Ovarian tissue cryopreservation is a vital fertility preservation technique. However, cryopreservation-induced injury significantly limits its efficacy in frozen-thawed ovarian tissue transplantation. This study investigated the therapeutic potential of umbilical cord mesenchymal stem cells (UC-MSCs) and their derived extracellular vesicles (MSC-EVs) for repairing cryopreservation-induced ovarian injury via ferroptosis in an in vitro culture system. RNA sequencing revealed ferroptosis as a key regulatory pathway in cryopreserved ovarian tissue. This was characterized by downregulation of GPX4, upregulation of ACSL4, elevated Fe2+ levels, glutathione depletion, and enhanced lipid peroxidation. UC-MSCs and MSC-EVs effectively suppressed ferroptosis, restored GPX4 expression, and reduced lipid peroxidation, mirroring the effects of the ferroptosis inhibitor ferrostatin-1. In an in vitro culture system, UC-MSCs and MSC-EVs enhanced follicular survival and angiogenesis, while reducing apoptosis. Transplantation experiments demonstrated improved graft vascularization, hormone secretion, and oocyte retrieval in treated groups. Notably, MSC-EVs demonstrated comparable efficacy to UC-MSCs, suggesting their potential as a nanoscale therapeutic strategy. These findings elucidate ferroptosis as a critical mechanism in cryopreservation-induced injury and highlight UC-MSCs and MSC-EVs as promising therapeutic agents for improving ovarian tissue cryopreservation outcomes.

TCAB1 (telomerase Cajal body protein 1), encoded by the WRAP53 gene on chromosome 17p13.1, is a molecular scaffold critical for protein-nucleic acid interactions. In normal cells, TCAB1 plays a pivotal role in localizing telomerase to Cajal bodies, thereby ensuring proper telomere maintenance and genomic stability. In cancer cells, however, TCAB1 is frequently overexpressed, which supports unchecked proliferation and therapy resistance. Conversely, knockdown of TCAB1 triggers multiple tumor-suppressive mechanisms, including G1 cell cycle arrest - mediated by impaired p21 ubiquitination and subsequent Cyclin E/CDK2 inactivation - as well as telomere shortening and genomic instability due to mitochondrial dysfunction and defective DNA repair. Notably, the induction of cellular senescence emerges as a key anticancer mechanism upon TCAB1 depletion, particularly in early-stage tumors retaining wild-type p53. This review delineates the dual roles of TCAB1, highlighting its function as a context-dependent oncoprotein and the therapeutic potential of targeting it to induce senescence.

Background: Systemic inflammatory response syndrome (SIRS) is a systemic nonspecific inflammatory response to severe injury caused by infectious or noninfectious factors. Methods: The impact method was used to make unilateral femoral comminuted fractures in tree shrews and lipopolysaccharide was injected intravenously to create a traumatic systemic inflammatory response syndrome model. The treatment group was infused with tree shrew umbilical cord mesenchymal stem cells via the tail vein. Result: After 10 days of treatment, 3 tree shrews were killed in each of the control group, the model group and the treatment group and pancreatic tissue was taken for RNA sequencing analysis. The results showed statistically significant changes in some genes. The two genes with the most significantly changed expression levels were ENSTBEG00000013408 (gene name TMEM211) and ENSTBEG00000006837 (gene name C1QL3). Their expression levels were increased in the model group and decreased after treatment. The changes in inflammatory factors indicate that the model group has an obvious inflammatory response and the treatment of umbilical cord mesenchymal stem cells has the effect of reducing inflammatory factors, which further proves the anti-inflammatory effect of umbilical cord mesenchymal stem cells. There was also an obvious change in the expression of the ENSTBEG00000021330 gene (gene name ATP8), which decreased in the model group and increased after treatment, indicating that ATP8 is a gene that contributes to the recovery of systemic inflammatory response syndrome. In the future, we may use this gene to treat systemic inflammatory response syndrome.

OBJECTIVE:The regulatory ability of bone marrow stem cells (BMSC) to chemokines and inflammatory factors has a significant effect in a variety of diseases. It is very important to delay the aging of BMSC and restore the function of aging BMSC. METHODS:Mouse BMSC was prepared and identified. TCAB1 gene interference (Sh-TCAB1), interference control (Sh-NC), overexpression (OE-TCAB1) and overexpression control (OE-NC) stable cell lines of BMSC were established, and the relationship between TCAB1 expression and senescence of BMSC cells was analyzed. Transcriptome high-throughput sequencing was performed to further analyze the mechanism of TCAB1 in BMSC aging. RESULTS:The phenotype of BMSC was normal by flow cytometry, and the cultured cells were identified as BMSC by osteogenic lipogenic differentiation staining. The fluorescence transfection efficiency of TCAB1-interfered and overexpressed stable strain was 90%, and the stable strain of interfered and overexpressed TCAB1 gene was successfully constructed. Overexpression of TCAB1 inhibits BMSC senescence, and TCAB1 interferes with and accelerates BMSC senescence. Transcriptome sequencing results show that TCAB1 can regulate signaling pathways related to BMSC metabolism and cell cycle to play an anti-BMSC senescence role. CONCLUSION:Transcriptome sequencing suggests that the mechanism of TCAB1 inhibiting BMSC senescence is related to cell cycle and metabolic process, and exerts anti-BMSC senescence function by regulating the expression of key factors Slc2a1 and Egln3. This study confirmed that the expression of TCAB1 is closely related to the senescence of BMSC through molecular level, which provides a new technique for the clinical treatment of cell senescence.

Background: Natural killer (NK) cells are an important part of the immune system and play a central role in cell-mediated immune responses. Recent studies have shown that NK cells promote organ rejuvenation, thus achieving anti-aging effects from the inside to out. Methods: We amplified and cultured human peripheral blood NK cells and the positive rate reached 43.91%. After purification, the positive rate reached 92.65%. The cultured NK showed a good killing effect on tumor cells and the NK cells reached the standard preparation scale. The NK cells cultured by us were co-cultured with the PBMC of aging macaques and the quantitative PCR and WB results showed that the expression of aging gene and aging protein of the PBMC of aging macaques decreased significantly after co-culture, with statistical significance (P less than 0.01). Result: These results indicate that NK cells prepared with standard preparation have anti-aging effect, which will have extensive significance in preclinical research.

Background: The impact method was used to make unilateral femoral comminuted fractures in tree shrews, which were then intravenously injected with lipopolysaccharide to create a traumatic systemic inflammatory response syndrome model. Methods: The treatment group was infused with tree shrew umbilical cord mesenchymal stem cells via the tail vein. After 10 days of treatment, 6 tree shrews were killed in the control, model and treatment groups and pancreatic tissue was collected for metabonomic analysis. Result: The 25 differential metabolites in the normal, model and stem cell treatment groups showed a trend of first decreasing and then increasing and 11 metabolites showed a trend of first increasing and then decreasing. These findings are in line with the recovery of the curative effect of the stem cell therapy model group.

Background: Stem cell protein chips can detect stem cell-associated proteins expressed in cells. Methods: We isolated and cultured rabbit peripheral blood mononuclear cells (R-PBMCs) in two bottles, one containing medium and one containing medium with 40% chicken egg white extract. Similarly, 293T cells were also divided into two vials: one vial contained medium and one vial contained medium with 40% chicken egg white extract. After 3 days of culture, proteins were extracted from the cells and used for the stem cell protein chip. Result: Protein chip detection showed that 10 stem cell-associated proteins were increased in the rabbit peripheral blood mononuclear cells induced by chicken egg white extract. There were also 4 stem cell-associated proteins increased in the 293T cells induced by chicken egg white extract. The results of KEGG enrichment analysis indicated that peripheral blood mononuclear cells of rabbits were involved in the regulation of stem cell pluripotency before and after induction. Moreover, 293T cells were involved in the regulation of stem cell pluripotency before and after induction.

Background Recent studies have shown that umbilical cord mesenchymal stem cells have an anti-aging effect in ovaries, but the cellular and molecular mechanisms of HA-MSC ovarian anti-aging remain to be studied. Therefore, we conducted a 10X Genomics single-nucleus transcriptome sequencing experiment on the ovaries of macaque monkeys after HA-MSC treatment. Methods The results of cell subgroup classification were visualized by 10X Genomics single nuclear transcriptome sequencing. The aging model of hGCs was established, and the migration ability of the cells was determined after coculture of HA-MSCs and aging hGCs. The genes screened by single nuclear transcriptional sequencing were verified in vitro by qPCR. Results Compared with the aging model group, the number of cell receptor pairs in each subgroup of the HA-MSC-treated group increased overall. Treatment with 200 μmol/L H 2 O 2 for 48 h was used as the optimum condition for the induction of hGC senescence. After coculture of noncontact HA-MSCs with senescent hGCs, it was found that HA-MSCs can reverse the cell structure, proliferation ability, senescence condition, expression level of senescence-related genes, and expression level of key genes regulating the senescence pathway in normal hGCs. Conclusions HA-MSC therapy can improve the tissue structure and secretion function of the ovary through multiple cellular and molecular mechanisms to resist ovarian aging. In vitro validation experiments further supported the results of single-cell sequencing, which provides evidence supporting a new option for stem cell treatment of ovarian senescence.

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.

Ovarian senescence is a special type of organ senescence and the ovaries are the earliest aging organs. The ovaries have approximately 1 million to 2 million follicles at birth, but only approximately 1000 primordial follicles are left in menopause. Ovarian function also decreases with age. Women’s fertility also declines. The ovaries are the core female reproductive organs and are of great significance for maintaining reproductive system function and endocrine stability. Ovarian aging is also considered an indicator of female body aging, which drives the aging of many organs of the body. Therefore, how to prevent and treat ovarian aging has become a research question that has been widely studied by biomedical scientists and geriatric researchers in recent years. Recently, studies have shown that bone marrow mesenchymal stem cells (BMSCs) can prevent and treat ovarian aging. This article reviews the characteristics of ovarian aging, the advantages and disadvantages of various clinical treatment measures and the advantages of bone marrow mesenchymal stem cell therapy, aiming to provide references for the prevention and treatment of ovarian aging. My article was written at the Basic Medical Laboratory of the 920th Hospital of the Joint Logistics Support Force of PLA and written from 2021 to 2022.

Background: Umbilical cord mesenchymal stem cell transplantation can treat metabolic syndrome, but the tracing of cells in the body after transplantation has always been a problem. Tree shrew umbilical cord mesenchymal stem cells were labeled with the dark red fluorescent dye DIR and a metabolic syndrome model in tree shrew was generated. The migration, distribution, colonization and survival of the cells were observed. Methods: Tree shrew umbilical cord mesenchymal stem cells were labeled with the dark red fluorescent dye DIR. Three days after the tree shrew model was generated, the pancreas, kidney and liver were placed in a small animal live imager to observe the distribution of the labeled cells. Result: The labeled cells showed deep red fluorescence in the live imager. After treatment with the transplanted cells, dark red fluorescent signals were observed in the liver, kidney and pancreas of the tree shrews but not in the untreated group and no dark red fluorescent signal was observed in the cell distribution.

Abstract Background The tumorigenesis of infused umbilical cord mesenchymal stem cells (UC-MSCs) is being preclinically evaluated. Methods We observed tumor formation in NOD SCID mice after a single subcutaneous injection of hUC-MSCs and the effect of these cells on tumor growth in tumor-bearing mice. Three generations (P5, P7, and P10) of hUC-MSCs (1 × 107) from two donors (hUC-MSC1 and hUC-MSC2) were inoculated subcutaneously into NOD SCID mice. Subcutaneous transplantation models were established in NOD SCID mice with human cervical cancer HeLa cells (solid tumor) and human B cell lymphoma Raji cells (hematological tumor). Then, the animals were euthanized, gross dissection was performed, and tissues were collected. Various organs were observed microscopically to identify pathological changes and tumor metastasis. Results In the tumorigenesis experiment, no general anatomical abnormalities were observed. In the tumor promotion experiment, some animals in the HeLa groups experienced tumor rupture, and one animal died in each of the low- and medium-dose hUC-MSC groups. The results may have occurred due to the longer feeding time, and the tumor may have caused spontaneous infection and death. Pathological examination revealed no metastasis to distant organs in any group. In the Raji tumor model, some animals in each group experienced tumor rupture, and one animal in the medium-dose hUC-MSC group died, perhaps due to increased tumor malignancy. Thus, hUC-MSCs neither promoted nor inhibited tumor growth. No cancer cell metastasis was observed in the heart, liver, spleen, lungs, kidneys or other important organs, except that pulmonary venule metastasis was observed in 1 animal in the model group. Conclusions Injected hUC-MSCs were not tumorigenic and did not significantly promote or inhibit solid or hematological tumor growth or metastasis in NOD SCID mice.

背景:战创伤防治研究一直是军事医学领域的核心内容,建立可重复的针对战创伤性感染、全身炎症反应综合征、休克、多器官衰竭动物模型及以该动物模型为基础的新型治疗手段具有重大军事及科学意义.目的:探讨脐带间充质干细胞治疗创伤性全身炎症反应综合征的疗效.方法:50只树鼩中选取40只采用撞击法制作单侧股骨粉碎性骨折模型后静脉注射脂多糖创建创伤性全身炎症反应综合征模型,另外10只做为正常对照组.造模后10 d,选取20只经尾静脉回输GFP标记的树鼩脐带间充质干细胞,18只不回输细胞做为模型对照组.细胞移植后2,10 d时,取材检测GFP标记细胞在各组树鼩各脏器内的分布;细胞移植后20 d,取材制备病理切片观察各脏器病变情况,采外周静脉血检测肝功能、肾功能和心功能各指标变化.结果与结论:①移植后2,10 d时,各脏器中均观察到GFP阳性细胞分布,其中移植后2 d时胰腺组织中荧光细胞分布较明显,移植后10 d时肝脏组织中GFP阳性细胞分布较多;②移植后20 d时,模型对照组各脏器苏木精-伊红染色结果可见细胞坏死,炎症细胞浸润,治疗组接近正常对照组;治疗组肝功能、肾功能和心功检测结果恢复正常;③结果表明,树鼩脐带间充质干细胞治疗创伤性全身炎症反应综合征有一定的疗效,为现代武器创伤治疗提供了新技术方案.

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.

Based on the characteristics of modern weapon injury, a repetitive model of traumatic systemic inflammatory response syndrome (SIRS) and an evaluation system were established. The models were treated with GFP-labeled tree shrew umbilical cord mesenchymal stem cells (UCMSCs). Forty out of 50 tree shrews were used to make a unilateral femoral comminuted fracture. Lipopolysaccharide was injected intravenously to create a traumatic SIRS model. The other 10 shrews were used as normal controls. After the model was established for 10 days, 20 tree shrews were injected intravenously with GFP-labeled UCMSCs, and 18 tree shrews were not injected as the model control group. The distribution of GFP-labeled cells in vivo was measured at 2 and 10 days after injection. Twenty days after treatment, the model group, the normal control group, and the treatment group were taken to observe the pathological changes in each tissue, and blood samples were taken for the changes in liver, renal, and heart function. Distribution of GFP-positive cells was observed in all tissues at 2 and 10 days after injection. After treatment, the HE staining results of the treatment group were close to those of the normal group, and the model group had a certain degree of lesions. The results of liver, renal, and heart function tests in the treatment group were returned to normal, and the results in the model group were abnormally increased. UCMSCs have a certain effect on the treatment of traumatic SIRS and provide a new technical solution for modern weapon trauma treatment.

背景:系统性红斑狼疮是一种原因不明的自身免疫疾病,建立系统性红斑狼疮树鼩模型,有利于弄清其发病机制,并为干细胞移植治疗自身免疫疾病提供依据.目的:建立系统性红斑狼疮树鼩模型以及脐带间充质干细胞移植的治疗效果评估.方法:树鼩分组腹腔注射降植烷、脂多糖和联合注射,注射后3周筛选成模树鼩12只分为治疗组和模型对照组各6只,另选6只为正常对照组.治疗组每只树鼩尾静脉回输1×106个DiR标记的树鼩脐带间充质干细胞,回输后2周取树鼩的心、肝、脾、肺、肾做病理切片进行苏木精-伊红染色,肾组织进行Masson染色和免疫复合物检测,同时取3组树鼩的心、肝、脾、肺、肾进行体外成像.结果与结论:①苏木精-伊红染色表明模型对照组心、肝、脾、肺、肾有不同程度的病理改变,肾组织有大量免疫复合物沉积,治疗组病理改变好转,结构恢复到接近正常对照组;②体外成像表明DiR标记细胞主要分布于治疗组树鼩的肺、肝和脾,治疗组树鼩的荧光强度值明显大于正常对照组和模型对照组,差异有显著性意义(P<0.05);③结果表明,腹腔联合注射降植烷和脂多糖是诱导树鼩发生系统性红斑狼疮病理改变的最佳方法,用脐带间充质干细胞治疗后病理改变发生好转,说明脐带间充质干细胞对树鼩系统性红斑狼疮模型有一定的治疗效果.

背景:目前代谢综合征的治疗主要是改善生活和药物为主的综合性治疗,这些治疗要求患者有较强的随访性且不良反应多,尚不能从根本上改善、延缓代谢综合征发展.目的:探讨树鼩脐带间充质干细胞移植治疗树鼩代谢综合征的疗效,为临床应用干细胞移植治疗代谢综合征提供理论基础及参考方法.方法:实验采用贴壁培养法获得树鼩脐带间充质干细胞,鉴定其符合脐带间充质干细胞的生物学特性,深红色荧光碘化物DIR标记树鼩脐带间充质干细胞.32只树鼩给予高糖高胆固醇高盐饲料及糖水饮食,联合注射链脲佐菌素诱导代谢综合征模型,随机分为模型对照组(n=10)和脐带间充质干细胞治疗组(n=22).治疗组树鼩经尾静脉注射体外标记的脐带间充质干细胞,模型对照组树鼩于同一时间注入等体积的生理盐水.移植后进行血液生化指标、葡萄糖耐量、胰岛素抵抗指数和动脉血压检测.结果与结论:①成功构建树鼩代谢综合征模型,表现出明显的胰岛素抵抗、高血糖、脂代谢紊乱、高血压,符合代谢综合征诊断标准;②脐带间充质干细胞移植能够显著降低代谢综合征树鼩血糖、血脂,改善胰岛素抵抗,调节胰岛素分泌;③移植的脐带间充质干细胞可归巢至代谢综合征树鼩肝脏、肾脏及胰腺组织内,并产生一定的修复作用.

Ischemia-reperfusion injury is an important contributor to acute kidney injury and a major factor affecting early functional recovery after kidney transplantation. We conducted this experiment to investigate the protective effect of induced multipotent stem cell transplantation on renal ischemia-reperfusion injury. Forty rabbits were divided into four groups of 10 rabbits each. Thirty rabbits were used to establish the renal ischemia-reperfusion injury model, and ten rabbits served as the model group and were not treated. Among the 30 rabbits with renal ischemia-reperfusion injury, 10 rabbits were treated with induced peripheral blood mononuclear cells (PBMCs), and 10 other rabbits were treated with noninduced PBMCs. After three weekly treatments, the serum creatinine levels, urea nitrogen levels and urine protein concentrations were quantified. The kidneys were stained with hematoxylin-eosin (HE), periodic acid-Schiff (PAS) and Masson’s trichrome and then sent for commercial metabolomic testing. The kidneys of the rabbits in the model group showed different degrees of pathological changes, and the recovery of renal function was observed in the group treated with induced cells. The results indicate that PBMCs differentiate into multipotent stem cells after induction and exert a therapeutic effect on renal ischemia-reperfusion injury.

BACKGROUND:To study the effect of allogeneic umbilical cord mesenchymal stem cell transplantation on the structure and function of the thymus in aged C57 mice and provide a new method for the treatment of senile thymic atrophy.RESULTS:The changes in the thymus cortex and medulla volume and the lymphocyte ratio were analyzed by immunofluorescence. For thymus tissue sections, immunohistochemical staining was performed to detect p16, p53, SOD, becline1, LC3b, p62, sirt1, and sirt3. Changes in CK5, CK8, CD4 and CD8 expression were observed. Treatment with mUCMSCs could promote hair regeneration in aging mice and regenerate the thymus structure.CONCLUSIONS:mUCMSCs inhibited senescence of the thymus and promoted structural and functional thymus regeneration by downregulating the senescence genes p53 and p16 and upregulating the SOD, Sirt1 and Sirt3 genes, but the mechanism requires further research.METHODS:C57 mice were obtained and met the requirements of thymic aging. mUCMSCs were infused via the tail vein at a dose of 1×107 cells/kg twice per week for 3 weeks. Six weeks after the last transplantation, the thymus was weighed, and the thymus-to-body weight ratio was calculated. The thymus tissue was stained with HE.