Advances in mesenchymal stem cell-derived extracellular vesicles therapy for Sjogren's syndrome-related dry eye disease.

Sjogren's syndrome (SS) is a chronic autoimmune disorder that affects exocrine glands, particularly lacrimal glands, leading to dry eye disease (DED). DED is a common ocular surface disease that affects millions of people worldwide, causing discomfort, visual impairment, and even blindness in severe cases. However, there is no definitive cure for DED, and existing treatments primarily relieve symptoms. Consequently, there is an urgent need for innovative therapeutic strategies based on the pathophysiology of DED. Mesenchymal stem cells (MSCs) have emerged as a promising therapeutic tool for various autoimmune disorders, including SS-related DED (SS-DED). A particularly intriguing facet of MSCs is their ability to produce extracellular vesicles (EVs), which contain various bioactive components such as proteins, lipids, and nucleic acids. These molecules play a key role in facilitating communication between cells and modulating a wide range of biological processes. Importantly, MSC-derived EVs (MSC-EVs) have therapeutic properties similar to those of their parent cells, including immunomodulatory, anti-inflammatory, and regenerative properties. In addition, MSC-EVs offer several notable advantages over intact MSCs, including lower immunogenicity, reduced risk of tumorigenicity, and greater convenience in terms of storage and transport. In this review, we elucidate the underlying mechanisms of SS-DED and discuss the relevant mechanisms and targets of MSC-EVs in treating SS-DED. In addition, we comprehensively review the broader landscape of EV application in autoimmune and corneal diseases. This review focuses on the efficacy of MSC-EVs in treating SS-DED, a field of study that holds considerable appeal due to its multifaceted regulation of immune responses and regenerative functions.

The mediating role of organizational commitment between workplace bullying and turnover intention among clinical nurses in China: a cross-sectional study.

BACKGROUND: Workplace bullying experienced by clinical nurses is a critical and pervasive issue that not only detrimentally impacts nurses but also poses a significant threat to the overall quality of nursing services and patient care. This study aimed to determine the mediating role of organizational commitment in the relationship between workplace bullying and turnover intention among clinical nurses in China. METHODS: Participants were recruited from 40 hospitals in various provinces of China from December 2, 2021 to February 25, 2023, using convenience sampling. After obtaining hospital ethical approval and participants' informed consent, clinical nurses (n = 585) from different nursing departments in different hospitals completed the questionnaire. The Socio-demographic Questionnaire, Negative Acts Qestionnaire, Chinese Workers' Organizational Commitment Scale and Turnover Intention Questionnaire were used to collect general demographic data of nurses and assess workplace bullying they experienced, their level of organizational commitment and turnover intention. Descriptive statistics, Pearson correlation analyses and structural equation model were adopted to analyze the data. RESULTS: Pearson's correlation analysis showed that that workplace bullying was significantly negatively correlated with organizational commitment (r = - 0.512, P<0.01) and significantly positively correlated with turnover intention (r = 0.558, P<0.01), organizational commitment was significantly negatively correlated with turnover intention (r = - 0.539, P<0.01). Mediation analysis indicated organizational commitment partially mediated the association between workplace bullying and turnover intention. The total effect (ß = 0.69) of workplace bullying on turnover intention consisted of its direct effect (ß = 0.41) and the indirect effect mediated through organizational commitment (ß = 0.280), with the mediating effect accounting for 40.58% of the total effect. CONCLUSION: Organizational commitment mediated the associations of workplace bullying and turnover intention. Therefore, healthcare organizations and nursing managers should develop appropriate strategies to enhance nurses' organizational commitment in order to reduce their turnover intention.

Quantitative haplotype-resolved analysis of mitochondrial DNA heteroplasmy in Human single oocytes, blastoids, and pluripotent stem cells.

Maternal mitochondria are the sole source of mtDNA for every cell of the offspring. Heteroplasmic mtDNA mutations inherited from the oocyte are a common cause of metabolic diseases and associated with late-onset diseases. However, the origin and dynamics of mtDNA heteroplasmy remain unclear. We used our individual Mitochondrial Genome sequencing (iMiGseq) technology to study mtDNA heterogeneity, quantitate single nucleotide variants (SNVs) and large structural variants (SVs), track heteroplasmy dynamics, and analyze genetic linkage between variants at the individual mtDNA molecule level in single oocytes and human blastoids. Our study presented the first single-mtDNA analysis of the comprehensive heteroplasmy landscape in single human oocytes. Unappreciated levels of rare heteroplasmic variants well below the detection limit of conventional methods were identified in healthy human oocytes, of which many are reported to be deleterious and associated with mitochondrial disease and cancer. Quantitative genetic linkage analysis revealed dramatic shifts of variant frequency and clonal expansions of large SVs during oogenesis in single-donor oocytes. iMiGseq of a single human blastoid suggested stable heteroplasmy levels during early lineage differentiation of naïve pluripotent stem cells. Therefore, our data provided new insights of mtDNA genetics and laid a foundation for understanding mtDNA heteroplasmy at early stages of life.

Semi-supervised segmentation of coronary DSA using mixed networks and multi-strategies.

The coronary arteries supply blood to the myocardium, which originate from the root of the aorta and mainly branch into the left and right. X-ray digital subtraction angiography (DSA) is a technique for evaluating coronary artery plaques and narrowing, that is widely used because of its time efficiency and cost-effectiveness. However, automated coronary vessel classification and segmentation remains challenging using a little data. Therefore, the purpose of this study is twofold: one is to propose a more robust method for vessel segmentation, the other is to provide a solution that is feasible with a small amount of labeled data. Currently, there are three main types of vessel segmentation methods, i.e., graphical- and statistical-based; clustering theory based, and deep learning-based methods for pixel-by-pixel probabilistic prediction, among which the last method is the mainstream with high accuracy and automation. Under this trend, an Inception-SwinUnet (ISUnet) network combining the convolutional neural network and Transformer basic module was proposed in this paper. Considering that data-driven fully supervised learning (FSL) segmentation methods require a large set of paired data with high-quality pixel-level annotation, which is expertise-demanding and time-consuming, we proposed a Semi-supervised Learning (SSL) method to achieve better performance with a small amount of labeled and unlabeled data. Different from the classical SSL method, i.e., Mean-Teacher, our method used two different networks for cross-teaching as the backbone. Meanwhile, inspired by deep supervision and confidence learning (CL), two effective strategies for SSL were adopted, which were denominated Pyramid-consistency Learning (PL) and Confidence Learning (CL), respectively. Both were designed to filter the noise and improve the credibility of pseudo labels generated by unlabeled data. Compared with existing methods, ours achieved superior segmentation performance over other FSL and SSL ones by using data with a small equal number of labels. Code is available in https://github.com/Allenem/SSL4DSA.

Single-cell individual full-length mtDNA sequencing by iMiGseq uncovers unexpected heteroplasmy shifts in mtDNA editing.

The ontogeny and dynamics of mtDNA heteroplasmy remain unclear due to limitations of current mtDNA sequencing methods. We developed individual Mitochondrial Genome sequencing (iMiGseq) of full-length mtDNA for ultra-sensitive variant detection, complete haplotyping, and unbiased evaluation of heteroplasmy levels, all at the individual mtDNA molecule level. iMiGseq uncovered unappreciated levels of heteroplasmic variants in single cells well below the conventional NGS detection limit and provided accurate quantitation of heteroplasmy level. iMiGseq resolved the complete haplotype of individual mtDNA in single oocytes and revealed genetic linkage of de novo mutations. iMiGseq detected sequential acquisition of detrimental mutations, including large deletions, in defective mtDNA in NARP/Leigh syndrome patient-derived induced pluripotent stem cells. iMiGseq identified unintended heteroplasmy shifts in mitoTALEN editing, while showing no appreciable level of unintended mutations in DdCBE-mediated mtDNA base editing. Therefore, iMiGseq could not only help elucidate the mitochondrial etiology of diseases, but also evaluate the safety of various mtDNA editing strategies.

Acupuncture-related therapies for the poststroke cognitive impairment patients: A protocol for systematic review and network meta-analysis.

BACKGROUND: To compare and rank the clinical effects of different acupuncture and acupuncture-related therapies on patients with poststroke cognitive impairment. METHODS: We evaluated the direct and indirect evidence from relevant studies using network meta-analysis. Eight databases were examined in order to find randomized controlled trials of acupuncture-related therapies for individuals with poststroke cognitive impairment. After 2 researchers independently scanned the literature, extracted the data, and assessed the risk of bias in the included studies, the data were analyzed using RevMan5.4, Stata15.0, and WinBUGS1.4.3 software. RESULTS: We assess the benefits and drawbacks of various acupuncture-related therapies, rank the efficacy of various acupuncture-related therapies in the treatment of poststroke cognitive impairment, and describe the best acupuncture intervention approaches or combinations based on the available data. CONCLUSION: This study will contribute to the existence of data on the safety and efficacy of acupuncture-related therapies in the treatment of poststroke cognitive impairment, and it may aid clinical guideline makers in selecting the best acupuncture treatment for poststroke cognitive impairment. REGISTRATION NUMBER: INPLASY2021120117.

Histone demethylase KDM4A overexpression improved the efficiency of corrected human tripronuclear zygote development.

Human zygotes are difficult to obtain for research because of limited resources and ethical debates. Corrected human tripronuclear (ch3PN) zygotes obtained by removal of the extra pronucleus from abnormally fertilized tripronuclear (3PN) zygotes are considered an alternative resource for basic scientific research. In the present study, eight-cell and blastocyst formation efficiency were significantly lower in both 3PN and ch3PN embryos than in normal fertilized (2PN) embryos, while histone H3 lysine 9 trimethylation (H3K9me3) levels were much higher. It was speculated that the aberrant H3K9me3 level detected in ch3PN embryos may be related to low developmental competence. Microinjection of 1000 ng/µl lysine-specific demethylase 4A (KDM4A) mRNA effectively reduced the H3K9me3 level and significantly increased the developmental competence of ch3PN embryos. The quality of ch3PN zygotes improved as the grading criteria, cell number and pluripotent expression significantly increased in response to KDM4A mRNA injection. Developmental genes related to zygotic genome activation (ZGA) were also upregulated. These results indicate that KDM4A activates the transcription of the ZGA program by enhancing the expression of related genes, promoting epigenetic modifications and regulating the developmental potential of ch3PN embryos. The present study will facilitate future studies of ch3PN embryos and could provide additional options for infertile couples.

Melatonin protects against oxybenzone-induced deterioration of mouse oocytes during maturation.

Oxybenzone (OBZ), an ultraviolet light filter that is widely used in sunscreens and cosmetics, is an emerging contaminant found in humans and the environment. Recent studies have shown that OBZ has been detected in women's plasma, urine, and breast milk. However, the effects of OBZ exposure on oocyte meiosis have not been addressed. In this study, we investigated the detrimental effects of OBZ on oocyte maturation and the protective roles of melatonin (MT) in OBZ-exposed mouse models. Our in vitro and in vivo results showed that OBZ suppressed oocyte maturation, while MT attenuated the meiotic defects induced by OBZ. In addition, OBZ facilitated H3K4 demethylation by increasing the expression of the Kdm5 family of genes, elevating ROS levels, decreasing GSH, impairing mitochondrial quality, and disrupting spindle configuration in oocytes. However, MT treatment resulted in significant protection against OBZ-induced damage during oocyte maturation and improved oocyte quality. The mechanisms underlying the beneficial roles of MT involved reduction of oxidative stress, inhibition of apoptosis, restoration of abnormal spindle assembly and up-regulation of H3K4me3. Collectively, our results suggest that MT protects against defects induced by OBZ during mouse oocyte maturation in vitro and in vivo.

Generation of Artificial Gamete and Embryo From Stem Cells in Reproductive Medicine.

In addition to the great growing need for assisted reproduction technologies (ART), additional solutions for patients without functional gametes are strongly needed. Due to ethical restrictions, limited studies can be performed on human gametes and embryos; however, artificial gametes and embryos represent a new hope for clinical application and basic research in the field of reproductive medicine. Here, we provide a review of the research progress and possible application of artificial gametes and embryos from different species, including mice, monkeys and humans. Gametes specification from adult stem cells and embryonic stem cells (ESCs) as well as propagation of stem cells from the reproductive system and from organized embryos, which are similar to blastocysts, have been realized in some nonhuman mammals, but not all achievements can be replicated in humans. This area of research remains noteworthy and requires further study and effort to achieve the reconstitution of the entire cycle of gametogenesis and embryo development in vitro.

Selenium Biofortification of Crop Food by Beneficial Microorganisms.

Selenium (Se) is essential for human health, however, Se is deficient in soil in many places all around the world, resulting in human diseases, such as notorious Keshan disease and Keshin-Beck disease. Therefore, Se biofortification is a popular approach to improve Se uptake and maintain human health. Beneficial microorganisms, including mycorrhizal and root endophytic fungi, dark septate fungi, and plant growth-promoting rhizobacteria (PGPRs), show multiple functions, especially increased plant nutrition uptake, growth and yield, and resistance to abiotic stresses. Such functions can be used for Se biofortification and increased growth and yield under drought and salt stress. The present review summarizes the use of mycorrhizal fungi and PGPRs in Se biofortification, aiming to improving their practical use.

Antibacterial activity and mechanism of moxifloxacin nanoparticles against drug-resistant Pseudomonas aeruginosa / 药学学报

We prepared moxifloxacin (MXF) loaded nanoparticles by nano-precipitation/self-assembly method, then compared the antibacterial activity of MXF and MXF loaded nanoparticles, and investigated the antibacterial mechanism of MXF loaded nanoparticles against Pseudomonas aeruginosa in vitro. The physicochemical properties such as particle size and zeta potential were investigated by laser particle size analyzer. The in vitro release characteristics were investigated by high performance liquid chromatography (HPLC). The effect of nanoparticles on HBE cells viability was investigated by CCK-8 assay. In addition, the in vitro antibacterial activity was investigated by minimum inhibitory concentration (MIC) assay, biofilm formation assays and transmission electron microscope (TEM) observation, then the antibacterial mechanism was initially explored. The particle size measurement showed that the nanoparticles had a size of 332.5 ± 2.7 nm, a polymer dispersion index (PDI) of 0.125 ± 0.053, a zeta potential of -24.3 ± 1.7 mV, and a uniform particle size distribution, drug loading content was (6.02 ± 1.27) %, encapsulation efficiency was (16.69 ± 1.17) %. The TEM results show that the nanoparticles have a spheroidal structure, and the particle size and distribution are consistent with the particle size measurement results. The nanoparticles can be effectively and rapidly released in phosphate buffer saline (PBS), releasing about 70% in 24 h, and releasing 87% in 72 h, and almost completely releasing the MXF at 120 h. At the same time, compared with moxifloxacin free drug, its MIC value is 8 μg·mL-1, which is 1/2 of MXF solution, and can significantly inhibit the formation of bacterial biofilms. It has well antibacterial activity in vitro and can be targeted to the surface of bacteria to exert its efficacy and improve the antibacterial effect. The moxifloxacin nanoparticles prepared in this study has a uniform particle size distribution, well drug release performance and antibacterial effect, and provides new ideas and strategies for the treatment of bacterial lung infection and the development of new antibacterial nanoformulations.

Precise Personalized Medicine in Gynecology Cancer and Infertility.

Since the conception of precision medicine has been put forward in oncology, this idea has been popularized and applied in many specialties. Significant progress has been made toward personalizing the entire process, including diagnosis, treatment planning, and embryo identification, and combining large-scale genetic information data and knowledge discovery can offer better prospects in reproductive medicine. This work reviews the application of precision medicine and possibilities in reproductive medicine and gynecologic cancer diagnosis and treatment. The limitations and challenges of precision medicine in this area remain to be discussed.

Determination of Clopidogrel and Its Three Metabolites Concentrations in Human Plasma by LC-MS/MS / 中国药房

OBJECTIVE： To establish the method for simultaneous determination of clopidogrel （CLP）， its intermediate metabolite （2-O-CLP）， inactive metabolite （CLPCA） and active metabolite （CLPTM） in human plasma. METHODS： Totally 90 patients diagnosed as stroke were selected from the First Affiliated Hospital of Army Medical University. They were given one CLP tablet （75 mg/tablet） orally on an empty stomach in the morning. Blood samples were collected 2 h after taking the tablet. CLPTM- D was formed by derivation of CLPTM with 2-bromo-3’-methoxyacetophenone and extracted by precipitation of acetonitrile protein together with the other three substances to be measured. LC-MS/MS method was adopted. The determination was performed on Agilent poroshell 120 EC-C18 column with mobile phase consisted of acetonitrile （0.1％ formic acid） and water （0.1％ formic acid） （90 ∶ 10， V/V）. The quantitation analysis was performed using multiple reaction monitoring at the specific ion transitions of m/z 308.1→198.1 （CLPCA）， 322.3→212.0 （CLP）， 338.3→155.0 （2-O-CLP）， 504.4→354.1 （CLPTM-D） and 264.0→154.1 （ticlopidine， internal standard）， respectively. RESULTS： The retention time of CLPCA， CLP， 2-O-CLP， CLPTM-D and internal standard were 2.01， 3.32， 2.83， 2.68， 1.87 min， respectively. The linear range of CLPCA， CLP， 2-O-CLP and CLPTM-D were 100-10 000， 0.2-20， 0.3-30， 0.5-50 ng/mL （all r≥0.999 5）. The intra-day and inter-day RSD were all less than 9.5％ （n＝5）. Accuracy ranged from 93.5％-98.9％ （n＝5）， and extraction recovery was from 85.4％ to 95.9％ （n＝5）. The matrix effect ranged from 2.7％-6.2％ （n＝5）. In stability tests （storing at －80 ℃ for 3 months， 3 freeze-thaw cycles， storing at 4 ℃ for 8 h）， RE of CLP， CLPCA and CLPTM-D were all lower than 10.0％ （n＝5）. CONCLUSIONS： Established LC-MS/MS method has the advantages of high specificity， accuracy and reliability， and can be used to detect the concentration of CLP and its three metabolites in human plasma.