Background Circulating metabolites have been associated with cross-sectional renal function in population-based research. Nevertheless, there is currently little proof to support the idea that metabolites either cause or prevent renal function. New treatment targets and ways to screen individuals with impaired renal function will be made possible via an in-depth analysis of the causal relationship between blood metabolites and renal function.Methods We assessed the causal relationship between 452 serum metabolites and six renal phenotypes (CKD, rapid progression to CKD [CKDi25], rapid eGFR decline [CKD rapid3], dialysis, estimated glomerular filtration rate, and blood urea nitrogen) using univariate Mendelian randomization, primarily employing the inverse variance weighted method with robust sensitivity analyses. Heterogeneity and pleiotropy were examined via Cochrane’s Q test and MR-Egger regression, and statistical significance was adjusted using Bonferroni correction. To assess potential adverse effects of metabolite modulation, we conducted a phenome-wide Mendelian randomization analysis, followed by multivariate Mendelian randomization to adjust for confounders.Results We identified glycine and N-acetylornithine as potential causal mediators of CKD and renal dysfunction. Notably, lowering glycine levels may increase the risk of cholelithiasis and cholecystitis, while reducing N-acetylornithine could have unintended effects on tinnitus.Conclusion Glycine and N-acetylornithine represent promising therapeutic targets for CKD and renal function preservation, but their modulation requires careful risk-benefit assessment to avoid adverse effects.

Recent epigenetic studies have revealed a strong association between DNA methylation and aging and lifespan, which changes (increases or decreases) with age. Based on these, the construction of age prediction models associated with DNA methylation levels can be used to infer biological ages closer to the functional state of the organism. We downloaded methylation data from the Gene Expression Omnibus (GEO) public database for normal peripheral blood samples from people of different ages. We grouped the samples according to age (18-35 years and >50 years), screened the methylation sites that differed between the two groups, identified 44 differentially methylated sites, and subsequently obtained 11 age-related characteristic methylation sites using the random forest method. Then, we constructed an age classification model with these 11 characteristic methylation sites using an artificial neural network and evaluated its efficacy. The age classification model was constructed by an artificial neural network and its efficacy was evaluated. The model predicted an area under the curve (AUC) of 0.97 in the validation set and accurately distinguished between those aged 18-35 and >50 years. Furthermore, the levels of these 11 characteristic methylation sites also differed significantly between the two sets of samples in the validation set, including six newly identified age-related methylation sites (P<0.001). Finally, we constructed a multifactor regulatory network based on the corresponding genes of age-related methylation sites to reveal the transcriptional and post-transcriptional regulation patterns. As a result of the increasing problem of aging, the age classification model we constructed allows us to accurately distinguish different age groups at the molecular level, which will be more predictive than chronological age for assessing individual aging and future health status.

BackgroundA decrease in the number and activity of thymic epithelial cells (TECs) is an important factor in thymic degeneration. Mesenchymal stem cells (MSCs) treating thymic ageing is a promising strategy, but the DNA methylation modification mechanism in TECs remains unclear.MethodsAged rhesus monkeys were treated with MSCs to establish a thymic senescence model, and hematoxylin-eosin (HE) staining, immunofluorescence staining, and ELISA were performed to observe the structure and function of the thymus. TEC aging model and MSCs co-culture system were established to detect DNA methylation modification and transcriptomic changes, correlation analysis between transcription factor methylation and mRNA expression, and q-PCR, immunofluorescence staining, and Western blot were used to identified key genes.ResultsMSCs improved the structure and function of thymus in elderly macaque monkeys; reduced the expression levels of β-Gal, P16, and P21; and increased the activity of aging TECs. There were 501 genes with increased methylation in the promoter region in the treated group compared with the untreated group, among which 23 genes were involved in the negative regulation of cell growth, proliferation and apoptosis, while 591 genes had decreased methylation, among which 37 genes were associated with promoting cell growth and proliferation and inhibiting apoptosis. Furthermore, 66 genes showed a negative correlation between promoter methylation levels and gene transcription; specifically, PDE5A, DUOX2, LAMP1 and SVIL were downregulated with increased methylation, inhibiting growth and development, while POLR3G, PGF, CHTF18, KRT17, FOXJ1, NGF, DYRK3, LRP8, CDT1, PRELID1, F2R, KNTC1 and TRIM3 were upregulated with decreased methylation, promoting cell growth.ConclusionMSCs improve the structure and function of aged thymus, which involves the regulation of DNA methylation profiles and a decrease in the methylation level of the transcription factor NGF to specifically upregulate KRT17 and FOXJ1 to promote the proliferation of TECs.

Background: Protein extracts from chicken egg whites were prepared by utilizing different solvents and we studied the differences in proliferation. Methods: The egg white extract was prepared by using lysate, phosphate-buffered saline (PBS), saline and pure water. The in vitro mixture experiment was carried out to observe the effect of different egg white preparations on the proliferation of cells. The samples were divided into the following groups: the control group with media, the original lysate group, the new lysate group, the PBS group, the saline group and the pure water group. Result: The study found that at final concentrations of 10%, 20%, 30%, 40% and 50%, the differences among the six groups were statistically significant (P less than 0.01, F greater than 100). The results of pairwise comparison showed that the proliferative effect of chicken egg albumin extract prepared by PBS was significantly higher than that of the medium at final concentrations of 20%, 30% and 50% (P less than 0.05). When the fetal bovine serum concentration was only 8%, 7% and 5%, the cell proliferation effect was better than that of the control group with 10% fetal bovine serum, indicating that the chicken egg white extract promoted cell proliferation. This result indicated that the best chicken egg albumin extract was obtained through PBS addition. The quantity of expensive fetal bovine serum could be considerably reduced by supplementing the media with chicken egg albumin extract. Among the solvents tested, PBS was the best solvent for preparing chicken egg albumin extract.

Abstract Background: A decreased number of thymic epithelial cells and the development of dysfunction of this population have been reported to be important factors in thymic degeneration, which can lead to thymus degeneration, proliferation defects and peripheral T-cell dysfunction. Previous research showed that umbilical cord mesenchymal stem cells can restore the structure and function of the aging thymus in vivo, but the specific mechanism is still unclear.Methods: We treated thymic epithelial cells with H2O2 to establish a cellular senescence model. We assessed the effect of umbilical cord mesenchymal stem cells on thymic epithelial cells and investigated the lncRNA, miRNA and mRNA profiles of these cells. Gene Ontology and Kyoto Genome Encyclopedia analyses were performed on these RNAs to identify key pathways.Results: Umbilical cord mesenchymal stem cells significantly reversed the H2O2-induced senescence of thymic epithelial cells. We identified 172 differentially expressed lncRNAs, 23 differentially expressed miRNAs and 272 differentially expressed mRNAs associated with the reversal of thymic epithelial senescence by umbilical cord mesenchymal stem cells. In addition, the PI3K-Akt signaling pathway was identified as a key signaling pathway that promotes cell proliferation by regulating the cell cycle. Finally, we constructed a lncRNA-associated competing endogenous RNA (ceRNA) network using matched miRNA, lncRNA, and mRNA expression profiles in the model dataset and altered miRNA expression profiles in umbilical cord mesenchymal stem cells.Conclusion: Umbilical cord mesenchymal stem cells have a protective effect against H2O2-induced thymic epithelial cell senescence. Our study provides new insights into the ceRNA-mediated gene regulation of thymic senescence progression and the mechanism of action of umbilical cord mesenchymal stem cells.

Multiple organ dysfunction syndrome (MODS) is a clinical syndrome characterized by the dysfunction of two or more systems or organs. This internal environment disorder occurs simultaneously 24 h after severe trauma, shock, or infection. MODS has a high fatality rate ranging from 20% to 100%.[1] In the development of MODS following severe trauma or infection, multiple organ system fail sequentially, involving the lungs, kidneys, liver, cardiovascular, central nervous system, gastrointestinal, immune, and the blood coagulation system. MODS has complex pathogenic factors, and there are four main pathogenesis hypotheses: (1) uncontrolled inflammation hypothesis, (2) ischemia–reperfusion injury, (3) the gastrointestinal hypothesis, and (4) the biphasic pre-excitation theory. Although the current clinical treatment methods of MODS, such as inflammatory factor antibodies, highly effective anti-coagulants, renal function replacement therapy, and other treatment methods, have been improved continuously, their effectiveness in MODS treatment is not ideal. Therefore, clinical MODS treatment is faced with great challenges [Figure 1].Figure 1: MSCs therapy for MODS. After the organ is severely hitted by trauma and infection, it produces cytokine storm, and then develops into MODS. MSCs therapy regulates immune balance and repairs organ damage through paracrine. The picture is drawn by BioRender software (https://app.biorender.com). AKI: Acute kidney injury; CD81: Cluster of differentiation 81; DNA: Deoxyribonucleic acid; M1: M1 macrophage; M2: M2 macrophage; miR: Micro ribonucleic acid; MODS: Multiple organ dysfunction syndrome. MSC: Mesencymal stromal/stem cells; MSC-Exos:Mesencymal stromal/stem cells exocrine; MSC-EVs: Mesencymal stromal/stem cells extracellular vesicles; IL-1β: Interlukin 1β; IL-10: Interlukin 10; ROS: Reactive oxygen species.Every organ in the body is made up of stem cell-derived cells. Theoretically, the stem cells and their derivatives can repair any tissue in the body that has been lost or damaged by disease or injury.[2] Mesenchymal stromal/stem cells (MSCs) have the potential for self-renewal and multi-lineage differentiation. They are favored for their strong paracrine ability, two-way immune regulatory ability, and tissue repair potential.[3] In MODS, the protective effect of beneficial local inflammation is transformed into harmful systemic inflammatory damage, and studies have shown that MSC has positive effects on the immune system, coagulation, and other systems. MSC therapy is thought to be an ideal remedy for such multifactorial complex systemic inflammatory disease.[4] Furthermore, MSCs and their derivatives have been shown to be effective in treating organ injury and dysfunction caused by trauma, infection, and other causes. Therefore, we reviewed the efficacy and mechanism of MSCs and their derivatives in treating different organ dysfunction to investigate MSCs’ prospects in treating MODS. MSCs and their derivatives therapy can secrete immunomodulatory factors, growth factors, and chemokines, promoting stem cell colonization in damaged tissue and tissue repair and inhibiting the body's inflammatory response. As a subtype of CD4+ T cells, helper T cell 17 (Th17)/regulatory T cell (Treg) can regulate pulmonary inflammation and alleviate lung injury by regulating the imbalance of Th17/Treg in the lungs of acute respiratory distress syndrome mice after transforming growth factor β-1 overexpression in MSCs.[5] The expression of pro-inflammatory cytokines in knee synovial fluid decreased after treatment with bone marrow (BM)-derived MSCs- exosomes (MSC-Exos) in a rat osteoarthritis model, while the release of anti-inflammatory cytokines increased, inflammatory M1 macrophage production decreased, and anti-inflammatory M2 macrophage production increased; cartilage injury was alleviated, and osteophyte formation and synovial macrophage infiltration decreased, thereby relieving osteoarthritis.[6] Lung tissue injury is repaired by MSCs through different mechanisms, and respiratory function is improved. Researchers discovered that the expression of miR-193b-5p target gene tight junction protein antibody (Ocln) increased in lung tissue after treatment of a septic mouse model with MSC.[7] The MSC-Exos-mediated miR-23a-3p and miR-182-5p can prevent lipopolysaccharide-induced lung injury in mice by silencing Ikbkb and IKK β and inhibiting nuclear factor kappa-B and hedgehog pathways.[8] miR-27A-3p is an important regulator of M2 macrophage polarization. MSCs and MSCs-extracellular vesicles (MSC-EVs) can transfer miR-27A-3p to alveolar macrophages, increasing the miR-27A-3p level in alveolar macrophages, promoting M2 cell polarization, and reducing acute lung injury (ALI).[9] Most current research on improving MSCs function in the brain is focused on the neural circuit repair, neural plasticity, and astrocyte correlation. MSC-Exos can significantly improve sensorimotor and cognitive impairment, reduce hippocampal nerve cell loss, promote angiogenesis and nerve regeneration, and reduce neuroinflammation.[10] In the scratch injury model of human astrocytes (T98G cells), human adipose mesenchymal stem cells (CM-hMSCA) were found to regulate the expression of different proteins in AKT/pAKT and ERK1/2/pERK signal pathways, mediate the cellular localization of Ngb, decrease cytosolic calcium concentration, regulate mitochondrial dynamics and the respiratory chain, and decrease expression of astrocyte activation regulator cluster of differentiation 81 (CD81).[11] MSCs and their derivatives may become an important treatment option for acute kidney injury (AKI). MSCs have been shown to regulate heat shock proteins 20 and phosphatidylinositol-3-kinase/Akt signal pathway in the treatment of AKI.[12] MSC-EVs act on the mitochondrial transcription factor A (TFAM) pathway in a mouse AKI model, restoring the stability of TFAM protein and the TFAM-mtDNA complex in damaged renal tubular cells, reversing mitochondrial DNA deletion and mitochondrial oxidative phosphorylation (OXPHOS) deficiency, and reducing mitochondrial DNA damage and inflammation.[13] BM-derived MSC-EVs can increase microRNA-200a-3p expression in proximal renal tubular epithelial cells of mice with renal ischemia–reperfusion injury and activate Keap1-Nrf2 signal pathway, stimulate mitochondrial antioxidant defense and adenosine triphosphate production, reduce oxidative damage of proximal renal tubular epithelial cells, and achieve a therapeutic effect on AKI.[14] The in vivo evolution of stem cells and the safety of stem cell therapy remain the focus of research. Although researchers have discovered that coagulopathy, thromboprophylaxis, and mode of delivery play an important role in the safety and efficacy of treatment,[15] the safety of MSCs transplantation under certain conditions remains unknown. Some researchers have investigated various cell therapy delivery platforms and biomaterials to increase survival rate and time, and improve treatment efficiency. Simultaneously, researchers also focused on its large-scale and standardized production. Overall, MSC and their derivatives’ high proliferation characteristics, treatment timing, and cell microenvironment conditions need to be explored further [Figure 1]. This study focuses primarily on the stem cell mechanism in treating trauma and inflammation-induced MODS. MSCs and their derivatives are a good option for reducing MODS morbidity and mortality. Therefore, incorporating stem cell therapy into existing clinical treatment methods may become an effective breakthrough in reducing MODS mortality. Funding This work was supported by grants from the all-army laboratory animal project (No. SYDW[2020]19), the General Project of Yunnan Applied Basic Research Program (No. 202101AT070212), and the 920th Hospital of the PLA Joint Logistics Support Force In-Hospital Technology Plan (No. 2020YGD05). Conflicts of interest None.

背景:卵巢作为女性生殖系统的核心器官,通过排卵与分泌激素行使女性生殖功能并影响全身的组织器官,随着人口老龄化加速以及多种诱因导致卵巢衰老人群不断增加,但卵巢衰老在整体上还处于初步研究阶段,依旧是尚待解决的科学问题.目的:探索骨髓间充质干细胞对老年猕猴卵巢衰老的修复作用.方法:健康老年雌猴10只,年龄23-27岁,体质量4.5-8.0 kg,随机分为老年对照组(n=4),老年治疗组(n=6).第4代幼年猕猴骨髓间充质干细胞经股静脉输注到老年猕猴体内,1次/d,隔日输注,连续输注3次;老年对照组猕猴同一时间输注等体积生理盐水.在输注细胞的第8个月,安乐死处理猕猴取卵巢组织,称质量,40 g/L多聚甲醛固定,苏木精-伊红染色观察各级卵泡和卵巢结构变化;Tunel染色统计细胞凋亡率;Masson染色分析胶原蛋白所占面积比;免疫组织化学法观察CD34标记阳性血管数量.结果 与结论:①老年对照组卵巢脏器指数为1.8×10-5,老年治疗组卵巢脏器指数为6.0×10-5,两组比较差异有显著性意义(P<0.05);②苏木精-伊红染色结果显示老年治疗组可见原始、初级、次级、闭锁卵泡,未见成熟卵泡,髓质与间质明显,有少量钙结节;老年对照组基本无卵泡结构,局部只见闭锁卵泡,有大量脂肪组织填充;③Masson染色结果显示老年治疗组卵巢组织胶原纤维所占面积比明显低于老年对照组,两组比较差异有显著性意义(P<0.05);④Tunel染色结果显示老年治疗组细胞凋亡率明显低于老年对照组,两组比较差异有显著性意义(P<0.05);⑤免疫组织化学结果显示老年治疗组血管数明显多于老年对照组,两组比较差异有显著性意义(P<0.05).⑥结果表明,骨髓间充质干细胞能够促进卵泡再生、改善卵巢组织结构、减少卵巢细胞凋亡、抑制卵巢纤维化进展、促进血管再生,以延缓甚至逆转卵巢衰老.

tRNA源性片段(tRNA-derived fragments,tRFs)和tRNA源性应激诱导RNAs(tRNA-derived stress-induced RNAs,tiRNAs)是tRNAs的衍生片段,属于短的非编码RNA家族,通过转录、翻译、信号通路等途径参与复杂的生物反应.该文旨在验证鸡卵清提取液诱导293T细胞后升高的3个tRFs&tiRNAs分子的细胞功能.将293T细胞加于6孔板中,3个孔加普通培养基,3个孔加50％鸡卵清提取液的培养基,共培养3天.对照组3个样本和诱导组3个样本进行高通量测序检测tRFs&tiRNAs分子在两组中的差异表达.经检测验证诱导后的细胞有3个tRFs&tiRNAs分子稳定升高.这3个分子上调表达有统计学意义.合成这3个分子转染293T细胞,WB检测多能因子OCT4和NANOG的变化,定量PCR检测多能基因OCT4和NANOG的变化和端粒的相对表达量,流式检测多能因子OCT4和NANOG的变化.同时检测这3个分子转染293T细胞后细胞增殖、细胞凋亡和细胞周期的变化情况.结果 表明,3个分子转染293T细胞后,WB检测到多能因子OCT4和NANOG表达对比未转染细胞明显升高,定量PCR检测多能基因OCT4和NANOG相对表达量对比未转染细胞明显升高,端粒对比未转染细胞明显增长.流式检测到多能因子OCT4和NANOG阳性表达细胞对比未转染细胞明显增多.这3个分子转染293T细胞后细胞活性增强,细胞凋亡减少,细胞周期也发生了一定的改变.该研究证明了这3个分子过表达可促进293T细胞多能因子OCT4和NANOG表达升高,促进端粒增长,使细胞年轻化.同时这3个分子过表达可使细胞活性增强,细胞凋亡减少.

Islet transplantation is arguably one of the most promising strategies to treat patients suffering with diabetes mellitus. However, a combination of a lack of donors and chronic immune rejection limit clinical applications. Here, we evaluated the efficacy of cell therapy using islet-like cells differentiated from umbilical cord mesenchymal stem cells (UC-MSCs) of tree shrews for the treatment of type 2 diabetes. Enhanced green fluorescent protein (eGFP) labeled UC-MSCs were directly injected into type 2 diabetic tree shrews, where UC-MSC differentiated into functional islet-like cells and alleviated disease severity, as evidenced by improved biochemical features and reduced concentrations of inflammatory cytokines. We also demonstrated that in vitro culture of UC-MSCs for six days in a high-glucose environment (40 mmol/L or 60 mmol/L glucose) resulted in significant gene methylation. The potency of UC-MSCs differentiated into insulin-secreting cells was attributed to the activation of Notch signal pathways. This study provides evidence that cell therapy of islet-like cells differentiated from UC-MSCs is a feasible, simple and inexpensive approach in the treatment of type 2 diabetes.

Background: Chronic kidney disease (CKD) is an independent risk factor for cardiovascular disease (CVD). However, the association between CKD and CVD risk in patients with type 2 diabetes mellitus (T2DM) in China has not yet been well investigated. This study aimed to determine the association of CKD with the risks of coronary heart disease (CHD) and stroke in a Chinese population with T2DM. Methods: A total of 1401 inpatients with T2DM at the Second Affiliated Hospital of Zhejiang University School of Medicine between April 2008 and November 2013 were included in this study. The CKD-Epidemiology Collaboration equation for Asians was used to classify CKD. The UK Prospective Diabetes Study risk engine was used to estimate the risks of CHD and stroke. Results: CHD risk was significantly increased with CKD stage (20.1%, 24.8%, and 34.3% in T2DM patients with no CKD, CKD Stage 1–2, and Stage 3–5, respectively; P < 0.001 for all). The stroke risk was also increased with CKD stage (8.6%, 12.7%, and 25.4% in T2DM patients with no CKD, CKD Stage 1–2, and Stage 3–5, respectively; P < 0.001 for all). Compared with no-CKD group, the odds ratios (OR s) for high CHD risk were 1.7 (P < 0.001) in the CKD Stage 1–2 group and 3.5 (P < 0.001) in the CKD Stage 3–5 group. The corresponding OR s for high stroke risk were 1.9 (P < 0.001) and 8.2 (P < 0.001), respectively. Conclusion: In patients with T2DM, advanced CKD stage was associated with the increased risks of CHD and stroke.

Carotid atherosclerosis (CA) and carotid plaque (CP) are highly correlated with cardiovascular disease. We aimed to determine the prevalence of CA and CP and their relationship with 10-year risks of stroke and coronary heart disease (CHD) in type 2 diabetes mellitus (T2DM). We studied 1584 T2DM patients aged 20 years and older. CA and CP were detected using ultrasonography. Ten-year stroke and CHD risk were determined using the United Kingdom Prospective Diabetes Study (UKPDS) risk engine. The prevalence of CA and CP increased gradually with age. Men had a higher prevalence of CA than women (CA: 58.18% vs 51.54%, P<.01). The 10-year CHD risk (27.9% vs 15.4%, P<.001) and stroke risk (15.2% vs 5.70%, P<.001) were higher in patients with CA than that of those without CA. Compared with patients without CA, the odds ratios (ORs) of CHD in CA and CP group were 4.47 and 10.78 for men, and 4.19 and 5.20 for women, respectively; in the case of stroke, the OR in CA and CP group were 8.83 and 12.07 for men, and 4.35 and 4.90 for women, respectively (P<.001 for all). Multivariate binary logistic regression analysis showed that CA was an independent risk factor for CHD [OR=2.66, 95% confidence interval (95% CI), 2.05-3.46, P<.001] and stroke (OR=3.11, 95% CI, 2.38-4.07, P<.001). CA and CP were prevalent in patients with T2DM and positively correlated with 10-year CHD and stroke risk. CA was an independent risk factor for 10-year CHD risk.

Coronary heart disease (CHD) and stroke are common complications of type 2 diabetes mellitus (T2DM). We aimed to explore the differences in the risks of CHD and stroke between Chinese women and men with T2DM and their association with metabolic syndrome (MS). This study included 1514 patients with T2DM. The Asian Guidelines of ATPIII (2005) were used for MS diagnosis, and the UKPDS risk engine was used to evaluate the 10-year CHD and stroke risks. Women had lower CHD risk (15.3% versus 26.3%), fatal CHD risk (11.8% versus 19.0%), stroke risk (8.4% versus 10.3%), and fatal stroke risk (1.4% versus 1.6%) compared with men with T2DM (p < 0.05–0.001). The CHD risk (28.4% versus 22.6%, p < 0.001) was significantly higher in men with MS than in those without MS. The CHD (16.2% versus 11.0%, p < 0.001) and stroke risks (8.9% versus 5.8%, p < 0.001) were higher in women with MS than in those without MS. In conclusion, our findings indicated that Chinese women with T2DM are less susceptible to CHD and stroke than men. Further, MS increases the risk of both these events, highlighting the need for comprehensive metabolic control in T2DM.