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To better understand cellular communication driving diverse behaviors, we need to uncover the molecular mechanisms of within-cell-type functional heterogeneity. While single-cell RNA sequencing (scRNAseq) has advanced our understanding of cell heterogeneity, linking individual cell phenotypes to transcriptomic data remains challenging. Here, we used a phenotypic cell sorting technique to ask whether phenotypically supervised scRNAseq analysis (pheno-scRNAseq) can provide more insight into heterogeneous cell behaviors than unsupervised scRNAseq. Using a simple 3D in vitro breast cancer (BRCA) model, we conducted pheno-scRNAseq on invasive and non-invasive cells and compared the results to phenotype-agnostic scRNAseq analysis. Pheno-scRNAseq identified unique and more selective differentially expressed genes than unsupervised scRNAseq analysis. Functional studies validated the utility of pheno-scRNAseq in understanding within-cell-type functional heterogeneity and revealed that migration phenotypes were coordinated with specific metabolic, proliferation, stress, and immune phenotypes. This approach lends new insight into the molecular systems underlying BRCA cell phenotypic heterogeneity.

This article introduces the clinical experience of Professor Jian-qiao FANG in treatment of postherpetic neuralgia (PHN) with acupuncture and moxibustion. Jian-qiao FANG believes that PHN is characterized by "stagnation" and "deficiency" in pathogenesis and it is treated by "resolving stasis" and "replenishing for deficiency". Electroacupuncture is predominated in treatment and the combined regimen of various techniques of acupuncture and moxibustion is adopted in therapeutic strategy. According to the pain characteristics of patients, the acupuncture and moxibustion prescription is drafted for resolving stasis and masses, replenishing for deficiency and regulating the mind. The acupoints include Ashi points, Jiaji (EX-B2), Hegu (LI4), Waiguan (TE5), Zusanli (ST36), Sanyinjiao (SP6), Shenting (GV24), Baihui (GV20) and Sishencong (EX-HN1). In treatment with electroacupuncture, the stimulation parameters are selected flexibly according to different pain characteristics. The frequency of 2/100 Hz is the first option, 100 Hz is used for the acute seizure and 2 Hz is for mild pain. After electric stimulation, the treatment with roller type of dermal needle and moxibustion can be applied to ensure clinical efficacy. [ABSTRACT FROM AUTHOR]

Despite significant progress in tissue engineering, the full regeneration of chronic wounds persists as a major challenge, with the immune response to tissue damage being a key determinant of the healing process's quality and duration. Post-injury, a crucial aspect is the transition of macrophages from a pro-inflammatory state to an anti-inflammatory. Thus, this alteration in macrophage polarization presents an enticing avenue within the realm of regenerative medicine. Recent advancements have entailed the integration of a myriad of cellular and molecular signals into hydrogel-based constructs, enabling the fine-tuning of immune cell activities during different phases. This discussion explores modern insights into immune cell roles in skin regeneration, underscoring the key role of immune modulation in amplifying the overall efficacy of wounds. Moreover, a comprehensive review is presented on the latest sophisticated technologies employed in the design of immunomodulatory hydrogels to regulate macrophage polarization. Furthermore, the deliberate design of hydrogels to deliver targeted immune stimulation through manipulation of chemistry and cell integration is also emphasized. Moreover, an overview is provided regarding the influence of hydrogel properties on immune traits and tissue regeneration process. Conclusively, the accent is on forthcoming pathways directed toward modulating immune responses in the milieu of chronic healing., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)