A stratified perspective is essential to understand the divergent impacts of plant-based diets on osteoarthritis risk in older populations.

Objective To investigate the role of RANKL in the initiation and progression of bone metastasis in thyroid cancer. Methods We first retrospectively analyzed clinical data to identify factors associated with bone metastasis. In vitro, we compared the expression of RANK/RANKL signaling components and downstream targets in thyroid cancer cells, and verified the effects by pharmacologic inhibition or genetic knockdown of RANKL. RANKL-overexpressing and RANKL-silenced cell lines were generated to assess DNA damage responses. Endoplasmic reticulum stress (ER stress) was induced, and the expression of ER stress-related proteins was evaluated under different RANKL levels. DNA damage signaling was then inhibited to determine its impact on ER stress. Finally, Hedgehog pathway activity was measured under varying RANKL expression and ER stress conditions. Results In patients, RANKL, γ-H2AX, Ki-67, and ATF6 were markedly higher in bone-metastatic than in non-metastatic thyroid tumors, and tumor tissues exhibited more pronounced DNA damage and ER stress than adjacent non-neoplastic tissues. In cell lines, RANKL overexpression accelerated proliferation, increased the S-phase population, and reduced apoptosis. RANKL knockdown had the opposite effects. Transcriptomic analysis showed that RANKL levels correlated with the expression of DNA damage, ER stress, and Hedgehog genes. Electron microscopy confirmed that RANKL suppression attenuated ER stress and Hedgehog signaling. In vivo, HDAC inhibition reduced tumor volume, and the combination of an HDAC inhibitor with RANKL knockdown further improved antitumor efficacy. Conclusion RANKL may promote bone metastasis of thyroid cancer by triggering DNA damage via the ER stress–Hedgehog signaling axis, and targeting this network may offer a novel therapeutic strategy.

BACKGROUND:Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by dysregulated T cell responses and metabolic disturbances. Mesenchymal stromal cells (MSCs) have shown therapeutic promise, but their mechanisms, particularly concerning T cell metabolism, remain incompletely defined. This study investigated whether human umbilical cord-derived MSCs (hUC-MSCs) ameliorate collagen-induced arthritis (CIA) by modulating T cell metabolism and differentiation. METHODS:CIA was induced in DBA/1 mice. Animals received PBS or hUC-MSCs on day 28. Arthritis index (AI), joint histology, serum cytokines (TNF-α, IL-6, IL-17, and TGF-β), and metabolites (lactate and pyruvate) were assessed. Splenic T cell transcription factors (FOXP3, RORγt, and PU.1) and glycolytic genes (GLUT1, G6PD, and PFKFB3) were analyzed by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. In vitro, human CD4+ T cells were cocultured with hUC-MSCs under T-helper 17 (Th17)-polarizing conditions. T cell subsets, glycolytic metabolites, and gene/protein expression were evaluated by flow cytometry, colorimetric assays, RT-qPCR, and western blot. RESULTS:MSC treatment significantly attenuated arthritis severity, joint destruction, and splenomegaly in CIA mice. It reduced serum pro-inflammatory cytokines and normalized elevated lactate and pyruvate levels. In the spleen, MSCs suppressed RORγt and PU.1 while enhancing FOXP3 expression, and downregulated GLUT1 and G6PD mRNA. Positive correlations were found between glycolytic markers (GLUT1 and G6PD) and pro-inflammatory transcription factors (RORγt and PU.1), and between serum lactate and inflammatory cytokines. In vitro, hUC-MSCs directly inhibited Th17 differentiation and promoted Treg generation in human CD4+ T cells. This metabolic reprogramming was functionally coupled to a shift in T cell differentiation: a suppression of pro-inflammatory Th17 cells and a promotion of regulatory T (Treg) generation in human CD4+ T cells. This was accompanied by reduced lactate production and significant downregulation of GLUT1, G6PD, and PFKFB3 at both mRNA and protein levels. CONCLUSIONS:hUC-MSCs ameliorate CIA by restoring the Th17/Treg balance through metabolic reprogramming of T cells, specifically by suppressing glycolysis. This immunometabolic mechanism highlights the therapeutic potential of MSCs in RA.

Background and purpose:Patients with rheumatoid arthritis (RA) face an elevated risk of sepsis and its associated neurological complications, most notably sepsis-associated encephalopathy (SAE). Nonetheless, early identification of SAE in this population remains a substantial clinical challenge. To address this gap, we aimed to develop and validate a predictive model that incorporates the neutrophil-to-albumin ratio (NAR) to estimate SAE risk in RA patients with sepsis. Methods:This retrospective multicenter cohort study included a derivation cohort of 89 patients with RA and sepsis from two centers and an independent external validation cohort of 37 patients from a third center. Patients in the derivation cohort were classified into SAE and non-SAE groups. Three machine learning algorithms (LASSO, random forest, and XGBoost) were applied for feature selection, and the optimal model was selected based on area under the receiver operating characteristic curve (AUC). Model performance was evaluated using bootstrap resampling, calibration curves, and decision curve analysis. The final XGBoost model was subsequently evaluated in the external validation cohort. A web-based dynamic prediction tool was developed for clinical application. Kaplan-Meier analysis and Cox regression were performed to evaluate 28-day survival. Results:SAE occurred in 23.6% of patients and was associated with significantly higher 28-day mortality (61.9% vs. 33.8%, p = 0.04). Six consensus predictors (SOFA score, procalcitonin, platelet count, length of stay, NAR, and age) were identified. The XGBoost model achieved a bootstrap-corrected AUC of 0.859 (95% CI: 0.751-0.946) in the derivation cohort. In the independent external validation cohort, the final XGBoost model achieved an AUC of 0.849 (95% CI: 0.674-1.000). SHAP analysis demonstrated that higher SOFA, procalcitonin, length of stay, age, and NAR values increased SAE risk, whereas higher platelet count was protective. Kaplan-Meier analysis showed significantly lower 28-day survival in the high NAR group (p = 0.023), and elevated NAR was associated with increased mortality risk (HR = 2.178, 95% CI: 1.095-4.332). Conclusion:The NAR-integrated XGBoost model provides a robust and interpretable tool for early SAE prediction in RA patients with sepsis, showing potential clinical utility for bedside risk stratification.

Mesenchymal stromal cells (MSCs) have emerged as a promising disease-modifying therapy for the complications of diabetes mellitus (DM), including diabetic retinopathy (DR). However, the optimal treatment regimen remains unclear, and challenges persist regarding the timing, route of delivery and the mechanisms underlying the therapeutic effects. This study focused on human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs), to elucidate their retinal protective effects, and investigate the underlying mechanisms by which a single intravenous injection might ameliorate the pathological alterations of DR. Two time points after the development of DM were chosen for the in vivo experiments to study the effects of the intervention after different times of exposure to hyperglycemia. hUC-MSCs were injected via the tail vein at 8 and 16 weeks after STZ injection. Retinal samples were collected 2 weeks post-treatment to analyze the therapeutic effect of MSCs on DR. In vitro experiments were conducted using a Müller cell line and a retinal microvascular endothelial cell line cultured under high-glucose conditions, with treatment by hUC-MSCs conditioned media (MSC-CM), to explore the underlying mechanisms. After a single intravenous injection of hUC-MSCs at week 16 and not 8 weeks post-STZ injection, retinal tissue showed improved thickness of the inner nuclear layer. There was also an increase in the number of acellular capillaries observed in retinal flat mounts of diabetic animals which was improved in the DM and MSC treatment group. MSC treatment reduced high glucose induced activation markers (GFAP and Vimentin) of Müller cells and alleviated endoplasmic reticulum (ER) stress. VEGF expression was also reduced in the retina. MSC-conditioned media also reversed high glucose-induced expression of VEGF in Müller cells. Finally, in a retinal microvascular endothelial cell line, high glucose concentrations, demonstrated increased ER stress which was reduced by MSC conditioned media. Single Intravenous injection of hUC-MSC to DM animals could alleviate DR via reducing Müller cell and endothelial cell activation and ER stress, and thus might represent a promising therapy for DR.

ABSTRACT Objective Treatment with human umbilical cord mesenchymal stem cells (hUC‐MSCs) attenuated the clinical manifestations of systemic lupus erythematosus (SLE). We investigated the metabolic mechanism whereby hUC‐MSCs modify CD4 + T cell cytokine secretion in lupus. Methods The study enrolled 30 untreated lupus patients and 20 sex, age, and body mass index matched healthy controls (HCs). CD4 + T cells were isolated by magnetic sorting, and stimulated with anti‐CD3/CD28. The hUC‐MSCs treatment (MSCT) groups were coculturing hUC‐MSCs to CD4 + T cells from moderate and severe SLE (SLE‐MS) groups for 72 h at ratios of 1:25 (T1), 1:10 (T2), and 1:5 (T3). Cytokine concentration and proliferation of the CD4 + T cells were measured by Luminex liquid chip assay and cell counting kit‐8, respectively. Glucose metabolic capacity was measured by Seahorse real‐time metabolic analysis. The role of hUC‐MSCs on cytokine secretion was analyzed by transcriptome sequencing. Glucose enzymes levels and HSP90AA1/PI3K/AKT pathway activity were analyzed by real‐time quantitative PCR and western blot. The CD4 + T cell subsets were detected by flow cytometry. Results Compared with HCs, the enhanced glycolysis and mitochondrial oxygen consumption of SLE‐CD4 + T cells were positively associated with disease activity. Treatment with hUC‐MSCs proportionally decreased glucose metabolism and proliferation of SLE‐CD4 + T cells. The hUC‐MSCs treatment significantly diminished supernatant concentrations of interferon‐γ, tumor necrosis factor‐α, interleukin (IL)‐4, and IL‐17 in SLE‐MS group, as well as inhibited HSP90AA1 in the glucose‐activated PI3K‐AKT pathway. In animal experiment, the systemic administration of hUC‐MSCs and inhibition of HSP90AA1 resulted in a reduction of glucose metabolites, enzymes, pro‐inflammatory factor levels, and HSP90AA1 / PI3K/AKT signaling pathway activity. Conclusions The hUC‐MSCs treatment inhibited overactive glucose metabolism of SLE‐CD4 + T cells. HSP90AA1 in the PI3K‐AKT pathway induced by the glucose metabolism may be involved in the anti‐inflammatory function of hUC‐MSCs treatment.

Systemic lupus erythematosus (SLE) is a chronic, multi-organ autoimmune disease with complex pathogenesis and unclear causes. Elevated levels of IgD have been observed in the peripheral blood of SLE patients, suggesting a potential role for IgD through its interaction with the IgD Fc receptor (FcδR). This study aimed to explore the impact of IgD on T cell function in SLE and evaluate the therapeutic potential of targeting the IgD-FcδR pathway using an IgD-Fc-Ig fusion protein. In SLE patients, biomarkers such as BAFF, ESR, anti-dsDNA and SLEDAI-2k, which are used to assess disease activity and clinical presentations, were significantly correlated with sIgD levels. As an IgD-FcδR blocker, IgD-Fc-Ig effectively suppressed the activation and proliferation of CD4+ T cells stimulated by IgD, restored the balance between Th17 and Treg cell subsets, and reduced the expression and interaction of phosphorylated Lck (p-Lck) and JAK2 (p-JAK2). Moreover, in vivo study demonstrated that IgD-Fc-Ig may also ameliorates disease manifestations in MRL/lpr mice with lupus nephritis. IgD-Fc-Ig could reduce serum IgD levels, proteinuria level and the kidney deposition of immune complex C3, ameliorate histopathological changes in kidney and spleen tissue. Additionally, it reversed the state of excessive activation and imbalance of Th17/Treg cell subsets, reduced cytokine levels, and downregulated p-JAK2 and p-STAT3 expression. In conclusion, our study revealed a correlation between abnormally increased sIgD and SLE pathogenesis, IgD-FcδR-Lck-JAK2-STAT3 may act as an important mechanism contributing to T cell activation in SLE. IgD-Fc-Ig fusion protein may represent a promising targeted therapy for SLE.

Objective: Postprandial hyperglycemia is a major risk factor for type 2 diabetes and cardiovascular disease. Inhibition of α-amylase and α-glucosidase can attenuate postprandial glycemic response (PPGR). This study aimed to investigate the inhibitory effects of mulberry leaf and corn silk on these enzymes in vitro and their impact on postprandial glucose (PG) levels in prediabetic individuals using milk-based matrices. Research Design and Methods: In vitro, enzyme inhibition was assessed using the DNS method (α-amylase) and pNPG method (α-glucosidase). A randomized crossover trial was conducted in 11 prediabetic individuals with four interventions: pure milk; lactose-hydrolyzed milk; lactose-hydrolyzed milk with mulberry leaf, corn silk, and resistant dextrin; and GOS milk with mulberry leaf and corn silk. PPGR was assessed by area under the glucose curve, 1 and 2 h PG, maximum PG, and 2 h glucose excursion. Paired Wilcoxon signed-rank tests were used for comparisons. Results: Mulberry leaf and corn silk extracts inhibited both enzymes dose-dependently, with synergistic effects. No significant differences in PPGR indices were observed across interventions in the overall prediabetic individuals. However, in the overweight subgroup, the combination of GOS milk supplemented with mulberry leaf and corn silk significantly reduced 1 h PG (median difference [P25, P75]: −0.84 mmol/L [−1.05, −0.49]), maximum PG (−0.54 mmol/L [−0.75, −0.25]), and glucose excursion (−0.62 mmol/L [−0.75, −0.24]) compared to pure milk. Conclusions: Mulberry leaf and corn silk extracts inhibit α-amylase and α-glucosidase in vitro and may attenuate postprandial glucose excursions in overweight prediabetic individuals when delivered in a GOS milk matrix.

T cells play a crucial role in the pathogenesis of systemic lupus erythematosus (SLE), with their functions regulated by various metabolic pathways. This study explores SLE pathogenesis and the therapeutic effects of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) via metabolic reprogramming. Clinical data and peripheral blood samples were collected from 15 SLE patients and matched healthy controls. CD4+ T cells were isolated and activated in vitro with anti-CD3/CD28. Following 72 h of co-culture with hUC-MSCs, CD4+ T cell viability was assessed using the CCK-8 assay. The oxygen consumption rate (OCR) and glycolytic proton efflux rate (glycoPER) were measured with a Seahorse analyzer. Cytokine levels were detected by multiplex assay, and transcriptome sequencing was performed. Western blotting analyzed glucose metabolism-related enzymes and signaling pathways in lupus model mice. Compared to healthy controls, activated CD4+ T cells from SLE patients exhibited significantly increased OCR and glycoPER levels (P < 0.05). Following 72 h of co-culture with hUC-MSCs, OCR, glycoPER, cell viability, and pro-inflammatory factors in SLE-CD4+ T cells decreased markedly (P < 0.01). Upregulation of 434 genes and downregulation of 172 genes was observed, particularly in the JAK-STAT and PI3K-Akt pathways. hUC-MSCs inhibited the expression of glucose metabolism-related enzymes and the JAK-STAT and PI3K-Akt signaling pathways in lupus model mice. hUC-MSCs inhibited the proliferation and function of aberrant CD4+ T cells in SLE patients by modulating glycometabolism and the JAK-STAT and PI3K-Akt signaling pathways, providing new insights into the therapeutic mechanisms of MSCs based on metabolic reprogramming.

T cell immuno-metabolic regulation plays a key role in the development of systemic lupus erythematosus (SLE). This study aimed to analyze the role of CD4+ T cell glucose metabolism in SLE development. Clinical data and blood samples were collected from 20 untreated SLE patients and healthy controls (HCs) matched for age, sex, and body mass index. After being isolated by magnetic sorting and cultured with anti-CD3/CD28 for 72 h, CD4+ T cells were subjected to real-time metabolic analysis. CD4+ T cell proliferation and cytokines were measured with cell counting kit-8 and Luminex liquid chip assay, respectively. Compared to HCs, SLE-CD4+ T cells exhibited significantly higher glycolytic capacity and mitochondrial oxidative phosphorylation (OXPHOS) (both p < 0.001). Additionally, SLE-CD4+ T cells demonstrated increased proliferation rates and elevated cytokine levels in both plasma and culture supernatants (both p < 0.05). OXPHOS and glycolysis of SLE-CD4+ T cells were positively correlated with SLE disease activity index-2000 (SLEDAI-2K) and cytokines, and negatively correlated with SLE-CD4+ T cell numbers (all p < 0.05). CD4+ T cells from SLE patients showed higher glucose metabolic activity than those from HCs, and the enhanced glucose metabolism of SLE-CD4+ T cells was strongly correlated with disease activity, suggesting that glucose metabolic reprogramming plays an essential role in the pathogenesis of SLE.

BACKGROUND:Diabetic cardiomyopathy (DCM) is a serious health-threatening complication of diabetes mellitus characterized by myocardial fibrosis and abnormal cardiac function. Human umbilical cord mesenchymal stromal cells (hUC-MSCs) are a potential therapeutic tool for DCM and myocardial fibrosis via mechanisms such as the regulation of microRNA (miRNA) expression and inflammation. It remains unclear, however, whether hUC-MSC therapy has beneficial effects on cardiac function following different durations of diabetes and which mechanistic aspects of DCM are modulated by hUC-MSC administration at different stages of its development. This study aimed to investigate the therapeutic effects of intravenous administration of hUC-MSCs on DCM following different durations of hyperglycemia in an experimental male model of diabetes and to determine the effects on expression of candidate miRNAs, target mRNA and inflammatory mediators. METHODS:A male mouse model of diabetes was induced by multiple low-dose streptozotocin injections. The effects on severity of DCM of intravenous injections of hUC-MSCs and saline two weeks previously were compared at 10 and 18 weeks after diabetes induction. At both time-points, biochemical assays, echocardiography, histopathology, polymerase chain reaction (PCR), immunohistochemistry and enzyme-linked immunosorbent assays (ELISA) were used to analyze blood glucose, body weight, cardiac structure and function, degree of myocardial fibrosis and expression of fibrosis-related mRNA, miRNA and inflammatory mediators. RESULTS:Saline-treated diabetic male mice had impaired cardiac function and increased cardiac fibrosis after 10 and 18 weeks of diabetes. At both time-points, cardiac dysfunction and fibrosis were improved in hUC-MSC-treated mice. Pro-fibrotic indicators (α-SMA, collagen I, collagen III, Smad3, Smad4) were reduced and anti-fibrotic mediators (FGF-1, miRNA-133a) were increased in hearts of diabetic animals receiving hUC-MSCs compared to saline. Increased blood levels of pro-inflammatory cytokines (IL-6, TNF, IL-1β) and increased cardiac expression of IL-6 were also observed in saline-treated mice and were reduced by hUC-MSCs at both time-points, but to a lesser degree at 18 weeks. CONCLUSION:Intravenous injection of hUC-MSCs ameliorated key functional and structural features of DCM in male mice with diabetes of shorter and longer duration. Mechanistically, these effects were associated with restoration of intra-myocardial expression of miRNA-133a and its target mRNA COL1AI as well as suppression of systemic and localized inflammatory mediators.

Mesenchymal stromal cells (MSCs) a potentially effective disease-modulating therapy for diabetic nephropathy (DN) but their clinical translation has been hampered by incomplete understanding of the optimal timing of administration and in vivo mechanisms of action. This study aimed to elucidate the reno-protective potency and associated mechanisms of single intravenous injections of human umbilical cord-derived MSCs (hUC-MSCs) following shorter and longer durations of diabetes. A streptozotocin (STZ)-induced model of diabetes and DN was established in C57BL/6 mice. In groups of diabetic animals, human (h)UC-MSCs or vehicle were injected intravenously at 8 or 16 weeks after STZ along with vehicle-injected non-diabetic animals. Diabetes-related kidney abnormalities was analyzed 2 weeks later by urine and serum biochemical assays, histology, transmission electron microscopy and immunohistochemistry. Serum concentrations of pro-inflammatory and pro-fibrotic cytokines were quantified by ELISA. The expression of autophagy-related proteins within the renal cortices was investigated by immunoblotting. Bio-distribution of hUC-MSCs in kidney and other organs was evaluated in diabetic mice by injection of fluorescent-labelled cells. Compared to non-diabetic controls, diabetic mice had increases in urine albumin creatinine ratio (uACR), mesangial matrix deposition, podocyte foot process effacement, glomerular basement membrane thickening and interstitial fibrosis as well as reduced podocyte numbers at both 10 and 18 weeks after STZ. Early (8 weeks) hUC-MSC injection was associated with reduced uACR and improvements in multiple glomerular and renal interstitial abnormalities as well as reduced serum IL-6, TNF-α, and TGF-β1 compared to vehicle-injected animals. Later (16 weeks) hUC-MSC injection also resulted in reduction of diabetes-associated renal abnormalities and serum TGF-β1 but not of serum IL-6 and TNF-α. At both time-points, the kidneys of vehicle-injected diabetic mice had higher ratio of p-mTOR to mTOR, increased abundance of p62, lower abundance of ULK1 and Atg12, and reduced ratio of LC3B to LC3A compared to non-diabetic animals, consistent with diabetes-associated suppression of autophagy. These changes were largely reversed in the kidneys of hUC-MSC-injected mice. In contrast, neither early nor later hUC-MSC injection had effects on blood glucose and body weight of diabetic animals. Small numbers of CM-Dil-labeled hUC-MSCs remained detectable in kidneys, lungs and liver of diabetic mice at 14 days after intravenous injection. Single intravenous injections of hUC-MSCs ameliorated glomerular abnormalities and interstitial fibrosis in a mouse model of STZ-induced diabetes without affecting hyperglycemia, whether administered at relatively short or longer duration of diabetes. At both time-points, the reno-protective effects of hUC-MSCs were associated with reduced circulating TGF-β1 and restoration of intra-renal autophagy.

Objective The efficacy of sirolimus in treating severe or refractory systemic lupus erythematosus (SLE) has been confirmed by small-scale clinical trials. However, few studies focused on mild or moderate SLE. Therefore, in this study we elucidated clinical efficacy of add-on sirolimus in patients with mild or moderate SLE.Methods Data of 17 consecutive patients with SLE were retrospectively collected. SLE Disease Activity Index-2000 (SLEDAI-2K), clinical manifestation, laboratory data and peripheral T lymphocyte subsets with cytokines were collected before and 6 months after sirolimus add-on treatment. T cell subsets were detected by flow cytometry and cytokines were determined by multiplex bead-based flow fluorescent immunoassay simultaneously. Twenty healthy controls matched with age and sex were also included in our study.Results (1) The numbers of peripheral blood lymphocytes, T cells, T helper (Th) cells, regulatory T (Treg) cells, Th1 cells, Th2 cells and Treg/Th17 ratios in patients with SLE were significantly lower, while the numbers of Th17 cells were evidently higher than those of healthy control (p<0.05). (2) After 6 months of sirolimus add-on treatment, urinary protein, pancytopenia, immunological indicators and SLEDAI-2K in patients with SLE were distinctively improved compared with those before sirolimus treatment (p<0.05). (3) The numbers of peripheral blood lymphocytes, T cells, Th cells, Treg cells, Th2 cells and the ratios of Treg/Th17 in patients with SLE after treatment were clearly higher than those before (p<0.05). (4) The levels of plasma interleukin (IL)-5, IL-6 and IL-10 in patients with SLE decreased notably, conversely the IL-4 levels increased remarkably compared with pretreatment (p<0.05).Conclusions (1) Patients with SLE presented imbalanced T cell subsets, especially the decreased ratio of Treg/Th17. (2) Sirolimus add-on treatment ameliorated clinical involvement, serological abnormalities and disease activity without adverse reactions in patients with SLE. (3) The multi-target therapy facilitates the enhanced numbers of Treg cells, Treg/Th17 imbalance and anti-inflammatory cytokines, simultaneously, reducing inflammatory cytokines.

Background Spinal cord injury (SCI) is a serious clinical condition that has pathological changes such as increased neuroinflammation and nerve tissue damage, which eventually manifests as fibrosis of the injured segment and the development of a spinal cord cavity leading to loss of function. Cell-based therapy, such as mesenchymal stem cells (MSCs) and neural stem cells (NSCs) are promising treatment strategies for spinal cord injury via immunological regulation and neural replacement respectively. However, therapeutic efficacy is rare reported on combined transplantation of MSC and NSC in acute mice spinal cord injury even the potential reinforcement might be foreseen. Therefore, this study was conducted to investigate the safety and efficacy of co-transplanting of MSC and NSC sheets into an SCI mice model on the locomotor function and pathological changes of injured spinal cord. Methods To evaluate the therapeutic effects of combination cells, acute SCI mice model were established and combined transplantation of hiPSC-NSCs and hMSCs into the lesion site immediately after the injury. Basso mouse scale was used to perform the open-field tests of hind limb motor function at days post-operation (dpo) 1, 3, 5, and 7 after SCI and every week after surgery. Spinal cord and serum samples were collected at dpo 7, 14, and 28 to detect inflammatory and neurotrophic factors. Hematoxylin–eosin (H&E) staining, masson staining and transmission electron microscopy were used to evaluate the morphological changes, fibrosis area and ultrastructure of the spinal cord. Result M&N transplantation reduced fibrosis formation and the inflammation level while promoting the secretion of nerve growth factor and brain-derived neurotrophic factor. We observed significant reduction in damaged tissue and cavity area, with dramatic improvement in the M&N group. Compared with the Con group, the M&N group exhibited significantly improved behaviors, particularly limb coordination. Conclusion Combined transplantation of hiPSC-NSC and hMSC could significantly ameliorate neuroinflammation, promote neuroregeneration, and decrease spinal fibrosis degree in safe and effective pattern, which would be indicated as a novel potential cell treatment option. Graphical abstract

Background: Diabetic cardiomyopathy (DCM) is a complication of diabetes mellitus that endangers human health. DCM results in cardiac dysfunction, which eventually progresses to heart failure. Mesenchymal stromal cells (MSCs), a type of multipotent stem cell, have shown promising therapeutic effects in various cardiovascular diseases and diabetic complications in preclinical studies due to their immunomodulatory and regenerative abilities. However, there is still a lack of evidence to summarize the effectiveness of MSCs in the treatment of DCM. Therefore, a meta-analysis and systematic review are warranted to evaluate the therapeutic potential of MSCs for DCM in preclinical studies.Methods: A comprehensive literature search in English or Chinese was conducted in PubMed, EMBASE, web of Science, Cochrane Library, and China National Knowledge Internet from inception to June 30, 2022. The summarized outcomes included echocardiography, morphology, and pathology. Data were independently extracted and analyzed by two authors. The software we adopted was Review Manager5.4.1. This systematic review was written in compliance with PRISMA 2020 and the review protocol was registered on PROSPERO, registration no. CRD42022350032. Results: We included 20 studies in our meta-analysis to examine the efficacy of MSCs in the treatment of DCM. The MSC-treated group showed a statistically significant effect on left ventricular ejection fraction (WMD=12.61, 95% CI 4.32 to 20.90, P=0.003) and short axis fractional shortening (WMD=6.84, 95% CI 4.09 to 9.59, P < 0.00001). The overall effects on the ratio of early to late diastolic mitral annular velocity, left ventricular end-diastolic pressure, maximum positive pressure development, maximum negative pressure development, left ventricular relaxation time constant, heart weight to body weight ratio, fibrosis area, and arteriole density were analyzed, suggesting that MSCs represent an effective therapy for the treatment of DCM. Conclusion: Our results suggest a therapeutic role for MSCs in the treatment of DCM, and these results provide support for the use of MSCs in clinical trials of patients with DCM.

Spinal cord injury (SCI) is a catastrophic event that incurs substantial personal and social costs. The complex pathophysiology associated with SCI often limits the regeneration of nerve tissue at the injured site and leads to permanent nerve damage. With advances in stem cell biology, the field of regenerative medicine offers the hope of solving this challenging problem. Neural stem/progenitor cells (NSPCs) possess nerve regenerative and neuroprotective effects, and transplanting NSPCs in their optimized form into an injured area holds promising therapeutic potential for SCI. In this review, we summarize the advantages and disadvantages of NSPCs derived from different sources while highlighting the utility of NSPCs derived from induced pluripotent stem cells, an NSPC source with superior advantages, according to data from in vivo animal models and the latest clinical trials.

Background:Astrocytes can be involved in motor neuron toxicity in amyotrophic lateral sclerosis (ALS) induced by noncell autonomous effects, and inflammatory cytokines may play the main role in mediating this process. However, the etiology of aberrant cytokine secretion is unclear. The present study assessed possible involvement of the mTOR-autophagy pathway in aberrant cytokine secretion by ALS patient iPSC-derived astrocytes. Method and Results. PBMCs from sporadic ALS patients and control subjects were reprogrammed into iPSCs, which were then differentiated into astrocytes and/or motor neurons. Comparison with control astrocytes indicated that conditioned medium of ALS astrocytes reduced the viability of the control motor neurons (p < 0.05) assessed using the MTT assay. The results of ELISA showed that the concentrations of TNFα, IL1β, and IL6 in cell culture medium of ALS astrocytes were increased (p < 0.05). ALS astrocytes had higher p62 and mTOR levels and lower LC3BII/LC3BI ratio and ULK1 and p-Beclin-1 (Ser15) levels (p < 0.05), indicating defective autophagy. Exogenous inhibition of the mTOR-autophagy pathway, but not the activation of the pathway in control subject astrocytes, increased the levels of p62 and mTOR and concentration of IL-1β, TNF-α, and IL-6 in cell culture medium and decreased the LC3BII/LC3BI ratio and levels of ULK1 and p-Beclin-1 (Ser15), and these changes were comparable to those in ALS astrocytes. After 48 h of rapamycin (autophagy activator) and 3-methyladenine (autophagy inhibitor) treatments, the exogenous activation of the mTOR-autophagy pathway, but not inhibition of the pathway, in ALS astrocytes significantly reduced the concentrations of TNFα, IL1β, and IL6 in cell culture medium and reduced the levels of p62, while increasing the levels of LC3B-II/LC3B-I, ULK1, and p-Beclin-1 (Ser15), and these changes were comparable to those in control subject astrocytes.Conclusion:Alteration in the mTOR/ULK1/Beclin-1 pathway regulated cytokine secretion in ALS astrocytes, which was able to lead to noncell autonomous toxicity. Autophagy activation mitigated cytokine secretion by ALS astrocytes.

Background Systemic sclerosis (SSc) is an autoimmune disease with high morbidity and mortality characterized by fibrosis of the skin and internal organs. Some studies have investigated the use of stem cells to treat SSc. Herein, a systematic review and meta-analysis was conducted to determine the efficacy and safety of mesenchymal stem cells (MSCs) in the treatment of SSc. Methods PubMed, Embase, Cochrane Library, Web of Science, OVID, China National Knowledge Infrastructure and Wanfang databases were searched up to February 1, 2021. Literature screening, data extraction and quality assessment were conducted independently by two researchers in according to the inclusion and exclusion criteria. The discrepancies were resolved by a third researcher. Results A total of 9 studies encompassing 133 SSc patients were included in the study. Compared to the baseline after treatment with MSCs: 1. The modified Rodnan skin score (mRSS) was significantly reduced in patients with SSc ( P < 0.00001). 2. MSCs decreased the number of digital ulcer, mouth handicap scale, and visual analog scale of hand pain in SSc patients ( P = 0.0007 and P = 0.03, respectively). 3. No statistical differences were detected in Raynaud's condition score and Cochin hand function scale score at 6 months of MSCs therapy ( P = 0.5 and P = 0.62). 4. After 12 months of follow-up, MSCs improve carbon monoxide diffusing capacity and forced vital capacity of SSc patients ( P < 0.05). 5. Overall, MSCs application was safe; a few cases exhibited swelling at the injection site, diarrhea and arthralgia, which had self-recovery, and no severe adverse events occurred in the included trials. Conclusions MSC therapy improves the degree of skin thickening, lung function, and mouth opening and relieves finger ulcers and pain in patients with SSc without severe adverse events. Thus, MSCs or MSCs combined with plasma and traditional medicine might be an effective and promising treatment of SSc patients. PROSPERO registration number : CRD42020200350

Abstract Background: Dysregulated lipid metabolism, aetiology of which is unknown, causes astrocyte dysfunction in Alzheimer's disease (AD). This study assessed the potential of amyloid-beta 42 (Aβ42) to alter lipid metabolism-related variables in non-AD iPSC-derived APOE ε3/ε3 astrocytes (n=3), and the effects of exogenous activation and inhibition of mTOR-autophagy pathway on the lipid variables in Aβ-treated astrocytes. Methods and results: 24h and 48h Aβ treatments of astrocytes increased cellular GFAP and complement 3 protein and the culture medium concentrations of TNF-α, IL-1β, and IL-6. Lipid droplet (LD) sizes and number, and unesterified cholesterol levels were increased but fatty acid uptake was reduced. Cellular ApoE level was higher and lower after 24h and 48h Aβ treatments, respectively. 24h and 48h Aβ treatments increased cellular LC3B-II/LC3B-I ratio and p62. The modulatory effect of the mTOR-autophagy pathway on Aβ-induced lipids was tested. Relative to the 48h Aβ-treated astrocytes, treating the 24h Aβ-treated astrocytes with fresh medium for 24h increased the LC3BII/LC3BI ratio and p62, reduced LD number without altering the size, reduced ApoE, and increased fatty acid uptake and cholesterol. Autophagy activation further increased p62, reduced LC3B-II/LC3B-I ratio, decreased LD number, increased LD size and cholesterol, and reduced ApoE and fatty acid uptake. Autophagy inhibition further increased LC3B-II/LC3B-I ratio without altering p62, increased LD number, cholesterol and ApoE, reduced fatty acid uptake, but not LD size. Conclusion: Aβ42 alters inflammatory-state, lipid-related variables, and autophagy concurrently in astrocytes. The withdrawal of Aβ medium activates autophagy that could modulate the effects of exogenous mTOR-autophagy pathway activation and inhibition on lipids in Aβ-treated astrocytes.

Background Diabetes mellitus as a chronic metabolic disease is threatening human health seriously. Although numerous clinical trials have been registered for the treatment of diabetes with stem cells, no articles have been published to summarize the efficacy and safety of mesenchymal stem cells (MSCs) in randomized controlled trials (RCTs). Methods and findings The aim of this study was to systematically review the evidence from RCTs and, where possible, conduct meta-analyses to provide a reliable numerical summary and the most comprehensive assessment of therapeutic efficacy and safety with MSCs in diabetes. PubMed, Web of Science, Ovid, the Cochrane Library and CNKI were searched. The retrieval time was from establishment of these databases to January 4, 2020. Seven RCTs were eligible for analysis, including 413 participants. Meta-analysis results showed that there were no significant differences in the reduction of fasting plasma glucose (FPG) compared to the baseline [mean difference (MD) = -1.05, 95% confidence interval (CI) (-2.26,0.16), P<0.01, I-2 = 94%] and the control group [MD = -0.62, 95%CI (-1.46,0.23), P<0.01, I-2 = 87%]. The MSCs treatment group showed a significant decrease in hemoglobin (Hb) A1c [random-effects, MD = -1.32, 95%CI (-2.06, -0.57), P<0.01, I-2 = 90%] after treatment. Additionally, HbA1c reduced more significantly in MSC treatment group than in control group [random-effects, MD = -0.87, 95%CI (-1.53, -0.22), P<0.01, I-2 = 82%] at the end of follow-up. However, as for fasting C-peptide levels, the estimated pooled MD showed that there was no significant increase [MD = -0.07, 95%CI (-0.30, 0.16), P<0.01, I-2 = 94%] in MSCs treatment group compared with that in control group. Notably, there was no significant difference in the incidence of adverse events between MSCs treatment group and control group [relative risk (RR) = 0.98, 95%CI (0.72, 1.32), P = 0.02, I-2 = 70%]. The most commonly observed adverse reaction in the MSC treatment group was hypoglycemia (29.95%). Conclusions This meta-analysis revealed MSCs therapy may be an effective and safe intervention in subjects with diabetes. However, due to the limited studies, a number of high-quality as well as large-scale RCTs should be performed to confirm these conclusions.