ß-Amyloid in blood neuronal-derived extracellular vesicles is elevated in cognitively normal adults at risk of Alzheimer's disease and predicts cerebral amyloidosis.

BACKGROUND: Blood biomarkers that can be used for preclinical Alzheimer's disease (AD) diagnosis would enable trial enrollment at a time when the disease is potentially reversible. Here, we investigated plasma neuronal-derived extracellular vesicle (nEV) cargo in patients along the Alzheimer's continuum, focusing on cognitively normal controls (NCs) with high brain ß-amyloid (Aß) loads (Aß+). METHODS: The study was based on the Sino Longitudinal Study on Cognitive Decline project. We enrolled 246 participants, including 156 NCs, 45 amnestic mild cognitive impairment (aMCI) patients, and 45 AD dementia (ADD) patients. Brain Aß loads were determined using positron emission tomography. NCs were classified into 84 Aß- NCs and 72 Aß+ NCs. Baseline plasma nEVs were isolated by immunoprecipitation with an anti-CD171 antibody. After verification, their cargos, including Aß, tau phosphorylated at threonine 181, and neurofilament light, were quantified using a single-molecule array. Concentrations of these cargos were compared among the groups, and their receiver operating characteristic (ROC) curves were constructed. A subset of participants underwent follow-up cognitive assessment and magnetic resonance imaging. The relationships of nEV cargo levels with amyloid deposition, longitudinal changes in cognition, and brain regional volume were explored using correlation analysis. Additionally, 458 subjects in the project had previously undergone plasma Aß quantification. RESULTS: Only nEV Aß was included in the subsequent analysis. We focused on Aß42 in the current study. After normalization of nEVs, the levels of Aß42 were found to increase gradually across the cognitive continuum, with the lowest in the Aß- NC group, an increase in the Aß+ NC group, a further increase in the aMCI group, and the highest in the ADD group, contributing to their diagnoses (Aß- NCs vs. Aß+ NCs, area under the ROC curve values of 0.663; vs. aMCI, 0.857; vs. ADD, 0.957). Furthermore, nEV Aß42 was significantly correlated with amyloid deposition, as well as longitudinal changes in cognition and entorhinal volume. There were no differences in plasma Aß levels among NCs, aMCI, and ADD individuals. CONCLUSIONS: Our findings suggest the potential use of plasma nEV Aß42 levels in diagnosing AD-induced cognitive impairment and Aß+ NCs. This biomarker reflects cortical amyloid deposition and predicts cognitive decline and entorhinal atrophy.

Risk factors for relapse and nomogram for relapse probability prediction in patients with minor ischemic stroke.

BACKGROUND: The identification of risk factors for recurrence in patients with minor ischemic stroke (MIS) is a critical medical need. AIM: To develop a nomogram for individualized prediction of in-hospital recurrence in MIS patients. METHODS: Based on retrospective collection, a single-center study was conducted at the First Affiliated Hospital of Anhui Medical University from January 2014 to December 2019. Univariate and multivariate logistic regression analyses were used to determine the risk factors associated with MIS recurrence. The least absolute shrinkage and selection operator regression was performed for preliminary identification of potential risk factors. Uric acid, systolic blood pressure, serum total bilirubin (STBL), and ferritin were integrated for nomogram construction. The predictive accuracy and calibration of the nomogram model were assessed by the area under the receiver operating characteristic curve (AUC-ROC) and Hosmer-Lemeshow test, respectively. RESULTS: A total of 2216 MIS patients were screened. Among them, 155 were excluded for intravascular therapy, 146 for unknown National Institutes of Health Stroke Scale score, 195 for intracranial hemorrhage, and 247 for progressive stroke. Finally, 1244 patients were subjected to further analysis and divided into a training set (n = 796) and a validation set (n = 448). Multivariate logistic regression analysis revealed that uric acid [odds ratio (OR): 0.997, 95% confidence interval (CI): 0.993-0.999], ferritin (OR: 1.004, 95%CI: 1.002-1.006), and STBL (OR: 0.973, 95%CI: 0.956-0.990) were independently associated with in-hospital recurrence in MIS patients. Our model showed good discrimination; the AUC-ROC value was 0.725 (95%CI: 0.646-0.804) in the training set and 0.717 (95%CI: 0.580-0.785) in the validation set. Moreover, the calibration between nomogram prediction and the actual observation showed good consistency. Hosmer-Lemeshow test results confirmed that the nomogram was well-calibrated (P = 0.850). CONCLUSION: Our present findings suggest that the nomogram may provide individualized prediction of recurrence in MIS patients.

A Novel Probable Pathogenic PSEN2 Mutation p.Phe369Ser Associated With Early-Onset Alzheimer's Disease in a Chinese Han Family: A Case Report.

The pathogenesis of Alzheimer's disease is complex, and early-onset Alzheimer's disease (EOAD) is mostly influenced by genetic factors. Presenilin-1, presenilin-2 (PSEN2), and amyloid precursor protein are currently known as the three main causative genes for autosomal dominant EOAD, with the PSEN2 mutation being the rarest. In this study, we reported a 56-year-old Chinese Han proband who presented with prominent progressive amnesia, aphasia, executive function impairment, and depression 5 years ago. The 3-year follow-up showed that the patient experienced progressive brain atrophy displayed on magnetic resonance imaging (MRI) and dramatic cognitive decline assessed by neuropsychological evaluation. This patient was clinically diagnosed as EOAD based on established criteria. A heterozygous variant (NM_000447.2: c.1106T>C) of PSEN2 was identified for the first time in this patient and her two daughters. This mutation causing a novel missense mutation (p.Phe369Ser) in transmembrane domain 7 encoded by exon 11 had not been reported previously in 1000Genomes, ExAC, or ClinVar databases. This mutation was predicted by four in silico prediction programs, which all strongly suggested that it was damaging. Our results suggest that this novel PSEN2 Phe369Ser mutation may alter PSEN2 protein function and associate with EOAD.

Hemorrhagic transformation of ischemic cerebral proliferative angiopathy: A case report.

BACKGROUND: Cerebral proliferative angiopathy (CPA) is a rare vascular disease characterized by the presence of diffuse vascular proliferation, progressive vascular hyperflow and vasodilation of multiple vessels in the normal brain parenchyma. Unlike cerebral arteriovenous malformations, CPA has a mixed appearance between that of lesions with cell proliferation and endothelial proliferation. To date, the pathogenesis of CPA is unclear, in which changes induced by cortical ischemia in the elastic layer of the blood supply artery and smooth muscle cells may be involved. CASE SUMMARY: In this article, we retrospectively analyzed a case of hemorrhagic transformation of ischemic CPA diagnosed by digital subtraction angiography and reviewed the related literature for further exploration of its pathogenesis, diagnosis and treatment. CONCLUSION: The information in the present case report may facilitate further clinical research on this cerebrovascular disease.

[Effects of vertical rotary subsoiling with combined organic and inorganic fertilization on water use efficiency and yield of forage maize in a semi-arid area.] / åŠå¹²æ—±åŒºç«‹å¼æ·±æ—‹è€•å’Œæœ‰æœºæ— æœºè‚¥é æ–½å¯¹é¥²ç”¨çŽ‰ç±³æ°´åˆ†åˆ©ç”¨æ•ˆçŽ‡å’Œäº§é‡çš„å½±å“.

Both reasonable soil tillage and fertilization management play critical roles in improving the yield and water use efficiency (WUE) of forage maize in the semi-arid area of Loess Plateau. A field experiment was conducted at Dingxi experimental station of Gansu Academy of Agricultural Sciences between 2017 and 2019. We explored the effects of tillage method and fertilization type on yields and WUE of forage maize, as well as the economic benefits. There were four treatments in the experiment, including traditional rotary tillage + organic-inorganic fertilization (TOF), deep rotary tillage + organic-inorganic fertilization (DOF), and vertical rotary subsoiling + organic-inorganic fertilization (VROF), and the traditional rotary tillage + inorganic fertilization as the control (TF). Our results showed that, compared with DOF, TOF, TF, and VROF all decreased soil water storage in 0-300 cm soil layer at flowering stage, ranging from 16.9 mm to 79.9 mm, but they all increased soil water consumption by 9.7-22.4 mm during vegetative growing stages, 11.0-19.8 mm during reproductive stage in the dry years. Due to significant improvement in water absorption, VROF increased dry matter weight at maturity by 3.9%-13.4% compared to other treatments. Similarly, plant height, ear length, grain number per ear, 100-grain weight, and double ear rate under VROF were significantly increased, while bald head length was decreased significantly, when compared with other treatments. As a result, over the three experimental seasons, VROF increased the grain and biological yield by 4.3%-51.5% and 4.3%-25.7% compared to other treatments, respectively. Accordingly, WUE calculated by grain and biomass yields were increased by 2.7%-36.9% and 3.6%-13.5% under VROF, compared to other treatments. VROF increased the unit gross total output value and the net income by 5.1%-32.9% and 6.9%-80.5% respectively, compared to other treatments. These results demonstrated that VROF is a drought-resistant and yield-increasing farming technology for sustainable forage maize production in the semi-arid area of the Loess Plateau, Northwest China.

[Effects of organic fertilizer application on flag leaf C/N ratio, photosynthetic characteristics and yield of spring wheat with full plastic film mulching]. / å ¨è†œè¦†åœŸä¸‹æ–½æœ‰æœºè‚¥å¯¹æ˜¥å°éº¦æ——å¶ç¢³æ°®æ¯”ã€å ‰åˆç‰¹æ€§å’Œäº§é‡çš„å½±å“.

A field experiment was conducted in the rain-fed semi-arid region of central Gansu in 2016 and 2017, with the treatments 1) hill-drop flat planting with full plastic film mulching (PMS), 2) hill-drop flat planting with full plastic film mulching plus organic fertilizers (PMO), and 3) hill-drop flat planting without soil mulching (CK). We investigated the relations among soil moisture, photosynthetic rate (Pn), stomatal conductance (gs) and transpiration rate (Tr), C/N ratio, and total nitrogen of flag leaf from the heading stage to the seed-filling stage in different treatments to probe into their effects on the yield and yield components of spring wheat variety 'Longchun 27'. The results showed that organic fertilizer application could increase soil moisture at the middle and late growth stages of spring wheat. PMO increased soil water storage in 0-300 cm depth from the heading stage to the seed filling stage by 4.6% and 8.5%, decreased population canopy temperature by 0.1-1.3 ℃ and 1.4-4.9 ℃, increased net photosynthetic rate of flag leaf by 9.3% and 29.7%, stomatal conductance by 30.9% and 103.8%, transpiration rate by 5.1% and 55.0%, total nitrogen content by 6.6% and 18.9%, and decreased C/N ratio by 6.4% and 22.8%, respectively. Compared with PMS and CK, PMO significantly improved grain number per spike and 1000-grain weight, and increased grain yield by 9.1% and 53.7%, respectively. From the heading stage to filling stage, the Pn and gs of flag leaf had negative correlation with C/N, while C/N was negatively correlated with grain yield. Consequently, PMO could improve soil water storage and promote photosynthesis of flag leaf, reduce the intensity of physiological drought stress and the limitations of nitrogen absorption and assimilation in flag leaf from the heading stage to the seed-filling stage, and increase grain number and grain weight and consequently the yield of spring wheat in semi-arid region.

[Effects of optimal nitrogen fertilizer management on water and fertilizer utilization efficiency and yield under double-ridge-furrow sowing with the whole plastic film mulching in maize in a semi-arid area]. / åŠå¹²æ—±åŒºæ°®è‚¥è¿ç­¹å¯¹å ¨è†œåŒåž„æ²Ÿæ’­çŽ‰ç±³æ°´è‚¥åˆ©ç”¨å’Œäº§é‡çš„å½±å“.

Improper fertilization style is one of the main reasons for low water and fertilizer use efficiency of double-ridge-furrow sowing with the whole plastic film mulching in maize production in the semi-arid area. Understanding the effects of reduction, postponing, and organic fertilizer substitution of nitrogen fertilizer on water and fertilizer use efficiency and yield of maize can provide theore-tical basis for effective management of water and fertilizer in maize production. Based on a 4-year field experiment with three treatments: all fertilizers as base fertilizer under double-ridge-furrow sowing with the whole plastic film mulching (CK), nitrogen fertilizer reduced by 15% and topdres-sing in tasseling stage (RN), 30% of the chemical fertilizer replaced by organic fertilizer and topdressing in tasseling stage (RNM), we measured water consumption characteristics, growth and development, water and fertilizer utilization efficiency of maize. The results showed that fertilization pattern significantly affected water and fertilizer utilization efficiency and yield of maize, which was dependent on annual rainfall. In dry and normal rainfall year, water consumption in pre-flowering stage of RN was decreased by 16.1%-18.8% and that in post-flowering stage was increased by 18.0%-22.2%, while water consumption in pre-flowering and post-flowering stages of RNM did not differ from that in CK. In wet year, water consumption in pre-flowering stage of RN and RNM was decreased by 16.7% and 6.3%, while that in post-flowering stage was increased by 11.4% and 29.7%, respectively. Compared with CK, RN significantly increased the relative content of chlorophyll (SPAD) of maize leaves after topdressing, the biomass in post-flowering stage was increased by 15.6%-44.9%, the ear length, the number and weight of grains per spike and the 100-grain weight were increased significantly, yield was increased by 9.8%-17.0%, and water use efficiency (WUE) was increased by 6.3%-21.4%, with the partial productivities of fertilizer (PEPT), N (PEPTN), P (PEPTP) and K (PEPTK) were all increased significantly. In conclusion, RN could improve water consumption and the SPAD value in post-flowering stage of maize in different precipitation years, increase post-flowering biomass, and optimize the ear character, obviously improve yield, water and fertilizer use efficiency. It was a effective fertilizer management mode with high-efficiency utilization of water and fertilizer under double-ridge-furrow sowing with the whole plastic film mulching in maize in the semi-arid area.

[Effects of annual whole-film mulching on freezing-thawing characteristics, moisture, and temperature distribution of maize soil in semi-arid area]. / åŠå¹²æ—±åŒºå‘¨å¹´å ¨è†œè¦†ç›–å¯¹çŽ‰ç±³ç”°åœŸå£¤å†»èžç‰¹æ€§å’Œæ°´çƒ­åˆ†å¸ƒçš„å½±å“.

Based on a 3-year field experiment (2015-2017) with two treatments, annual whole-film mulching (PM) and uncovered (CK), we analyzed the relationship between soil temperature, moisture, and soil hydrothermal movement in semi-arid area. The results showed that freezing-thawing processes under both PM and CK were one-way freezing and two-way melting. Compared with CK, the freezing period in PM treatment was lagged, freezing rate was slowed down, freezing depth was 20 cm shallower, but melting rate was faster, and melting period was shortened by 6-7 days. In freezing period, soil temperature gradients of PM and CK were positive, with heat being transmitted toward top soil layer, and the conduction strength in PM treatment was greater than CK. During the melting period, soil temperature gradient of PM was also positive, with heat being transmitted toward upper soil layer, and that of CK was conversed. Soil water in PM treatment transported to upper soil layer during freezing-thawing period, but it appeared a "down-up-down" movement mode under CK in freezing period, "up-down" in thawing period. There was positively correlation between temperature and moisture gradient in the freezing period under both PM and CK treatments, with closer correlation in PM than CK. During melting period, soil temperature and moisture gradient was positively correlated in PM treatment with soil heat and moisture moved upward synchronously, while that in CK was negatively correlated with soil heat and moisture simultaneously moved to the lower layer soil. Driven by soil temperature and moisture gradient, soil temperature in 0-10 cm, 10-20 cm and 20-30 cm layers increased by 1.13-1.34 ℃, 0.96-1.24 ℃ and 0.89-1.32 ℃, while average soil water content increased by 3.4%-5.6%, 1.4%-2.2% and 6.7%-7.8%, respectively in PM treatment before sowing. Our results indicated that PM could provide water and heat protection for re-greening of winter crop and sowing, emergence and seedling of spring-sown crops in semi-arid areas.

Regulation of mulch conditions on photosynthesis and water utilization of spring wheat in northwest semi-arid area of China.

A field experiment was conducted from 2015 to 2016 in the northwestern Loess Plateau, China, to analyze the relationships among flag-leaf photosynthetic characteristics, water-consumption characteristics, and yield components of spring wheat (Triticum aestivum 'Longchun 35'). There were three treatments: whole-field plastic mulching (PMS), sand mulching (SM), and uncovered (CK). The results showed that soil-water storage levels at 0-300 cm of soil profile before wheat filling under PMS and SM treatments were greater than that in CK by 47.8 and 31.6 mm, respectively, while that under PMS was lower than CK by 15.6 mm at the filling stage. Water consumption under PMS and SM increased in the flagging-heading and flowering-filling stages compared with the CK. Leaf area indices under PMS and SM were increased by 59.0%-73.7% and 40.1%-52.7%, respectively, and leaf SPAD values were increased by 3.5%-28.4% and 2.9%-23.9%, respectively, compared with CK. The net photosynthetic rate of PMS was increased by 23.5%, 33.0% and 17.7% at the flagging, heading, and flowering stages. The corresponding stoma-tal conductance rate was increased by 32.6%, 76.4% and 66.9%, respectively. Net photosynthetic and stomatal conductance rates at the filling stage were decreased by 26.2% and 16.4%, respectively. At the heading, flowering, and filling stages, stomatal limitation values in PMS were decreased by 14.6%, 23.9% and 22.3%, respectively, and by 25.7%, 29.8% and 17.4%, respectively in SM. The instantaneous water-use efficiency of spring wheat in PMS was increased by 57.8% at the flagging stage and decreased by 11.2% at the flowering stage. At the heading and flowering stages, the apparent quantum efficiency was increased by 22.6% and 18.7% in PMS, and by 26.8% and 14.3% in SM, respectively. Plant height and yield component indices in PMS and SM were significantly greater than that in CK, with the enhancement being greater than that in dry years. Grain yield was increased by 36.2% and 8.7% and water-use efficiency increased by 9.4% and 3.4% in PMS and SM, respectively. PMS and SM treatments increased soil water storage before the pre-filling stage of wheat, aggravated water consumption during the flagging-shooting and flowering-filling stages, resulting in greater SPAD values and leaf area indices, which promoted the photosynthetic functions of flag leaves, facilitated sink formation and photosynthetic assimilate transportation, resulting in increased grain yields and water use efficiency of spring wheat. The effects of PMS treatment were more remarkable than those of SM in terms of increasing spring wheat's yield potential in wet years and adaptability in dry years.

Effects of vertically rotary sub-soiling tillage on water utilization and yield of potato in semi-arid area of northwest China.

To examine the effects of vertically rotary sub-soiling tillage (VRT) on stage water consumption, individual and population development, yield, water use efficiency, and economic profit of potato is helpful to find the optimized tillage method which could increase crop drought-resistant, yield, and resource use efficiency. With randomized block design, the field experiment involved in three treatments, 1) vertically rotary sub-soiling tillage for 40 cm depth (VRT), 2) deep loosing tillage for 40 cm (DLT), and 3) rotary tillage for 15 cm (TT). The soil water storage in 0-200 cm soil profile in different growth stages of potato, foliar SPAD value, leaf area index (LAI), plant dry matter content, tuber yield were recorded, and stage water consumption, water use efficiency (WUE), tuber commodity rate, tuber commodity yield, and profit were calculated, to investigate the effects of VRT on production efficiency and economic profit. The results showed that VRT increased water consumption in flowering and tuber expanding stage by 46.7, 35.7 mm in 2016 and 27.2, 47.3 mm in 2017, as compared with DLT and TT. Based on the increased stage water consumption, foliar SPAD value, dry matter content, and LAI increased significantly, suggesting the VRT promoted individual and population development. The higher individual and population biomass resulted in significant increase in tuber yield which increased by 156.8%, 47.8% in 2016 and 24.8%, 41.0% in 2017 respectively, as compared with DLT and TT. Accordingly, WUE increased by 92.3%, 19.2% and 18.9%, 26.6%. The tuber commodity yield and profit significantly increased in VRT treatment, as well as the profit which reached to 12631.9, 11019.1 yuan·hm-2 in 2016 and 29498.3, 18245.5 yuan·hm-2 in 2017, respectively. VRT promoted potato water consumption in flowering and tuber expanding stages, resulted in significant increases of foliar SPAD value, plant dry matter, and LAI, with positive consequences on the tuber yield, WUE, as well as the tuber commodity yield and economic profit. These indicated that the VRT was helpful to increase potato drought resistance, yield and profit, and thus would be the appropriate tillage method on semi-arid northwest Loess Plateau.

Computational Prediction of Alzheimer's and Parkinson's Disease MicroRNAs in Domestic Animals.

As the most common neurodegenerative diseases, Alzheimer's disease (AD) and Parkinson's disease (PD) are two of the main health concerns for the elderly population. Recently, microRNAs (miRNAs) have been used as biomarkers of infectious, genetic, and metabolic diseases in humans but they have not been well studied in domestic animals. Here we describe a computational biology study in which human AD- and PD-associated miRNAs (ADM and PDM) were utilized to predict orthologous miRNAs in the following domestic animal species: dog, cow, pig, horse, and chicken. In this study, a total of 121 and 70 published human ADM and PDM were identified, respectively. Thirty-seven miRNAs were co-regulated in AD and PD. We identified a total of 105 unrepeated human ADM and PDM that had at least one 100% identical animal homolog, among which 81 and 54 showed 100% sequence identity with 241 and 161 domestic animal miRNAs, respectively. Over 20% of the total mature horse miRNAs (92) showed perfect matches to AD/PD-associated miRNAs. Pigs, dogs, and cows have similar numbers of AD/PD-associated miRNAs (63, 62, and 59). Chickens had the least number of perfect matches (34). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses suggested that humans and dogs are relatively similar in the functional pathways of the five selected highly conserved miRNAs. Taken together, our study provides the first evidence for better understanding the miRNA-AD/PD associations in domestic animals, and provides guidance to generate domestic animal models of AD/PD to replace the current rodent models.

Autophagy and ubiquitin-mediated proteolysis may not be involved in the degradation of spermatozoon mitochondria in mouse and porcine early embryos.

The mitochondrial genome is maternally inherited in animals, despite the fact that paternal mitochondria enter oocytes during fertilization. Autophagy and ubiquitin-mediated degradation are responsible for the elimination of paternal mitochondria in Caenorhabditis elegans; however, the involvement of these two processes in the degradation of paternal mitochondria in mammals is not well understood. We investigated the localization patterns of light chain 3 (LC3) and ubiquitin in mouse and porcine embryos during preimplantation development. We found that LC3 and ubiquitin localized to the spermatozoon midpiece at 3 h post-fertilization, and that both proteins were colocalized with paternal mitochondria and removed upon fertilization during the 4-cell stage in mouse and the zygote stage in porcine embryos. Sporadic paternal mitochondria were present beyond the morula stage in the mouse, and paternal mitochondria were restricted to one blastomere of 4-cell embryos. An autophagy inhibitor, 3-methyladenine (3-MA), did not affect the distribution of paternal mitochondria compared with the positive control, while an autophagy inducer, rapamycin, accelerated the removal of paternal mitochondria compared with the control. After the intracytoplasmic injection of intact spermatozoon into mouse oocytes, LC3 and ubiquitin localized to the spermatozoon midpiece, but remnants of undegraded paternal mitochondria were retained until the blastocyst stage. Our results show that paternal mitochondria colocalize with autophagy receptors and ubiquitin and are removed after in vitro fertilization, but some remnants of sperm mitochondrial sheath may persist up to morula stage after intracytoplasmic spermatozoon injection (ICSI).

[Effects of fertilizer application on water consumption characteristics and yield of potato cultured under ridge-furrow and whole filed plastic mulching in rain-fed area.] / æ–½è‚¥å¯¹æ—±åœ°å ¨è†œè¦†ç›–åž„æ²Ÿç§æ¤é©¬é“ƒè–¯è€—æ°´ç‰¹å¾åŠäº§é‡çš„å½±å“.

Chemical fertilizer reduction and organic manure substitution are the useful methods to increase potato water-and nutrient use efficiency, which is cultured under ridge-furrow and whole soil surface mulched by plastic film in semiarid rain-fed area. A 4-year field experiment was carried out from 2011 to 2014 with three treatments: 1) traditional chemical fertilizer application (F), 2) chemical nitrogen fertilizer reduced by 25% and dressing at flowering stage (DF), and 3) chemical nitrogen fertilizer reduced by 50% and organic manure substitution (OF). The soil moisture and potato yield were investigated, and seasonal water consumption, water use efficiency (WUE) were calculated to study the regulations of different nutrient management methods on potato water use process, as well as its effects on potato tuber yield and WUE. The results showed that soil water storage in potato flowering stage was the highest under DF treatment, but there were no significant difference among these three treatments. The depth of soil water depletion in DF and OF showed an increasing trend at post-flowering stage. Potato water consumption decreased significantly at pre-flowering stage, but increased by 36.2%, 23.2%, 24.8% and 19.0% respectively at post-flowering stage in 2011-2014 under DF treatment, as compared with those under F treatment. OF treatment increased potato water consumption by 20.7% and 16.3% than that under F treatment at post-flowering stage from 2011 to 2012, respectively, but exerted no significant effect at pre-flowering stage. Compared with F, DF increased potato tuber yield averagely by 2595.1 kg·hm-2 from 2012 to 2014 and significantly increased the WUE by 14.4% and 6.3% in 2013 and 2014, respectively; OF significantly increased potato tuber yield averagely by 2945 kg·hm-2 tuber yield in 4 experimental years and WUE was significantly higher than that under F from 2012 to 2014. It was indicated that both DF and OF could regulate water consumption between pre-flowering and post-flowering stages, and increase potato tuber yield and WUE. OF showed more significant effects than DF on the increment of tuber yield and WUE.

A comprehensive expression profile of micrornas in rat's pituitary.

MicroRNAs (miRNAs) are a class of small, non-coding RNA molecules that act as a negative regulator of most mRNAs. miRNAs influence the gene expression as transcriptional regulators and play an important role in many fundamental biological processes. It is generally acknowledged that miRNAs have a very important affection on mammalian pituitary. However, the answers of which role miRNAs play in the development of sexual function or how much they contribute to the pituitary function are not exactly. In our study, we used three female 21-day-old rats and three female 12-month-old rats to analysis the function of miRNAs. By the analyses of microarray data, we finished the stem-loop real-time RT-PCR for the differentially expressed miRNAs. We detected a total of 93 differentially expressed miRNAs between 21-day-old rats' pituitary and 12-month-old rats'. Stem-loop real-time RT-PCR suggests that the obtained data is of high credibility. Among these miRNAs, 7 miRNAs' expression (rno-miR-880, rno-miR-503, rno-miR-125a-3p, rno-miR-3596b, rno-miR-30e, rno-miR-214 and rno-miR-22) are significant different (P≤0.05). In a word, this study identified a number of specific changes in the expression of miRNAs, in rats by detecting the expression profile of miRNAs in rat's pituitary, and all of that lay the foundation for elucidating the regulatory mechanisms of miRNAs in rat's reproduction process. These differentially expressed miRNAs may play a very important role in rat's reproduction process.

Expression of microRNA­26b and identification of its target gene EphA2 in pituitary tissues in Yanbian cattle.

microRNAs (miRNAs/miRs) are a class of single­stranded non­coding RNA molecules of 19­24 nucleotides (nt) in length. They are widely expressed in animals, plants, bacteria and viruses. Via specific mRNA complementary pairing of target genes, miRNAs are able to regulate the expression of mRNA levels or inhibit protein translation following transcription. miRNA expression has a time­ and space specificity, and it is involved in cell proliferation and differentiation, apoptosis, development, tumor metastasis occurrence and other biological processes. miR­26b is an miRNA of 22 nt and is important in the regulation of cellular processes. With the advancement of molecular biology techniques in recent years, there have been extensive investigations into miR­26b. Numerous studies have observed that miR­26b is involved in early embryonic development, cell proliferation regulation, pituitary hormone secretion and other physiological activities. miRNAs are associated with the function of propagation. The present study used reverse transcription quantitative polymerase chain reaction to detect the relative expression levels of miR­26b in the pituitary tissue of Yanbian cattle at different developmental stages. The 2­∆∆Ct method was used to calculate the relative gene expression levels. The miRNA target gene database TargetScan and RNA22 were used for prediction of the miR­26b target gene and selective recognition was also performed. The results demonstrated that miR­26b is expressed in the pituitary tissues of Yanbian cattle at 6 and 24 months of age. The relative expression levels of miR­26b in the pituitary tissues of 24­month­old Yanbian cattle were 2.41 times that of those in the six­month­old Yanbian cattle, demonstrating significant differences in the relative expression (P<0.01). The relative expression of the candidate target genes, EphA2 and miR­26b, exhibited the opposite expression pattern. The relative expression levels in the pituitary tissues of six­month­old Yanbian cattle were 3.34 times that of those in 24­month­old Yanbian cattle (P<0.01). There are miR­26b binding sites in the 3'­untranslated region (3'­UTR) of EphA2 in bovine, human, murine and other mammalian mRNAs, suggesting that the EphA2 gene may be a target gene of miR­26b. The results of a Luciferase reporter system assay revealed that miR­26b is able to suppress EphA2 expression at the transcription level. Following the site­directed mutagenesis of plasmid EphA2 3'­UTR pmirGLO­MUT­ and miR­26b mimic­transfected HeLa cells, the dual­luciferase reporter gene assay revealed that there were three consecutive nucleotide mutations in the 3'­UTR, binding with the predicted seed region. This may have caused the miR­26b inhibition of luciferase activity to decrease from 60% in the wild­type to 26%, suggesting that miR­26b achieved its function via binding with the TACTTGAA sequence of the 3'­UTR in EphA2. In conclusion, the present study successfully assessed the expression pattern of miR­26b in the pituitary tissue of Yanbian cattle, and also confirmed that EphA2 was a target gene of miR­26b in Yanbian cattle in vitro. The present study provided the theoretical basis to further investigate the role of miR­26b in early embryonic development, pituitary hormone secretion and other reproductive functions.

Cat fertilization by mouse sperm injection.

Interspecies intracytoplasmic sperm injection has been carried out to understand species-specific differences in oocyte environments and sperm components during fertilization. While sperm aster organization during cat fertilization requires a paternally derived centriole, mouse and hamster fertilization occur within the maternal centrosomal components. To address the questions of where sperm aster assembly occurs and whether complete fertilization is achieved in cat oocytes by interspecies sperm, we studied the fertilization processes of cat oocytes following the injection of cat, mouse, or hamster sperm. Male and female pronuclear formations were not different in the cat oocytes at 6 h following cat, mouse or hamster sperm injection. Microtubule asters were seen in all oocytes following intracytoplasmic injection of cat, mouse or hamster sperm. Immunocytochemical staining with a histone H3-m2K9 antibody revealed that mouse sperm chromatin is incorporated normally with cat egg chromatin, and that the cat eggs fertilized with mouse sperm enter metaphase and become normal 2-cell stage embryos. These results suggest that sperm aster formation is maternally dependent, and that fertilization processes and cleavage occur in a non-species specific manner in cat oocytes.

[Effects of nitrogen application and elevated atmospheric CO2 on electron transport and energy partitioning in flag leaf photosynthesis of wheat].

Wheat (Triticum aestivum) plants were pot-cultured in open top chambers at the nitrogen application rate of 0 and 200 mg x kg(-1) soil and the atmospheric CO2 concentration of 400 and 760 micromol x mol(-1). Through the determination of flag leaf nitrogen and chlorophyll contents, photosynthetic rate (Pn)-intercellar CO2 concentration (Ci) response curve, and chlorophyll fluorescence parameters at heading stage, the photosynthetic electron transport rate and others were calculated, aimed to investigate the effects of nitrogen application and elevated atmospheric CO2 concentration on the photosynthetic energy partitioning in wheat flag leaves. Elevated atmospheric CO2 concentration decreased the leaf nitrogen and chlorophyll contents, compared with the ambient one, and the chlorophyll a/b ratio increased at the nitrogen application rate of 200 mg x kg(-1). With the application of nitrogen, no evident variations were observed in the maximal photochemical efficiency (Fv/Fm), maximal quantum yield under irradiance (Fv'/Fm') of PS II reaction center, photochemical fluorescence quenching coefficient (q(p)), and actual PS II efficiency under irradiance (phi(PS II) at elevated atmospheric CO2 concentration, and the total photosynthetic electron transport rate (J(F)) of PS II reaction center had no evident increase, though the non-photochemical fluorescence quenching coefficient (NPQ) decreased significantly. With no nitrogen application, the Fv'/Fm', psi(PS II), and NPQ at elevated atmospheric CO2 concentration decreased significantly, and the J(F) had a significant decrease though the Fv/Fm and q(p) did not vary remarkably. Nitrogen application increased the J(F) and photochemical electron transport rate (Jc); while elevated atmospheric CO2 concentration decreased the photorespiration electron transport rate (J0), Rubisco oxidation rate (V0), ratio of photorespiration to photochemical electron transport rate (J0/Jc) , and Rubisco oxidation/carboxylation rate (Vo/Vc), but increased the photochemical electron transport rate (Jc) and Rubisco carboxylation rate (Vc). It was concluded that elevated atmospheric CO2 concentration decreased the leaf nitrogen and chlorophyll contents, while nitrogen application increased the photosynthetic electron transport rate of PS II reaction center significantly, and promoted the photosynthetic electron flow towards photochemistry, making more photosynthetic electron take part in Rubisco carboxylation and leading to the significant increase of Pn.

[Effects of whole field-surface plastic mulching and planting in furrow on soil temperature, soil moisture, and corn yield in arid area of Gansu Province, Northwest China].

Taking spring corn (Zea mays) cultivar Shendan 16 as test material, a field experiment was conducted to study the effects of the treatments whole-field surface plastic mulching and planting in furrow (PMF), whole-field surface sand mulching and flat planting (SM), and uncovered and flat planting (CK) on the soil temperature, soil moisture, and corn yield on the dry land of arid area (annual average precipitation 415 mm) in middle Gansu Province. Comparing with CK, treatments PMF and SM increased the average temperature in 0-25 cm soil layer before tasselling stage, with the highest increment in treatment PMF. As for the soil water consumption, its depth in the three treatments increased with increasing years of planting. In the first year of planting, the soil water consumption was the most in 20-120 cm soil layer; whereas in the second year, the consumption was the most in 120-200 cm soil layer, with the soil water loss being the highest in treatment PMF. Treatment PMF had the highest grain number, grain weight per spike, and 100-grain weight, followed by treatment SM, and CK. In 2009 and 2010, the average grain number, average grain weight per spike, and average 100-grain weight in treatment PMF were increased by 13.5% and 114.2%, 29.8% and 321.1%, and 14.4% and 95.4% respectively, as compared to treatments SM and CK, and the grain yield and water use efficiency in treatments PMF and SM were increased by 333.1% and 240.2%, and 290.6% and 227.6%, respectively, as compared to CK. After two years continuous cropping of corn, the soil water loss in 120-200 cm soil layer in treatment PMF was up to 72 mm, being significantly higher than that in treatments SM (45 mm) and CK (40 mm). It was suggested that PMF could increase the soil temperature at seedling-tasselling stage, promote the corn growth in its early growth period, improve the soil water use by corn, and consequently, increase the grain number per spike and 100-grain weight, manifesting a good effect in improving corn yield and water use efficiency. However, PMF also induced more soil water consumption in 100-200 cm soil layer, which was not beneficial to the water balance through years.

Enhanced histone acetylation in somatic cells induced by a histone deacetylase inhibitor improved inter-generic cloned leopard cat blastocysts.

The objective was to determine whether alterations of histone acetylation status in donor cells affected inter-generic SCNT (igSCNT)-cloned embryo development. Leopard cat cells were treated with trichostatin A (TSA; a histone deacetylase inhibitor) for 48 h, and then donor cells were transferred into enucleated oocytes from domestic cats. Compared to non-treated cells, the acetylated histone 3 at lysine 9 (AcH3K9) and histone 4 at lysine 5 (AcH4K5) in the TSA group increased for up to 48 h (P < 0.05). The AcH3K9 signal ratios of igSCNT group was higher than control group 3 h after activation (P < 0.05). Treatment with TSA significantly increased total cell number of blastocysts (109.1 ± 6.9 vs. 71.8 ± 2.9, mean ± SEM), with no significant effects on rates of cleavage or blastocyst development (71.1 ± 2.8 vs. 67.6 ± 2.9 and 12.2 ± 2.6 vs. 11.0 ± 2.6, respectively). When igSCNT cloned embryos were transferred into a domestic cat oviduct and recovered after 8 d, blastocyst development rates and total cell numbers were greater in the TSA-igSCNT group (20.7 ± 3.0% and 2847.6 ± 37.2) than in the control igSCNT group (5.7 ± 2.2% and 652.1 ± 17.6, P < 0.05). Average total cell numbers of blastocysts were approximately 4.4-fold higher in the TSA-igSCNT group (2847.6 ± 37.2, n = 10) than in the control group (652.1 ± 17.6, n = 8; P < 0.05), but were ∼2.9-fold lower than in vivo cat blastocysts produced by intrauterine insemination (8203.8 ± 29.6, n = 5; P < 0.001). Enhanced histone acetylation levels of donor cells improved in vivo developmental competence and quality of inter-generic cloned embryos; however, fewer cells in blastocysts derived from igSCNT than blastocysts produced by insemination may reduce development potential following intergeneric cloning (none of the cloned embryos were maintained to term).

[Effects of nitrogen fertilization on wheat leaf photosynthesis under elevated atmospheric CO2 concentration].

In this paper, the effects of nitrogen (N) fertilization on the wheat leaf photosynthesis under long-term elevated atmospheric CO2 concentration (760 micromol x mol(-1)) was studied, based on the measurements of photosynthetic gas exchange parameters and light intensity-photosynthetic rate response curves at jointing stage. Under the long-term elevated atmospheric CO2 concentration, applying sufficient N could increase the wheat leaf photosynthetic rate (Pn), transpiration rate (Tr), and instantaneous water use efficiency (WUEi). Comparing with those under ambient atmospheric CO2 concentration, the Po and WUEi under the elevated atmospheric CO2 concentration increased, while the stomatal conductance (Gs) and intercellular CO2 concentration (Ci) decreased. With the increase of light flux intensity, the Pn and WUEi under the elevated atmospheric CO2 concentration were higher those under ambient atmospheric CO2 concentration, Gs was in adverse, while Ci and Tr had less change. At high fertilization rate of N, the Gs was linearly positively correlated with Pn, Tr, and WUEi, and the Gs and Ci had no correlation with each other under the elevated atmospheric CO2 concentration but negatively correlated under ambient atmospheric CO2 concentration. At low fertilization rate of N, the Gs had no correlations with Pn and WUEi but linearly positively correlated with Ci and Tr. It was suggested that under the elevated atmospheric CO2 concentration, the wheat leaf Pn at low N fertilization rate was limited by non-stomatal factor.