A multi-channel collagen scaffold loaded with neural stem cells for the repair of spinal cord injury

Collagen scaffolds possess a three-dimensional porous structure that provides sufficient space for cell growth and proliferation,the passage of nutrients and oxygen,and the discharge of metabolites.In this study,a porous collagen scaffold with axially-aligned luminal conduits was prepared.In vitro biocompatibility analysis of the collagen scaffold revealed that it enhances the activity of neural stem cells and promotes cell extension,without affecting cell differentiation.The collagen scaffold loaded with neural stem cells improved the hindlimb motor function in the rat model of T8 complete transection and promoted nerve regeneration.The collagen scaffold was completely degraded in vivo within 5 weeks of implantation,exhibiting good biodegradability.Rectal temperature,C-reactive protein expression and CD68 staining demonstrated that rats with spinal cord injury that underwent implantation of the collagen scaffold had no notable inflammatory reaction.These findings suggest that this novel collagen scaffold is a good carrier for neural stem cell transplantation,thereby enhancing spinal cord repair following injury.This study was approved by the Animal Ethics Committee of Nanjing Drum Tower Hospital(the Affiliated Hospital of Nanjing University Medical School),China(approval No.2019AE02005)on June 15,2019.

Induced pluripotent stem cell technology for spinal cord injury: a promising alternative therapy

Spinal cord injury has long been a prominent challenge in the trauma repair process. Spinal cord injury is a research hotspot by virtue of its difficulty to treat and its escalating morbidity. Furthermore, spinal cord injury has a long period of disease progression and leads to complications that exert a lot of mental and economic pressure on patients. There are currently a large number of therapeutic strategies for treating spinal cord injury, which range from pharmacological and surgical methods to cell therapy and rehabilitation training. All of these strategies have positive effects in the course of spinal cord injury treatment. This review mainly discusses the problems regarding stem cell therapy for spinal cord injury, including the characteristics and action modes of all relevant cell types. Induced pluripotent stem cells, which represent a special kind of stem cell population, have gained impetus in cell therapy development because of a range of advantages. Induced pluripotent stem cells can be developed into the precursor cells of each neural cell type at the site of spinal cord injury, and have great potential for application in spinal cord injury therapy.

Generation of male germ cells from induced pluripotent stem cells （iPS cells）： an in vitro and in vivo study

最近的研究报导了从老鼠和人的房间能区分进初发的细菌房间的那导致的 pluripotent 茎(iPS ) 。然而， iPS 房间是否能够生产男细菌房间，不被知道。这研究的目的进 spermatogonial 干细胞和迟了阶段的男细菌房间的老鼠 iPS 房间是调查区别潜在的。我们使用了在 vitro 区别并且在 vivo 移植联合的一条途径。胚胎植物或动物身体(EB ) 用白血球过多症禁止者因素(LIF ) 从 iPS 房间被获得免费媒介。量的 PCR 表明在 Oct4 表示的减少和 Stra8 的增加和在 EB 的 Vasa mRNA 源于 iPS 房间。iPS 导出房间的 EB 被区分进 spermatogonial 干细胞(SSC ) ，，的酸由他们 VASA，以及 CDH1 和 GFR&#x003b1 的表示证实了的 retinoic 导致； 1，它是 SSC 的标记。而且，从 iPS 房间导出的这些细菌房间被移植进与 busulfan 被预先对待的老鼠的接受者睾丸。尤其是， iPS 导出房间的 SSC 能区分进从 spermatogonia 到圆 spermatids，由 VASA 和 SCP3 表示出现的男细菌房间。这研究证明 iPS 房间有潜力区分进迟了阶段的男细菌房间。从 iPS 房间的男细菌房间的推导在男不孕的处理有潜在的应用程序并且为揭开提供一个模型位于男细菌房间开发下面的分子的机制。

Automated closed-cell production platform solves dilemma of industrial-scale manufacturing for mesenchymal stromal cell-based therapy

Mesenchymal stromal cells(MSCs),known for their therapeutic bioactivity,find widespread application as cellular drugs for treating various diseases.MSCs obtained from patients or donors require extensive large-scale expansion for clinical applications.However,the conventional method of cultivating MSCs involves several manual processes and yields inconsistent batch-to-batch quality.Consequently,it has not been scalable as a cell therapy[1].To overcome the limitations of conventional planar cell culture,van Wezel initially proposed a system for culturing cells in suspension using microcarriers and successfully proliferated rabbit embryonic skin cells and human embryonic lung cells[2].Subsequently,microcarrier technology has been employed across various pharmaceutical applications,leading to the development and commercialization of a diverse array of microcarriers with distinct physicochemical properties.

Dynamic changes in the systemic immune responses of spinal cord injury model mice

Intraspinal inflammatory and immune responses are considered to play central roles in the pathological development of spinal cord injury.This study aimed to decipher the dynamics of systemic immune responses,initiated by spinal cord injury.The spinal cord in mice was completely transected at T8.Changes in the in vivo inflammatory response,between the acute and subacute stages,were observed.A rapid decrease in C-reactive protein levels,circulating leukocytes and lymphocytes,spleen-derived CD4~+interferon-γ+T-helper cells,and inflammatory cytokines,and a marked increase in neutrophils,monocytes,and CD4~+CD25~+FOXP3~+regulatory T-cells were observed during the acute phase.These systemic immune alterations were gradually restored to basal levels during the sub-acute phase.During the acute phase of spinal cord injury,systemic immune cells and factors showed significant inhibition;however,this inhibition was transient,and the indicators of these serious disorders gradually returned to baseline levels during the subacute phase.All experiments were performed in accordance with the institutional animal care guidelines,approved by the Institutional Animal Care and Use Committee of Experimental Animal Center of Drum Tower Hospital,China(approval No.2019 AE01040)on June 25,2019.

Vitamin E in the management of pancreatic cancer: A scoping review

Pancreatic cancer is the leading cause of cancer mortality worldwide.Research investigating effective management strategies for pancreatic cancer is ongoing.Vitamin E,consisting of both tocopherol and tocotrienol,has demonstrated debatable effects on pancreatic cancer cells.Therefore,this scoping review aims to summarize the effects of vitamin E on pancreatic cancer.In October 2022,a literature search was conducted using PubMed and Scopus since their inception.Original studies on the effects of vitamin E on pancreatic cancer,including cell cultures,animal models and human clinical trials,were considered for this review.The literature search found 75 articles on this topic,but only 24 articles met the inclusion criteria.The available evidence showed that vitamin E modulated proliferation,cell death,angiogenesis,metastasis and inflammation in pancreatic cancer cells.However,the safety and bioavailability concerns remain to be answered with more extensive preclinical and clinical studies.More in-depth analysis is necessary to investigate further the role of vitamin E in the management of pancreatic cancers.

Role and prospects of regenerative biomaterials in the repair of spinal cord injury

Axonal junction defects and an inhibitory environment after spinal cord injury seriously hinder the regeneration of damaged tissues and neuronal functions. At the site of spinal cord injury, regenerative biomaterials can fill cavities, deliver curative drugs, and provide adsorption sites for transplanted or host cells. Some regenerative biomaterials can also inhibit apoptosis, inflammation and glial scar formation, or further promote neurogenesis, axonal growth and angiogenesis. This review summarized a variety of biomaterial scaffolds made of natural, synthetic, and combined materials applied to spinal cord injury repair. Although these biomaterial scaffolds have shown a certain therapeutic effect in spinal cord injury repair, there are still many problems to be resolved, such as product standards and material safety and effectiveness.

Targeting ferroptosis suppresses osteocyte glucolipotoxicity and alleviates diabetic osteoporosis

Diabetic osteoporosis(DOP) is the leading complication continuously threatening the bone health of patients with diabetes. A key pathogenic factor in DOP is loss of osteocyte viability. However, the mechanism of osteocyte death remains unclear. Here, we identified ferroptosis, which is iron-dependent programmed cell death, as a critical mechanism of osteocyte death in murine models of DOP. The diabetic microenvironment significantly enhanced osteocyte ferroptosis in vitro, as shown by the substantial lipid peroxidation, iron overload, and aberrant activation of the ferroptosis pathway. RNA sequencing showed that heme oxygenase-1(HO-1) expression was notably upregulated in ferroptotic osteocytes. Further findings revealed that HO-1 was essential for osteocyte ferroptosis in DOP and that its promoter activity was controlled by the interaction between the upstream NRF2 and c-JUN transcription factors. Targeting ferroptosis or HO-1 efficiently rescued osteocyte death in DOP by disrupting the vicious cycle between lipid peroxidation and HO-1 activation, eventually ameliorating trabecular deterioration. Our study provides insight into DOP pathogenesis, and our results provide a mechanism-based strategy for clinical DOP treatment.

Prevention against diffuse spinal cord astrocytoma: can the Notch pathway be a novel treatment target?

This study was designed to investigate whether the Notch pathway is involved in the development of diffuse spinal cord astrocytomas. BALB/c nude mice received injections of CD133+ and CD133- cell suspensions prepared using human recurrent diffuse spinal cord astrocytoma tissue through administration into the right parietal lobe. After 7–11 weeks, magnetic resonance imaging was performed weekly. Xenografts were observed on the surfaces of the brains of mice receiving the CD133+ cell suspension, and Notch-immunopositive expression was observed in the xenografts. By contrast, no xenografts appeared in the identical position on the surfaces of the brains of mice receiving the CD133- cell suspension, and Notch-immunopositive expression was hardly detected either. Hematoxylin-eosin staining and immunohistochemical staining revealed xenografts on the convex surfaces of the brains of mice that underwent CD133+ astrocytoma transplantation. Some sporadic astroglioma cells showed pseudopodium-like structures, which extended into the cerebral white matter. However, it should be emphasized that the subcortex xenograft with Notch-immunopositive expression was found in the fourth mouse received injection of CD133- astrocytoma cells. However, these findings suggest that the Notch pathway plays an important role in the formation of astrocytomas, and can be considered a novel treatment target for diffuse spinal cord astrocytoma.

Plasticity of male germline stem cells and their applications in reproductive and regenerative medicine

Spermatogonial 干细胞(SSC ) ，也已知的同样男的 germline 干细胞，是小 subpopulation 与自强的潜力打 spermatogonia 在哺乳动物的睾丸维持干细胞水池和区别进 spermatids。自从他们能仅仅在生精的小管以内产生精子， SSC 以前被认为是 unipotent 干细胞。然而，因为众多的研究证明了与生长因素有教养的 SSC 能获得 pluripotency 成为胚胎的像茎的细胞，这个概念最近被质问了。在里面 vivo 并且在 vitro，从同伴和我们的研究清楚地表明了那 SSC 直接装 transdifferentiate 进词法，另外的系的 phenotypic，和功能的房间。到另外的纸巾的房间的直接变换为再生药有重要意义。从精子缺乏病人的 SSC 能被导致与授精和发展潜力区分进 spermatids。因此， SSC 能由于他们的唯一、大的潜力在繁殖、再生的药有重要应用。在这评论，我们探讨 SSC 的重要粘性，与他们的自强，区别， dedifferentiation， transdifferentiation，和翻译的药研究的焦点。

Three-dimensional bioprinting sodium alginate/gelatin scaffold combined with neural stem cells and oligodendrocytes markedly promoting nerve regeneration after spinal cord injury

Accumulating research has indicated that the transplantation of combined stem cells and scaffolds is an effective method for spinal cord injury(SCI).The development of three-dimensional(3D)bioprinting technology can make the 3D scaffolds combined with cells more accurate and effective for SCI treatment.However,unmyelinated newborn nerve fibers have no nerve signaling conduction,hampering recovery of motor function.In this study,we designed and printed a type of sodium alginate/gelatin scaffold loaded with neural stem cells and oligodendrocytes,which were involved in the formation of the myelin sheaths of neural cell axons.In order to observe the effectiveness of this 3D bioprinting scaffold,we transplanted it into the completely transected rat spinal cord,and then immunofluorescence staining,hematoxylin–eosin staining and behavioral assessment were performed.The results showed that this 3D bioprinting scaffold markedly improved the hindlimb motor function and promoted nerve regeneration.These findings suggested that this novel 3D bioprinting scaffold was a good carrier for cells transplantation,thereby enhancing spinal cord repair following injury.

Characterization, isolation, and culture of spermatogonial stem cells in Macaca fascicularis

Spermatogonial stem cells(SSCs)have great applications in both reproductive and regenerative medicine.Primates including monkeys are very similar to humans with regard to physiology and pathology.Nevertheless,little is known about the isolation,the characteristics,and the culture of primate SSCs.This study was designed to identify,isolate,and culture monkey SSCs.Immunocytochemistry was used to identify markers for monkey SSCs.Glial cell line-derived neurotrophic factor family receptor alpha-1(GFRAl)-enriched spermatogonia were isolated from monkeys,namely Macaca fascicularis(M.fascicularis),by two-step enzymatic digestion and magnetic-activated cell sorting,and they were cultured on precoated plates in the conditioned medium.Reverse transcription-polymerase chain reaction(RT-PCR),immunocytochemistry,and RNA sequencing were used to compare phenotype and transcriptomes in GFRAl-enriched spermatogonia between 0 day and 14 days of culture,and xenotransplantation was performed to evaluate the function of GFRAl-enriched spermatogonia.SSCs shared some phenotypes with rodent and human SSCs.GFRAl-enriched spermatogonia with high purity and viability were isolated from M.fascicularis testes.The freshly isolated cells expressed numerous markers for rodent SSCs,and they were cultured for 14 days.The expression of numerous SSC markers was maintained during the cultivation of GFRAl-enriched spermatogonia.RNA sequencing reflected a 97.3%similarity in global gene profiles between 0 day and 14 days of culture.The xenotransplantation assay indicated that the GFRAl-enriched spermatogonia formed colonies and proliferated in vivo in the recipient c-Kitw/w(W)mutant mice.Collectively,GFRAl-enriched spermatogonia are monkey SSCs phenotypically both in vitro and in vivo.This study suggests that monkey might provide an alternative to human SSCs for basic research and application in human diseases.

Targeting pancreatic cancer immune evasion by inhibiting histone deacetylases

The immune system plays a vital role in maintaining the delicate balance between immune recognition and tumor development.Regardless,it is not uncommon that cancerous cells can intelligently acquire abilities to bypass the antitumor immune responses,thus allowing continuous tumor growth and development.Immune evasion has emerged as a significant factor contributing to the progression and immune resistance of pancreatic cancer.Compared with other cancers,pancreatic cancer has a tumor microenvironment that can resist most treatment modalities,including emerging immunotherapy.Sadly,the use of immunotherapy has yet to bring significant clinical breakthrough among pancreatic cancer patients,suggesting that pancreatic cancer has successfully evaded immunomodulation.In this review,we summarize the impact of genetic alteration and epigenetic modification(especially histone deacetylases,HDAC)on immune evasion in pancreatic cancer.HDAC overexpression significantly suppresses tumor suppressor genes,contributing to tumor growth and progression.We review the evidence on HDAC inhibitors in tumor eradication,improving T cells activation,restoring tumor immunogenicity,and modulating programmed death 1 interaction.We provide our perspective in targeting HDAC as a strategy to reverse immune evasion in pancreatic cancer.

PUMA facilitates EMI 1-promoted cytoplasmic Rad51 ubiquitination and inhibits DNA repair in stem and progenitor cells

Maintenance of genetic stability via proper DNA repair in stem and progenitor cells is essential for the tissue repair and regeneration,while preventing cell transformation after damage.Loss of PUMA dramatically increases the survival of mice after exposure to a lethal dose of ionizing radiation(IR),while without promoting tumorigenesis in the long-term survivors.This finding suggests that PUMA(p53 upregulated modulator of apoptosis)may have a function other than regulates apoptosis.Here,we identify a novel role of PUMA in regulation of DNA repair in embryonic or induced pluripotent stem cells(PSCs)and immortalized hematopoietic progenitor cells(HPCs)after IR.We found that PUMA-deficient PSCs and HPCs exhibited a significant higher doublestrand break(DSB)DNA repair activity via Rad51-mediated homologous recombination(HR).This is because PUMA can be associated with early mitotic Inhibitor 1(EMI1)and Rad51 in the cytoplasm to facilitate EMI1-mediated cytoplasmic Rad51 ubiquitination and degradation,thereby inhibiting Rad51 nuclear translocation and HR DNA repair.Our data demonstrate that PUMA acts as a repressor for DSB DNA repair and thus offers a new rationale for therapeutic targeting of PUMA in regenerative cells in the context of DNA damage.

Ginsenoside Rb1 inhibits oxidative stress-induced ovarian granulosa cell injury through Akt-FoxO1 interaction

Ginsenoside Rb1 shows a strong antioxidant effect and has potential activation effects on Akt.The aim of the present study was to investigate the protective effect of Rb1 on age-related ovarian granulosa cell injury.Ovarian granulosa cells(GCs)were obtained from 50 young women(≤30 years)and 50 aged women(≥38 years)at an IVF center.Young and aged ICR mice were administered with or without Rb1(10 mg kg^(-1),i.p.)for 2 weeks.The protective effects of Rb1 were investigated and the role of Rb1 on the modulation of Akt-FoxO1 interaction was determined with immunofluorescence,Western blotting,immunoprecipitation,si RNA silencing and pharmacological inhibitor.Rb1 effectively decreased LDH and MDA,and reversed the apoptotic-related protein levels in h GL cells from old patients.Similar results were found in mice.In addition,the mitochondrial membrane potential was restored and the overaccumulation of ROS was reversed by Rb1.Rb1 preserved peroxide-impaired Akt activation,to some extent,by increasing phosphorylation at Ser473.Rb1 also facilitated p-Akt binding to FoxO1 and promoted the phosphorylation of FoxO1.Si RNA silencing of Akt,Akt inhibitor LY294002,and FoxO1 inhibitor AS1842856 attenuated the effects of Rb1.Ginsenoside Rb1 inhibits age-related GCs oxidative damage by activating Akt phosphorylation at Ser473 and by further interaction with FoxO1.

Non-catalytic roles for TET1 protein negatively regulating neuronal differentiation through srGAP3 in neuroblastoma cells

methylcytosine dioxygenases TET 蛋白质(TET1， TET2，和 TET3 ) 在神经功能起重要规章的作用。在这研究，我们作为一个模型用 Neuro2a 房间在 neuronal 区别调查了 TET 蛋白质的角色。我们观察了那支持的 TET1， TET2 或 TET3 击倒 Neuro2a 房间的 neuronal 区别，和他们的 overexpression 禁止了导致的 VPA (valproic 酸) neuronal 区别，建议所有三 TET 蛋白质否定地调整 Neuro2a 房间的 neuronal 区别。有趣地， TET 蛋白质的导致的活动独立于它的酶的活动。我们的以前的研究证明了 srGAP3 能否定地调整 Neuro2a 房间的 neuronal 区别。而且，我们表明 TET1 能断然调整它的催化活动的 srGAP3 表示独立人士，并且 srGAP3 为 Neuro2a 房间的调停 TET 的 neuronal 区别被要求。这里介绍的结果可以在 neuronal 区别便于 TET 蛋白质的角色的更好的理解，并且为 neuroblastoma 提供一个可能的治疗目标。

Aging hallmarks of the primate ovary revealed by spatiotemporal transcriptomics

The ovary is indispensable for female reproduction,and its age-dependent functional decline is the primary cause of infertility.However,the molecular basis of ovarian aging in higher vertebrates remains poorly understood.Herein,we apply spatiotemporal transcriptomics to benchmark architecture organization as well as cellular and molecular determinants in young primate ovaries and compare these to aged primate ovaries.From a global view,somatic cells within the non-follicle region undergo more pronounced transcriptional fluctuation relative to those in the follicle region,likely constituting a hostile microenvironment that facilitates ovarian aging.Further,we uncovered that inflammation,the senescent-associated secretory phenotype,senescence,and fibrosis are the likely primary contributors to ovarian aging(PCOA).Of note,we identified spatial co-localization between a PCOA-featured spot and an unappreciated MT2(Metallothionein 2)highly expressing spot(MT2^(high))characterized by high levels of inflammation,potentially serving as an aging hotspot in the primate ovary.Moreover,with advanced age,a subpopulation of MT2^(high)accumulates,likely disseminating and amplifying the senescent signal outward.Our study establishes the first primate spatiotemporal transcriptomic atlas,advancing our understanding of mechanistic determinants underpinning primate ovarian aging and unraveling potential biomarkers and therapeutic targets for aging and age-associated human ovarian disorders.

PRPS2 mutations drive acute lymphoblastic leukemia relapse through influencing PRPS1/2 hexamer stability

Tumor relapse is the major cause of treatment failure in childhood acute lymphoblastic leukemia(ALL),yet the underlying mechanisms are still elusive.Here,we demonstrate that phosphoribosyl pyrophosphate synthetase 2(PRPS2)mutations drive ALL relapse through influencing PRPS1/2 hexamer stability.Ultra-deep sequencing was performed to identify PRPS2 mutations in ALL samples.The effects of PRPS2 mutations on cell survival,cell apoptosis,and drug resistance were evaluated.In vitro PRPS2 enzyme activity and ADP/GDP feedback inhibition of PRPS enzyme activity were assessed.Purine metabolites were analyzed by ultra-performance liquid-chromatography tandem mass spectrometry(UPLC–MS/MS).Integrating sequencing data with clinical information,we identified PRPS2 mutations only in relapsed childhood ALL with thiopurine therapy.Functional PRPS2 mutations mediated purine metabolism specifically on thiopurine treatment by influencing PRPS1/2 hexamer stability,leading to reduced nucleotide feedback inhibition of PRPS activity and enhanced thiopurine resistance.The 3-amino acid V103-G104-E105,the key difference between PRPS1 and PRPS2,insertion in PRPS2 caused severe steric clash to the interface of PRPS hexamer,leading to its low enzyme activity.In addition,we demonstrated that PRPS2 P173R increased thiopurine resistance in xenograft models.Our work describes a novel mechanism by which PRPS2 mutants drive childhood ALL relapse and highlights PRPS2 mutations as biomarkers for relapsed childhood ALL.