A Novel Semisynthetic Molecule Icaritin Stimulates Osteogenic Differentiation and Inhibits Adipogenesis of Mesenchymal Stem Cells

Background We previously reported that the constitutional flavonoid glycosides derived from herb Epimedium (EF, composed of seven flavonoid compounds with common nuclear stem) exerted beneficial effects on the bone, including promoting bone formation and inhibiting bone marrow fat deposition. Recent in vivo study showed that Icaritin was a common metabolite of these constitutional flavonoid glycosides, indicating that Icaritin is a bioactive compound. The present study was designed to investigate whether Icaritin could promote osteogenic differentiation and suppress adipogenic differentiation of marrow mesenchymal stem cells (MSCs).Methods Primary MSCs were harvested from adult mice and exposed to Icaritin to evaluate whether it could promote osteogenesis and suppress adipogenesis using the following assays: determination of alkaline phosphatase (ALP) activity and mineralization; mRNA expression of osteogenic differentiation marker Runx2; osteocalcin and bone sialoprotein (BSP) by RT-PCR; quantification of adipocyte-like cells by Oil Red O staining assay and mRNA expression for adipogenic differentiation markers peroxisome proliferator-activated receptor gamma (PPARγ); adipocyte fatty acid binding protein (aP2) and lipoprotein lipase (LPL) by RT-PCR. For the underlying mechanism, glycogen synthase kinase-3beta (GSK3β) and β-catenin were also explored by western blotting.Results Icaritin promoted osteogenic differentiation and maturation of MSCs as indicated by increased mRNA expression for Runx2, osteocalcin and BSP, and enhanced ALP activity and mineralization; Icaritin inhibited adipogenic differentiation, as indicated by decreased mRNA expression for PPARγ, LPL, aP2, and suppressed formation of adipocyte-like cells; Icaritin inactivated GSK3β and suppressed PPARγ expression when promoting osteogenesis and suppressing adipogenesis of MSCs.Conclusion This was the first study demonstrating that the novel semisynthetic molecule Icaritin could stimulate osteogenic differentiation and inhibit adipogenesis of MSCs, which was associated with the suppression of GSK3β and PPARγ.     

Enhanced reductive removal of ciprofloxacin in pharmaceutical wastewater using biogenic palladium nanoparticles by bubbling H2

To treat waste with waste and efficiently remove the organic pollutant, waste palladiums(ii) were adsorbed and reduced on microorganism surface to catalyze the reductive removal of ciprofloxacin in pharmaceutical wastewater. By optimizing conditions such as pH and temperature, the amount of biogenic palladium adsorbed and reduced on E. coli reached 139.48 mg g⁻¹ (Pd/microorganisms). Moreover, most of the Pd(ii) was reduced to nanometer-sized Pd(0) as characterized by TEM and SEM with EDXA. Using the obtained biogenic palladium, the reductive removal of ciprofloxacin is up to 87.70% at 25 °C, 3.03 folds of that achieved in the absence of H₂. The results show that waste E. coli microorganisms can efficiently adsorb and remove waste Pd(ii) and produce Bio-Pd nanoparticle catalysts in the presence of H₂. This biogenic palladium presents high catalytic activity and great advantages in the reductive degradation of ciprofloxacin. Our method can also be applied to other waste metal ions to prepare the biogenic metals, facilitate their recovery and reuse in degrading organic pollutants in wastewater to achieve “treating waste using waste”.

A solution has been successfully introduced to three key challenges from the wastewater containing waste microorganisms, metal and ciprofloxacin, respectively.     

Magnetic core–shell Fe3O4@Cu2O and Fe3O4@Cu2O–Cu materials as catalysts for aerobic oxidation of benzylic alcohols assisted by TEMPO and N-methylimidazole

In this work, core–shell Fe₃O₄@Cu₂O and Fe₃O₄@Cu₂O–Cu nanomaterials for aerobic oxidation of benzylic alcohols are reported with 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) and N-methylimidazole (NMI) as the co-catalysts. To anchor Cu₂O nanoparticles around the magnetic particles under solvothermal conditions, the magnetic material Fe₃O₄ was modified by grafting a layer of l-lysine (l-Lys) to introduce –NH₂ groups at the surface of the magnetic particles. With amine groups as the anchor, Cu(NO₃)₂ was used to co-precipitate the desired Cu₂O by using ethylene glycol as the reducing agent. Prolonging the reaction time would lead to over-reduced forms of the magnetic materials in the presence of copper, Fe₃O₄@Cu₂O–Cu. The nanomaterials and its precursors were fully characterized by a variety of spectroscopic techniques. In combination with both TEMPO and NMI, these materials showed excellent catalytic activities in aerobic oxidation of benzylic alcohols under ambient conditions. For most of the benzylic alcohols, the conversion into aldehydes was nearly quantitative with aldehydes as the sole product. The materials were recyclable and robust. Up to 7 repeat runs, its activity dropped less than 10%. The over-reduced materials, Fe₃O₄@Cu₂O–Cu, exhibited slightly better performance in durability. The magnetic properties allowed easy separation after reaction by simply applying an external magnet.

Robust core–shell magnetic materials catalyse quantitatively the aerobic oxidation of a wide range of benzylic alcohols into corresponding aldehydes at room temperature showing excellent tolerance towards the substituents on the phenyl ring.     

Highly crystalline nickel hexacyanoferrate as a long-life cathode material for sodium-ion batteries

Prussian blue analogs (PBAs) are attractive cathode candidates for high energy density, including long life-cycle rechargeable batteries, due to their non-toxicity, facile synthesis techniques and low cost. Nevertheless, traditionally synthesized PBAs tend to have a flawed crystal structure with a large amount of [Fe(CN)₆]⁴⁻ openings and the presence of crystal water in the framework; therefore the specific capacity achieved has continuously been low with poor cycling stability. Herein, we demonstrate low-defect and sodium-enriched nickel hexacyanoferrate nanocrystals synthesized by a facile low-speed co-precipitation technique assisted by a chelating agent to overcome these problems. As a consequence, the prepared high-quality nickel hexacyanoferrate (HQ-NiHCF) exhibited a high specific capacity of 80 mA h g⁻¹ at 15 mA g⁻¹ (with a theoretical capacity of ∼85 mA h g⁻¹), maintaining a notable cycling stability (78 mA h g⁻¹ at 170 mA g⁻¹ current density) without noticeable fading in capacity retention after 1200 cycles. This low-speed synthesis strategy for PBA-based electrode materials could be also extended to other energy storage materials to fabricate high-performance rechargeable batteries.

A low-speed synthesis strategy was designed to fabricate Prussian blue analog based electrode materials for high-performance rechargeable batteries.     

A novel strategy to alleviate medium acidosis for simultaneously yielding more bacterial cellulose and electricity

Bacterial cellulose (BC), a fascinating and renewable polymer, can be applied widely in various bio-based materials. However, its synthesis is generally limited by medium acidosis. Herein, we demonstrated a built-in galvanic cell within the BC fermentation medium to alleviate the acidosis, by which BC yield was promoted by 191%, and simultaneously the yield of electrical power of 0.68 W to 8.10 W during the incubation.

Bacterial cellulose (BC), a fascinating and renewable polymer, can be applied widely in various bio-based materials.

Bacterial cellulose (BC) is biologically synthesized by several bacteria, notably, the strain of Acetobacter xylinum.¹ This fascinating and renewable polymer possesses remarkable chemical and physical attributes (e.g. high porosity, excellent mechanical strength, and large surface area, etc.) that can be applied in a wide range of bio-based materials and products.^2–4 Its wide applications have motivated researchers to focus on the fermentation process, especially to promote the BC yield. However, the fermentation process is extensively related to many aspects, such as the employed strains, medium conditions (carbon source, nutrients, pH, and dissolved oxygen), and incubation conditions (the employed medium volume, duration, surface area).^5–8Among these factors, the medium acidosis caused by the released organic acids, such as glucuronic acid and acetic acid, in the medium will negatively affect the BC biosynthesis during bacterial metabolism, which could weaken the bacteria activities and reduce the BC yield as a consequence. Generally, the pH of the BC fermentation medium can be dramatically decreased from 5.0 to lower than 2.0 during the incubation if no pH adjustment strategy was employed.^5,9,10 Current methods for BC incubation include static, agitated, and bioreactor cultures.¹¹ For agitated and bioreactor cultures, the regulation of pH is relatively easy because of the culture system is under stirring. However, the static culture of BC is still an ideal way to obtain high-quality BC membranes for many novel applications.^12–14 Unfortunately, it is quite difficult to adjust the pH of medium during the incubation because the formation of BC film in static culture will be seriously disturbed by the mechanical agitation. Therefore, to find an ingenious way to control the medium pH in a suitable range will be very critical to the BC production in practice.The essence of medium acidosis sources from the ionized hydrogen ions (H⁺) of the released organic acids in bacterial metabolic activity. If there is a proper approach to remove the H⁺ from culture medium in a controllable way, the BC production may significantly be promoted. Nowadays, the application of batteries has greatly facilitated people''s life. This revolutionary device was originated from the prototype of “galvanic cell” (or “voltaic piles”), which was invented by Alessandro Volta in 1800.¹⁵ Even now, the galvanic cell still be employed to demonstrate the mechanism of battery for the educational aim, such as the fruit battery.^16,17 Using lemons, oranges, grapefruits, potatoes, or apples which riches in citric acid, phosphoric acid, or malic acid as the electrolyte, and two different metallic electrodes were plugged into the fruit to form a battery. The fruit battery could provide continuous electricity to run various small devices, such as light-emitting diode (LED) and a digital clock. When the two metallic electrodes was connected with those devices by wire, the ionized H⁺ of organic acids inside of the fruit will be slowly consumed in anode and producing electric energy to drive those devices.^18–20 Inspired by the principle of a galvanic cell, we assumed that the ionized H⁺ of accumulated organic acids during the BC fermentation can be consumed in the anode of the built-in galvanic cell to form H₂, which will greatly weaken the acidified conditions of culture medium. Theoretically, a suitable pH condition for bacteria synthesis will promote the BC yield in a consequent. Meanwhile, the liberated metal ions from the cathode can serve as a suitable activator to the biochemical process of bacteria if the electrodes can be selected approximately, which also can potentially promote the BC yield.²¹ Based on this conception, the BC fermentation integrated a couple of electrodes to construct a galvanic cell system as its schematic diagram in Fig. 1 (the details of this built-in galvanic cell see ESI, page S2^†). To check the possibility of this first attempt, the BC yield of this newly constructed system (GC-medium) was compared with that of the normal medium, and the performance of electricity release was investigated as well.Open in a separate windowFig. 1Schematic diagram of the fermentation apparatus and the data acquisition system.The medium pH was monitored at every 12 h during static incubation as shown in Fig. 2.²² It could be easily found that the pH of the normal medium displayed a continuous decrease as the fermentation time was prolonged to 144 h. By contrast, the pH in the GC-medium firstly increased to 4.6 after 24 h of incubation, then maintained in the range from 4.2 to 4.4. Obviously, the medium pH for BC fermentation can be stabilized via loading the galvanic cell to consume the extra acids from the medium process.Open in a separate windowFig. 2Changes of medium pH during the incubation. Roman numerals of (I), (II), and (III) and Arabic numerals of (1) and (2) represent the stage of pH changes during the incubation in GC-medium and normal medium, respectively.Correspondingly, the BC yield from the GC-medium was 0.358 g L⁻¹, which was 2.9-fold higher than that of the normal medium (Fig. 3). Apparently, the pH adjustment by the galvanic cell can work well to yield more BC, verifying the possibility of the proposed conception. Besides, the residual glucose in the GC-medium was lower than that of the normal medium, as well as the residual glucuronic acid. These results again proved that more carbon source was converted into BC. When the BC film started to form at the air–liquid interface, the bacteria would be embedded into the film and fixed, which can result in a clearer medium. The relatively lower OD₆₁₀ in the GC-medium indicated that the more bacteria were fixed result from the thicker BC film that produced in GC-medium.²³ Moreover, the relatively higher DO (dissolved oxygen) was remained in the GC-medium after 6 days incubation also demonstrated that the conditions for cellulose biosynthesis of bacteria were better than that of the normal medium,²⁴ which also suggested more sugar will be consumed for BC synthesis once the galvanic cell was built during the fermentation process.Open in a separate windowFig. 3Medium conditions after 6 days incubation. BC referred to the yield of bacterial cellulose (g L⁻¹); OD₆₁₀ is the optical density of medium at 610 nm; DO is dissolved oxygen (mg L⁻¹); RG is residual glucose (g L⁻¹); GA is glucuronic acid (g L⁻¹). Analysis of variance (ANOVA): ***p < 0.001, * 0.01 < p < 0.05.The formed current with 2 Ω-load of the built-in galvanic cell can be detected by the digital multi-meter. As shown in Fig. 4a, the current decreased sharply from 2013 μA to 878 μA in the first 2 h, this result was related to the rapid consumption of the H⁺ that contained in the strain when the built-in galvanic cell was activated. When the bacteria started to self-replicate in the first 72 h, more organic acid was produced and released to the medium (refer to Fig. 2), which slowed down the rate of current reduction in the following 22 h. The prominent increase of current appeared in 24–72 h, proving that the medium was still rich in H⁺, meanwhile, the number of bacteria grew exponentially. These results also could be observed in Fig. 2, the changes of pH were well matched with the adjustment of the built-in galvanic cell. This correlativity between the medium pH and the formed current of GC has the potential to be designed as a fully automatic pH control system (ESI, Fig. S1^†).Open in a separate windowFig. 4Electricity generation performance during the incubation. (a) Current monitoring with 2 Ω resistor; (b) cumulative energy output in the whole BC fermentative production.Afterward, the current was maintained around 1163 μA during the following 3 days, which can demonstrate the amount of Acetobacter xylinum in GC-medium reach a stable period. Meanwhile, the pH of GC-medium was still within a suitable range of 4.2–4.4 while the pH of the normal medium has been reduced to around 2.7–2.8, which was already negative to the bacterial metabolic activities (Fig. 2). Generally, the death period of bacteria was mainly attributed to the medium acidosis.¹⁰ Hence, the mechanism of improving the BC yield by this built-in galvanic cell was to prevent the medium acidosis and maintained the activity of bacteria, which can stabilize the BC biosynthesis successfully.The cumulative energy output was calculated and showed in Fig. 4b, although the initial pH of GC-medium was relatively lower than that after 48 h, the energy output had a remarkable promotion. This result proved the major factors for the energy output was not only affected by the apparent medium pH but also related to the ability to provide H⁺ within bacteria metabolism. This ability was mainly contributed by the number of bacteria and the bacterial activity, which would be in a better condition if there was a built-in galvanic cell. Previous studies have suggested that the H⁺ concentration in the electrolyte, metal type of electrode, and the number of galvanic cells to be assembled in series are the key factors that influenced the output voltage of the galvanic cell.¹⁷ In this study, the magnesium (Mg) anode, copper (Cu) cathode, and the H⁺-rich culture medium formed an integrated galvanic cell system. The voltage was determined as 1.55–1.65 V for this built-in galvanic cell during the BC production, which can light the LED bulb (ESI, Fig. S2^†). Of course, it also can provide enough electricity to drive digital clocks, sensors, and other low-power devices in the whole incubation time. These results substantially proved that the conception of this work to produce electricity and promote BC production can be achieved via loading the built-in galvanic cell.The schematic mechanism diagram of acidification, pH adjustment, and electricity generation of the galvanic cell were illustrated with Fig. 5, the metabolic by-products, such as citric acid (CA), acetic acid (AC), gluconic acid (GLCA), and pyruvic acid (PA), are generally derived from the tricarboxylic acid (TCA) cycle, pentose phosphate pathway (HMP) and glycolytic pathway (EMP).²⁵ At the beginning of fermentation, glucose (GLC) in the medium was high enough to support the cell self-replication through the aerobic respiration of bacteria in the TCA cycle, which results in an amount of GLCA released to the medium. In this process, the by-products, such as CO₂, CA, and H₂O, also could be generated correspondingly. In addition, a considerable amount of AC and some PA can be released from the HMP and EMP, respectively.²⁴ These by-products, including GLCA, AC, CA, and PA or CO₂, were mainly responsible for medium acidosis, which inhibited bacteria activity and reduced the BC production as a consequence. Besides, the produced GLCA would inhibit the Acetobacter xylinum biosynthesis for BC.²⁶ The dissolved CO₂ in the medium also limited the aerobic respiration of bacteria, which will make the bacteria produce more CA and PA as a feedback regulation. This could create a vicious circle to deteriorate the medium pH and cease in BC production eventually. However, when the galvanic cell was built inside of the culture medium, those organic acids could provide the ionized H⁺ to maintain the galvanic reaction. The H⁺ would move to the Cu-electrode and be reduced as hydrogen gas by the acceptance of the electrons. The Mg-electrode would lose electrons and be oxidized to Mg²⁺, which can involve several cell life activities and potentially promote the BC yield (the effect of Mg²⁺ on BC production shown in ESI, Fig. S3^†). During this process, the directional transfer of electrons in the external circuit created the current, and the electric energy will be continuously output throughout the fermentation. It can image that this built-in galvanic cell can yield considerable power once a large-scale BC production was considered. Moreover, the conception of built-in galvanic cell in bioconversion processes that need pH control, such as anaerobic digestion of easy-acidification substrates, and ethanol fermentation by yeast can also be considered to apply this conception to maintain the stable and suitable pH.Open in a separate windowFig. 5The schematic mechanism diagram of acidification, pH adjustment, and electricity generation of the galvanic cell. GLC (glucose), GLCA (gluconic acid), CA (citric acid), AC (acetic acid), PA (pyruvic acid), GLC6P (glucose-6-phosphate), GLC1P (glucose-1-phosphate), UDPG (uridine diphosphoglucose), HMP (pentose phosphate pathway), FRU6P (fructose-6-phosphate), GAP (glyceraldehyde-3-phosphate), G3P (3-phosphoglycerate), PEP (phosphoenolpyruvate), PYR (pyruvate), ATP (adenosine triphosphate).     

超声心动图自动心肌运动定量技术对尿毒症透析患者早期左心室收缩功能的评估

目的探讨超声心动图自动心肌运动定量技术（aCMQ）评估不同透析时间段的尿毒症患者左心室收缩功能的价值。 方法选择2018年5月至2019年2月在浙江省杭州市萧山区第一人民医院血液透析室接受透析治疗的尿毒症患者52例。根据其接受血透时间的不同分为3组,A组为透析时间≤3年的患者,共27例;B组为透析时间＞3年且≤7年的患者,共16例;C组为透析时间＞7年的患者,共9例;正常对照组共60例,既往均无心脏及肾疾病病史,常规心电图及超声心动图检查均未见明显异常。首先运用M型超声心动图,通过Teichholz法获得病例组及对照组左心室射血分数（LVEF）,然后启动X-Plane技术采集4个心动周期的四腔心及两腔心,根据心电图指示分别描记左心室舒张末期及左心室收缩末期,运用双平面Simpson法分别计算出病例组及对照组LVEF,最后运用aCMQ分别获取病例组及对照组研究对象的左心室整体长轴应变（LVGLS）、心尖两腔心长轴应变（LVAP2LS）、心尖四腔心长轴应变（LVAP4LS）及心尖三腔心长轴应变（LVAP3LS）,分析不同透析年限组尿毒症患者左心室长轴应变变化。多组间比较采用方差分析,组间两两比较采用LSD-t检验。 结果Teichholz法测得病例组与对照组LVEF分别为：A组（67.21±6.63）%;B组（64.73±6.47）%;C组（64.58±8.38）%;对照组（67.02±3.62）%。Simpson法测得病例组与对照组LVEF分别为：A组（64.71±4.93）%;B组（64.08±6.02）%;C组（63.91±7.49）%;对照组（66.17±3.14）%。病例组与对照组LVEF比较以及病例组间LVEF比较,差异均无统计学意义（P均＞0.05）。运用aCMQ得到病例组与对照组LVGLS分别为：A组（-20.79±2.70）%、B组（-20.03±3.58）%、C组（-18.32±3.71）%、对照组（-24.39±2.05）%;LVAP4LS分别为：A组（-22.09±2.76）%、B组（-20.11±3.94）%、C组（-19.49±3.73）%、对照组（-24.61±2.37）%;LVAP3LS分别为：A组（-19.32±3.85）%、B组（-19.28±4.37）%、C组（-16.61±4.40）%、对照组（-23.53±6.18）%;LVAP2LS分别为：A组（-20.09±2.53）%、B组（-19.57±2.65）%、C组（-18.09±4.01）%、对照组（-23.51±7.52）%。病例组LVGLS、LVAP2LS、LVAP4LS及LVAP3LS的测值均较对照组减低,差异具有统计学意义（A组 vs 对照组：t=-5.949、-3.844、-6.117、-4.863,P均＜0.001;B组 vs 对照组：t=-5.883、-5.619、-5.036、-4.650,P均＜0.001;C组 vs 对照组：t=-6.541、-5.081、-6.130、-4.854,P均＜0.001）,其中A组LVGLS、LVAP4LS和LVAP3LS测值比C组减低,差异具有统计学意义（t=-2.493、-2.405、-2.012,P=0.014、=0.018、=0.047）,A组与C组LVAP2LS测值比较,差异无统计学意义（P＞0.05）,A组与B组间以及B组与C组间在LVGLS、LVAP2LS、LVAP3LS、LVAP4LS测值比较,差异也均无统计学意义（P均＞0.05）。 结论aCMQ能早期发现尿毒症透析患者左心室收缩功能的异常,为临床早期预防及治疗心功能衰竭提供了新的途径。     

自拟增液育胎方治疗妊娠晚期羊水过少的临床效果观察

目的:探讨自拟增液育胎方治疗妊娠晚期羊水过少的临床效果。方法:将2018年2月~2019年2月在某院产科治疗的120例妊娠晚期羊水过少患者随机分为观察组和对照组。对照组采用常规西医治疗,在此基础上观察组使用自拟增液育胎方治疗,比较两组患者的临床疗效、羊水和脐血流变化、分娩方式、妊娠结局。结果:观察组治疗有效率为83.33%,高于对照组的73.33%;观察组治疗后AFI、AFD明显高于对照组,而S/D明显低于对照组(P<0.05);观察组早产、新生儿窒息、新生儿低体重、新生儿Apgar评分≤7分等不良妊娠结局发生率明显低于对照组(P<0.05);观察组自然分娩率明显高于对照组,而剖宫产率明显低于对照组(P<0.05)。结论:自拟增液育胎方治疗妊娠晚期羊水过少的临床效果显著,能有效增加羊水量,改善胎盘血液循环,降低不良妊娠结局的发生率,提高自然分娩率,具有积极的临床意义。     

关于磷酸氯喹治疗新型冠状病毒肺炎给药方案的安全性刍议

国家卫生健康委员会和国家中医药管理局2020年2月18日发布的《新型冠状病毒肺炎诊疗方案(试行第六版)》将磷酸氯喹作为抗病毒试用药物之一,其后又于同月26日发布了《关于调整试用磷酸氯喹治疗新冠肺炎用法用量的通知》,2020年3月3日发布的《新型冠状病毒肺炎诊疗方案(试行第七版)》对磷酸氯喹给药方案进行了修订。磷酸氯喹半衰期长、全身分布广泛,易在体内蓄积,且不良反应与剂量相关。本文结合磷酸氯喹的药代动力学特点对给药方案进行了安全性分析,供临床医师和药师在诊治新型冠状病毒肺炎患者时参考,以降低发生不良反应的风险。     

酒石酸布托啡诺、地佐辛与盐酸托烷司琼在镇痛泵内配伍的稳定性考察

目的:考察酒石酸布托啡诺、地佐辛、盐酸托烷司琼在0.9%氯化钠注射液中的配伍稳定性。方法:模拟临床实际用药的配伍浓度和使用环境,检测0,1,4,6,24,48,72 h外观、pH、相对百分含量的变化。结果:酒石酸布托啡诺、地佐辛与盐酸托烷司琼的配伍液在72 h内pH未出现明显变化,外观稳定,各药品成分相对百分含量也没有明显变化。结论:在室温条件下,酒石酸布托啡诺、地佐辛与盐酸托烷司琼在0.9%氯化钠注射液中72 h内保持稳定。因此,酒石酸布托啡诺(6 mg)、地佐辛(20 mg)与盐酸托烷司琼(10 mg)可以配伍使用,在镇痛泵内配伍稳定。