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Existing approaches to evaluate cell viability involve cell staining with chemical reagents. However, the step of exogenous staining makes these methods undesirable for rapid, nondestructive, and long-term investigation. Here, we present an instantaneous viability assessment of unlabeled cells using phase imaging with computation specificity. This concept utilizes deep learning techniques to compute viability markers associated with the specimen measured by label-free quantitative phase imaging. Demonstrated on different live cell cultures, the proposed method reports approximately 95% accuracy in identifying live and dead cells. The evolution of the cell dry mass and nucleus area for the labeled and unlabeled populations reveal that the chemical reagents decrease viability. The nondestructive approach presented here may find a broad range of applications, from monitoring the production of biopharmaceuticals to assessing the effectiveness of cancer treatments. Common methods for characterising cell viability involve cell staining with chemical reagents. Here the authors report a method for cell viability assessment that does not require labelling; this uses quantitative phase imaging combined with deep learning.

In recent years, diagnostic and therapeutic approaches for rheumatoid arthritis (RA) have continued to improve. However, in the advanced stages of the disease, patients are unable to achieve long-term clinical remission and often suffer from systemic multi-organ damage and severe complications. Patients with RA usually have no overt clinical manifestations in the early stages, and by the time a definitive diagnosis is made, the disease is already at an advanced stage. RA is diagnosed clinically and with laboratory tests, including the blood markers C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) and the autoantibodies rheumatoid factor (RF) and anticitrullinated protein antibodies (ACPA). However, the presence of RF and ACPA autoantibodies is associated with aggravated disease, joint damage, and increased mortality, and these autoantibodies have low specificity and sensitivity. The etiology of RA is unknown, with the pathogenesis involving multiple factors and clinical heterogeneity. The early diagnosis, subtype classification, and prognosis of RA remain challenging, and studies to develop minimally invasive or non-invasive biomarkers in the form of biofluid biopsies are becoming more common. Non-coding RNA (ncRNA) molecules are composed of long non-coding RNAs, small nucleolar RNAs, microRNAs, and circular RNAs, which play an essential role in disease onset and progression and can be used in the early diagnosis and prognosis of RA. In this review of the diagnostic and prognostic approaches to RA disease, we provide an overview of the current knowledge on the subject, focusing on recent advances in mRNA–ncRNA as diagnostic and prognostic biomarkers from the biofluid to the tissue level.

During mammalian gastrulation, germ layers arise and are shaped into the body plan while extraembryonic layers sustain the embryo. Human embryonic stem cells, cultured with BMP4 on extracellular matrix micro-discs, reproducibly differentiate into gastruloids, expressing markers of germ layers and extraembryonic cells in radial arrangement. Using single-cell RNA sequencing and cross-species comparisons with mouse, cynomolgus monkey gastrulae, and post-implantation human embryos, we reveal that gastruloids contain cells transcriptionally similar to epiblast, ectoderm, mesoderm, endoderm, primordial germ cells, trophectoderm, and amnion. Upon gastruloid dissociation, single cells reseeded onto micro-discs were motile and aggregated with the same but segregated from distinct cell types. Ectodermal cells segregated from endodermal and extraembryonic but mixed with mesodermal cells. Our work demonstrates that the gastruloid system models primate-specific features of embryogenesis, and that gastruloid cells exhibit evolutionarily conserved sorting behaviors. This work generates a resource for transcriptomes of human extraembryonic and embryonic germ layers differentiated in a stereotyped arrangement.

Treatment of blood smears with Wright’s stain is one of the most helpful tools in detecting white blood cell abnormalities. However, to diagnose leukocyte disorders, a clinical pathologist must perform a tedious, manual process of locating and identifying individual cells. Furthermore, the staining procedure requires considerable preparation time and clinical infrastructure, which is incompatible with point-of-care diagnosis. Thus, rapid and automated evaluations of unlabeled blood smears are highly desirable. In this study, we used color spatial light interference microcopy (cSLIM), a highly sensitive quantitative phase imaging (QPI) technique, coupled with deep learning tools, to localize, classify and segment white blood cells (WBCs) in blood smears. The concept of combining QPI label-free data with AI for the purpose of extracting cellular specificity has recently been introduced in the context of fluorescence imaging as phase imaging with computational specificity (PICS). We employed AI models to first translate SLIM images into brightfield micrographs, then ran parallel tasks of locating and labelling cells using EfficientNet, which is an object detection model. Next, WBC binary masks were created using U-net, a convolutional neural network that performs precise segmentation. After training on digitally stained brightfield images of blood smears with WBCs, we achieved a mean average precision of 75% for localizing and classifying neutrophils, eosinophils, lymphocytes, and monocytes, and an average pixel-wise majority-voting F1 score of 80% for determining the cell class from semantic segmentation maps. Therefore, PICS renders and analyzes synthetically stained blood smears rapidly, at a reduced cost of sample preparation, providing quantitative clinical information.

Objective To explore the complex interactions between gut microbiota and immune cell phenotypes in rheumatoid arthritis development and identify potential therapeutic targets within the gut microbiota–immune cell axis. Methods We conducted a Mendelian randomization analysis to explore the causal relationship between gut microbiota and rheumatoid arthritis, including the role of immune cell mediators. Sensitivity analyses assessed pleiotropy and heterogeneity, while mediation analysis identified pathways through which immune cells mediate gut microbiota effects on rheumatoid arthritis development. Key microbial taxa and their effects on rheumatoid arthritis were quantified. Results Our analysis identified 27 gut microbiota taxa significantly associated with rheumatoid arthritis, with Provencibacterium massiliense showing the strongest protective effect (odds ratio = 0.807, 95% confidence interval: 0.700–0.911, P = 0.003). Additionally, 20 immune cell phenotypes with IgD+ CD38dim AC were significantly linked to rheumatoid arthritis (odds ratio = 1.064, 95% confidence interval: 1.027–1.102). Mediation analysis uncovered 13 significant gut microbiota–immune cell pathways, with the UBA8517–CCR2 monocyte pathway mediating 10.1% of the total effect (beta1 = −0.595, beta12 = 0.027, mediation proportion = 10.1%). Conclusion This study offers novel insights into the gut microbiota–immune cell axis in rheumatoid arthritis, identifying Provencibacterium massiliense, IgD+ CD38dim AC and the UBA8517—CCR2 monocyte pathway as potential therapeutic targets for rheumatoid arthritis treatment.

Purpose To investigate the surgical efficacy of Percutaneous Endoscopic Lumbar Discectomy (PELD) versus Interlaminar Fenestration Decompression (IFD) in treating male patients with Lumbar Disc Herniation (LDH) complicated with Cauda Equina Syndrome (CES), and to evaluate their impact on sexual function. Methods A retrospective analysis was conducted on male patients with LDH complicated with CES who underwent PELD or IFD at Shenzhen Traditional Chinese Medicine Hospital from January 2014 to December 2022. The patients were divided into PELD group and IFD group based on the surgical procedure. Surgical efficacy was assessed using Visual Analogue Scale (VAS), Japanese Orthopaedic Association (JOA) score, and Oswestry Disability Index (ODI). Sexual function was evaluated using the International Index of Erectile Function-5 (IIEF-5), Premature Ejaculation Diagnostic Tool (PEDT), and the International Spinal Cord Injury Male Sexual Function Basic Data Set. Results  A total of 98 patients were included (47 in the PELD group and 51 in the IFD group). Perioperative Indicators: The PELD group showed significant advantages over the IFD group in intraoperative blood loss (15.84 ± 5.31 ml vs. 42.56 ± 12.79 ml, P  < 0.001), incision length (0.74 ± 0.17 cm vs. 4.53 ± 0.81 cm, P  < 0.001), and time to ambulation (24.52 ± 6.28 h vs. 46.84 ± 10.57 h, P  < 0.001), and did not require drainage. Clinical Efficacy: At 1 month postoperatively, the PELD group had lower low back pain and leg pain VAS scores (3.12 ± 1.24/3.57 ± 1.36 vs. IFD group), higher JOA scores (15.84 ± 2.81), and lower ODI indices (35.57 ± 5.81%) ( P  < 0.05). There was no significant difference in pain and function scores between the two groups at 2 years postoperatively ( P  > 0.05). Impact on Sexual Function: At 3 months postoperatively, the PELD group had significantly higher IIEF-5 scores (15.88 ± 3.21), psychogenic erection normal rate (38.30%), and orgasm normal rate (40.43%) than the IFD group ( P  < 0.05). Sexual function recovered to similar levels in both groups at 2 years postoperatively (IIEF-5 score > 20, P  > 0.05). Conclusion PELD demonstrates advantages of minimal trauma, faster early pain relief, and quicker functional recovery compared with IFD in the treatment of LDH complicated with CES, and exerts more significant early protective effects on male sexual function, while the long-term efficacy is comparable between the two procedures. This technique can be an option for male CES patients seeking rapid recovery and sexual function protection.

γ-Tocotrienol, a kind of isoprenoid phytochemical, has antitumor activity. However, there is limited evidence that it has an effect on cervical cancer. In this study, the capacity to inhibit proliferation and induce apoptosis in human cervical cancer HeLa cells and the mechanism underlying these effects were examined. The results indicated that a γ-tocotrienol concentration over 30 μM inhibited the growth of HeLa cells with a 50% inhibitory concentration (IC50) of 46.90 ± 3.50 μM at 24 h, and significantly down-regulated the expression of proliferative cell nuclear antigen (PCNA) and Ki-67. DNA flow cytometric analysis indicated that γ-tocotrienol arrested the cell cycle at G0/G1 phase and reduced the S phase in HeLa cells. γ-tocotrienol induced apoptosis of HeLa cells in a time- and dose-dependent manner. γ-tocotrienol-induced apoptosis in HeLa cells was accompanied by down-regulation of Bcl-2, up-regulation of Bax, release of cytochrome from mitochondria, activation of caspase-9 and caspase-3, and subsequent poly (ADP-ribose) polymerase (PARP) cleavage. These results suggested that γ-tocotrienol could significantly inhibit cell proliferation through G0/G1 cell cycle arrest, and induce apoptosis via the mitochondrial apoptotic pathway in human cervical cancer HeLa cells. Thus, our findings revealed that γ-tocotrienol may be considered as a potential agent for cervical cancer therapy.

Purpose This study aimed to investigate the impact of sarcopenia on the clinical outcomes of patients undergoing posterior surgical treatment for degenerative lumbar scoliosis. Methods A retrospective analysis was conducted on clinical data from 76 patients with degenerative lumbar scoliosis, who meet the selection criteria between January 2019 and December 2023. The patients were categorized into a sarcopenia group (31 cases) and a non-sarcopenia group (45 cases) based on the diagnostic criteria of the European Working Group on Sarcopenia in Older People (EWGSOP). Operative time, intraoperative blood loss, hospital stay duration, incision length, incision healing time, and complications were compared between the sarcopenia and non-sarcopenia groups. The improvement in clinical symptoms was evaluated using the Visual Analog Scale (VAS) and the Oswestry Disability Index (ODI). To assess the scoliosis correction effect, measurements were taken for the coronal Cobb angle (CCA), C7 plumb line-center sacral vertical line (C7-CSVL), lumbar lordosis (LL), and C7 plumb line-sagittal vertical axis (C7-SVA). Results All patients successfully underwent the surgical procedure. The average operation time in the sarcopenia group was 256.27 ± 28.09 (180–350) min, which was not significantly different from the 249.82 ± 24.35(185–320) min in the non-sarcopenia group ( p  > 0.05). The average intraoperative blood loss in the sarcopenia group was 786.25 ± 38.19 (420–1365) mL, compared to 810.62 ± 45.47 (456–1780) mL in the non-sarcopenia group ( p  > 0.05). The average incision length in the sarcopenia group was 12.57 ± 1.29 (10-16)cm, compared to 12.83 ± 2.03 (9-20)cm in the non-sarcopenia group ( p  > 0.05). The incision healing time in the sarcopenia group required an average of 15.72 ± 1.74 (12-25)d, which longer than the 10.18 ± 1.05 (10–14 ) d in the non-sarcopenia group ( p  < 0.05). The average hospital stay in the sarcopenia group was 13.46 ± 1.37 (8-26) d, which was also longer than the 8.33 ± 0.92 (6-12)d in the non-sarcopenia group ( p  < 0.05). The complication rate in the sarcopenia group was 29.03% (9/31), which was higher than the 13.33% (6/45) in the non-sarcopenia group ( p  < 0.05). The VAS and ODI scores of both groups at the last follow-up were significantly improved compared to preoperative levels ( p  < 0.05). At the last follow-up, the ODI in the non-sarcopenia group was better than that in the sarcopenia group ( p  < 0.05). There was no statistically significant difference between the two groups in terms of VAS scores at the last follow-up ( p  > 0.05). Both groups demonstrated significant improvements in CCA, C7-CSVL, LL, and C7-SVA compared to preoperative levels ( p  < 0.05). However, no statistically significant differences were observed between the groups at the final follow-up ( p  > 0.05). Conclusion Sarcopenia does not significantly affect the radiological outcomes of patients with degenerative lumbar scoliosis undergoing posterior surgical procedures, ODI was better in the non-sarcopenia group. In addition, patients with sarcopenia typically require a longer postoperative recovery period and are more susceptible to various complications compared to those without sarcopenia.

This study evaluated the prevalence and factors associated with sleep disturbance in a large cohort of HIV-infected patients across China. A cross-sectional study was conducted among HIV-infected patients on antiretroviral therapy at 20 AIDS clinics. The Pittsburgh Sleep Quality Index was self-administered by subjects. Socio-demographic characteristics, medical history and HIV-related clinical data were collected. 4103 patients had complete data for analysis. Sleep disturbances were observed in 43.1% of patients. Associated factors in multivariable analysis included psychological factors: anxiety (odds ratio [OR], 3.13; 95% confidence interval [CI], 2.44–4.00; P < 0.001), depression (OR, 2.09; 95% CI, 1.70–2.57; P < 0.001), and both anxiety and depression (OR, 5.90; 95% CI, 4.86–7.16; P < 0.001); sociodemographic factors: MSM (OR, 1.26; 95% CI, 1.04–1.52; P = 0.018), being single (OR, 1.45; 95%CI 1.21–1.74; P < 0.001), higher education (OR, 1.25; 95% CI, 1.03–1.53; P = 0.025); and clinical factors: suboptimal adherence (OR,1.51; 95% CI,1.23–1.85; P < 0.001), regimen-switching (OR, 1.94; 95% CI, 1.12–3.35; P = 0.018), and antidepressant use (OR, 1.98; 95% CI, 1.47–2.67; P = 0.044). Prevalence of sleep disturbance is high in this large Chinese cohort. Associated factors appear related to psychological and social-demographic factors. Health workers may consider routinely assessing sleep disturbances among HIV-infected patients, especially in the first three months after HIV diagnosis, and referring for mental health services, which may positively impact adherence to treatment.