Effects of Nanofiber Scaffolds Coated with Nanoparticulate and Microparticulate Freeze Dried Bone Allograft on the Morphology,Adhesion, and Proliferation of Human Mesenchymal Stem Cells

Background:Freeze dried bone allograft nanoparticles on a nanofiber membrane may serve as an ideal scaffold for bone regeneration. This study aimed to assess the biological behavior of human MSCs in terms of proliferation and adhesion to nanoparticulate and microparticulate FDBA scaffolds on PLLA nanofiber membrane. Methods:In this experimental study, PLLA nanofiber scaffolds were synthesized by the electrospinning method. The FDBA nanoparticles were synthesized mechanically. The FDBA nanoparticles and microparticles were loaded on the surface of PLLA nanofiber membrane. A total of 64 scaffold samples in four groups of n-FDBA/PLLA, FDBA/PLLA, PLLA and control were placed in 24-well polystyrene tissue culture plates; 16 wells were allocated to each group. Data were analyzed using one-way ANOVA and Bonferroni test. Results:The proliferation rate of MSCs was significantly higher in the nanoparticulate group compared to the microparticulate group at five days (p = 0.034). Assessment of cell morphology at 24 hours revealed spindle-shaped cells with a higher number of appendages in the nanoparticulate group compared to other groups. Conclusion:MSCs on n-FDBA/PLLA scaffold were morphologically more active and flatter with a higher number of cellular appendages, as compared to FDBA/PLLA. It seems that the nanoparticulate scaffold is superior to the microparticulate scaffold in terms of proliferation, attachment, and morphology of MSCs in vitro.Key Words: Allografts, Bone regeneration, Mesenchymal stem cells, Nanofibers     

Characteristics of Human Endometrial Stem Cells in Tissue and Isolated Cultured Cells: An Immunohistochemical Aspect

Background:

The aim of this study was to investigate the percentage of the stem cells population in human endometrial tissue sections and cultured cells at fourth passage.

Methods:

Human endometrial specimens were divided into two parts, one part for morphological studies and the other part for in vitro culture. Full thickness of human normal endometrial sections and cultured endometrial cells at fourth passage were analyzed via immunohistochemistry for CD146 and some stemness markers such as Oct4, Nanog, Sox2, and Klf4 and the expression of typical mesenchymal stem cell markers CD90, CD105.

Results:

11.88±1.29% of human endometrial cells within tissue sections expressed CD146 marker vs. 28±2.3% of cultured cells, CD90 and CD105 were expressed by functionalis stroma (85±2.4 and 89±3.2%) than basalis stroma (16±1.4 and 17±1.9%), respectively (P<0.05). Oct4 and Nanog-expressing cells comprise 1.43±0.08 and 0.54±0.01% of endometrial stromal cells in endometrial sections vs. 12±3.1% and 8±2.9% of cultured cells, respectively. They reside near the glands in the basal layer of endometrium. Sox2 and Klf4 were not commonly expressed in tissue samples and cultured cells. CD9 and EpCAM were expressed by epithelial cells of the endometrium, rather than by stroma or perivascular cells.

Conclusion:

The human endometrial stem cells and pluripotency markers may be localized more in basalis layer of endometrium. The immunostaining observations of endometrial cells at fourth passage were correlated with the immunohistochemistry data.Key Words: Endometrium, Immunohistochemistry, Mesenchymal stem cells     

The Effect of Human Platelet-Rich Plasma on Adipose-Derived Stem Cell Proliferation and Osteogenic Differentiation

Background: The cultured mesenchymal stem cells (MSC) have been used in many clinical trials; however, there are still some concerns about the cultural conditions. One concern is related to the use of FBS as a widely used xenogeneic supplement in the culture system. Human platelet-rich plasma (hPRP) is a candidate replacement for FBS. In this study, the effect of hPRP on MSC proliferation and osteogenic differentiation has been evaluated. Methods: Human adipose-derived stem cells (hADSC) were expanded. Cells from the third passage were characterized by flow cytometric analysis and used for in vitro experiments. Resazurin and alizarin red stains were used for cell proliferation and osteogenic differentiation assays, respectively. Results: Treatment with hPRP resulted in a statistically significant increase in cell proliferation compare to the negative control group (P<0.001). Cell proliferation in the 15% hPRP group was also significantly higher than that in the 10% hPRP group (P<0.05). Additionally, it caused less osteogenic differentiation of the hADSC compared to the FBS (P<0.001), but in comparison to negative control, it caused acceptable mineralization (P<0.001). Conclusion: These findings indicate that hPRP not only improves the proliferation but also it can be a suitable substitution in osteogenic differentiation for clinical purposes. However, the clinical application value of hPRP still needs more investigation. Key Words: Platelet-Rich Plasma, Adipose tissue, Stem Cells, Cell differentiation, Cell proliferation     

砖茶氟的防龋功效及其应用

龋齿是世界性疾病,通过对青砖茶的防龋试验,表明它有明显的防龋效果,且价廉味浓,便于普及推广。     

茶黄素对高糖诱导的人晶状体上皮细胞氧化损伤的影响

研究茶黄素(Theaflavin,TF)对高糖(High glucose,HG)诱导的人晶状体上皮细胞(Human lens epithelial cells,HLECs)氧化损伤的影响。采用体外培养人晶状体上皮细胞,用葡萄糖诱导细胞成氧化损伤模型,外加一定浓度的茶黄素干预,比色法检测细胞体内SOD、CAT、GSH-Px、MDA活力和含量的变化。结果表明相对于正常培养的细胞,高糖诱导细胞体内SOD、CAT、GSH的活性降低,MDA含量上升;加入茶黄素后,提高了细胞内SOD、CAT、GSH-Px的活力,降低了细胞内MDA的含量,与模型组细胞相比,具有显著差异(P0.01)。     

Fucoidan Independently Enhances Activity in Human Immune Cells and Has a Cytostatic Effect on Prostate Cancer Cells in the Presence of Nivolumab

Fucoidan compounds may increase immune activity and are known to have cancer inhibitory effects in vitro and in vivo. In this study, we aimed to investigate the effect of fucoidan compounds on ex vivo human peripheral blood mononuclear cells (PBMCs), and to determine their cancer cell killing activity both solely, and in combination with an immune-checkpoint inhibitor drug, Nivolumab. Proliferation of PBMCs and interferon gamma (IFNγ) release were assessed in the presence of fucoidan compounds extracted from Fucus vesiculosus, Undaria pinnatifida and Macrocystis pyrifera. Total cell numbers and cell killing activity were assessed using a hormone resistant prostate cancer cell line, PC3. All fucoidan compounds activated PBMCs, and increased the effects of Nivolumab. All fucoidan compounds had significant direct cytostatic effects on PC3 cells, reducing cancer cell numbers, and PBMCs exhibited cell killing activity as measured by apoptosis. However, there was no fucoidan mediated increase in the cell killing activity. In conclusion, fucoidan compounds promoted proliferation and activity of PBMCs and added to the effects of Nivolumab. Fucoidan compounds all had a direct cytostatic effect on PC3 cells, as shown through their proliferation reduction, while their killing was not increased.     

化控剂对玉米光合、激素及茎秆力学特性的影响

以东农253为试验材料,在玉米9叶期喷施密高和KP(主要成分DCPTA和ETH),在玉米5个生育时期测定叶片光合酶活、内源激素及茎秆力学特性参数,并进行测产及相关性分析,探究大田条件下化控剂对玉米生长发育的影响。结果表明,喷施密高和KP后叶片SPAD、P_n、RuBPc和PEPc显著提高,增加叶片IAA、GA和ZR含量并降低ABA含量。玉米茎秆单位节间长度干物重、穿刺强度、横折强度和田间致倒伏推力有所增大,降低扁率及子粒含水量,密高和KP处理下玉米产量分别增加6.47%和9.23%。相关性分析表明,除ABA含量与产量呈显著负相关外,光合和其他激素指标呈显著正相关,PEPc、SPAD在完熟期与产量相关性最大,RuBPc在乳熟期与产量相关性最大,P_n在灌浆初期与产量相关性最大。     

Lxx侵染甘蔗后在蔗茎中的积累及对蔗茎组织结构的影响

以接种病原菌 Leifsonia xyli subsp. Xyli（Lxx）的‘拔地拉’（‘Badila’）甘蔗为研究材料,利用实时定量PCR技术研究病原菌Lxx在蔗茎中的积累情况,采用半薄切片技术研究茎维管束的病变规律。结果表明,在接种Lxx的Badila中,Lxx菌量积累随蔗茎由基层到尾层呈现递减趋势。半薄切片结果表明,接种Lxx蔗茎基部维管组织中的韧皮部和木质部损坏严重,甘蔗茎节处相对茎间的基本细胞小得多,且具有木质部异形、细胞大小不一、细胞整体较为密集。接种Lxx蔗茎维管组织中的韧皮部和木质部损坏程度与Lxx菌量分布呈正相关。甘蔗茎节与茎间的组织结构差异是Lxx菌量由基层到尾层呈现递减的原因之一。本研究为进一步研究Lxx与甘蔗寄主的互作提供了理论依据。     

浸种和外源激素处理对毛马齿苋种子发芽的影响

为提高毛马齿苋种子播种出苗率,分析不同时间浸种以及采用GA3、NAA和6-BA 3种外源激素处理对其种子发芽的影响。结果表明：浸种处理没有提高毛马齿苋种子的发芽势和发芽率的效果;GA3、NAA和6-BA三种激素浸种处理均能提高毛马齿苋种子的发芽势和发芽率;最适宜的浸种浓度分别为GA3 30 mg/L、10～20 mg/L NAA或6-BA 30 mg/L。其中GA3有效浓度范围较宽泛,建议采用,但GA3处理不能解除该种子发芽的需光性。     

水分胁迫对大豆苗期叶片内源激素含量与保护酶活性的影响

在盆栽条件下,以大豆(Glycine max)绥农10号为材料,通过人为控制水分(干旱和水渍处理),比较叶片中几种内源激素含量变化的差异,研究水分胁迫对大豆幼苗保护酶活性的影响.结果表明:干旱处理提高了大豆苗期叶片内赤霉素(GA)、脱落酸(ABA)的含量但也降低了玉米素核苷(ZR)和生长素(IAA)的含量,同时干旱处理提高了丙二醛(MDA)含量以及超氧化物歧化酶(SOD)活性;水渍处理降低了ABA含量和SOD活性,提高了IAA、GA以及MDA的含量.     

马铃薯应用外源激素及叶面微肥的增产效果

为了探索马铃薯应用外源激素及叶面微肥的增产效果,选取农户普遍应用的马铃薯块茎膨大剂6种,新面世的产品3种,加对照共10个处理,进行小区试验。经方差和效益分析表明:欧罗壮叶面肥较对照增产13.0%,增加纯效益2 289元.hm-2,投入产出比1:16.9;狮王花宝叶面肥增产11.7%,增加效益2 123元.hm-2,投入产出比1:17;凯普克增产10.3%,增加效益1 791元.hm-2,投入产出比1:12.4;三禾绿丰增产6.1%,增加效益1 058元.hm-2,投入产出比1:9.6;金农富增产3.2%,增加效益497元.hm-2,投入产出比1:4.7。其它产品增产不明显,有些产品出现减产现象,在生产上应小心选用。     

in vitro Anti-Proliferative Effect of Adiponectin on Human Endometriotic Stromal Cells through AdipoR1 and AdipoR2 Gene Receptor Expression

Background:

Endometriosis is a complex disorder in reproductive age women which consist of stromal and epithelial cells implantation outside the uterine cavity. Adiponectin is a member of cytokine family with various metabolic roles and proliferation inhibition of many cancer cells. The aim of the present research was to determine adiponectin effect on human endometriotic stromal cells (ESCs) proliferation and their expression of adiponectin receptors.

Methods:

In this experimental study, endometrial biopsies (n=7) were taken. ESCs isolation was done by enzymatic digestion and cell filtrations. ESCs of each biopsy were divided into four groups: 0 (control), 10, 100, and 200 ng/ml adiponectin concentrations in three different times (24, 48, or 72 h). The effect of adiponectin on ESC viability and expression of mRNA Adipo receptor1 (R1) and Adipo receptor2 (R2) was determined by Trypan blue staining and semi-quantitative RT-PCR, respectively. Data were analyzed by one-way ANOVA and unpaired student’s t-test, and P<0.05 was considered statistically significant.

Results:

Adiponectin inhibited human endometriotic stromal cell proliferation in time- and dose-dependent manners significantly (P=0.001). Expression of AdipoR1 and AdipoR2 gene receptors was increased in human ESCs significantly (P<0.05).

Conclusions:

Adiponectin can suppress endometriosis by inhibiting ESC proliferation and increased AdipoR1 and AdipoR2 expression.Key Words: Adiponectin, Adiponectin receptors, Endometriosis, Stromal cells     

外源激素与蔗糖对冬小麦穗粒数和粒重的调控效应

为阐明外源激素和蔗糖调控冬小麦穗粒数和粒重的机理,以冬小麦农大211为材料,设置不同浓度蔗糖和激素,通过离体穗培养及盆栽不同水分条件下喷施激素处理,考察了冬小麦穗粒数、穗粒重、千粒重以及穗部糖含量对外源蔗糖和激素的响应。离体穗培养试验表明,蔗糖浓度为40 g·L^-1,IAA、GA、6-BA和ABA浓度分别为10^-4 mol·L^-1、10^-5 mol·L^-1、10^-5 mol·L^-1和10^-6 mol·L^-1时取得了较高的穗粒数,且穗粒数以穗上部强势粒数与穗中部弱势粒数增幅较大;适当增加培养基GA和ABA浓度也能提高粒重,从而提升穗粒重。盆栽试验表明,与水分适宜处理相比,干旱使成熟期穗粒数、穗粒重和单株生物量显著下降,千粒重无显著变化,干旱处理的幼穗中CTK和ABA含量显著增加。干旱条件下,喷施IAA较其对照显著增加了穗粒重和单株生物量以及幼穗中可溶性糖和IAA含量,显著降低了幼穗ABA含量;喷施GA较其对照显著增加了单株生物量和幼穗GA含量,显著降低了幼穗ABA含量;喷施6-BA较其对照显著增加了单株生物量和幼穗CTK含量,显著降低了幼穗可溶性糖和ABA含量;喷施ABA较其对照显著增加了穗粒重和单株生物量,显著降低了幼穗蔗糖和可溶性糖含量。水分适宜条件下,喷施IAA较其对照显著降低了穗粒数、穗粒重和单株生物量,但显著增加了千粒重和幼穗可溶性糖、CTK和ABA含量;喷施GA较其对照显著增加了幼穗可溶性糖、GA、CTK和ABA含量,但显著降低了幼穗蔗糖含量;喷施6-BA较其对照显著增加了幼穗可溶性糖、CTK和ABA含量;喷施ABA较其对照显著降低了穗粒数,显著增加了幼穗可溶性糖、IAA、CTK和ABA含量。总之,外源蔗糖与激素对冬小麦穗粒数、穗粒重和千粒重都有显著影响,适宜蔗糖和激素处理提高穗粒数,主要由于增加了穗上部强势粒和穗中部弱势粒数。外源喷施激素可缓解干旱胁迫对冬小麦穗粒数和穗粒重的影响,以喷施IAA效果最好,主要通过调控幼穗糖含量和激素含量来调控穗粒数和粒重。     

甲基氨草磷对橡胶树悬浮细胞同步化及微核诱导的影响

植物微原生质体融合(microprotoplast fusion)是将部分基因组转移而又避免染色体损伤的不对称融合技术,微原生质体成功诱导是微原生质体融合的先决条件。本研究在摸索甲基氨草磷(amiprophos-methyl,APM)对橡胶树悬浮细胞有丝分裂中期指数和染色体形态影响基础上,研究了酶解时间及酶解过程中添加APM对微原生质体产量的影响。结果发现,悬浮细胞随APM处理浓度和时间的增加,有丝分裂中期指数先升高再下降,处理12 h时达到最大值;染色体聚集程度也先增加再降低,处理16 h时染色体开始解聚。在酶解过程中添加APM和延长酶解时间会大大降低微原生质体的产量。悬浮细胞经过1 μmol/L APM处理10 h后,酶解10 h,中期细胞可达到79.4%,微核化细胞指数达到7.3%,从而建立了APM诱导橡胶树悬浮细胞同步化及高效获得橡胶树微化细胞的方法。     

低温胁迫对稻茬小麦根系抗氧化酶活性及内源激素含量的影响

为探索低温对小麦根系的伤害机理,在分蘖期和拔节期模拟低温,研究低温胁迫对稻茬不同小麦品种(烟农19,抗寒性较强;郑麦9023,抗寒性较差)根系抗氧化酶活性及内源激素含量的影响。结果表明,低温胁迫对小麦根系中抗氧化酶活性和内源激素含量的影响均达到显著水平(P0.05)。2个小麦品种受到低温胁迫后,根系中超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性均显著增强,丙二醛含量(MDA)显著下降,且拔节期低温处理小麦的抗氧化酶活性增强幅度更大,MDA含量下降幅度更小。抗寒性较强的小麦品种(烟农19)根系中脱落酸(ABA)、赤霉素(GA3)和生长素(IAA)含量在低温胁迫后均表现为上升趋势。抗寒性较差的品种(郑麦9023)根系中GA3含量在低温胁迫后增加;ABA含量在低温胁迫后下降;IAA含量在分蘖期低温胁迫后表现为增加,拔节期表现为下降。以上结果表明,分蘖期和拔节期遭受低温后,不同品种稻茬小麦根系中抗氧化酶活性及激素含量受影响的程度不同,抗寒性差的品种根系受低温影响较大,且以拔节期影响更为严重。     

剪叶对不同穗型小麦品种光合速率及单茎产量的影响

为了探讨高产条件下不同穗型小麦品种叶片对产量的贡献,选用两种穗型小麦品种为试验材料,研究了抽穗期剪叶对剩余叶片光合速率、茎秆干重及籽粒结实与粒重的影响.结果表明,剪叶可提高剩余叶片光合速率,以多穗型品种豫麦49剩余叶片净光合速率增幅较大,但不足以弥补剪叶造成的损失,表现为结实粒数减少、粒重降低;随被剪叶片数的增加和叶位的升高,结实粒数和粒重降低更多.剪叶后大穗型品种宿2001茎秆干重明显减轻.抽穗期剪叶对多穗型品种豫麦49籽粒产量的影响大于大穗型品种宿2001.     

施用ABT5号增产灵对甜菜农艺性状和内源激素含量的影响

在框栽和田间条件下研究了ABT5号增产灵浸种处理和叶喷处理对甜菜农艺性状及对甜菜叶片和根系内源激素含量变化的影响。结果表明,ABT5号处理明显增加根中赤毒素(GA_(1、3、4、7))、生长素(IAA)含量。明显增加根及叶中细胞分裂素(DHZR_S)含量,降低脱落酸(ABA)含量,导致刺激生长的激素(DHZR_S GA_(1、3、4、7) IAA)与抑制生长激素(ABA)的比例在根组织中表现出浸种处理>叶喷处理>CK的结果。这是ABT5号处理后甜菜块根增产的生理机制之一。本实验证实,ABT5号处理仅促进主根生长,不形成簇状叉根。浸种处理在框栽和田间试验中分别比对照增产12.58％和13.10％,叶喷处理比对照增产9.85％和10.67％,ABT5号处理对块根含糖率没有影响。     

氧化应激对Vero细胞的损伤及EGCG的保护作用

研究了氧化应激对肾细胞Vero细胞的损伤及EGCG的保护作用。用MTT、吖啶橙染色和DNA凝胶电泳等方法研究了H2O2和Cr6+应激对Vero细胞的损伤,及EGCG对凋亡细胞的保护作用及其机理。结果表明,H2O2和Cr6+剂量效应地抑制Vero细胞活性,IC50分别为175.6和9.8mgL-1;其中50 mgL-1 H2O2和400 mgL-1/2h Cr6+可诱导Vero细胞凋亡。0~60 mgL-1 EGCG有效抑制H2O2和Cr6+应激引起的细胞活性下降,且40 mgL-1 EGCG显著抑制细胞凋亡。对于Cr6+所诱导的细胞凋亡,EGCG的保护作用与EDTA和Vc的协同作用效果相当,表明清除活性氧和络合金属离子都有助于减轻Cr6+应激损伤。可见,EGCG通过清除活性氧和络合离子有效地保护了肾细胞免受应激损伤,这对易遭受活性氧损伤的肾脏无疑具有一定的临床价值。