Long non-coding RNA H19 regulates neurogenesis of induced neural stem cells in a mouse model of closed head injury

Stem cell-based therapies have been proposed as a potential treatment for neural regeneration following closed head injury.We previously reported that induced neural stem cells exert beneficial effects on neural regeneration via cell replacement.However,the neural regeneration efficiency of induced neural stem cells remains limited.In this study,we explored differentially expressed genes and long non-coding RNAs to clarify the mechanism underlying the neurogenesis of induced neural stem cells.We found that H19 was the most downregulated neurogenesis-associated lnc RNA in induced neural stem cells compared with induced pluripotent stem cells.Additionally,we demonstrated that H19 levels in induced neural stem cells were markedly lower than those in induced pluripotent stem cells and were substantially higher than those in induced neural stem cell-derived neurons.We predicted the target genes of H19 and discovered that H19 directly interacts with mi R-325-3p,which directly interacts with Ctbp2 in induced pluripotent stem cells and induced neural stem cells.Silencing H19 or Ctbp2 impaired induced neural stem cell proliferation,and mi R-325-3p suppression restored the effect of H19 inhibition but not the effect of Ctbp2 inhibition.Furthermore,H19 silencing substantially promoted the neural differentiation of induced neural stem cells and did not induce apoptosis of induced neural stem cells.Notably,silencing H19 in induced neural stem cell grafts markedly accelerated the neurological recovery of closed head injury mice.Our results reveal that H19 regulates the neurogenesis of induced neural stem cells.H19 inhibition may promote the neural differentiation of induced neural stem cells,which is closely associated with neurological recovery following closed head injury.

Induced neural stem cells regulate microglial activation through Akt-mediated upregulation of CXCR4 and Crry in a mouse model of closed head injury

Microglial activation that occurs rapidly after closed head injury may play important and complex roles in neuroinflammation-associated neuronal damage and repair.We previously reported that induced neural stem cells can modulate the behavior of activated microglia via CXCL12/CXCR4 signaling,influencing their activation such that they can promote neurological recovery.However,the mechanism of CXCR4 upregulation in induced neural stem cells remains unclear.In this study,we found that nuclear factor-κB activation induced by closed head injury mouse serum in microglia promoted CXCL12 and tumor necrosis factor-αexpression but suppressed insulin-like growth factor-1 expression.However,recombinant complement receptor 2-conjugated Crry(CR2-Crry)reduced the effects of closed head injury mouse serum-induced nuclear factor-κB activation in microglia and the levels of activated microglia,CXCL12,and tumor necrosis factor-α.Additionally,we observed that,in response to stimulation(including stimulation by CXCL12 secreted by activated microglia),CXCR4 and Crry levels can be upregulated in induced neural stem cells via the interplay among CXCL12/CXCR4,Crry,and Akt signaling to modulate microglial activation.In agreement with these in vitro experimental results,we found that Akt activation enhanced the immunoregulatory effects of induced neural stem cell grafts on microglial activation,leading to the promotion of neurological recovery via insulin-like growth factor-1 secretion and the neuroprotective effects of induced neural stem cell grafts through CXCR4 and Crry upregulation in the injured cortices of closed head injury mice.Notably,these beneficial effects of Akt activation in induced neural stem cells were positively correlated with the therapeutic effects of induced neural stem cells on neuronal injury,cerebral edema,and neurological disorders post–closed head injury.In conclusion,our findings reveal that Akt activation may enhance the immunoregulatory effects of induced neural stem cells on microglial activation via upregulation of CXCR4 and Crry,thereby promoting induced neural stem cell–mediated improvement of neuronal injury,cerebral edema,and neurological disorders following closed head injury.

Molecular signaling in cancer stem cells of tongue squamous cell carcinoma:Therapeutic implications and challenges

Head and neck squamous cell carcinoma is the seventh most common cancer worldwide with high mortality rates.Amongst oral cavity cancers,tongue carcinoma is a very common and aggressive oral cavity carcinoma.Despite the implementation of a multimodality treatment regime including surgical intervention,chemo-radiation as well as targeted therapy,tongue carcinoma shows a poor overall 5-year survival pattern,which is attributed to therapy resistance and recurrence of the disease.The presence of a rare population,i.e.,cancer stem cells(CSCs)within the tumor,are involved in therapy resistance,recurrence,and distant metastasis that results in poor survival patterns.Therapeutic agents targeting CSCs have been in clinical trials,although they are unable to reach into therapy stage which is due to their failure in trials.A more detailed understanding of the CSCs is essential for identifying efficient targets.Molecular signaling pathways,which are differentially regulated in the CSCs,are one of the promising targets to manipulate the CSCs that would provide an improved outcome.In this review,we summarize the current understanding of molecular signaling associated with the maintenance and regulation of CSCs in tongue squamous cell carcinoma in order to emphasize the need of the hour to get a deeper understanding to unravel novel targets.

Targeting head and neck tumoral stem cells： From biologicalaspects to therapeutic perspectives

Head and neck squamous cell cancer(HNSCC) is the sixth most common cancer in the world. Effective therapeutic modalities such as surgery, radiation, chemotherapy and combinations of each are used in the management of the disease. In most cases, treatment fails to obtain total cancer cure. In recent years, it appears that one of the key determinants of treatment failure may be the presence of cancer stem cells(CSCs) that escape currently available therapies. CSCs form a small portion of the total tumor burden but may play a disproportionately important role in determining outcomes. CSCs have stem features such as self-renewal, high migration capacity, drug resistance, high proliferation abilities. A large body of evidence points to the fact that CSCs are particularly resistant to radiotherapy and chemotherapy. In HNSCC, CSCs have been increasingly shown to have an integral role in tumor initiation, disease progression, metastasis and treatment resistance. In the light of such observations, the present review summarizes biological characteristics of CSCs in HNSCC, outlines targeted strategies for the successful eradication of CSCs in HNSCC including targeting the self-renewal controlling pathways, blocking epithelial mesenchymal transition, niche targeting, immunotherapy approaches and highlights the need to better understand CSCs biology for new treatments modalities.

Protective Effect of Human Umbilical Cord Mesenchymal Stem Cells in Glucocorticoid-induced Osteonecrosis of Femoral Head

Objective:To evaluate the effect of human umbilical cord mesenchymal stem cells(hUC-MSCs)on preventing rats from glucocorticoid-induced osteonecrosis of femoral head(GC-ONFH)in the early stage in vivo and to investigate the possible mechanism of hUC-MSCs in regulating the balance of osteogenesis and adipogenesis.Methods:All rats were randomly divided into 3 groups:control group(C group),model group(M group),and intervention group(Ⅰ group).The model of GC-ONFH was developed by a sequential administration of lipopolysaccharide and methylprednisolone.The rats in the Ⅰ group were treated with caudal vein injection of hUC-MSCs.Six weeks later,the blood samples were obtained to measure the activity of alkaline phosphatase(ALP)and the content of triglyceride(TG)in serum,and the femoral heads were harvested and observed by hematoxylin-eosin staining,Micro-CT,Western blot and real-time quantitative polymerase chain reaction.Results:After intervention of hUC-MSCs,the necrosis rate of femoral head decreased from 83%(10/12)to 33%(4/12),the rate of empty bone lacuna was significantly decreased,the activity of ALP increased significantly,the content of TG decreased significantly,the bone density increased obviously,the expression of RUNX2 and Col Ⅰ increased significantly and the expression of PPARγ decreased significantly.Conclusion:These results revealed that caudal vein injection of hUC-MSCs can effectively reduce the incidence of GC-ONFH in rats by increasing ALP activity and reducing TG content in serum,increasing bone mineral density,promoting the expression of RUNX2 and Col I,and inhibiting the expression of PPARγ.

Co-culture of Mesenchymal Stem Cells with Umbilical Vein Endothelial Cells under Hypoxic Condition

By co-culturing humm mesenchymal stem cells (hMSCs) and human umbilical rein endothelial cells (HUVECs) under hypoxia and creating a microenvironment similar to that of transplanted hMSCs for the treatment of avascular ni ANFH, the effect of hMSCs on survival, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) under the hypoxic condition were investigated in vitro. hMSCs and HUVECs were cultured and identified in vitro. Three kinds of conditioned media, CdM-CdMNOR, CdM-CdMHYP and HUVEC-CdMHYP were prepared. HUVECs were cultured with these conditioned media under hypoxia. The survival rate, apoptosis rate, migration and angiogenesis of HUVECs were respectively detected by CCK-8, flow cytometry, Transwell and tube formation assay. The content of SDF-1α, VEGF and IL-6 in CdM was determined by ELISA. Our results showed that hMSCs and HUVECs were cultured and identified successfully. Compared with MSC-CdMNOR and HUVEC-CdMHYP groups, the survival rate, migra-tion and angiogenesis of HUVECs in MSC-CdMHYP group were significantly increased while the apoptosis rate was declined (P<0.05). Moreover, the expression of SDF-1α, VEGF and IL-6 in MSC-CdMHYP group was up-regulated. Under hypoxia, the apoptosis of HUVECs was inhibited while survival, migration and angiogenesis were improved by co-culture of hMSCs and HUVECs. The underlying mechanism may be that hMSCs could secrete a number of cytokines and improve niche, which might be helpful in the treatment of femoral head necrosis.

Immunocytochemical identification and localization of APUD cells in the gut of seven stomachless teleost fishes

AIM To study the cell types,localization,distribution density and morphology of APUDcells in the intestinal mucosa of stomachlessteleost fishes.METHOD By using the peroxidase-antiperoxidase complex(PAP)immunocytochemical staining technique theidentification,localization and morphology ofimmunoreactive(IR)endocrine cells seattered inthe intestinal mucosa of grass carp(Cyenopharyngodon idellus),black carp(Mylopharyngodon piceus)and common carp(Cyprinus carpio)were investigated with 20kinds of antisera prepared against mammalianpeptide hormones of APUD cells,and likewise byusing avidin-biotin-peroxidase complex(ABC)method those of silver carp(Hypophthalmichthys molitrix),bighead(Aristichthys nobilis),silver crucian carp(Carassius gibelio)and bluntnose black bream(Megalobrama amblyocephala)were alsostudied with 5 different antisera.Thereplacement of the first antiserum by phosphatebuffered saline(PBS)was employed as a control.IR endocrine cells were counted with asquare-mesh ocular micrometer from 10 fieldsselected randomly in every section of each partof the intestine specimen.The average numberof IR endocrine cells per mm2 was counted toquantify their distribution density.RESULT Gastrin(GAS)-,Gastric inhibitorypeptide(GIP)-,glucagon(GLU)-,glucagon-likeimmunoreactants(GLI)-,bovine pancreaticpolypeptide(BPP)-,leucine-enkephalin(ENK)-and substance P(SP)-IR endocrine cells werefound in the gut of grass carp,black carp andcommon carp,and somatostatin(SOM)-IRendocrine cells were only seen in common carp.GAS-,GIP-and GLU-IR endocrine cells werefound in the intestinal mucosa of silver carp,bighead,silver crucian carp and bluntnose blackbream.Most of IR endocrine cells had the higherdistribution density in the foregut and midgut,and were longer in shape.They had a long apicalcytoplasmic process extended to the gut lumenand a basal process extended to adjacent cellsor basement membrane and touched with it.Sometimes,the basal cytoplasmic processformed an enlarged synapse-like structure in thecontiguous part with basement membrane.Thisphenomenon provided new morphologicalevidence for neuroendocrine and paracrinesecretory function of these enteroendocrinecells.CONCLUTION At least 8 kinds of IR endocrinecells were found in the gut of stomachlessteleost species for the first time in China.TheseIR endocrine cells scattering in the gut mucosabelong to the APUD system.Among them,thehormones secreted by SP-,ENK-,SOM-and GLU-IR endocrine cells belong to the peptides of dualdistribution in the brain and gut.This providednew evidence for the concept of brain-gutpeptide.According to the cell types,distribution density,morphologicalcharacteristics and variety in shape of APUDcells in the gut of stomachless teleost fishes,itis deemed that the digestive tract of fishes isalso an endocrine organ of great importance andcomplexity.

On the cell biology of pit cells,the liver-specific NK cells

INTRODUCTION Natural killer (NK) cells are functionally defined by their ability to kill certain tumor cells and virus-infected cells without prior

PGRMC1 Elevation in Multiple Cancers and Essential Role in Stem Cell Survival

Cancer is one of the leading causes of death in America, and there is an urgent need for new therapeutic approaches. The progesterone receptor membrane component 1 (PGRMC1) is a cytochrome b5 related protein that binds heme and is associated with signaling, apoptotic suppression and autophagy. PGRMC1 is essential for tumor formation, invasion and metastasis, and is upregulated in breast, colon, lung and thyroid tumors. In the present study, we have analyzed PGRMC1 levels in over 600 tumor sections, including a larger cohort of lung tumors than in previous studies, and report the first clinical analysis of PGRMC1 levels in human oral cavity and ovarian tumors compared to corresponding nonmalignant tissues. PGRMC1 was highly expressed in lung and ovarian cancers and correlated with patient survival. PGRMC1 has been previously associated with drug resistance, a characteristic of cancer stem cells. The stem cell theory proposes that a subset of cancerous stem cells contribute to drug resistance and tumor maintenance, and PGRMC1 was detected in lung-tumor derived stem cells. Drug treatment with a PGRMC1 inhibitor, AG-205, triggered stem cell death whereas treatment with erlotinib and the ERK inhibitor, PD98059, did not, suggesting a specific role for PGRMC1 in cancer stem cell viability. Together, our data demonstrate PGRMC1 as a potential tumor biomarker across a variety of tumors, as well as a therapeutic target for cancer stem cells.

组织工程技术在股骨头坏死治疗中的应用及前景

背景:股骨头坏死早期多采用植骨类保髋手术治疗,其中自体骨与异体骨是常用的植骨材料,但自体骨移植创伤性大且供骨来源有限,异体骨虽来源丰富,但存在严重的免疫排斥反应和吸收风险。近年来,以间充质干细胞为基础的组织工程技术是治疗股骨头坏死的新方法,经过基础实验和临床应用的检验,正逐渐得到广泛应用。目的:综述组织工程技术在股骨头坏死治疗中的应用及前景,为临床治疗股骨头坏死提供新的选择。方法:由第一作者检索2013-2023年的PubMed数据库和中国知网,英文检索词为“tissue engineering,mesenchymal stem cells,biological scaffolds,cytokines,osteonecrosis of the femoral head,bone graft,hip preservation”,中文检索词为“组织工程,间充质干细胞,生物支架,细胞因子,股骨头坏死,植骨,保髋”,选择运用组织工程技术治疗股骨头坏死方面的文献,根据纳入与排除标准对所有文章进行初筛后,共纳入55篇代表性文献进行综述。结果与结论:①随着生物技术与材料学的不断发展,骨组织工程技术治疗股骨头坏死己取得较大进展,例如基因修饰的间充质干细胞修复骨坏死、基因重组技术及表面改性技术与骨组织工程的结合在股骨头坏死治疗中的应用等;②组织工程技术应用于坏死股骨头时可促进坏死骨组织再生和血管系统的修复,同时为坏死区域提供生物力学稳定性,并且利用生物活性因子促进种子细胞的修复作用,完成坏死区新生骨的再生;③但目前这些研究大多数尚处于动物实验阶段,骨组织工程研究仍存在许多未解决的问题和挑战,随着纳米技术、组织工程学和临床医学的快速发展,性能完善的仿生替代植骨材料将有望诞生;④在未来应用骨组织工程技术治疗股骨头坏死有望成为一种令人满意的治疗方式,给保髋患者带来福音。

组织工程治疗股骨头坏死软骨下分离的适用技术

背景:股骨头坏死出现新月征是病情进程的“分水岭”,修复和稳定骨-软骨界面对阻止病情继续进展和预防股骨头塌陷尤为重要。利用组织工程学同步修复、整合骨-软骨界面具有潜在优势。目的:综述探讨解决股骨头坏死软骨下分离的潜在适宜技术。方法:检索1970年1月至2023年4月PubMed、Web of Science及中国知网、万方数据库中发表的相关文献,英文检索词:“Femoral head necrosis,Avascular necrosis of femoral head,Osteonecrosis of femoral head”等,中文检索词:“股骨头坏死,软骨下骨,软骨,软骨与软骨下骨整合”等,最终纳入114篇文献进行综述分析。结果与结论:①结构缺陷、缺血缺氧环境、炎症因素和应力集中可能造成股骨头坏死软骨下分离现象,软骨下骨分离会造成塌陷进展,并且可能与保髋手术失败相关,利用组织工程支架实现支架与骨-软骨界面的整合是治疗股骨头坏死软骨下分离的潜在方法之一。②目前的文献研究表明,多相、梯度支架和复合材料在促进骨、软骨细胞黏附与增殖,骨软骨基质的沉积方面均有提升,有助于支架与骨-软骨界面的整合,对治疗股骨头坏死软骨下分离有参考价值。③通过对支架表面进行修饰可以提高与界面整合的效率,但有各自不同的优缺点,提供不同环境的支架能够诱导同种间充质干细胞差异分化,有助于不同界面之间的整合。④未来有望应用于股骨头坏死软骨下分离的支架应为复合材料,具有梯度化和差异化的仿生结构,通过表面修饰和干细胞加载促进骨-软骨界面与支架的整合以实现治疗目的,但仍需进一步研究验证,而支架的降解速率与修复进度同步和不同界面之间的稳定性是未来需要解决的主要问题。

miR-124-3p调控激素性股骨头坏死患者骨髓间充质干细胞成骨分化作用的研究

目的探讨miRNAs和CTNNB1在激素性股骨头坏死患者中的表达水平,以及miR-124-3p对骨髓间充质干细胞成骨分化的影响。方法选择2020年9月至2022年5月就诊于新疆医科大学第五附属医院行髋关节置换术的患者30例,激素性股骨头坏死患者15例作为实验组,股骨颈骨折患者15例作为对照组。术中扩髓时收集患者的骨髓组织,提取总mRNA,应用qRT-PCR检测各组miR-322-5p、miR-124-3p、miR-125a-3p及CTNNB1的表达量,用Western Blot检测CTNNB1蛋白的表达量。在骨髓间充质干细胞(BMSCs)中转染miR-124-3p的模拟物和抑制剂来检测其对BMSCs分化的影响作用,并验证miR-124-3p与CTNNB1的靶标关系。结果在实验组BMSCs中miR-125a-3p、CTNNB1表达量高于对照组(P<0.05);miR-124-3p的表达量低于对照组(P<0.001),miR-322-5p的表达量在两组中无明显差异。转染miR-124-3p+mimics组的miR-124-3p表达量高于其他组,CTNNB1表达量低于其他组(P<0.001),转染miR-124-3p inhibitor组的结果则相反。分化能力评估发现过表达miR-124-3p可以促进BMSCs成骨分化能力,抑制表达则结果相反。双荧光素酶报告系统结果显示,miR-124-3p与CTNNB1有靶向结合关系。结论miR-124-3p可以靶向CTNNB1,并且miR-124-3p可以调控骨髓间充质干细胞并促进其成骨分化。

慢病毒沉默Piezo1蛋白与人骨髓间充质干细胞成骨分化及TAZ的表达

背景:Piezo1作为机械敏感蛋白与成骨分化密切相关,TAZ也被证明参与调节成骨分化,但Piezo1调控人骨髓间充质干细胞成骨分化时TAZ是否参与其中目前尚不明确,故研究其具体机制对防治股骨头坏死具有重要意义。目的:探讨Piezo1对人骨髓间充质干细胞成骨分化及TAZ表达的影响。方法:构建靶向Piezo1的siRNA,转染至293T细胞,通过RT-qPCR检测其沉默效率并筛选出Piezo1-Homo-2337进行包装,使用荧光染色检测其最佳感染复数值。Piezo1沉默重组慢病毒进一步被转染至人骨髓间充质干细胞中,通过RT-qPCR、Western blot检测其沉默效果;通过茜素红染色、碱性磷酸酶活性分析、免疫荧光染色、RT-qPCR及Western blot检测分析沉默Piezo1对人骨髓间充质干细胞成骨分化能力及TAZ表达的影响。结果与结论:①与正常组、阴性对照组比较,转染si-Piezo1后人骨髓间充质干细胞中Piezo1的mRNA及蛋白水平明显降低;②与阴性对照组比较,si-Piezo1组碱性磷酸酶活性明显降低,钙沉积明显减少;③与阴性对照组比较,si-Piezo1组成骨相关基因Runx2、OPN、DLX5、osteocalcin、β-catenin及TAZ的mRNA水平明显降低。si-Piezo1组TAZ、β-catenin的蛋白表达明显降低;相反,si-Piezo1组p-TAZ、p-β-catenin的蛋白表达明显增加;④与阴性对照组比较,免疫荧光染色结果提示si-Piezo1组人骨髓间充质干细胞中TAZ、β-catenin的表达较少;⑤结果表明Piezo1可促进人骨髓间充质干细胞的成骨分化,沉默Piezo1后人骨髓间充质干细胞的成骨能力明显降低,同时TAZ的表达也同样降低。

骨痹通消颗粒对激素型股骨头坏死人骨髓间充质干细胞成骨与成脂分化的影响

目的 探究骨痹通消颗粒对激素性股骨头坏死人骨髓间充质干细胞(h BMMSC)成骨与成脂分化的影响。方法 取激素性股骨头坏死患者骨髓,体外进行h BMMSC的培养,通过细胞形态学观察、成骨及成脂分化潜能来鉴定h BMMSC。以完培组(基础培养基+10%的胎牛血清)作为对照,采用CCK-8法测定1%、2%、5%、8%、10%体积分数的骨痹通消含药血清A组作用24 h后对细胞增殖的影响。细胞在96孔板的培养过程中,每孔加入地塞米松溶液0.52μl,以完培组(基础培养基+10%的胎牛血清)和损伤组(基础培养组+10%的胎牛血清+0.52μl地塞米松溶液)作为对照,采用CCK-8法测定2%、5%、8%体积分数的骨痹通消含药血清B组对激素环境中细胞活力的影响。以无激素诱导细胞作为对照组(基础培养基+10%的胎牛血清),将体外激素诱导的h BMMSC细胞分为模型组(基础培养基+10%的胎牛血清+10-5mol/L地塞米松溶液)、实验组(基础培养基+5%的骨痹通消含药血清+10-5mol/L地塞米松溶液)。茜素红染色试剂盒测定各组成骨分化后矿化结节的形成;油红O染色试剂盒测定各组成脂分化后脂滴的形成;RT-q PCR测定成脂相关标志基因PPARγ、C/EBP-α与Fabp4中m RNA的表达,Western blot检测成骨相关蛋白BMP-2、Runx2及β-catenin的蛋白表达含量。结果 原代细胞培养7 d后,可见梭形、纺锤形或多角形的贴壁细胞。第三代细胞生长均匀,平行排列,透光率好。成骨诱导21 d,细胞发生改变,局部细胞聚集,并有矿化结节产生,茜素红染色呈阳性;成脂诱导21 d,脂滴与油红O染液结合变为红色,油红O染色呈阳性。与完培组比较,10%体积分数的含药血清A组细胞活力下降(P<0.05)。与完培组比较,损伤组细胞活力下降(P<0.05);与损伤组比较,2%、5%、8%体积分数的含药血清B组细胞活力升高(P<0.05)。与对照组比较,模型组染色面积降低(P<0.05);与模型组比较,实验组中矿化结节与沉积升高,染色范围也更广(P<0.05)。与对照组比较,模型组细胞内脂质积累增多(P<0.05);与模型组比较,实验组较细胞内脂质积累降低(P<0.05)。与对照组比较,模型组PPARγ、C/EBP-α和Fabp4的表达量升高(P<0.01),与模型组比较,实验组PPARγ、C/EBP-α和Fabp4的表达量降低(P<0.05)。与对照组比较,模型组BMP-2、Runx2及β-catenin蛋白表达含量升高(P<0.01),与模型组比较,实验组BMP-2、Runx2及β-catenin蛋白表达含量降低(P<0.05)。结论 骨痹通消颗粒能够促进激素性股骨头坏死h BMMSC的增殖及其向成骨分化的能力,抑制成脂分化,为临床上防治激素性股骨头坏死提供了理论基础。

Mechanisms involved in cancer stem cell resistance in head and neck squamous cell carcinoma

Despite scientific advances in the Oncology field,cancer remains a leading cause of death worldwide.Molecular and cellular heterogeneity of head and neck squamous cell carcinoma(HNSCC)is a significant contributor to the unpredictability of the clinical response and failure in cancer treatment.Cancer stem cells(CSCs)are recognized as a subpopulation of tumor cells that can drive and maintain tumorigenesis and metastasis,leading to poor prognosis in different types of cancer.CSCs exhibit a high level of plasticity,quickly adapting to the tumor microenvironment changes,and are intrinsically resistant to current chemo and radiotherapies.The mechanisms of CSC-mediated therapy resistance are not fully understood.However,they include different strategies used by CSCs to overcome challenges imposed by treatment,such as activation of DNA repair system,anti-apoptotic mechanisms,acquisition of quiescent state and Epithelial-mesenchymal transition,increased drug efflux capacity,hypoxic environment,protection by the CSC niche,overexpression of stemness related genes,and immune surveillance.Complete elimination of CSCs seems to be the main target for achieving tumor control and improving overall survival for cancer patients.This review will focus on the multi-factorial mechanisms by which CSCs are resistant to radiotherapy and chemotherapy in HNSCC,supporting the use of possible strategies to overcome therapy failure.

脂肪间充质干细胞来源外泌体改善放射性口腔黏膜炎

背景:头颈肿瘤放疗极易引起放射性口腔黏膜炎,严重影响患者的健康及肿瘤的治疗计划。间充质干细胞在多种疾病中表现出治疗潜力,外泌体是其发挥功能的重要因素之一。目前尚无脂肪间充质干细胞来源外泌体应用于放射性口腔黏膜炎的相关研究。目的:探讨脂肪间充质干细胞来源外泌体在放射性口腔黏膜炎中的作用。方法:提取脂肪间充质干细胞及脂肪间充质干细胞来源外泌体并进行鉴定。通过3 Gy X射线辐射口腔黏膜上皮细胞诱导放射性口腔黏膜炎体外模型,在造模前给予脂肪间充质干细胞或脂肪间充质干细胞来源外泌体预处理48 h,EdU实验和克隆形成实验检测口腔黏膜上皮细胞的增殖能力。通过3 Gy X射线辐射构建放射性口腔黏膜炎小鼠模型,将脂肪间充质干细胞和脂肪间充质干细胞来源外泌体分别注入放射性口腔黏膜炎小鼠尾静脉,采用苏木精-伊红染色和免疫组化评估口腔黏膜上皮组织的炎症变化。结果与结论:(1)与对照组相比,脂肪间充质干细胞及脂肪间充质干细胞来源外泌体均可促进口腔黏膜上皮细胞克隆形成,增加口腔黏膜上皮细胞EdU阳性率;(2)脂肪间充质干细胞及脂肪间充质干细胞来源外泌体均能够缓解辐射处理小鼠口腔黏膜上皮组织的炎症;口腔黏膜上皮组织中CD45阳性细胞均减少,PCNA阳性细胞均增加。结果表明,脂肪间充质干细胞来源外泌体可促进口腔黏膜上皮细胞增殖,对放射性口腔黏膜炎小鼠口腔黏膜炎症具有缓解作用。

FVFG术与髓芯干细胞植入术治疗股骨头坏死的临床价值比较

目的对比分析吻合血管游离腓骨移植(FVFG)术、髓芯干细胞植入术治疗股骨头坏死(ONFH)的临床价值。方法收集南阳文和骨科医院2021年3月至2023年5月103例ONFH患者临床资料,依照手术方案差异分为两组,对照组(51例)行髓芯干细胞植入术治疗,研究组(52例)行FVFG术治疗,比较两组围术期情况、术后并发症发生率以及术前、术后3个月Harris髋关节功能评分、血液流变学指标、骨代谢指标。结果研究组手术时间为(98.41±10.37)min,长于对照组(47.62±5.19)min(P<0.05);术后3个月,研究组血清25羟基维生素D3[25-(OH)D3]、Harris髋关节功能评分、血清骨碱性磷酸酶(BAP)分别为(24.16±2.55)ng/mL、(91.28±4.57)分、(0.75±0.12)pg/mL,均高于对照组(20.38±2.14)ng/mL、(88.51±6.02)分、(0.64±0.09)pg/mL,全血高切黏度、血清Ⅱ型胶原C端肽(CTX-Ⅱ)、血浆黏度、全血低切黏度分别为(4.27±0.65)mPa·s、(132.71±18.69)ng/L、(1.84±0.52)mPa·s、(7.36±1.80)mPa·s,均低于对照组(4.80±0.71)mPa·s、(161.80±21.43)ng/L、(2.16±0.47)mPa·s、(8.94±1.72)mPa·s(P<0.05);研究组术后并发症发生率5.77%较对照组19.61%低(P<0.05)。结论与髓芯干细胞植入术比较,FVFG术治疗ONFH的手术时间较长,但对患者髋关节功能、血液流变学指标、骨代谢指标水平的改善效果更好。

自体骨髓间充质干细胞移植在老年股骨头坏死髓芯减压术中的应用价值

目的观察自体骨髓间充质干细胞(BMSCs)移植在老年股骨头坏死(ONFH)髓芯减压术中的应用效果。方法该研究为前瞻性研究,选择平顶山市第一人民医院2022年1月至2023年5月收治的113例ONFH老年患者为研究对象,以计算机随机分组法将其分为试验组(57例)和对照组(56例),对照组予以传统髓芯减压术治疗,试验组经自体BMSCs移植联合髓芯减压术治疗,比较两组治疗前后的BMSCs表面分子、骨代谢水平、骨密度、骨坏死面积、髋关节功能及疼痛症状改善情况。结果在不同治疗方案下,试验组的BMSCs表面分子CD34、CD105、CD166表达量分别为(2.48±0.76)%、(13.36±3.27)%、(37.49±5.28)%,均高于对照组[(1.92±0.36)%、(11.45±2.13)%、(34.23±5.16)%](P<0.05);试验组骨保护素(OPG)、骨钙素(BGP)、成纤维细胞生长因子(FGF-2)分别为(5.45±1.36)pg/mL、(6.27±1.45)μg/mL、(43.35±10.46)ng/L,均高于对照组[(4.41±1.36)pg/mL、(5.52±1.44)μg/mL、(37.25±10.41)ng/L](P<0.05);试验组术后1个月、术后3个月的骨密度分别为(0.32±0.11)g/cm^(2)、(0.49±0.23)g/cm^(2),均高于对照组[(0.25±0.11)g/cm^(2)、(0.35±0.16)g/cm^(2)](P<0.05);骨坏死面积分别为(26.44±5.17)%、(20.45±5.23)%,均低于对照组[(29.12±5.33)%、(23.32±5.18)%](P<0.05);试验组术后1个月、术后3个月的视觉模拟疼痛量表(VAS)评分分别为(4.25±1.27)分、(2.82±0.46)分,均低于对照组[(5.33±1.77)分、(3.21±0.52)分](P<0.05);试验组术后1个月、术后3个月的髋关节Harris量表评分分别为(70.33±10.32)分、(85.66±10.45)分,均高于对照组[(65.22±10.36)分、(80.33±10.71)分](P<0.05)。结论自体BMSCs能通过上调骨髓干细胞表面分子而改善老年ONFH患者的骨代谢水平,自体BMSCs移植联合髓芯减压术可有效提升患者骨密度水平,并减少骨坏死面积,对缓解患者髋关节疼痛、促进髋关节功能恢复均有积极意义。

Current concepts on osteonecrosis of the femoral head

It is estimated that 20000 to 30000 new patients are diagnosed with osteonecrosis annually accounting for approximately 10% of the 250000 total hip arthroplasties done annually in the United States. Thelack of level 1 evidence in the literature makes it difficult to identify optimal treatment protocols to manage patients with pre-collapse avascular necrosis of the femoral head, and early intervention prior to collapse is critical to successful outcomes in joint preserving procedures. There have been a variety of traumatic and atraumatic factors that have been identified as risk factors for osteonecrosis, but the etiology and pathogenesis still remains unclear. Current osteonecrosis diagnosis is dependent upon plain anteroposterior and frog-leg lateral radiographs of the hip, followed by magnetic resonance imaging(MRI). Generally, the first radiographic changes seen by radiograph will be cystic and sclerotic changes in the femoral head. Although the diagnosis may be made by radiograph, plain radiographs are generally insufficient for early diagnosis, therefore MRI is considered the most accurate benchmark. Treatment options include pharmacologic agents such as bisphosphonates and statins, biophysical treatments, as well as joint-preserving and joint-replacing surgeries. the surgical treatment of osteonecrosis of the femoral head can be divided into two major branches: femoral head sparing procedures(FHSP) and femoral head replacement procedures(FHRP). In general, FHSP are indicated at pre-collapse stages with minimal symptoms whereas FHRP are preferred at post-collapse symptomatic stages. It is difficult to know whether any treatment modality changes the natural history of core decompression since the true natural history of core decompression has not been delineated.

Effect of HuoguⅡFormula(活骨Ⅱ方) with Medicinal Guide Radix Achyranthis Bidentatae on Bone Marrow Stem Cells Directional Homing to Necrosis Area after Osteonecrosis of the Femoral Head in Rabbit

Objective： To investigate the effect of Huogu II Formula （活骨 II方 ） with medicinal guide Radix Achyranthis Bidentatae （Ach） on bone marrow stem cells （BMSCs） homing to necrosis area after osteonecrosis of the femoral head （ONFH） frozen by liquid nitrogen in rabbit as well as to explore the mechanism of prevention and treatment for ONFH. Methods： The animal model of ONFH was established by liquid nitrogen frozen on the rabbit left hind leg. Forty-eight Japanese White rabbits were randomly assigned to sham-operated group, model group, Huogu II group, and Huogu II plus Ach group, with 12 rabbits in each. During the course of ONFH animal model establishment, all rabbits were subcutaneously injected with recombinant human granulocyte colony-stimulating factor [rhG-CSF, 30 μg/（kg.day） for continuous 7 days]. Meanwhile, normal saline and decoction of the two formulae were administrated by gavage, respectively. White blood cells （WBC） were counted in peripheral blood before and after injection of rhG-CSF. Materials were drawn on the 2rid and 4th weeks after model built; bone glutamine protein （BGP） and bone morphogenetic protein 2 （BMP2） levels in serum were tested. Histopathologic changes were observed by hematoxylin and eosin （HE） staining. BMP2 mRNA levels were detected with in situ hybridization （iSH） staining. 5-Bromo-2＇-deoxyuridine （BrdU） and stromal cell derived factor 1 （SDF-1） were measured by immunohistochemical assay in femoral head of the left hind leg. Results： Compared with the shamoperated group, the ratio of empty lacuna, serum BGP, and SDF-1 level in the model group increased significantly, and BMP2 in both serum and femoral head decreased significantly. However, in comparison with the model group, the empty lacuna ratio of Huogu II group and Huogu II plus Ach group decreased obviously in addition to the levels of serum BGP and BMP2, and the expressions of BMP2 mRNA, BrdU, and SDF-1 increased significantly. Above changes were particularly obvious in Huogu II plus Ach group. BGP and SDF-1 on the 2nd week and empty lacuna rate and serum BMP2 level on the 4th week in Huogu II group significantly exceeded their counterparts. On the 2nd week, only in Huogu II plus Ach group that the BrdU counting rose significantly. On the 4th week, empty lacuna rate and serum BMP2 level in Huogu II plus Ach group exceeded those in Huogu II group distinctively. Conclusions： To a certain extent, the medicinal guide Ach improves the preventive and therapeutic effects of Huogu II Formula on expedmental ONFH model. The possible mechanism of this is related to its promoting effect on directional homing of BMSCs to the necrosis area.