Clonal isolation of endothelial colony-forming cells from early gestation chorionic villi of human placenta for fetal tissue regeneration

BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an essential physiological process that occurs during embryonic development and tissue regeneration.Angiogenesis is the growth of new capillaries from pre-existing blood vessels,which is observed both prenatally and postnatally.The placenta is an organ composed of a variety of fetal-derived cells,including ECFCs,and therefore has significant potential as a source of fetal ECFCs for tissue engineering.AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi(CV-ECFCs)of the placenta,and assess their potential for tissue engineering.METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion.Cells expressing CD31 were selected using magnetic-activated cell sorting,and plated in endothelial-specific growth medium.After 2-3 wks in culture,colonies displaying cobblestone-like morphology were manually picked using cloning cylinders.We characterized CV-ECFCs by flow cytometry,immunophenotyping,tube formation assay,and Dil-Ac-LDL uptake assay.Viral transduction of CVECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector,and transduction efficiency was tested by fluorescent microscopy and flow cytometry.Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved,small intestinal submucosa extracellular matrix scaffold.RESULTS After four passages in 6-8 wks of culture,we obtained a total number of 1.8×107 CV-ECFCs using 100 mg of early gestational chorionic villus tissue.Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31,CD144,CD146,CD105,CD309,only partially expressed CD34,and did not express CD45 and CD90.CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation,similar to cord blood-derived ECFCs(CB-ECFCs).CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%.Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold.CONCLUSION In summary,we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs.A substantial number of CV-ECFCs can be obtained within a short time frame,representing a promising novel source of ECFCs for fetal treatments.

Hierarchy of mesenchymal stem cells: Comparison of multipotentmesenchymal stromal cells with fibroblast colony forming units

The organization of the compartment of mesenchymal stem cells is still obscure. Two types of human stromal precursor cells are known. Both of them are analyzed in in vitro system: mesenchymal multipotent stromal cells (MMSC) and fibroblast colony forming units (CFU-F). The aim of this study was to compare the main characteristics of MMSC and CFU-F derived from the bone marrow of 24 healthy donors. Growth and differentiation parameters, as well as relative expression levels of different genes were analyzed in MMSC and CFU-F. MMSC were cultivated for 5 passages. CFU-F concentration was determined for each bone marrow sample. The data obtained demonstrated the heterogeneity and hierarchical organization of both studied populations of stromal precursor cells-MMSC and CFU-F. These two types of stromal precursor cells turned to be different in most parameters studied. Altogether MMSC seemed to be more immature cells than CFU-F and took up the higher position in hierarchical tree of mesenchymal stem cells. The rate of differentiation and proliferative potential decreased with the donor’s age in both populations MMSC and CFU-F.

FEA technique of hot plate forming process using cell-typed die with cooling device

Hot plate forming using a cell-typed die is a process for forming a large thick plate with a spherical shape for the manufacture of a large spherical LNG tank.Cell-typed upper and lower dies made of a framework of steel plates fitted to make a grid pattern are used in this process,and an air-cooling device is separately installed inside the lower die.A finite element analysis (FEA) technique was developed,which included hot forming,air flow,cooling and thermal deformation analysis for the hot plate forming process using the cell-typed die.Further,the convective and interface heat transfer coefficients were used to reproduce analytically the effects of the cooling device in the hot plate forming analysis.A small-scale model test of the process was conducted to verify the FEA technique.The analysis results show that the curvature of the final plate agrees well with that of the designed experiment within a maximum relative error of 0.03% at the corner of the plate.

Biofilm forming and leaching mechanism during bioleaching chalcopyrite by Thiobacillus ferrooxidans

The mechanism of attachment and leaching of thiobacillus ferrooxidans（T.f.） on chalcopyrite were studied. The shaking flasks with bacteria were observed by SEM. The process of T.f attached to the surface of the mineral sample and the biofilm forming were described. The promoting role of the biofilm for bioleaching was discussed. The existence of Fe2+ in the exopolysaccharide layer of T.f was demonstrated by EM（electronic microscope）cell-chemistry analysis. These results show that under the proper growth condition of bacteria, bioleaching of chalcopyrite results in the formation of complete biofilm after 23 weeks. There are iron ions in the outer layer polymer of T.f., which provides the micro-environment for themselves, and can guaruntee the energy needed for the bacteria growth in the biofilm. At the same time, Fe3+ions produced oxidize sulfide which brings about the increase of both growth rate of the bacterial and leaching rate of sulfide minerals.

Shape formation of closed-cell aluminum foam in solid–liquid–gas coexisting state

The mold pressing process was applied to investigate the formability of closed-cell aluminum foam in solid–liquid–gas coexisting state.Results show that the shape formation of closed-cell aluminum foam in the solid–liquid–gas coexisting state was realized through cell wall deformation and cell movement caused by primary α-Al grains that slid,rotated,deformed,and ripened within cell walls.During formation,characteristic parameters of closed-cell aluminum foam were almost unchanged.Under proper forming conditions,shaped products of closed-cell aluminum foam could be fabricated through mold pressing.

用Cell组件生成测试报表

介绍了一种 ＡｃｔｉｖｅＸ表格控件 Ｃｅｌｌ组件，并且详细地讨论了该组件在测试报表生成中的应用。

Effects of preservation time on proliferative potential of human limbal stem/progenitor cells

AIM: To determine the proliferative potential and the maintenance of stem cell activity in stored human limbal tissues, and correlate this with the preservation time, cell viability and the expression of stem cell markers. METHODS: Thirty limbal rims were split into 4 parts and stored in corneal preservation medium at 4 degrees C for 0, 1, 4, or 7 days. The limbal stem cell and mitotic markers P63, CK19, proliferating cell nuclear antigen (PCNA), and Ki67 were determined by immunohistochemical staining. The proliferative potential of limbal epithelial cells was assessed by cell viability, the ability of generating stratified epithelium, and colony forming assay. RESULTS: The stored tissues maintained limbal stratified structure to 7 days and exhibited comparable expression level of stem cell and mitotic markers. The proportion of viable cells decreased with the prolonged preservation time, while colony forming efficiency decreased from the 1st day and disappeared at the 4th day. When inoculated on amniotic membrane, the cells preserved for 1 day formed a stratified epithelium, while the cells from 4 days' preservation formed a discontinuous layer. CONCLUSION: The colony forming efficiency of limbal epithelial stem/progenitor cells decreased rapidly with the increasing preservation time, while the expression level of markers and capacity of forming epithelial monolayer on amniotic membrane decreased gradually. The limbal epithelial stem cells lost their function earlier than the lost expression level of stem cell markers. This may help us to better choose the appropriate preservation grafts for future limbal stem cell transplantation.

Mesenchymal stem cells and the neuronal microenvironment in the area of spinal cord injury

Cell-based technologies are used as a therapeutic strategy in spinal cord injury(SCI). Mesenchymal stem cells(MSCs), which secrete various neurotrophic factors and cytokines, have immunomodulatory, anti-apoptotic and anti-inflammatory effects, modulate reactivity/phenotype of astrocytes and the microglia, thereby promoting neuroregeneration seem to be the most promising. The therapeutic effect of MSCs is due to a paracrine mechanism of their action, therefore the survival of MSCs and their secretory phenotype is of particular importance. Nevertheless, these data are not always reported in efficacy studies of MSC therapy in SCI. Here, we provide a review with summaries of preclinical trials data evaluating the efficacy of MSCs in animal models of SCI. Based on the data collected, we have tried(1) to establish the behavior of MSCs after transplantation in SCI with an evaluation of cell survival, migration potential, distribution in the area of injured and intact tissue and possible differentiation;(2) to determine the effects MSCs on neuronal microenvironment and correlate them with the efficacy of functional recovery in SCI;(3) to ascertain the conditions under which MSCs demonstrate their best survival and greatest efficacy.

Mechanism of the DL-alpha-aminoadipic acid inhibitory effect on form-deprived myopia in guinea pig

AIM:To investigate the effect of intravitreal injection of DL-alpha-aminoadipic acid (DL-α-AAA) on ocular refractive state and retinal dopamine, transforming growth factor-β2 (TGFβ2 ), vasoactive intestinal polypeptide (VIP) in guinea pig form-deprived myopia. METHODS:Four-week-old pigmented guinea pigs were randomly assigned to 4 groups:normal control, deprivation, deprivation plus DL-α-AAA, deprivation plus saline. Form deprivation was induced with the self-made translucent eye shields, and lasted for 14 days. 8μg DL-α-AAA was injected into the vitreous chamber of deprived eyes. The corneal radius of curvature, refraction and axial length were measured. Retinal dopamine content was evaluated by the high-performance liquid chromatography with electrochemical detection, and TGFβ2 and VIP protein were detected by Western blotting. RESULTS:Fourteen days of eye occlusion caused the axial length to elongate and become myopic in the form-deprived eyes, with the decrease of retinal dopamine and the increase of TGFβ 2 and vasoactive intestinal polypeptide (VIP) protein. Intravitreal injection of DL-α-AAA could inhibit the myopic shift from (-3.65±1.06)D to (-1.48 ±0.63)D, P 【0.01 due to goggles occluding and cause the decrease of retinal TGFβ2 protein in the deprived eyes. However, intravitreal injection of DL-α-AAA had no significant effect on retinal dopamine and VIP protein in deprived eyes. Retinal TGFβ2 protein correlated highly with the ocular refraction (y =-3.34 + 0.31/x , F =74.75, P 【0.001) and axial length (y =8.39-0.02/ x , F =48.32, P 【0.001) in different treatment groups. ·CONCLUSION:Intravitreal injection of DL-α-AAA is effectively able to suppress the development of form deprivation myopia, which may be associated with retinalTGFβ2 protein in guinea pigs.

Neuronal-like cell differentiation of non-adherent bone marrow cell-derived mesenchymal stem cells

Non-adherent bone marrow cell-derived mesenchymal stem cells from C57BL/6J mice were sepa- rated and cultured using the ＂pour-off＂ method. Non-adherent bone marrow cell-derived mesen- chymal stem ceils developed colony-forming unit-fibroblasts, and could be expanded by supple- mentation with epidermal growth factor. Immunocytochemistry showed that the non-adherent bone marrow cell-derived mesenchymal stem cells exposed to basic fibroblast growth factor/epidermal growth factor/nerve growth factor expressed the neuron specific markers, neurofilament-200 and NeuN, in vitro. Non-adherent bone marrow cell-derived mesenchymal stem cells from 13-galactosidase transgenic mice were also transplanted into focal ischemic brain （right corpus striatum） of C57BL/6J mice. At 8 weeks, cells positive for LacZ and 13-galactosidase staining were observed in the ischemic tissues, and cells co-labeled with both 13-galactosidase and NeuN were seen by double immunohistochemical staining. These findings suggest that the non-adherent bone marrow cell-derived mesenchymal stem cells could differentiate into neuronal-like cells in vitro and in vivo.

Identification of circulating CD90^+ CD73^+ cells in cirrhosis of liver

AIM:To identify circulating CD90 + CD73 + CD45 cells and evaluate their in vitro proliferating abilities.METHODS:Patients with cirrhosis(n=43),and healthy volunteers(n=40)were recruited to the study.Mononuclear cells were isolated and cultured from the peripheral blood of controls and cirrhosis patients.Fibroblast-like cells that appeared in cultures were analyzed for morphological features,enumerated by flow cytometry and confirmed by immunocytochemistry(ICC).Colony forming efficiency(CFE)of these cells was assessed and expressed as a percentage.RESULTS:In comparison to healthy volunteers,cells obtained from cirrhotic patients showed a significantincrease(P<0.001)in the percentage of CD90+CD73+ CD45 cells in culture.Cultured cells also showed 10 fold increases in CFE.Flow cytometry and ICC confirmed that the proliferating cells expressed CD90 + CD73 + in the cultures from cirrhosis patients.CONCLUSION:These results indicate the presence of circulating CD90 + CD73 + CD45 cells in patients with liver cirrhosis that have the potential to proliferate at a higher rate.

Design and Development of an <i>in Vitro</i>Assay for Evaluation of Solid Vaginal Dosage Forms

Vaginal dosage forms are seen as a viable option for empowering women to protect themselves from the risk of HIV transmission. Because of limited research in the field, there is a lack of suitable dissolution methods established for determination of drug release from vaginal formulations inside the vaginal tract. The main aim of this study was to develop a simple, reliable and reproducible in vitro release method for evaluation of solid vaginal dosage forms (VDFs) which was hoped to exhibit a close in vitro-in vivo correlation. Dapivirine, a drug being developed as a microbicide and a well established marketed anti fungal drug, Clotrimazole were used as model drugs. Two doses (0.5 mg and 1.25 mg) of Dapivirine were prepared as novel rapidly disintegrating, bioadhesive tablets. Clotrimazole 100 mg, prepared in house as conventional release tablets and commercially available Canesten (Clotrimazole tablet 100 mg) were used. The in vitro drug release testing of these tablets was carried out using a designed system which consisted of modified USP dissolution Apparatus II in conjunction with Enhancer cell (as sample holder) in 150 ml capacity flasks instead of the standard 900 ml flasks. The suitability of the system was investigated for variable parameters such as formulation types, drug concentration, stirring speeds, media volume and comparison of in house product with marketed product. The method was successfully optimized at a volume of 100 ml and a low speed of 25 rpm at pH 4 and was found sensitive enough to distinguish between formulations and evaluate products of different strengths. A linear drug release profile (R2 = 0.99) was obtained in case of Dapivirine, indicating that drug release is controlled by diffusion. The developed dissolution system has a potential to exhibit a good in vitro-in vivo correlation in addition to carrying out routine dissolution tests for solid VDFs.

SOFC复合孔隙阳极材料的制备及性能研究

使用共流延工艺分别制备了以不同配比可溶性淀粉和碳纤维混合物作为造孔剂的阳极支撑型固体氧化物燃料电池(SOFC),阳极支撑层具有不规则的球型和直线型交错的复合孔隙,造孔剂总质量分数均为10%。电池以(Y_(2)O_(3))0.08(ZrO_(2))0.92(8YSZ)及Ni作为阳极材料,其中Ni与8YSZ的质量比为1∶1,以8YSZ作为电解质材料,以(La_(0.75)Sr_(0.25))0.95MnO_(3±δ)(LSM)及8YSZ作为阴极材料。使用氢气作为燃料测试了各电池在750℃下的电化学性能,结果表明,淀粉与碳纤维质量之比为1.5的复合造孔剂所制备电池的功率密度最高,可达0.199 W/cm^(2)。扫描电子显微镜法(SEM)图像显示,淀粉和碳纤维在阳极支撑层中所形成的复合孔隙相互交织,连通性较好,具有较理想的微观结构。

Binocular form deprivation influences the visual cortex

a-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors are considered to play a crucial role in synaptic plasticity in the developing visual cortex. In this study, we established a rat model of binocular form deprivation by suturing the rat binocular eyelids before eye-opening at postnatal day 14. During development, the decay time of excitatory postsynaptic currents mediated by a-amino-3- hydroxy-5-methyl-4-isoxazolepropionic acid receptors of normal rats became longer after eye- opening; however, the decay time did not change significantly in binocular form deprivation rats. The peak value in the normal group became gradually larger with age, but there was no significant change in the binocular form deprivation group. These findings indicate that binocular form deprivation influences the properties of excitatory postsynaptic currents mediated by a-amino-3- hydroxy-5-methyl-4-isoxazolepropionic acid receptors in the rat visual cortex around the end of the critical period, indicating that form stimulation is associated with the experience-dependent modification of neuronal synapses in the visual cortex.

血府逐瘀汤对狼疮性肾炎患者胸腺肽β4及Ea-RFC的影响

目的探讨血府逐瘀汤辅助治疗狼疮性肾炎(LN)的效果及对患者胸腺肽β4及Ea花环细胞(Ea-RFC)形成率的影响。方法选取河北北方学院附属第二医院2020年1月至2021年6月收治的60例狼疮性肾炎患者,按随机数字表法分为观察组(行血府逐瘀汤联合泼尼松与环磷酰胺治疗)30例与对照组(行泼尼松与环磷酰胺治疗)30例,治疗18个月。观察组治疗后临床效果、中医症状、胸腺肽β4表达、Ea-RFC形成率、肾功能及不良反应。结果观察组治疗总有效率为96.67%,高于对照组的80.00%,差异有统计学意义(P<0.05)。治疗后,观察组纳少腹胀、腰膝酸软、面色不华、口黏口苦等中医症状积分低于对照组,差异有统计学意义(P<0.05)。治疗后,观察组红细胞沉降率、血清肌酐及胸腺肽β4表达低于对照组,Ea-RFC形成率高于对照组,差异有统计学意义(P<0.05)。治疗后,观察组不良反应发生率与对照组比较差异无统计学意义(P>0.05)。结论血府逐瘀汤辅助治疗LN患者疗效确切,能减少患者体内胸腺肽β4表达,提高Ea-RFC,改善肾功能,安全性较高。

基于肠道菌群平衡分析微生态制剂联合浙贝黄芩汤对急性淋巴细胞白血病大剂量化疗后患者的临床影响

目的探讨微生态制剂联合浙贝黄芩汤对急性淋巴细胞白血病(ALL)大剂量化疗后患者粒细胞集落刺激因子受体(G-CSFR)、粒单系集落形成单位(CFU-GM)、肠道菌群及红系爆式集落形成单位(BFU-E)的影响。方法选取延安大学附属医院2019年6月至2022年12月收治的ALL患者130例作为研究对象,根据治疗方法将患者分为A组、B组、C组,3组患者均接受大剂量化疗,化疗结束48 h后A组患者实施常规治疗,B组患者单纯浙贝黄芩汤治疗,C组给予微生态制剂联合浙贝黄芩汤治疗,治疗12 d后,对3组患者G-CSFR、CFU-GM、BFU-E表达情况及血细胞数量进行检测。结果治疗后,C组血红蛋白、白细胞、血小板[(79±6)g/L、(3.8±0.4)×10^(9)/L、(66.4±3.6)×10^(9)/L]与A组[(59±7)g/L、(3.2±0.4)×10^(9)/L、(52.6±2.8)×10^(9)/L]、B组[(61±7)g/L、(3.1±0.3)×10^(9)/L、(52.8±2.6)×10^(9)/L]对比,差异有统计学意义(P<0.05)。C组G-CSFR(5.35±0.16)pg/ml和白细胞介素-11受体(IL-11R)(6.38±0.54)μg/kg水平均高于A组[(2.23±0.13)pg/ml和(1.49±0.24)μg/kg]和B组[(2.31±0.16)pg/ml和(2.31±0.49)μg/kg]差异有统计学意义(P<0.05)。治疗后,C组患者7 d CFU-GM(18.5±6.0)个和14 d BFU-E(83.5±7.5)个高于A组[7 d CFU-GM(9.5±2.0)个和14 d BFU-E(59.5±6.5)个]和B组[7 d CFU-GM(12.0±6.5)个和14 d BFU-E(63.5±5.0)个],差异有统计学意义(P<0.05)。7 d后,C组双歧杆菌(12.56±3.25)lgCFU/g、乳酸杆菌(13.56±2.58)lgCFU/g、肠杆菌(5.12±1.45)lgCFU/g、肠球菌(5.14±0.58)lgCFU/g高于A组[(9.26±1.03)lg CFU/g、(8.65±0.84)lg CFU/g、(8.08±0.64)lgCFU/g、(8.15±0.46)lgCFU/g]和B组[(11.35±1.36)lg CFU/g、(12.43±1.14)lgCFU/g、(6.49±0.55)lgCFU/g、(6.66±0.43)lgCFU/g],差异有统计学意义(P<0.05)。结论微生态制剂联合浙贝黄芩汤治疗可以有效提高ALL大剂量化疗后患者的G-CSFR、CFU-GM、BFU-E水平,可能更好地改善化疗引起的患者骨髓抑制情况,改善肠道菌群,具有临床研究价值。

苦参碱调节RhoA-ROCK信号通路对冠心病模型大鼠Th17/Treg细胞平衡的影响

目的探讨苦参碱(Matrine)对冠心病(coronary heart disease,CHD)大鼠辅助T细胞17(helper T cell 17,Th17)/调节性T细胞(regulatory T cells,Treg)细胞平衡及Ras同源基因家族成员A(RhoA)-Rho相关的卷曲螺旋激酶(ROCK)信号通路的影响。方法建立冠心病模型,将实验大鼠分为对照组、模型组、苦参碱低剂量(50 mg·kg^(-1))组、苦参碱高剂量(200 mg·kg^(-1))组及苦参碱高剂量(200 mg·kg^(-1))+LPA组(10 mg·kg^(-1))。超声心动图进行大鼠心功能检测;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)法进行白细胞介素17(IL-17)、转化生长因子β(TGF-β)水平检测;流式细胞术检测Th17、Treg数量及Th17/Treg比值;免疫组化进行内皮型一氧化氮合酶(eNOS)、内皮素1(ET-1)蛋白表达水平检测;Masson染色进行大鼠心肌组织的病理形态变化观察;TTC染色检测各组大鼠心肌梗死情况;TUNEL染色进行心肌组织中细胞凋亡情况检测;试剂盒检测RhoA活性;Western Blot法进行半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、B细胞淋巴瘤因子2(Bcl-2)、Bcl-2相关X蛋白(Bax)、RhoA、ROCK1、ROCK2蛋白表达水平检测。结果与对照组比较,模型组心肌组织有大量蓝色胶原纤维沉积,左室舒张末期容积(left ventricular end-diastolic volume,LVEDV)、左室收缩末期容积(left ventricular end-systolic volume,LVESV)、IL-17、Th17、Th17/Treg、ET-1、心肌梗死面积、细胞凋亡率、TUNEL阳性率、Bax、Caspase-3、RhoA活性、RhoA、ROCK1、ROCK2表达水平明显升高,左室射血分数(left ventricular ejection fraction,LVEF)、左室缩短分数(left ventricular shortening fraction,LVFS)、TGF-β、Treg、eNOS、Bcl-2表达水平明显降低(P<0.05)。与模型组比较,Matrine-L组、苦参碱高剂量组心肌组织蓝色胶原纤维逐渐减少,LVEDV、LVESV、IL-17、Th17、Th17/Treg、ET-1、心肌梗死面积、细胞凋亡率、TUNEL阳性率、Bax、Caspase-3、RhoA活性、RhoA、ROCK1、ROCK2表达水平依次明显降低,LVEF、LVFS、TGF-β、Treg、eNOS、Bcl-2表达水平依次明显升高(P<0.05)。与苦参碱高剂量组比较,苦参碱高剂量+LPA组心肌组织蓝色胶原纤维增多,LVEDV、LVESV、IL-17、Th17、Th17/Treg、ET-1、心肌梗死面积、细胞凋亡率、TUNEL阳性率、Bax、Caspase-3、RhoA活性、RhoA、ROCK1、ROCK2表达水平明显升高,LVEF、LVFS、TGF-β、Treg、eNOS、Bcl-2表达水平明显降低(P<0.05)。结论苦参碱通过抑制RhoAROCK信号通路调节Th17/Treg细胞平衡,改善冠心病大鼠心肌损伤。

大面积有机–无机杂化钙钛矿薄膜及其光伏应用研究进展

有机–无机杂化钙钛矿太阳能电池具有制备成本低、光电转换效率(Photoelectric Conversion Efficiency,PCE)高的巨大优势,显示出广阔的商业化前景。经过十几年的深入研究,钙钛矿太阳能电池(Perovskite Solar Cells,PSCs)的实验室器件(<1 cm^(2))、大面积器件(1~10 cm^(2))、迷你模组级器件(10~800 cm^(2))和模组级器件(>800 cm^(2))的最高认证PCE已分别提升至26.10%、24.35%、22.40%和18.60%。随着PSCs面积扩大,PCE急剧下降,这主要是因为制备方法的局限性,难以获得高质量的大面积钙钛矿薄膜。实验室器件常采用的旋涂法难以应用到实际生产中,目前大面积钙钛矿薄膜的制备方法主要有刮涂法和狭缝涂布法,但其存在薄膜成核结晶过程难以精确控制等问题。本文从大面积有机–无机杂化钙钛矿薄膜的制备方法入手,介绍了大面积钙钛矿层成膜机制及薄膜质量提升策略。最后,对未来高PCE、高稳定性的大面积PSCs的制备技术和应用进行了展望,旨在对高性能的大面积PSCs研究提供有益参考。

氮形态对小白菜根系生长、根区pH及细胞壁组分的影响

【目的】氮素(N)是调控蔬菜作物生长的重要因子,探究氮形态对小白菜根系生长及细胞壁组分的影响,明确根区pH变化与质膜(PM)H^(+)-ATPase的关系,为进一步研究氮素调控植物根系生长发育提供理论基础。【方法】以小白菜(‘华王’)为试材,设置3个N素形态(硝态氮(NO_(3)^(-)-N)、铵态氮(NH_(4)^(+)-N)、甘氨酸态氮(Gly-N)),采用营养液水培法,探究无机氮和有机氮调控小白菜根区pH和根细胞壁组分对根系生长的影响,同时探讨氮营养对小白菜根系生长素(IAA)和PM H^(+)-ATPase酶的影响。用根系扫描法和生理试验等方法,测定小白菜的总根长、总根体积、总根表面积和根尖数等根系形态指标以及介质pH、PM H^(+)-ATPase酶活性、IAA含量、细胞壁及其主要组分含量、果胶甲酯酶(PME)、扩张素(EXP)含量等生理指标,并分析根系形态和各生理指标之间的相关性。【结果】Gly-N和NO_(3)^(-)-N均碱化介质pH,且Gly-N碱化介质pH的能力高于NO_(3)^(-)-N处理,NH_(4)^(+)-N酸化介质pH。与NO_(3)^(-)-N处理相比,Gly-N处理显著降低小白菜地上部(40.23%)和根系干物质含量(21.74%)、主根长(55.15%)、总根长(58.63%)、总根表面积(53.12%)、总根尖数(73.07%)、根分叉数(65.39%)、内部链接数(40.91%)和外部链接数(73.94%),提高了根系细胞壁提取率(22.00%)、果胶(47.74%)、纤维素(25.30%)、EXP(109.53%)、PM H^(+)-ATPase酶活性(15.42%)、根冠比(30.33%)、拓扑指数(11.43%)和分形维数(4.32%);NH_(4)^(+)-N处理植株地上部干物质含量、主根长、根分叉数、内部链接数、外部链接数、根系PME活性和IAA含量分别显著降低22.98%、34.30%、35.32%、26.22%、29.02%、36.38%和8.74%,根系PM H^(+)-ATPase酶活性、细胞壁提取率、果胶和半纤维素分别显著提高14.60%、19.38%、22.98%、74.33%。根系形态指标与生理指标之间的相关分析结果表明,总根表面积与总根长、总根尖数、外部链接数、根分叉数呈极显著正相关(P<0.01),与主根长、内部链接数、叶干重呈显著正相关(P<0.05);总根表面积与根直径、扩张素、拓扑指数呈极显著负相关(P<0.01),与果胶、纤维素和分形维数呈显著负相关(P<0.05)。根直径与扩张素、纤维素、分形维数、拓扑指数呈极显著正相关(P<0.01),与果胶呈显著正相关(P<0.05)。【结论】Gly-N和NH_(4)^(+)-N均增加了小白菜根系细胞壁含量,抑制了小白菜根系的伸长生长;Gly-N更有利于果胶和纤维素的积累,NH_(4)^(+)-N有利于半纤维素的积累。

METTL3调控miR-301a促进缺氧环境下内皮集落形成细胞血管生成

目的:探讨缺氧环境对内皮集落形成细胞(ECFCs)血管生成能力的影响及甲基转移酶样3(METTL3)的调控作用。方法:分离、培养犬外周血ECFCs。实验分为正常对照组与缺氧实验组。缺氧实验组分别使用含50μmol/L、100μmol/L和200μmol/L氯化钴(CoCl_(2))的培养基处理细胞24 h,对照组CoCl_(2)浓度为0。CCK-8检测不同浓度CoCl_(2)对ECFCs增殖的影响,实时荧光定量PCR(RT-qPCR)检测缺氧诱导因子-1α(HIF-1α)的表达,筛选适宜浓度的CoCl_(2)用于缺氧环境构建。Transwell实验和管形成实验分别检测ECFCs迁移和血管生成能力。RT-qPCR和western blotting检测血管内皮生长因子(VEGF)、CD31、METTL3、miR-301a的表达情况。构建沉默METTL3慢病毒转染ECFCs,将ECFCs分为NC-shRNA组和METTL3-shRNA组。RT-qPCR和western blotting检测ECFCs细胞中VEGF、CD31、METTL3、miR-301a表达水平的变化。结果:与正常对照组比较,50μmol/L和100μmol/L的CoCl_(2)培养ECFCs 24 h后可促进细胞增殖(均P<0.0001),HIF-1α随着CoCl_(2)浓度升高而高表达(P<0.01)。100μmol/L CoCl_(2)组中ECFCs迁移能力和血管生成能力提高(均P<0.01),VEGF、CD31、METTL3、miR-301a mRNA水平表达升高(均P<0.01),VEGF、CD31、METTL3蛋白表达升高(均P<0.01)。与NC-shRNA组比较,METTL3-shRNA组中VEGF、CD31、METTL3、miR-301a mRNA表达降低(均P<0.05),VEGF、CD31、METTL3蛋白表达降低(均P<0.05)。结论:适宜的缺氧干预可提高ECFCs增殖、迁移和血管生成能力,METTL3/miR-301a轴可能参与调控血管生成。