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Abstract Background Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) hold great therapeutic potential in regenerative medicine. Therefore, it is crucial to establish a Good Manufacturing Practice (GMP)-compliant methodology for the isolation and culture of WJ-MSCs. Through comprehensive research, encompassing laboratory-scale experiments to pilot-scale studies, we aimed to develop standardized protocols ensuring the high yield and quality of WJ-MSCs manufacturing. Methods Firstly, optimization of parameters for the enzymatic digestion method used to isolate WJ-MSCs was conducted. These parameters included enzyme concentrations, digestion times, seeding densities, and culture media. Additionally, a comparative analysis between the explant method and the enzymatic digestion method was performed. Subsequently, the consecutive passaging of WJ-MSCs, specifically up to passage 9, was evaluated using the optimized method. Finally, manufacturing processes were developed and scaled up, starting from laboratory-scale flask-based production and progressing to pilot-scale cell factory-based production. Furthermore, a stability study was carried out to assess the storage and use of drug products (DPs). Results The optimal parameters for the enzymatic digestion method were a concentration of 0.4 PZ U/mL Collagenase NB6 and a digestion time of 3 h, resulting in a higher yield of P0 WJ-MSCs. In addition, a positive correlation between the weight of umbilical cord tissue and the quantities of P0 WJ-MSCs has been observed. Evaluation of different concentrations of human platelet lysate revealed that 2% and 5% concentrations resulted in similar levels of cell expansion. Comparative analysis revealed that the enzymatic digestion method exhibited faster outgrowth of WJ-MSCs compared to the explant method during the initial passage. Passages 2 to 5 exhibited higher viability and proliferation ability throughout consecutive passaging. Moreover, scalable manufacturing processes from the laboratory scale to the pilot scale were successfully developed, ensuring the production of high-quality WJ-MSCs. Multiple freeze-thaw cycles of the DPs led to reduced cell viability and viable cell concentration. Subsequent thawing and dilution of the DPs resulted in a significant decrease in both metrics, especially when stored at 20–27 °C. Conclusion This study offers valuable insights into optimizing the isolation and culture of WJ-MSCs. Our scalable manufacturing processes facilitate the large-scale production of high-quality WJ-MSCs. These findings contribute to the advancement of WJ-MSCs-based therapies in regenerative medicine.

Abstract Background Mesenchymal stem cells (MSCs) hold immense promise for use in immunomodulation and regenerative medicine. However, their inherent heterogeneity makes it difficult to achieve optimal therapeutic outcomes for a specific clinical disease. Primed MSCs containing a certain cytokine can enhance their particular functions, thereby increasing their therapeutic potential for related diseases. Therefore, understanding the characteristic changes and underlying mechanisms of MSCs primed by various cytokines is highly important. Results In this study, we aimed to reveal the cellular heterogeneity, functional subpopulations, and molecular mechanisms of MSCs primed with IFN-γ, TNF-α, IL-4, IL-6, IL-15, and IL-17 using single-cell RNA sequencing (scRNA-seq). Our results demonstrated that cytokine priming minimized the heterogeneity of the MSC transcriptome, while the expression of MSC surface markers exhibited only slight changes. Notably, compared to IL-6, IL-15, and IL-17; IFN-γ, TNF-α, and IL-4 priming, which stimulated a significantly greater number of differentially expressed genes (DEGs). Functional analysis, which included Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, indicated that IFN-γ, TNF-α, and IL-4-primed hUC-MSCs are involved in interferon-mediated immune-related processes, leukocyte migration, chemotaxis potential, and extracellular matrix and cell adhesion, respectively. Moreover, an investigation of various biological function scores demonstrated that IFN-γ-primed hUC-MSCs exhibit strong immunomodulatory ability, TNF-α-primed hUC-MSCs exhibit high chemotaxis potential, and IL-4-primed hUC-MSCs express elevated amounts of collagen. Finally, we observed that cytokine priming alters the distribution of functional subpopulations of MSCs, and these subpopulations exhibit various potential biological functions. Taken together, our study revealed the distinct regulatory effects of cytokine priming on MSC heterogeneity, biological function, and functional subpopulations at the single-cell level. Conclusions These findings contribute to a comprehensive understanding of the inflammatory priming of MSCs, paving the way for their precise treatment in clinical applications.

Peripheral blood mononuclear cell (PBMC) are recognized as a conveniently collected reprogramming resource. Several methods are available in academia to reprogram PBMC into induced pluripotent stem cells (iPSC). In this research, we reprogrammed PBMC of different genders by using non-integrative non-viral liposome electrotransfer containing the reprogramming factors OCT4, SOX2, KLF4, and c-MYC. The three obtained iPSC cell lines were karyotypically normal and showed significant tritiated differentiation potential in vitro and in vivo. Our study provided an efficient procedure for reprogramming PBMC into iPSC and obtained three well-functioning iPSC, that may contribute to advance personalized cell therapy in the future.

Abstract Coronavirus disease 2019 (COVID-19) treatments are still urgently needed for critically and severely ill patients. Human umbilical cord-mesenchymal stem cells (hUC-MSCs) infusion has therapeutic benefits in COVID-19 patients; however, uncertain therapeutic efficacy has been reported in severe patients. In this study, we selected an appropriate cytokine, IL-18, based on the special cytokine expression profile in severe pneumonia of mice induced by H1N1virus to prime hUC-MSCs in vitro and improve the therapeutic effect of hUC-MSCs in vivo. In vitro, we demonstrated that IL-18-primed hUC-MSCs (IL18-hUCMSC) have higher proliferative ability than non-primed hUC-MSCs (hUCMSCcon). In addition, VCAM-1, MMP-1, TGF-β1, and some chemokines (CCL2 and CXCL12 cytokines) are more highly expressed in IL18-hUCMSCs. We found that IL18-hUCMSC significantly enhanced the immunosuppressive effect on CD3+ T-cells. In vivo, we demonstrated that IL18-hUCMSC infusion could reduce the body weight loss caused by a viral infection and significantly improve the survival rate. Of note, IL18-hUCMSC can also significantly attenuate certain clinical symptoms, including reduced activity, ruffled fur, hunched backs, and lung injuries. Pathologically, IL18-hUCMSC transplantation significantly enhanced the inhibition of inflammation, viral load, fibrosis, and cell apoptosis in acute lung injuries. Notably, IL18-hUCMSC treatment has a superior inhibitory effect on T-cell exudation and proinflammatory cytokine secretion in bronchoalveolar lavage fluid (BALF). Altogether, IL-18 is a promising cytokine that can prime hUC-MSCs to improve the efficacy of precision therapy against viral-induced pneumonia, such as COVID-19.

Peripheral blood mononuclear cells (PBMCs) have been widely considered as a more convenient and almost unlimited reprogramming resource, while the reprogramming procedure and efficiency still need to be improved. We reprogrammed the PBMCs by using non-integrative non-viral vectors liposome electrotransfer containing the reprogramming factors OCT4, SOX2, KLF4, and c-MYC. The iPSC lines exhibited a normal karyotype with their corresponding PBMCs and exhibited significant cellular pluripotency. Teratoma formation assay revealed that the iPSCs we generated could differentiate into three embryonic germ layers. Our study provides a more effective procedure for peripheral blood monocyte reprogramming to iPSC, and promotes its future application.

Abstract Background Pulmonary fibrosis (PF) is a growing clinical problem with limited therapeutic options. Human umbilical cord mesenchymal stromal cell (hucMSC) therapy is being investigated in clinical trials for the treatment of PF patients. However, little is known about the underlying molecular and cellular mechanisms of hucMSC therapy on PF. In this study, the molecular and cellular behavior of hucMSC was investigated in a bleomycin-induced mouse PF model. Methods The effect of hucMSCs on mouse lung regeneration was determined by detecting Ki67 expression and EdU incorporation in alveolar type 2 (AT2) and lung fibroblast cells. hucMSCs were transfected to express the membrane localized GFP before transplant into the mouse lung. The cellular behavior of hucMSCs in mouse lung was tracked by GFP staining. Single cell RNA sequencing was performed to investigate the effects of hucMSCs on gene expression profiles of macrophages after bleomycin treatment. Results hucMSCs could alleviate collagen accumulation in lung and decrease the mortality of mouse induced by bleomycin. hucMSC transplantation promoted AT2 cell proliferation and inhibited lung fibroblast cell proliferation. By using single cell RNA sequencing, a subcluster of interferon-sensitive macrophages (IFNSMs) were identified after hucMSC infusion. These IFNSMs elevate the secretion of CXCL9 and CXCL10 following hucMSC infusion and recruit more Treg cells to the injured lung. Conclusions Our study establishes a link between hucMSCs, macrophage, Treg, and PF. It provides new insights into how hucMSCs interact with macrophage during the repair process of bleomycin-induced PF and play its immunoregulation function.

Programmed cell death ligand 1 (PD-L1) is widely expressed in a variety of human tumors, and inhibition of the PD-L1/PD-1 pathway represents one of the most promising therapy for many types of cancer. However, the physiological function of PD-L1 in tissue development is still unclear, although PD-L1 mRNA is abundant in many tissues. To address this puzzle, we investigated the function of PD-L1 in mammary gland development. Interestingly, we found that PD-L1 is enriched in protein C receptor (Procr)-expressing mammary stem cells (MaSCs), and PD-L1-expressing mammary basal cells (PD-L1+ basal cells) exhibit robust mammary regeneration capacity in transplantation assay. The lineage tracing experiment showed that PD-L1+ cells can differentiate into all lineages of mammary epithelium cells, suggesting that PD-L1+ basal cells have the activities of MaSCs. Furthermore, PD-L1 deficiency significantly impairs mammary development and reduces mammary regeneration capacity of mammary basal cells, suggesting that PD-L1 is not only enriched in MaSCs but also improves activities of MaSCs. In summary, these results demonstrated that PD-L1 is enriched in MaSCs and promotes mammary gland development and regeneration. Mechanistically, our data indicated that PD-L1 expression is induced by continuous activation of Wnt/ß-catenin signaling. In conclusion, these results demonstrated that PD-L1 is a marker of MaSCs, and PD-L1 is essential for mammary development. Our study provides novel insight into the physiological functions of PD-L1 in tissue development.

Mesenchymal stromal cells (MSCs) show potential for treating preclinical models of newborn bronchopulmonary dysplasia (BPD), but studies of their therapeutic effectiveness have had mixed results, in part due to the use of different media supplements for MSCs expansion in vitro. The current study sought to identify an optimal culture supplement of umbilical cord-derived MSCs (UC-MSCs) for BPD therapy. In this study, we found that UC-MSCs cultured with human platelet lysate (hPL-UCMSCs) were maintained a small size from Passage 1 (P1) to P10, while UC-MSCs cultured with fetal bovine serum (FBS-UCMSCs) became wide and flat. Furthermore, hPL was associated with lower levels of senescence in UC-MSCs during in vitro expansion compared with FBS, as indicated by the results of β-galactosidase staining and measures of senescence-related genes (CDKN2A, CDKN1A, and mTOR). In addition, hPL enhanced the proliferation and cell viability of the UC-MSCs and reduced their doubling time in vitro. Compared with FBS-UCMSCs, hPL-UCMSCs have a greater potential to differentiate into osteocytes and chondrocytes. Moreover, using hPL resulted in greater expression of Nestin and specific paracrine factors (VEGF, TGF-β1, FGF2, IL-8, and IL-6) in UC-MSCs compared to using FBS. Critically, we also found that hPL-UCMSCs are more effective than FBS-UCMSCs for the treatment of BPD in a rat model, with hPL leading to improvements in survival rate, lung architecture and fibrosis, and lung capillary density. Finally, qPCR of rat lung mRNA demonstrated that hPL-UCMSCs had lower expression levels of inflammatory factors (TNF-α and IL-1β) and a key chemokine (MCP-1) at postnatal day 10, and there was significant reduction of CD68+ macrophages in lung tissue after hPL-UCMSCs transplantation. Altogether, our findings suggest that hPL is an optimal culture supplement for UC-MSCs expansion in vitro, and that hPL-UCMSCs promote lung repair in rat BPD disease.

As a comprehensive analysis of all metabolites in a biological system, metabolomics is being widely applied in various clinical/health areas for disease prediction, diagnosis, and prognosis. However, challenges remain in dealing with the metabolomic complexity, massive data, metabolite identification, intra- and inter-individual variation, and reproducibility, which largely limit its widespread implementation. This study provided a comprehensive workflow for clinical metabolomics, including sample collection and preparation, mass spectrometry (MS) data acquisition, and data processing and analysis. Sample collection from multiple clinical sites was strictly carried out with standardized operation procedures (SOP). During data acquisition, three types of quality control (QC) samples were set for respective MS platforms (GC-MS, LC-MS polar, and LC-MS lipid) to assess the MS performance, facilitate metabolite identification, and eliminate contamination. Compounds annotation and identification were implemented with commercial software and in-house-developed PAppLineTM and UlibMS library. The batch effects were removed using a deep learning model method (NormAE). Potential biomarkers identification was performed with tree-based modeling algorithms including random forest, AdaBoost, and XGBoost. The modeling performance was evaluated using the F1 score based on a 10-times repeated trial for each. Finally, a sub-cohort case study validated the reliability of the entire workflow.

Coronary artery disease (CAD) is a major public health problem and the chief cause of morbidity and mortality worldwide. Panax notoginseng, a valuable herb in traditional Chinese medicine (TCM) with obvious efficacy and favorable safety, shows a great promise as a novel option for CAD and is increasingly recognized clinically. Firstly, this review introduced recent clinical trials on treatment with PNS either alone or in combination with conventional drugs as novel treatment strategies. Then we discussed the mechanisms of P. notoginseng and Panax notoginseng saponins (PNS), which can regulate signaling pathways associated with inflammation, lipid metabolism, the coagulation system, apoptosis, angiogenesis, atherosclerosis, and myocardial ischaemia.

The notion of a transposed Poisson $ n $-Lie algebra has been developed as a natural generalization of a transposed Poisson algebra. It was conjectured that a transposed Poisson $ n $-Lie algebra with a derivation gives rise to a transposed Poisson $ (n+1) $-Lie algebra. In this paper, we focus on transposed Poisson $ n $-Lie algebras. We have obtained a rich family of identities for these algebras. As an application of these formulas, we provide a construction of $ (n+1) $-Lie algebras from transposed Poisson $ n $-Lie algebras with derivations under a certain strong condition, and we prove the conjecture in these cases.

Abstract Objective To investigate the expression patterns and clinical significance of specific lymphocyte subsets in the peripheral blood of patients with severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection. Methods Between December 2022 and February 2023, a cohort of 165 patients from the First Affiliated Hospital of Guangzhou University of Chinese Medicine were analyzed. The participants represented various stages of coronavirus infection severity: mild, moderate, severe, and critical. Additionally, 40 healthy individuals constituted the control group. The FC 500MPL flow cytometer and associated reagents for flow cytometry. Results Compared with the healthy control group, activated B lymphocytes witnessed a pronounced increase (p .05). As the disease progressed, there was an uptick in midterm activated T lymphocytes (p .05). Conclusion Subsets of Treg and B‐cells could act as yardsticks for the trajectory of SARS‐CoV‐2 infection and might have potential in forecasting patient trajectories. A comprehensive evaluation of lymphocyte subsets, especially in real‐time, holds the key to discerning the clinical severity in those with COVID‐19. This becomes instrumental in monitoring treatment outcomes, tracking disease evolution, and formulating prognostications. Moreover, the results provide a deeper understanding of the cellular immune defense mechanisms against the novel coronavirus infection.

In order to further explore the effects of high salt intake on kidney of ordinary mice, C57BL/6J mice were randomly allocated into normal control group (NC), 4% high salt intake group (4% group) and 8% high salt intake group (8% group), which were fed with 0.5%NaCl normal diet, 4% or 8% high salt diet respectively. The renal function index, inflammatory factor secretion, pathological damage and fibrosis degree of mice from different groups were compared. The results indicated that high salt intake led to renal function injury in C57BL/6J mice. The contents of serum creatinine, serum urea nitrogen and urine Na were significantly increased, whereas the content of urine creatinine was significantly decreased. The 8% high salt diet showed further renal damage when compared to the 4% high salt diet, which include further decrease in urine creatinine and an increase in serum urea nitrogen. Compared with NC, 4% and 8% high salt intake also increased the secretion of inflammatory cytokines TNF-α、IL-6 and IL-1β in kidney, and the secretion of IL-6 and IL-1β was further increased in the 8% group compared with the 4% group. As the HE and Masson staining results shown, high salt intake caused renal fibrosis and pathological damage of cortex and medulla in C57BL/6J mice, and the 8% group was more severe than the 4% group. High salt diet induces kidney injury and fibrosis in C57BL/6J mice, and increasing salt intake can further aggravate the severity of injury and fibrosis.

In present study, Rhizopus chinensis 12 mixed fermentation with Bacillus sp. DU-106 were performed to improve the quality and flavor of Semen Sojae Praeparatum (SSP). The levels of physicochemical characteristics, bioactive compounds, free amino acids, and volatile components during the mixed fermentation process were monitored. Compared to SSP fermented only with R. chinensis 12, the levels of physicochemical characteristics were not obviously changed after mixed fermentation. However, thrombolytic enzyme activity and soy isoflavone content after mixed fermentation were significantly higher when compared to those of SSP fermented only with R. chinensis 12, and reached to 3927.84 IU/g and 3.91 mg/g, respectively. The amount of bitter amino acids and umami amino acids after mixed fermentation slightly lower than those of SSP fermented only with R. chinensis 12. The contents of flavor composition were considerably altered after mixed fermentation, of which, the pyrazine prevailed in SSP fermented only with R. chinensis 12, while, alkane predominated in SSP after mixed fermentation. Overall, the Rhizopus chinensis 12 mixed fermentation with Bacillus sp. DU-106 contributed to improve the thrombolytic activity, free amino acid content, and flavors of SSP.

Abstract Background Wnt/β-catenin signaling pathway is closely related to the pathogenesis Osteonecrosis of the femoral head (ONFH). β-catenin, as a major component of Wnt signaling pathway, plays a vital role in the proliferation of osteoblasts. But the effect of altering β-catenin level on the early diagnosis and staging of ONFH has not been studied. Our purpose is to investigate the role of β-catenin level in the progress of ONFH. Method One hundred and one patients with three stages of ONFH and fifty healthy controls were recruited between May 2016 and November 2016. We divided the patients into 32 cases of stage II, 41 cases of stage III and 28 cases of stage IV according to the Association Research Circulation Osseous (ARCO) classification. We evaluated the clinical bone histomorphology, expression position and level of β-catenin as well as the plasma β-catenin level. We investigated the level of β-catenin from the serum and tissue samples using ELISA and Western Blot assay. We also evaluated the expression of β-catenin in bone tissue by immunohistochemistry. Data were analyzed by independent t-test and ANOVA. Results We found that the mean (± SD) serum level of β-catenin was 66.99 ± 3.032 ng/ml in the ONFH patients, which was higher than 20.14 ± 1.715 ng/ml observed in the control group (P

Due to their excellent performance, superhydrophobic materials have received a lot of attention and research in friction reduction and wear resistance. However, the effect of different contact angles of superhydrophobicity on friction and wear properties has not been thoroughly studied. In this paper, a nanosecond pulsed laser was used to realize the preparation of a superhydrophobic nickel surface, which indeed reduced the coefficient of friction but also increased the wear volume when compared to the unprocessed surface. As the contact angle of the superhydrophobic nickel surface increased, the coefficient of friction gradually increased, and the wear volume decreased gradually in superhydrophobic nickel surfaces. When the laser energy density was 1 J/cm2, the contact angle was 150.3° and the minimum friction coefficient was 0.4. However, when the laser energy density was 50 J/cm2, the maximum contact angle was 156.4° and the minimum wear volume was 4.23 × 107 μm3. The friction direction also influenced the tribological properties of the superhydrophobic-textured surface. This method makes it possible to process superhydrophobic surfaces with more suitable friction and wear properties.

Friction is one of the main causes of mechanical component failure and material waste. It is known from the friction theory that changing the roughness of the surface of the material can change the friction property of the material. Laser surface texturing is a good method for constructing a rough surface. In the experiment, the texture of the circular pits with different areal densities was constructed on the nickel surface. The coefficient of friction of the circular pit under the water-lubricated friction condition was kept at about 0.17, while the untreated metal nickel under the same conditions exceeds 0.65. The circular pit texture improves the frictional properties of the nickel surface.

Plunger pumps are widely used in oil pumping units around the world. The water content of the wellbore is increasing along with the development progress, so the lubricating capacity of the well fluids between the plunger and barrel is decreasing correspondingly. Commonly, the substrate material of the plunger and barrel are stainless steel, and the plunger surface is usually covered with nickel-based coating. Therefore, the performance of the plunger and barrel has been affected due to poor lubrication and eccentric wear. Non-smooth surfaces have been proven to improve the tribology performance in many cases. A surface texturing plunger covered with specific dimples has been prepared by using laser surface texturing technology. The morphology of the surface texturing plunger was characterized and analyzed. The tribology performance of surface texturing plunger samples was tested using standard friction and wear test machines with oil and water lubrication, respectively. The results indicated that surface texturing could effectively reduce the coefficient of friction, and the wear resistance of the surface textured samples has been improved to some extent.

The superhydrophobic surface can change the friction property of the material, reduce the adhesion of the friction interface, and produce a certain slip, thereby reducing the friction coefficient. The laser has high energy, high density, and is especially suitable for the surface treatment of materials. The laser surface texturing is a good way to construct superhydrophobic surfaces. The experiment uses a nanosecond pulse laser to construct the groove texture on the nickel surface. The contact area between the air and the droplets retained on the rough surface is increased, effectively preventing the water droplets from entering the gully of the surface microstructure, reducing the water droplets and the solid surface. The contact area ultimately makes the surface exhibit excellent superhydrophobicity. A superhydrophobic nickel surface having an apparent contact angle of water (ACAW) of 160° and a sliding angle (SA) of less than 10° was prepared. The MM-W1B vertical universal friction and wear tester was used to test the groove texture samples with different depths. The surface texture can capture the wear debris generated by the wear and store the lubricant, which is beneficial to the formation of fluid dynamic pressure lubrication and improve the load. The friction coefficient is reduced from 0.65 of the unprocessed surfaces to 0.25 after the texturing, and the friction performance is greatly improved.