Co-S electrode with superior electrochemical performance with high specific capacitance (1830 F g-1/2434 F g-1 at 5 mV s-1/1 A g-1) and excellent rate capability (57.2%/72.3% retention at 1000 mV s-1/100 A g-1). The corresponding all-solid-state ASCs with CC/H-Ni@Al-Co-S and multilayer graphene/CNT film as cathode and anode, respectively, achieve a high energy density up to 65.7 W h kg-1 as well as superb cycling stability (90.6% retention after 10 000 cycles). Moreover, the ASCs also exhibit good flexibility and stability under different bending conditions. This work provides a general, effective route to prepare high-performance electrode materials for flexible all-solid-state energy storage devices.

hsa-miR-155 targeted NCSTN 3'UTR mutation promotes the pathogenesis and development of acne inversa.

OBJECTIVES: To investigate molecular mechanisms of nicastrin (NCSTN) mutations inducing acne inversa (AI). METHODS: New and old lesional and non-lesional skin samples were obtained from an AI patient. Healthy skin samples were obtained from the buttocks of 100 non-AI patients. Hematoxylin-eosin staining and immunohistochemistry of NCSTN protein were examined. All exon-intron and exon boundary sequences were polymerase chain reaction (PCR) -amplified and sequenced. Bioinformatic analyses of NCSTN 3'-untranslated regions (3'UTR) were conducted using RegRNA2.0. 3'UTR of NCSTN was cloned vector of psiCHECK-2 vector; the mutant 3'UTR NCSTN-psiCHECK-2 was constructed on a template of NCSTN 3'UTR. A dual-luciferase reporter gene assay, real-time reverse transcription (qRT)-PCR and Western blot analysis were conducted to evaluate functional changes associated with the mutation. RESULTS: We identified a novel deletion mutation of the NCSTN gene in the NCSTN 3'UTR region (designated c.2584-2585del CA) at the binding site of human micro-RNA-155 (hsa-miR-155). Levels of NCSTN protein were potently lower in epidermis and hair follicles of AI patients with lesions than in healthy skin. The hsa-miR-155+mutant NCSTN significantly downregulated in dual luciferase assay, qRT-PCR, and Western blot. The novel deletion mutation was confirmed to be a pathological cause of AI. CONCLUSIONS: miR-155 downregulates the expression of NCSTN by binding NCSTN 3'UTR, providing a possible new mechanism of loss of NCSTN function in AI patients. hsa-miR-155 functions as a promoter in AI, and is a potential therapy target for AI.

A phosphorylethanolamine-functionalized super-hydrophilic 3D graphene-based foam filter for water purification.

A phosphorylethanolamine-functionalized graphene foam (PNGF) has been proposed as an active filtration material for the capture and removal of heavy metal ions in water. Benefiting from its abundant hydrophilic portion of oxygen, nitrogen and phosphorus groups, the PNGF is super-hydrophilic. The selected heavy metal ions, Pb(II) and Cd(II), could be rapidly and efficiently absorbed within 10min using the PNGF through a filtration model, which is obviously less time compared with the several hours or even longer time when employing the traditional shaking or stirring model. In addition, the used PNGF filters can be easily reused after a simple, low-cost detachment using HCl to remove the heavy metals, providing a new approach for water purification.

Identification of 2 Novel Mutations in ATP2C1 Gene in Hailey-Hailey Disease and a Literature Review of Variations in a Chinese Han Population.

BACKGROUND Hailey-Hailey disease (HHD) is a rare autosomal dominant skin condition. The ATP2C1 gene was identified as the defective gene in HHD. To date, 166 pathogenic mutations in ATP2C1 have been observed worldwide. The aim of this study was to identify variations in HHD and summarize the features of the mutations identified in China. MATERIAL AND METHODS We examined 2 familial and 2 sporadic cases of HHD. Genomic DNA polymerase chain reaction and direct sequencing of the ATP2C1 were performed from HHD patients, unaffected family members, and 200 healthy individuals. We also searched the published literature for data about the ATP2C1 gene using PubMed and the Chinese Biological Medicine Database. RESULTS We detected 3 heterozygous mutations, including 2 novel frameshift mutations (c.819insA (273LfsX) and c.1264insTAGATGG (421LfsX)) and 1 recurrent nonsense mutation (c.115C>T (R39X)). To the best of our knowledge, 90 different mutations (including our current results) have been reported in China, all of which occurred in the Chinese Han population. CONCLUSIONS Our data may add to the existing list of ATP2C1 mutations and provide new insight into genetic variants of HHD in China.

MicroRNA­24­1­5p promotes malignant melanoma cell autophagy and apoptosis via regulating ubiquitin D.

The present study aimed to investigate the key roles and possible regulatory mechanism of microRNA (miR)­24­1­5p in regulating the autophagy, and apoptosis of malignant melanoma cells. The expression levels of miR­24­1­5p in malignant melanoma tissues were determined. Human melanoma A375 cells were transfected with miR­24­1­5p mimic and control. The effects of miR­24­1­5p overexpression on regulating the expressions of autophagy­related proteins [microtubule­associated protein 1A/1B­light chain 3 (LC3)­II, LC3­I and Beclin­1] and apoptosis­related proteins [apoptosis regulator Bcl­2 (Bcl­2) and (BCL2 like 1) Bcl­xL] were investigated. The percentage of apoptotic cells in different transfected cells was detected. In addition, luciferase reporter assays were performed to confirm whether ubiquitin D (UBD) was a target of miR­24­1­5p. The effects of UBD silencing on autophagy and apoptosis were also investigated. The expression levels of janus kinase (JNK), phosphorylated (P)­JNK, Jun proto­oncogene AP­1 transcription factor subunit (c­Jun) and p­c­Jun were determined following the overexpression of miR­24­1­5p, and UBD. In comparison with adjacent normal tissues, miR­24­1­5p was significantly downregulated in malignant melanoma tissues. Overexpression of miR­24­1­5p significantly increased the levels of LC3­II/I ratio and Beclin­1 expression, and decreased the expression levels of Bcl­2 and Bcl­xL. Flow cytometry also showed that miR­24­1­5p overexpression promoted cell apoptosis. Moreover, UBD was confirmed as a direct target of miR­24­1­5p. Silencing of UBD promoted melanoma cell autophagy and apoptosis via regulating the expression levels of related proteins. Besides, the levels of the p­JNK/JNK and p­c­Jun/Jun ratios were significantly increased following miR­24­1­5p overexpression, which were reversed following co­overexpression of miR­24­1­5p, and UBD. Overexpression of miR­24­1­5p may target UBD, and subsequently promote the autophagy and apoptosis of malignant melanoma cells through activation of the JNK signaling pathway.

Novel D323G mutation of DSG4 gene in a girl with localized autosomal recessive hypotrichosis clinically overlapped with monilethrix.

BACKGROUND: Localized autosomal recessive hypotrichosis (LAH) is an inherited rare disease caused by DSG4 mutations, characterized by short, sparse, brittle hair affecting restricted areas such as the scalp, trunk, and extremities. To date, DSG4 mutations have been reported in 14 pedigrees of LAH overlapping with monilethrix. METHODS: To clarify the etiology of hair defects for a 2-year-old Chinese girl, peripheral blood, skin, and hair samples were collected, and skin immunohistochemistry, electron microscopy (scanning and transmission types), Vivascope confocal microscopy, and DSG4 sequencing were investigated. RESULTS: The patient presented sparse hairs of various length and follicular hyperkeratotic papules. Eyebrows and lashes were also involved (broke or shed). The biopsy specimen revealed curled ingrown hair shafts within the hair follicle and keratin-filled hair follicles. Scanning electron microscopy revealed hair cuticle loosely and irregularly arranged, as well as a marked warping, curling, cracking, and detachment of hair cuticle. Transmission electron microscopy indicated notable dysadhesion between cells of the outer root sheath. A homozygous mutation A1103G in exon 8 of DSG4 was identified in the patient, resulting in the substitution of an aspartic acid by glycine (D323G) and reduced DSG4 expression in the affected scalp epidermis. CONCLUSIONS: The homozygous A1103G mutation in DSG4 was responsible for the disease development.