The effect of two cryopreservation methods on human sperm DNA damage

Several methods are currently available for selection when conducting sperm cryopreservation, however, these methods might cause different degrees of damage on sperm DNA. The aim of the this study is to compare the effects of storage at −80 °C (in ultra-low temperature refrigerator) and at −196 °C (in liquid nitrogen) on sperm DNA damage, thus to provide a reference for choosing the right method according to different aims. We randomly collected 28 semen samples from college students of Chongqing city. The samples stored at −80 °C were neat semen samples and the samples stored at −196 °C were mixed with additional cryoprotectants. Each sample was subjected to two freezing-thawing cycles, and the sperm DNA damage levels of fresh and thawed samples were measured by single cell gel electrophoresis (SCGE) and sperm chromatin structure assay (SCSA). Both SCGE and SCSA assays showed cryopreservation induced significant damage to sperm DNA. However, storage at −196 °C lead to more severe damage to sperm DNA than storage at −80 °C measured by SCSA. Sperm DNA damage increased simultaneously with the higher frequency of freezing-thawing cycles. We concluded that storage of neat semen samples at −80 °C had milder damage to sperm DNA than storage at −196 °C mixed with cryoprotectants. To avoid additional sperm DNA damage, repeated freezing and thawing should be prevented.     

3-Substituted-4-hydroxycoumarin as a new scaffold with potent CDK inhibition and promising anticancer effect: Synthesis,molecular modeling and QSAR studies

A new series of 3-substituted-4-hydroxycoumarin derivatives was designed, synthesized, and evaluated for CDK inhibiting and anticancer activities. All the synthesized target compounds showed remarkably high affinity and selectivity towards CDK1B, compared to flavopiridol, with Ki values in the low nanomolar range (Ki = 0.35–0.88 nM). Most of them elicited considerable inhibiting effect against CDK9T1 (Ki = 3.26–23.45 nM). Moreover, all the target compounds were tested in vitro against eighteen types of human tumor cell lines. The hydrazone 3a, N-phenylpyrazoline derivative 6b and 2-aminopyridyl-3-carbonitrile derivative 8c were the most potent anticancer agents against MCF-7 breast cancer cell line (IC₅₀ = 0.21, 0.21 and 0.23 nM, respectively). The target compounds 3a, 6b and 8c were further evaluated in MCF-7 breast cancer mouse xenograft model and showed in vivo efficacy at 10 mg/kg dose. The docking study confirmed a unique binding mode in the active site of CDK1B with better score than flavopiridol. Quantitative structure activity relationship study was done and revealed a highly predictive power R² of 0.81.     

Synthesis,characterization and thermoluminescence studies of Mn‐doped ZnS nanoparticles

ZnS:Mn nanoparticles were prepared by a chemical precipitation method and characterized by X‐ray diffraction (XRD), field emission gun scanning electron microscope (FEGSEM), and high resolution transmission electron microscopy (HRTEM). Capping agent (mercaptoethanol) concentrations used were 0 M, 0.005 M, 0.01 M, 0.015 M, 0.025 M, 0.040 M, and 0.060 M, and resulted in nanoparticles sizes of 2.98 nm, 2.9 nm, 2.8 nm, 2.7 nm, 2.61 nm, 2.2 nm and 2.1 nm, respectively. The thermoluminescence (TL) glow curve was recorded by heating the sample exposed to UV‐radiation, at a fixed heating rate 1°C sec^–1. The TL intensity initially increased with temperature, attained a peak value I_m for a particular temperature, and then decreased with further increase in temperature. The peak TL intensity increased with decreasing nanoparticle size, whereas the temperature corresponding to the peak TL intensity decreased slightly with reducing nanocrystal size. As a consequence of increase in surface‐to‐volume ratio and increased carrier recombination rates, the TL intensity increased with decreasing nanoparticle size. It was found that, whereas activation energy slightly decreased with decreasing nanoparticle size, the frequency factor decreased significantly with reduction in nanoparticle size. Copyright © 2015 John Wiley & Sons, Ltd.     

Responses of growth of four desert species to different N addition levels

Aims The increase in atmospheric N deposition has accelerated N cycling of ecosystems, thus altering the structure and function of ecosystems, especially in those limited by N availability. Studies on the response of plant growth to artificial N addition could provide basic data for a better understanding of how the structure of grasslands in northern China responds to increasing N deposition. Methods We investigated the seasonal dynamics of plant growth of four species after 2-year multi-level N addition in a field experiment conducted in a desert steppe of Ningxia in 2011. Plant biomass and the relative growth rate (RGR) of the studied species were measured and their relationships with C:N:P ratios of plants (community and leaf levels) and soils were analyzed. Important findings Results in 2012 showed that 2-year N addition promoted the growth of the four species and the effects were different among growth forms and were species-specific. In general, the plant biomass of the studied species was significantly correlated with leaf N concentration, leaf N:P ratio, community N pool, soil total N content and soil N:P ratio, while only weak relationships were observed between plant biomass and C:N and C:P ratios of plants and soils. In contrast, there was a significant linear relationship between RGR and N:P ratios both of plants and soils.Our results suggest that short-term N addition promoted the accumulation of plant biomass, and the species-specific responses to stimulated N addition can directly affect the structure of the desert steppe ecosystem. Plant N:P ratio and soil N:P ratio could indicate nutrient limitation of plant growth to a certain extent: N addition increased soil N content and N:P ratio, and thus relieved N limitation gradually. Once more N is available to plants, the growth of plants and the accumulation of community N was stimulated in turn.     

Foliar nitrogen and phosphorus stoichiometry of alien invasive plants and co-occurring natives in Xishuangbanna

Aims How alien invasive plants and co-occurring native plants utilize nutrients is one of major issues in invasion ecology. Foliar nitrogen (N) and phosphorus (P) contents and stoichiometry can elucidate the uptake ability and limitation status of nutrients in plants, which provides basic knowledge for understanding the invading ability and co-occurrence or disappearance of plants.
Methods Based on typical alien invasive plants (Chromolaena odorata, Ageratina adenophora) and native plants in southwestern China, this study focused on strategies of N and P utilization among invasive plants and native plants under different invasion conditions. The species compositions, aboveground biomass, leaf N and P contents and leaf N:P were investigated for plants in plots with no invasion and with different invasion extents (estimated by the plot-based percentage of invaders’ biomass in total community) at Mt. Kongming in Xishuangbanna region, Yunnan Province, China.
Important findings The species number decreased significantly with the invasion extent of both C. odorata and A. adenophora, although the aboveground biomass was greatly enhanced. Leaf N and P contents did not differ between the two studied invaders, but they showed significantly higher N and P levels than both co-occurring and only native species (p < 0.05). Besides, leaf N and P contents of invaders increased with the invasion extent, and leaf N of native plants also showed an increasing trend with the invasion extent. When the influence of invasion was checked for the same species, leaf P contents decreased, whereas leaf N and N:P increased for most native plants under invasion. Based on the absolute foliar N and P contents, N:P values, we inferred that native plants were still limited by N, although N availability might be enhanced by invasion. Both invasive plants had leaf N:P values lower than 10, suggesting a higher P uptake relative to N uptake. All above results highlighted a higher N and P uptake of typical alien invasive plants in southwestern China.     

8-Substituted 1,3-dimethyltetrahydropyrazino[2,1-f]purinediones: Water-soluble adenosine receptor antagonists and monoamine oxidase B inhibitors

Multitarget approaches, i.e., addressing two or more targets simultaneously with a therapeutic agent, are hypothesized to offer additive therapeutic benefit for the treatment of neurodegenerative diseases. Validated targets for the treatment of Parkinson’s disease are, among others, the A_2A adenosine receptor (AR) and the enzyme monoamine oxidase B (MAO-B). Additional blockade of brain A₁ ARs may also be beneficial. We recently described 8-benzyl-substituted tetrahydropyrazino[2,1-f]purinediones as a new lead structure for the development of such multi-target drugs. We have now designed a new series of tetrahydropyrazino[2,1-f]purinediones to extensively explore their structure–activity-relationships. Several compounds blocked human and rat A₁ and A_2AARs at similar concentrations representing dual A₁/A_2A antagonists with high selectivity versus the other AR subtypes. Among the best dual A₁/A_2AAR antagonists were 8-(3-(4-chlorophenyl)propyl)-1,3-dimethyl-6,7,8,9-tetrahydropyrazino[2,1-f]purine-2,4(1H,3H)-dione (41, K_i human A₁: 65.5 nM, A_2A: 230 nM; K_i rat A₁: 352 nM, A_2A: 316 nM) and 1,3-dimethyl-8-((2-(thiophen-2-yl)thiazol-4-yl)methyl)-6,7,8,9-tetrahydropyrazino[2,1-f]purine-2,4(1H,3H)-dione (57, K_i human A₁: 642 nM, A_2A: 203 nM; K_i rat A₁: 166 nM, A_2A: 121 nM). Compound 57 was found to be well water-soluble (0.7 mg/mL) at a physiological pH value of 7.4. One of the new compounds showed triple-target inhibition: (R)-1,3-dimethyl-8-(2,1,3,4-tetrahydronaphthalen-1-yl)-6,7,8,9-tetrahydropyrazino[2,1-f]purine-2,4(1H,3H)-dione (49) was about equipotent at A₁ and A_2AARs and at MAO-B (K_i human A₁: 393 nM, human A_2A: 595 nM, IC₅₀ human MAO-B: 210 nM) thus allowing future in vivo explorations of the intended multi-target approach.     

Influence of spectral wavelength and N:P ratio on the cell cycle of Synechococcus strain CSIRNIO1 isolated from the central Eastern Arabian Sea

The novel phycoerythrin-containing Synechococcus strain CSIRNIO1 belonging to phylogenetic clade II was isolated from the coastal Arabian Sea. Chromophore characteristics of this isolate revealed the presence of phycoerythrin I (PEI), which allows it to utilize green light efficiently. The DNA distribution data indicate a bimodal slow growth model synchronized with the light/dark cycle. The duration of the cell cycle was regulated by spectral wavelength and nutrient concentration. Nitrate and phosphate enrichment shortened G1 phase duration when cells were exposed to equal doses of photosynthetically usable radiation (PUR) of different spectral wavelengths. G2 phase duration was influenced by spectral quality and phosphate concentration. S phase duration was not affected by the spectral wavelength. However, a shorter doubling time corresponding to shortened G1 and S phases was observed under nitrate enrichment. Phosphate enrichment resulted in shortening of all three phases (G1, S and G2). More efficient utilization of green and red light than blue light regulated the duration of the cell cycle as well as the doubling time, suggesting spectral selectivity in this strain. The effects of spectral wavelengths under varying nutrient concentrations will determine the proliferation of Synechococcus and its adaptation to different environmental conditions.     

农村高血压患者协调性和连续性服务的利用现状研究

目的 分析高血压患者在农村医疗服务网络接受协调性和连续性服务的提供状况,为改进患者的整合服务利用提供决策参考。方法 根据国内外文献,以自行编制服务协调性和连续性提供的问卷为工具,调查数据采用描述性统计分析。结果 49.6%的患者伴有1种及以上疾病,约73.0%在乡镇卫生院有就医经历,到过2家及以上机构就医的占51.3%;在乡村两级治疗效果不佳的患者,能够获取村医和乡镇医生推荐转诊机构的比重最大,分别为28.4%和68.7%;上级医生能够根据下级机构相关诊疗信息连续诊疗的不到43.0%,下级医生能够根据前期诊疗信息继续诊疗的比例刚超4成;“上转容易、下转难”现象同样存在。 结论 农村慢性病患者的患病特点增加了在农村纵向医疗机构就诊的几率,但该网络提供连续性和协调性服务的程度却不高,应加强农村三级医疗机构间的全面整合。