2-苯乙醇对酿酒酵母生理生化特性影响

【目的】研究在不同浓度2-苯乙醇作用下,酵母生理生化特性的变化规律,为优化2-苯乙醇生物合成过程提供重要依据。【方法】透射电镜观察细胞形态;流式细胞术检测细胞膜渗透性、胞内ROS浓度、线粒体膜电位;实时荧光定量PCR检测关键酶基因表达。【结果】随着2-苯乙醇浓度增加(从0到4.0 g/L),酵母细胞分解代谢能力、细胞膜渗透性及aro10基因表达量逐渐降低;线粒体膜电位逐渐增加;胞内ROS浓度先增加后减少。当2-苯乙醇浓度从2.4 g/L增加到3.0 g/L,酵母的分解代谢能力、细胞膜渗透性、aro10基因表达水平等生理生化特性都发生较为显著的变化。【结论】产物原位转移过程中水相2-苯乙醇浓度可考虑控制在2.4 3.0 g/L。     

A multishear microfluidic device for quantitative analysis of calcium dynamics in osteoblasts

Reactive oxygen species (ROS) are an important factor in the development of skin lesions in diabetes. A new antioxidant, hydrogen, can selectively neutralize hydroxyl radicals (OH) and peroxynitrite (ONOO⁻) in cell-free systems, whereas it seldom reacts with other ROS. Fibroblasts are a key component of skin. In the present study, we investigated the protective effects of hydrogen-rich medium on human skin fibroblasts (HSFs) under oxidative stress. Confocal microscopy was used to assay both the intracellular superoxide anion () concentration and the mitochondrial membrane potential (ΔΨ). Cell viability was determined using the Cell Counting Kit-8 (CCK-8). The concentrations of cellular malonaldehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), 8-hydroxy-2′-deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) were also measured. The results revealed that both mannitol and high glucose could cause oxidative stress in HSFs. Interestingly, the use of a hydrogen-rich medium significantly reduced the level of intracellular , stabilized the ΔΨ and attenuated production of MDA, 8-OHdG and 3-NT which efficiently enhanced the antioxidative defense system and protected the HSFs from subsequent oxidative stress damage. In other words, hydrogen decreased the excessive generation of intracellular and elevated the cellular antioxidative defense. Based on our results, hydrogen may have applications in the treatment of skin diseases caused by diabetes.     

Hydrolytic Amino Acids Employed as a Novel Organic Nitrogen Source for the Preparation of PGPF-Containing Bio-Organic Fertilizer for Plant Growth Promotion and Characterization of Substance Transformation during BOF Production

Opportunity costs seriously limit the large-scale production of bio-organic fertilizers (BOFs) both in China and internationally. This study addresses the utilization of amino acids resulting from the acidic hydrolysis of pig corpses as organic nitrogen sources to increase the density of TrichodermaharzianumT-E5 (a typical plant growth-promoting fungi, PGPF). This results in a novel, economical, highly efficient and environmentally friendly BOF product. Fluorescence excitation-emission matrix (EEM) spectroscopy combined with fluorescence regional integration (FRI) was employed to monitor compost maturity levels, while pot experiments were utilized to test the effects of this novel BOF on plant growth. An optimization experiment, based on response surface methodologies (RSMs), showed that a maximum T-E5 population (3.72 × 10⁸ ITS copies g⁻¹) was obtained from a mixture of 65.17% cattle manure compost (W/W), 19.33% maggot manure (W/W), 15.50% (V/W)hydrolytic amino acid solution and 4.69% (V/W) inoculum at 28.7°C after a 14 day secondary solid fermentation. Spectroscopy analysis revealed that the compost transformation process involved the degradation of protein-like substances and the formation of fulvic-like and humic-like substances. FRI parameters (P_{I, n}, P_{II, n}, P_{III, n} and P_{V, n}) were used to characterize the degree of compost maturity. The BOF resulted in significantly higher increased chlorophyll content, shoot length, and shoot and root dry weights of three vegetables (cucumber, tomato and pepper) by 9.9%~22.4%, 22.9%~58.5%, 31.0%~84.9%, and 24.2%~34.1%, respectively. In summary, this study presents an operational means of increasing PGPF T-E5 populations in BOF to promote plant growth with a concomitant reduction in production cost. In addition, a BOF compost maturity assessment using fluorescence EEM spectroscopy and FRI ensured its safe field application.     

Novel HRPT2/CDC73 Gene Mutations and Loss of Expression of Parafibromin in Chinese Patients with Clinically Sporadic Parathyroid Carcinomas

Objective

It is widely recognized that the diagnosis of parathyroid carcinoma (PC) is often difficult because of the overlap of characteristics between malignant and benign parathyroid tumors, especially at an early stage. Based on the identification of tumor suppressor gene HRPT2/CDC73 and its association with hereditary and sporadic PC, screening of gene mutations and detection of parafibromin immunoreactivity have been suggested as diagnostic instruments of PC in Whites. There is little information about HRPT2/CDC73 mutations and its corresponding protein expression in patients with sporadic PC in Chinese population, and the long-term follow-up data is scarce.

Methods

Paraffin-embedded tissues were obtained from 13 patients with PC, 13 patients with parathyroid adenoma (PA) and 7 patients with parathyroid hyperplasia(PH), and 6 normal parathyroid (NP) tissues as controls. Peripheral blood from 11 patients with PC was collected. PCR products using Genomic DNA extracted from tumor tissues or blood as template was sequenced for HRPT2/CDC73 gene. Expression of parafibromin in tumor tissues was evaluated by immunohistochemical analysis.

Results

Six mutations in 6 of 13 patients with PC were identified, with three being novel. Four of them were germ-line mutations. Patients with mutations were susceptible to recurrence of the PC. Complete (8/13, 61.5%) or partial (5/13, 38.5%) loss of parafibromin expression was observed in PC tissues. All of tissue samples from normal parathyroid or benign parathyroid tumors displayed positive immunostaining of parafibromin except one adenoma.

Conclusions

The present study supplies information on the mutations and protein expression of HRPT2/CDC73 gene and phenotypes of parathyroid carcinoma in Chinese population. And the expanded mutation database of this gene may benefit patients in the diagnosis and treatment of this disease.     

基于MaxEnt模型预测中国兰属植物的分布格局及主导气候因子(附表)

兰属(Cymbidium)中，除了兔耳兰C.lancifolium以外的所有种均被列为国家重点保护野生植物。为探究其在未来气候条件下的潜在分布格局，该研究基于兰属植物已知的分布点和19个气候因子，利用最大熵(MaxEnt)模型和地理信息系统(ArcGIS)模拟兰属以及其中20种兰属植物在9种不同气候情景(当代以及未来2030s、2050s、2070s和2090s 4个时间段各两种温室气体排放情景)下的潜在分布格局。结果表明：(1)最干旱季降水量(Bio17)、年降水量(Bio12)和温度季节性变化(Bio4)是影响兰属植物地理分布格局的主导气候因子。(2)不同兰属植物在未来情景下的适生区表现出不同的变化趋势，并且影响其分布的主导气候因子也有所不同。其中，冬凤兰(C.dayanum)等8个物种的适生区面积整体呈扩张趋势，而西藏虎头兰(C.tracyanum)等12个物种的适生区面积整体则呈缩减趋势。该研究结果为兰属植物就地保护与迁地保护提供了重要参考，对兰属等濒危野生植物的保护具有积极意义。     

Synthesis and antitumor activities of novel α-aminophosphonates dehydroabietic acid derivatives

A series of novel α-aminophosphonate derivatives containing DHA structure were designed and synthesized as antitumor agents. In vitro antitumor activities of these compounds against the NCI-H460 (human lung cancer cell), A549 (human lung adenocarcinoma cell), HepG2 (human liver cancer cell) and SKOV3 (human ovarian cancer cell) human cancer cell lines were evaluated and compared with commercial anticancer drug 5-fluorouracil (5-FU), employing standard MTT assay. The pharmacological screening results revealed that many compounds exhibited moderate to high levels of antitumor activities against the tested cancer cell lines and that most demonstrated more potent inhibitory activities compared with the commercial anticancer drug 5-FU. The action mechanism of representative compound 7c was preliminarily investigated by acridine orange/ethidium bromide staining, Hoechst 33258 staining, JC-1 mitochondrial membrane potential staining and flow cytometry, which indicated that the compound can induce cell apoptosis in NCI-H460 cells. Cell cycle analysis showed that compound 7c mainly arrested NCI-H460 cells in G1 stage.     

力学载荷对骨整合影响的动物实验

植入体的骨整合使植入体与骨组织间形成牢固的接合, 可承受功能性生理载荷. 加载时间是决定植入体骨整合进程的关键因素. 然而, 关于初期载荷是否影响骨整合的进程, 或者, 无载荷愈合期是否可以缩短等问题现在还不清楚. 本文通过动物实验, 研究在骨愈合初期, 外部载荷如何影响骨整合的进程. 将钛植入体侧向植入山羊胫骨, 从术后4周开始, 对植入体加不同大小轴向载荷. 加载两周时, 取下连带植入体的胫骨; 设计专门的“拔出”力测试实验, 以检测不同分组(包括加载组和无载荷组)植入体的拔出力、骨-植入体界面的剪切强度. 并采用组织学染色、能量色散谱(energy dispersive spectroscopy, EDS)和扫描电子显微镜(scanning electron microscope, SEM)分析骨-植入体界面特征, 以评价骨整合状态. 结果表明, 术后4周时, 骨-植入体界面没有良好的骨整合, 在无载荷组样本的骨-植入体界面处可发现成纤维样组织, 而轴向10 N加载组样本的骨-植入体界面则发生良好的骨整合. 这表明, 植入手术后一定的载荷可以有利于骨整合的发生. 本研究提示, 术后初期的载荷会影响植入体骨整合的进程, 而适宜的载荷可缩短术后骨整合发生的时间.     

CD33‐targeting extracellular vesicles deliver antisense oligonucleotides against FLT3‐ITD and miR‐125b for specific treatment of acute myeloid leukaemia

IntroductionAcute Myeloid Leukaemia (AML) is the most common blood cancer in adults. Although 2 out of 3 AML patients go into total remission after chemotherapies and targeted therapies, the disease recurs in 60%–65% of younger adult patients within 3 years after diagnosis with a dramatically decreased survival rate. Therapeutic oligonucleotides are promising treatments under development for AML as they can be designed to silence oncogenes with high specificity and flexibility. However, there are not many well validated approaches for safely and efficiently delivering oligonucleotide drugs. This issue could be resolved by utilizing a new generation of delivery vehicles such as extracellular vesicles (EVs).MethodsIn this study, we harness red blood cell‐derived EVs (RBCEVs) and engineer them via exogenous drug loading and surface functionalization to develop an efficient drug delivery system for AML. Particularly, EVs are designed to target CD33, a common surface marker with elevated expression in AML cells via the conjugation of a CD33‐binding monoclonal antibody onto the EV surface.ResultsThe conjugation of RBCEVs with the CD33‐binding antibody significantly increases the uptake of RBCEVs by CD33‐positive AML cells, but not by CD33‐negative cells. We also load CD33‐targeting RBCEVs with antisense oligonucleotides (ASOs) targeting FLT3‐ITD or miR‐125b, 2 common oncogenes in AML, and demonstrate that the engineered EVs improve leukaemia suppression in in vitro and in vivo models of AML.ConclusionTargeted RBCEVs represent an innovative, efficient, and versatile delivery platform for therapeutic ASOs and can expedite the clinical translation of oligonucleotide drugs for AML treatments by overcoming current obstacles in oligonucleotide delivery.

In this study, we harness red blood cell‐derived EVs (RBCEVs) and engineer them with surface functionalization and exogenous drug loading to develop an efficient drug delivery system for AML. Anti‐CD33 antibody was conjugated to RBCEVs using an enzymatic method combined with the streptavidin‐biotin system. We load the antibody conjugated RBCEVs with ASOs targeting FLT3‐ITD or miR‐125b, 2 common oncogenes in AML, and demonstrate that the treatment with engineered EVs improve leukaemia suppression both in vitro and in vivo.