Optimizing injection time of GFP plasmid into sheep zygote

Microinjection of exogenous DNA into the cytoplasm of matured oocytes or zygotes is a promising technique to generate transgenic animals. However, the data about the microinjection time and procedure in sheep are limited and have not treated in detail. To obtain more in-depth information, the Sarda sheep oocytes from abattoir-derived ovaries were subjected to IVM and IVF. Then, the GFP plasmid as a reporter gene was injected into the cytoplasm of MII oocytes (n: 95) and zygotes at different post-insemination intervals (6–8 hpi, n: 120; 8–10 hpi, n: 122; 10–12 hpi, n: 110 and 12–14 hpi, n: 96). There were no significant differences in the cleavage rates between the groups. However, blastocyst rate of injected zygotes at all-time intervals was significantly lower than injected MII oocytes and control group (p < 0.05). Interestingly, the proportion of GFP-positive embryos was higher at 8–10 hpi compared with other injected groups (4 % versus 0 %, p  < 0.01). Among these, the proportion of mosaic embryos was high and two of those embryos developed to the blastocyst stage. In conclusion, we settled on the cytoplasmic microinjection of GFP plasmid at 8–10 hpi as an optimized time point for the production of transgenic sheep and subsequent experiments.     

黄瓜游离小孢子培养诱导成胚和植株再生

 以10个不同基因型黄瓜为试材进行游离小孢子培养,通过对成胚条件的系统研究,从‘7447’和‘Poinsett97’中获得了子叶形胚和再生植株。研究结果表明：基因型和小孢子发育时期是限制黄瓜游离小孢子成胚的关键因子。不同品种间胚状体产量差异显著,每皿产胚1.5～33.4个。单核靠边期是进行黄瓜游离小孢子培养的最佳时期。低温预处理有利于胚状体的诱导,4 ℃预处理2～4 d为宜,以处理2 d时胚状体产量最高。外源激素在诱导胚状体时并非必需,但低浓度的外源激素（0.5 mg·L^-1 2,4－D、0.2 mg·L^-1 6－BA）更能促进小孢子成胚。NLN和B5两种基础培养基在诱导胚状体上无显著差异。子叶形胚易分化成苗,而其它类型的胚状体不能获得再生植株。     

新疆阿尔泰山两河源国家级自然保护区树附生地衣群落物种分布与环境关系的研究

为了查明阿尔泰山两河源国家级自然保护区树附生地衣物种组成成分,以及探讨影响该地区树附生地衣群落分布格局与环境因素之间的关系,以附生地衣覆盖度为指标,应用除趋势对应分析(DCA)对样点进行分组,运用典范对应分析(CCA)分析附生地衣分布与环境因素的关系,结果表明:1)阿尔泰山两河源自然保护区树附生地衣共有49种隶属于15科30属。2)根据DCA分析将该地区树附生地衣划分为3个样点组,样点组1共有地衣22种,总盖度为2.639%,物种多样性指数为1.603;样点组2共有地衣27种,地衣总盖度为3.717%,物种多样性最大为1.972;样点组3:共有地衣12种,地衣总覆盖度为1.036%,物种多样性最小量为0.871。3)CCA排序结果表明:低海拔区,人为干扰强度高,地衣多样性较低;中海拔区,森林郁闭度高,湿度适中,干扰较少,地衣多样性较高;高海拔区,森林郁闭度较低,树种较少,光强度较强,干扰较少,主要以壳状地衣为主,多样性较低。     

Construction and immunogenicity studies of recombinant fowl poxvirus containing the S1 gene of Massachusetts 41 strain of infectious bronchitis virus

The spike 1 (S1) surface glycoprotein of infectious bronchitis virus (IBV) is the major inducer of the generation of virus neutralizing antibodies, and the administration of purified S1 has been shown to elicit a protective immune response against virulent virus challenge. On the basis of these observations, recombinant fowl poxvirus (rFPV) containing a cDNA copy of the S1 gene of IBV Mass 41 (rFPV-S1) was constructed and its immunogenicity and vaccine potential were evaluated. Initially, rFPV-S1 was shown to express the S1 in vito by indirect immunofluorescence staining and western blot analyses. Later, in vivo expression was demonstrated by the detection of IBV-specific serum immunoglobulin G and neutralization antibodies in the sera of chickens immunized with rFPV-S1. That the recombinant virus elicited anti-IBV protective immunity was indicated by the manifested, relatively mild clinical signs of disease, decreased titers of recovered challenge virus, and less severe histologic changes of the tracheas in virulent IBV Mass 41-challenged chickens previously receiving rFPV-S1 as compared with parental fowl poxvirus (FPV)-vaccinated control birds. In contrast, chickens immunized with either recombinant or parental FPV were resistant to a subsequent virulent FPV challenge. As to a preferred method of immunization, wing web administration appeared to be superior to the subcutaneous route because a greater percentage of birds vaccinated by the former protocol exhibited an anti-IBV humoral immune response. Thus, rFPV-S1 has potential as a poultry vaccine against both fowl pox and infectious bronchitis.     

Mesenchymal Stem Cells as an Alternative for Schwann Cells in Rat Spinal Cord Injury

Background: Spinal cord has a limited capacity to repair; therefore, medical interventions are necessary for treatment of injuries. Transplantation of Schwann cells has shown a great promising result for spinal cord injury (SCI). However, harvesting Schwann cell has been limited due to donor morbidity and limited expansion capacity. Furthermore, accessible sources such as bone marrow stem cells have drawn attentions to themselves. Therefore, this study was designed to evaluate the effect of bone marrow-derived Schwann cell on functional recovery in adult rats after injury. Methods: Mesenchymal stem cells were cultured from adult rats’ bone marrow and induced into Schwann cells in vitro. Differentiation was confirmed by immunocytochemistry and RT-PCR. Next, Schwann cells were seeded into collagen scaffolds and engrafted in 3 mm lateral hemisection defects. For 8 weeks, motor and sensory improvements were assessed by open field locomotor scale, narrow beam, and tail flick tests. Afterwards, lesioned spinal cord was evaluated by conventional histology and immunohistochemistry. Results: In vitro observations showed that differentiated cells had Schwann cell morphology and markers. In this study, we had four groups (n = 10 each): laminectomy, control, scaffold and scaffold + Schwann cells. Locomotor and sensory scores of cell grafted group were significantly better than control and scaffold groups. In histology, axonal regeneration and remyelination were better than control and scaffold groups. Conclusion: This study demonstrates that bone marrow-derived Schwann cells can be considered as a cell source for Schwann cells in SCI treatment. Key Words: Rats, Spinal cord injuries (SCI), Bone marrow, Schwann cells, Cell transdifferentiation     

Growth enhancement of rainbow trout (Oncorhynchus mykiss) by passive immunization against somatostatin-14

Juvenile rainbow trout (Oncorhynchus mykiss) were passively immunized by intraperitoneal immunization against somatostatin-14 (SS-14) using an antibody originating from egg-laying chicken (Gallus domesticus). Fish were immunized weekly (0, 7, 14, 21, 28, 35 days) with chicken egg yolk-derived immunoglobulin (IgY) against SS-14 (1:25 IgY, 5 mg mL^?1), and growth performance, feed utilization as well as plasma concentrations and mRNA levels of growth hormone (GH) and insulin-like growth factor I (IGF-I) were compared to the control group that received placebo immunization with PBS. Passive immunization significantly increased weight gain of treated fish (67.7 ± 7.4 g) compared to the control group (40.1 ± 2.0 g) after 35 days (p < 0.05). Feed conversion ratio (FCR) was significantly improved in the immunized fish (0.7 ± 0.08) compared to control group (1.2 ± 0.06) (p < 0.05). The concentrations of GH and IGF-I in the blood plasma showed no significant differences between the fish treated with anti-SS-14 and those of control during the treatment (p > 0.05). In both groups, GH levels decreased over the 35 days of the experiment (p < 0.05). However, IGF-I level during the period of treatment remained unchanged in both control and immunized fish with the anti-SS-14. Similarly, no changes were observed in pituitary GH and liver IGF-I mRNA levels between treatment and control at each sampling time (p > 0.05). There was no indication of a cumulative, long-lasting effect of repeated immunization on GH or IGF-I plasma concentrations or mRNA expression. The present study shows that a passive immunization of rainbow trout against SS-14 using a chicken egg yolk-derived SS-14 antibody could increase growth rate and improved FCR.     

Evaluation of efflux pump inhibitory activity of some plant extracts and using them as adjuvants to potentiate the inhibitory activity of some antibiotics against Staphylococcus aureus

Background:Antibiotic-resistant pathogens became a real global threat to human and animal health. This needs to concentrate the efforts to minimize and control these organisms. Efflux pumps are considered one of the important strategies used by bacteria to exclude harmful materials from the cell. Inhibition of these pumps can be an active strategy against multidrug resistance pathogens. There are two sources of efflux pump inhibitors that can be used, chemical and natural inhibitors. The chemical origin efflux pump inhibitors have many toxic side effects while the natural origin is characterized by a wide margin of safety for the host cell.Aim:In this study, the ability of some plant extracts like (propolis show rosemary, clove, capsaicin, and cumin) to potentiate the inhibitory activity of some antibiotics such as (ciprofloxacin, erythromycin, gentamycin, tetracycline, and ampicillin) against Staphylococcus aureus pathogen were tested.Methods:Efflux pump inhibitory activity of the selected plant extracts was tested using an ethidium bromide (EtBr) accumulation assay.Results:The results have shown that Propolis has a significant synergistic effect in combination with ciprofloxacin, erythromycin, and gentamycin. While it has no effect with tetracycline or ampicillin. Also, no synergic effect was noticed in a combination of the minimum inhibitory concentration for the selected plant extracts (rosemary, clove, capsaicin, and cumin) with any of the tested antibiotics. Interestingly, according to the results of the EtBr accumulation assay, Propolis has potent inhibitory activity against the S. aureus (MRS usa300) pump system.Conclusion:This study suggests that Propolis might act as a resistance breaker that is able to restore the activity of ciprofloxacin, erythromycin, and gentamycin against S. aureus strains, in case of the efflux-mediated antimicrobial resistance mechanisms.     

Determining suitability of thermal development models to estimate temperature parameters for embryonic development of <Emphasis Type="Italic">Sitona lepidus</Emphasis> Gyll. (Coleoptera: Curculionidae)

The clover root weevil, Sitona lepidus Gyll. is a pest of clovers (Trifolium spp.), particularly white clover (T. repens). Larva feeding severely impairs the capacity of white clover to fix atmospheric nitrogen, by attacking the nitrogen-fixing root nodules. Development of the egg stage was studied at six different constant temperatures ranging from 9 to 30°C to improve the basis for phenological forecasts. Development occurred over the entire range of temperatures, although hatching percentage at 30°C was only 24.4% compared to 90.1% at 28.5°C. Developmental time decreased with increase in temperature from 9 to 28.5°C but increased markedly at 30°C. At 28.5°C, the embryo development period was the shortest (10.35 days); whereas at 9°C, this parameter was the longest (55.08 days). Using linear regression, the lower threshold temperature was calculated 4.38°C and the thermal constant was 236.45 degree-days. The relationship between constant temperature and developmental rate was evaluated using nine models. The suitability of the models was evaluated based on eight indicators (R ², RSS, R _adj², AIC, BIC, AICC, W _j , and Z _i ). Of the nonlinear models, the Logan-6, Sharpe and DeMichele, and Lactin models were the most accurate at calculating t _opt of 27, 27.50, and 28°C, respectively. The results suggest that of the three models (Lactin, Briere, and Hilbert and Logan), that estimated critical temperatures, (t _min, t _opt, and t _max), Lactin gave the most suitable fit of data. This response of S. lepidus to temperature can be used for developing phonological models to predict the timing of egg hatch which are important for management programs targeting the weevil.     

用推力矢量控制技术改进超声速飞行器空气动力特性

为了提高某超声速飞机的空气动力学效率和机动性能，本文采用低阶的三维板块法和DATCOM半经验公式，在亚声速和超声速条件下，对不同马赫数和迎角情况计算了基本气动外形的飞机空气动力学特性、表面压力分布以及最大升力。此外还开发了一套软件以实现由引进的先进气动操纵面（如鸭翼等）控制的二维推力矢量技术。试验结果表明：气动操纵面结合推力矢量技术能够产生足够的低头力矩，且有能力满足高度机动飞行时的稳定性要求。此外，不论是亚声速还是超声速飞行，气动操纵面均可以提高气动效率5％-6％。     

Chemical Manipulation of Reproductive Efficiency of Chickpea (Cicer arietinnum L.) by Triadimefon in Relation to Changes in Ethylene

Reproductive efficiency was successfully manipulated by the application of different concentrations of triadimefon at pre‐flowering, flowering and post‐flowering stages in chickpea (Cicer arietinum L.). Triadimefon improved the parameters determining reproductive efficiency. Yield was significantly improved due to reductions in flower and pod abscission by the application of triadimefon, which also lowered the ethylene levels in leaves, flowers and pods and delayed senescence. These findings suggest a relationship between ethylene level and reproductive efficiency in chickpea. Further, they indicate that triadimefon can be used for chemical manipulation of the reproductive efficiency of chickpea under field conditions.