BACKGROUND Immature ovarian teratoma is a rare and aggressive neoplasm that affects young women.This report is the first to describe the development of immature teratoma after ovarian cystectomy for mature teratoma of...BACKGROUND Immature ovarian teratoma is a rare and aggressive neoplasm that affects young women.This report is the first to describe the development of immature teratoma after ovarian cystectomy for mature teratoma of the ovary in an adolescent female with a family history of ovarian teratoma.CASE SUMMARY A 16-year-old girl who had undergone bilateral ovarian cystectomy for mature teratomas 3 years ago showed bilateral adnexal tumors during her regular ultrasonography follow-up every 6 months.She received laparoscopic bilateral ovarian cystectomy,and final histopathology showed grade-1 immature teratoma of the left ovary and mature teratoma of the right ovary.Laparoscopic left salpingo-oophorectomy and staging procedures were performed again.Her mother,maternal aunt,and maternal grandmother had also received surgeries for mature ovarian teratomas.CONCLUSION It is important to have guidance on management of patient and family members with familial ovarian teratomas.展开更多
Common or bread wheat ( Triticum aestivum L., AABBDD, 2n=42) originated ca. 8 000 years ago from hybridization of tetraploid wheat ( Triticum turgidum L., AABB, 2n=28) and diploid Aegilops tauschii Coss. (DD...Common or bread wheat ( Triticum aestivum L., AABBDD, 2n=42) originated ca. 8 000 years ago from hybridization of tetraploid wheat ( Triticum turgidum L., AABB, 2n=28) and diploid Aegilops tauschii Coss. (DD, 2n=14). An essential prerequisite for this evolutionary step is that the natural hybrids between tetraploid wheat and diploid Aegilops tauschii can produce relatively many filled seeds which germinated well. In this study, without special techniques, e.g. immature embryo culture, out of 22 Ae. tauschii accessions, the genotype AS60 produced relatively many filled seeds which germinated well. The seed germination percentages in the crosses of Ae. tauschii ×tetraploid wheat, tetraploid wheat× Ae. tauschii and Ae. tauschii ×common wheat were, respectively, 50.0%, 57.1% and 45.5%. It seems that Ae. tauschii accession AS60 has a unique genotype which facilitate hybrid seed development and viability, and which meets with the prerequisite for wheat evolutionary. Furthermore, the significance of this finding for common wheat improvement and evolution was discussed.展开更多
In this experiment, floral development from tissue culture of bread wheat (Triticum aestivum L.) was investigated. Immature embryos of 45 wheat cultivars were cultured, and 11.1% of the genotypes regenerated floral or...In this experiment, floral development from tissue culture of bread wheat (Triticum aestivum L.) was investigated. Immature embryos of 45 wheat cultivars were cultured, and 11.1% of the genotypes regenerated floral organs from the calli near the bases of the green buds or plantlets regenerated. The floral buds were morphologically incomplete with the appearances of unisexual pistillate flowers which were naked, clustered with normal ovaries and exuberant feather-like stigmas, but without stamens, paleas, lemmas and glumes. The histological examination showed that the pistils originated from the meristematic cells near the green buds or plantlets, and the clustered pistils were formed by secondary pistillate regeneration. The development of the feather-like structures was earlier than that of the ovules. Biovule developed from an ovary besides normal uniovule. Statistical analysis by X 2 test for independency demonstrated highly significant difference of flower regeneration among the tested genotypes. Wheat cultivar YA-1 revealed higher percentage (44.4%) than other genotypes, and the response could well be repeated in different years. It was indicated that the floral regeneration of immature embryo explants of YA-1 is relatively stable. The frequency of floral regeneration was mainly regulated by the components in the subculture media, compared with the response of the dedifferentiation media, despite the obviously different components involving basal medium type, inorganic Fe2+ concentration and plant growth regulators. The results suggested the combination of 6-benzylaminopurine, alpha-naphthalene acetic-acid and doubled inorganic Fe2+ might be more beneficial to inducing the floral development than that of 2,4-D and normal inorganic Fe2+ concentration in subculture medium. However, both immature inflorescence and mature embryo, as cultured explants of YA-1, did not regenerate any flower organs. It is believed that the immature embryo culture of YA-1 can be used to establish ideal experimental system for the study of floral developmental mechanism in wheat.展开更多
The line of T240-6 was selected from 32 SC 2 lines of immature embryo culture of T240 (common wheat (Triticum aestivum L.)× Wheat-Haynaldia villosa (L.) Schur. 6D/6V substitution line) through powdery mildew ch...The line of T240-6 was selected from 32 SC 2 lines of immature embryo culture of T240 (common wheat (Triticum aestivum L.)× Wheat-Haynaldia villosa (L.) Schur. 6D/6V substitution line) through powdery mildew characterization, glutamate oxaloacetate transaminase (GOT) enzyme and gliadin (Gli) analyses and in situ hybridization. All of the individual plants resistant to powdery mildew lacked the locus of GOT at 6VL arm (GOT-V 2) and had gliadin locus at 6VS arm (Gli-V 2) of Haynaldia villosa. All the plants resistant to powdery mildew had one or two telocentric chromosomes that did not pair with wheat chromosomes but paired between themselves. T240-6 was identified as a telocentric line through in situ hybridization.展开更多
[Objective] The aim was to explore the conditions of high frequency induction of embryonic callus and plant regeneration of maize. [Method] Immature embryos of maize inbred lines were used as explants to study the eff...[Objective] The aim was to explore the conditions of high frequency induction of embryonic callus and plant regeneration of maize. [Method] Immature embryos of maize inbred lines were used as explants to study the effect of different 2,4-D concentrations on the induction of callus,the effect of different 6-BA concentrations on the differentiation of test-tube plantlet,as well as the effect of different IBA concentrations on the rooting of test-tube plantlet. [Result] 2,4-D showed obvious effect on the induction of inducement rate of maize,and the optimum induction medium was：N6 ＋ 2 mg/L of 2,4-D ＋ 500 mg/L of CH ＋ 500 mg/L of Pro ＋10 mg/L of AgNO3; the optimum differentiation medium was：N6 ＋ 0.5 mg/L of BA ＋ 500 mg/L of Pro; the optimum for the rooting of test-tube plantlet was 1/2 MS ＋ 0.5 mg/L of IBA. [Conclusion] The study had provided basis for the genetic transformation of maize.展开更多
Immature stages of 3 species of the tribe Ozophorini from China are described for the first time, they are Bryanellocoris orientalis Hidaka (3rd^5th instars), Primierus longispinus Zheng (4th^5th instars) and Vertoman...Immature stages of 3 species of the tribe Ozophorini from China are described for the first time, they are Bryanellocoris orientalis Hidaka (3rd^5th instars), Primierus longispinus Zheng (4th^5th instars) and Vertomannus brevicollum Zheng (4th^5th instars). Abrief study on fine structure of pale spots on head, pronotum and abdomen of 5th instar nymph of Bryanellocoris orientalis Hidaka was also given.展开更多
In this paper, the crawler, second instar female and male, prepupa, pupa and adult male of Cryptococcus ulmi are described and illustrated for the first time; adult female is redescribed; a key to all stages is gi...In this paper, the crawler, second instar female and male, prepupa, pupa and adult male of Cryptococcus ulmi are described and illustrated for the first time; adult female is redescribed; a key to all stages is given and the biology is briefly repor ted.展开更多
According to the study on the peripheral orogenic belts of the Junggar basin and combined with the interpretation of geophysical data, this paper points out that there is an Early Paleozoic basement of immature contin...According to the study on the peripheral orogenic belts of the Junggar basin and combined with the interpretation of geophysical data, this paper points out that there is an Early Paleozoic basement of immature continental crust in the Junggar area, which is mainly composed of Neoproterozoic-Ordovician oceanic crust and weakly metamorphosed covering sedimentary rocks. The Late Paleozoic tectonism and mineralization were developed on the basement of the Early Paleozoic immature continental crust. The Junggar metallogenic province is dominated by Cr, Cu, Ni and Au mineralization. Those large and medium-scale deposits are mainly distributed along the deep faults and particularly near the ophiolitic melange zones, and formed in the Late Paleozoic with the peak of mineralization occurring in the Carboniferous-Permian post-collisional stage. The intrusions related to Cu, Ni and Au mineralization generally have low Is, and positive εNd(t) values. The δ34S values of the ore deposits are mostly near zero, and the lead isotopes are mostly of normal lead. All these indicate that the ore-forming material comes either directly from the mantle-derived magma (for chromite and Cu-Ni deposits) or from recirculation of the basement material of the Early Paleozoic immature crust (for most Cu and Au deposits).展开更多
The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volum...The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volume and promote proliferation and differentiation of neural stem cells in the hippocampus and lateral ventricles. However, there is a lack of studies on whether total saponins of Panax notoginsertg have potential benefits on immature neuroblasts in the olfactory bulb following ischemia and reperfusion. This study established a rat model of global cerebral ischemia and reperfusion using four-vessel occlusion. Rats were administered total sa- ponins of Panax notoginseng at 75 mg/kg intraperitoneally 30 minutes after ischemia then once a day, for either 7 or 14 days. Total saponins of Panax notoginseng enhanced the number of dou- blecortin （DCX）＋ neural progenitor ceils and increased co-localization of DCX with neuronal nuclei and phosphorylated cAMP response element-binding/DCX＋ neural progenitor cells in the olfactory bulb at 7 and 14 days post ischemia. These findings indicate that following global brain ischemia/reperfusion, total saponins of Panax notoginseng promote differentiation of DCX＋ cells expressing immature neuroblasts in the olfactory bulb and the underlying mechanism is related to the activation of the signaling pathway of cyclic adenosine monophosphate response element binding protein.展开更多
Revascularization of immature necrotic teeth is a reliable treatment alternative to conventional apexogenesis or apexification. In case 1, a 12-year-old boy had his necrotic, immature mandibular left second premolar t...Revascularization of immature necrotic teeth is a reliable treatment alternative to conventional apexogenesis or apexification. In case 1, a 12-year-old boy had his necrotic, immature mandibular left second premolar treated with a revascularization technique. At a 24-month follow-up, periapical radiolucency had disappeared and thickening of the root wall was observed. In cases 2 and 3, a lO-year-old boy had his necrotic, immature, bilateral mandibular second premolars treated with the same modality. At 48-month （in case 2） and 42-month （in case 3） follow-ups, loss of periapical radiolucencies and increases in the root wall thickness were also observed.展开更多
Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues of...Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues often become severely necrotic after co-cultivation with Agrobacterium, which is one of the major obstacles in gene delivery. In this study, wheat varieties CB037, Kenong 199, Xinchun 9, Lunxuan 987, and Shi 4185 showed desirable culture potential or high infection ability in Agrobacterium-mediated transformation. Similarly, optimal regeneration conditions were determined by testing their ability to inhibit the cell necrosis and cell death phenotype. Two auxins of 2,4-dichlorophenoxyacetic acid （2,4-D） and 3,6-dichloro-o-anisic acid （dicamba） were evaluated for highly significant effect on both callus and plantlet production, although they were genotype-dependent in wheat. Substitution of 2,4-D by dicamba enhanced the growth and regeneration ability of callus from the immature embryos of most genotypes tested. The callus growth state couldn’t be modified by adding antioxidants such as ascorbic acid, cysteine, and silver nitrate or organic additives such as thiamine HCl and asparagine to the media. On the contrary, the best tissue statement and plant regeneration was achieved by employing the media containing the simplest MS （Murashige and Skoog） components and dicamba without organic components and vitamins. Thereby, our results are thought to inhibit wheat cell necrosis effectively and suggested to be used for more wheat genotypes.展开更多
Plants of the genus Taraxacum, commonly known as dandelions, are used to treat breast cancer in traditional folk medicine. However, their use has mainly been based on empirical findings without sufficient scientific e...Plants of the genus Taraxacum, commonly known as dandelions, are used to treat breast cancer in traditional folk medicine. However, their use has mainly been based on empirical findings without sufficient scientific evidence. Therefore, we hypothesized that dandelions would behave as a Selective estrogen receptor modulator （SERM） and be effective as hormone replacement therapy （HRT） in the postmenopausal women. In the present study, in vitro assay systems, including cell proliferation assay, reporter gene assay, and RT-PCR to evaluate the mRNA expression of estrogen-related genes （pS2 and progesterone receptor, PR）, were performed in human breast cancer cells. Dandelion ethanol extract （DEE） significantly increased cell proliferation and estrogen response element （ERE）-driven luciferase activity. DEE significantly induced the expression of estrogen related genes such as pS2 and PR, which was inhibited by tamoxifen at 1 gmol L-1. These results indicated that DEE could induce estrogenic activities mediated by a classical estrogen receptor pathway. In addition, immature rat uterotrophic assay was carried out to identify estrogenic activity of DEE in vivo. The lowest concentration of DEE slightly increased the uterine wet weight, but there was no significant effect with the highest concentration of DEE. The results demonstrate the potential estrogenic activities of DEE, providing scientific evidence supporting their use in traditional medicine.展开更多
The immature embryos （IEs） of wheat are the most widely used tissues for in vitro culture and genetic transformation due to its high regeneration competency. However, this explant can only be maintained in 4℃ daily...The immature embryos （IEs） of wheat are the most widely used tissues for in vitro culture and genetic transformation due to its high regeneration competency. However, this explant can only be maintained in 4℃ daily cooler for a short period time for its use in plant tissue culture or transformation experiments. This study aimed to investigate the effects of environmental temperature, cryopreservation storage temperature, and heat shock culture （HSC） temperature on the regeneration frequency of wheat IEs. Results indicated that environmental temperature significantly affected the induction of embryonic calli. The optimum total accumulated temperature （TAT） during the time of anthesis and sampling for regeneration of these tissues was around 280℃ for spring wheat type cv. CB037 and approximately 300℃ for winter wheat type cv. Kenong 199. Regeneration ability obviously declined when the highest environmental temperature was over 35℃ for 1 d or a high temperature between 30 and 33℃ lasted for 5 d during anthesis and sampling. This finding was verified by culturing the freshly isolated IEs under different temperatures from 29 to 37℃ in different controlled growth incubators for 5 d; the IEs almost completely lost regeneration ability when the temperature rose to 37℃. Cryopreservation of-20℃ caused the wheat samples lost ability of producing callus or embryonic callus in a few days, and cryopreservation of-10℃ more than 10 d made the regeneration potential of the tissues dramatically declined. Comparatively, the temperature that best maintained high regeneration ability was -5℃, at which the materials can be maintained for around 1 mon. In addition, the preservation of the immature samples at -5 or -10℃ inhibited the direct germination of the IEs, avoiding the embryo axis removing process. Our results are useful for ensuring that field collection and cryopreservation of the wheat IEs are done correctly to enable tissue culture and genetic transformation.展开更多
MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the ...MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically(P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased(P<0.001). In addition, the mRNA expression of miR-10b was negatively(P<0.01) correlated with DAZAP1 mRNA expression(r=–0.550). In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells. DAZAP1 mRNA and protein expressions were both regulated(P<0.05) by miR-10b. In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted(P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8(CCK-8) assay and the 5-Ethynyl-2′-deoxyuridine(EdU) assay. However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect(P>0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted(P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene. Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene.展开更多
·AIM: To study the role of immature dendritic cells (imDCs) on immune tolerance in rat penetrating keratoplasty (PKP) in high -risk eyes and to investigate the mechanism of immune hyporesponsiveness induced by do...·AIM: To study the role of immature dendritic cells (imDCs) on immune tolerance in rat penetrating keratoplasty (PKP) in high -risk eyes and to investigate the mechanism of immune hyporesponsiveness induced by donor-derived imDCs. ·METHODS: Seventy-five SD rats (recipient) and 39 Wistar rats (donor) were randomly divided into 3 groups: control, imDC and mature dendritic cell (mDC) group respectively. Using a model of orthotopic corneal transplantation in which allografts were placed in neovascularized high -risk eyes of recipient rat. Corneal neovascularization was induced by alkaline burn in the central cornea of recipient rat. Recipients in imDC group or mDC group were injected donor bone marrow-derived imDCs or mDCs of 1 ×10 6 respectively 1 week before corneal transplantation tail vein. Control rat received the same volume of PBS. In each group, 16 recipients were kept for determination of survival time and other 9 recipients were executed on day 3, 7 and 14 after transplantation. Cornea was harvested for hematoxylin eosin staining and acute rejection evaluation, Western blot was used to detect the expression level of Foxp3. ·RESULTS: The mean survival time of imDC group was significantly longer than that of control and mDC groups (all 【0.05). The expression level of Foxp3 on CD4 + CD25 + T cells of imDC group (2.24 ±0.18) was significantly higher than that in the control (1.68 ±0.09) and mDC groups (1.46±0.13) (all 【0.05).·CONCLUSION: Donor -derived imDC is an effective treatment in inducing immune hyporesponsiveness in rat PKP. The mechanism of immune tolerance induced by imDC might be inhibit T lymphocytes responsiveness by regulatory T cells. ·展开更多
BACKGROUND： α-asarone and acrous gramineus have been shown to play a necessary function in enhancing the reactivity and convulsant threshold to electric stimulation of immature rats. They have also been shown to eff...BACKGROUND： α-asarone and acrous gramineus have been shown to play a necessary function in enhancing the reactivity and convulsant threshold to electric stimulation of immature rats. They have also been shown to effectively suppress epileptic seizures induced by pentylenetetrazol in young rats. However, the mechanisms for these roles have been still unclear. OBJECTIVE： To observe the effects in immature rats of acrous gramineus and α -asarone on apoptosis of hippocampal neurons after epileptic seizure at the protein level, and to analyze the mechanism for these effects. DESIGN： A randomized controlled animal experiment. SETTINGS： Department of Pediatrics, First Hospital of Jilin University; Department of Histology and Embryology, Norman Bethune Medical School of Jilin University; Department of Internal Medicine, Children＇s Hospital of Changchun City; Department of Neurology, First Clinical Hospital affiliated to Harbin Medical University. MATERIALS： Fifty 3-week old Wistar rats, 34-40 g, irrespective of gender, were provided by Gaoxin Research Center of Medical Animal Experiment, Changchun. The animals were treated according to the animal ethical standards. The following chemicals were used for this study： acrous gramineus powders or infusion （Batch No, 0307113, Tianjiang Medicine Company Limited, Jiangyin）, α-asarone tablets （Batch No. 030219, Tianwei Pharmaceutical Factory, Shenyang）, and phenobarbital sodium tablets （Batch No. 020608, Xinya Medicine Company Limited, Shanghai）. The animals were divided into five groups randomly. First, ten rats were chosen as the normal controls. The remaining rats were treated with i.p. injections of pentylenetetrazol to stimulate an epileptic model. METHODS： The experiments were performed at the Neurological Laboratory of the First Hospital of Jilin University between October and December 2004. The rats were treated with i.p. injections of pentylenetetrazol （60 mg/kg） to establish an epileptic model. According to Racine＇ s standard, animals that reached stage 4 and 5 were chosen and randomly divided into 4 groups： model group, phenobarbital sodium, acrous gramineus, and a-asarone group. The normal control group was treated with an i.p. injection of physiological saline （0.5 mL）. After modeling, the model groups were intragastrically administrated 0.5 mL saline. The phenobarbital sodium, acrous gramineus, and α-asarone groups were intragastrically administrated 18 mg/kg/d phenobarbital sodium, 2 350 mg/kg/d acrous gramineus and 29 mg/kg/d α-asarone, respectively. The course of treatment was twice a day for 7 days. The normal group received intragastric administration of 0.5 mL saline at the same time. The rats were sacrificed and brain sections were prepared for light microscopy and electron microscopy. MAIN OUTCOME MEASURES： （1） Pathological changes of CA1 and CA3 hippocampal region neurons were observed by light microscopy and electronic microscopy. （2） Neuronal apoptosis in the CA1 and CA3 region was measured by TUNEL staining. （3） Bcl-2 and Bax expression in CA1 and CA3 region neurons was detected by immunohistochemistry and a ratio of Bcl-2/Bax was calculated. RESULTS： All 50 immature rats were included in the final analysis. （1） Pathological changes of CA1 and CA3 region hippocampal neurons： there were different pathological changes in all groups other than the normal control group. The number of damaged neurons in the model group was highest. The phenobarbital sodium, acrous gramineus, and α -asarone group exhibited different degrees of improvement. （2） Neuronal apoptosis in the CA1 and CA3 regions： there were less TUNEL-positive cells in the CA1 and CA3 regions in the normal control group. One week after PTZ-induced seizure, numerous TUNEL-positive cells were detected in the CA1 and CA3 regions in the remaining four groups. There was a significant difference between the normal control group and the remaining four groups （t = 12.089-19.162, P 〈 0.0 1）. The number of TUNEL-positive cells was less in the phenobarbital sodium, acrous gramineus, and α-asarone groups compared to the model group （t = 4.707-5.268, P 〈 0.01）. （3）Bcl-2 and Bax expression of neurons in the CA1 and CA3 regions： The number of Bcl-2- and Bax-positive cells was less in the normal control group. The Bax-positive cells exhibited a normal shape and had large round nuclei that were predominant. One week after PTZ-induced epilepsy, the number of Bcl-2- and Bax-positive cells in the CA1 and CA2 regions was significantly increased in the remaining four groups compared to the normal control group （t = 11.606-27.042, P 〈 0.01）. The Bax-positive cells exhibited a reduced size and nuclear pyknosis was predominant. However, there was no significant difference among the four groups （P 〉 0.05）. The number of Bcl-2-positive cells in the phenobarbital sodium, acrous gramineus, and a-asarone groups were significantly increased compared to the model group （t = 4.051-6.404, P 〈 0.01）. However, the number of Bax-positive cells was not significantly different among the four groups. The ratio of Bcl-2 to Bax expression was approximately 6.0 in the normal controls, 0.7 in the model group, and 1.0 in the remaining three groups. CONCLUSION： Acrous gramineus and a-asarone increased Bcl-2 expression and decreased Bax expression, and also reduced the number of apoptotic hippocampal neurons during PTZ-induced epileptic seizures in immature rats.展开更多
Since maize is one of the most important cereal crops in the world,establishment of an efficient genetic transformation system is critical for its improvement.In the current study,several elite corn lines were tested ...Since maize is one of the most important cereal crops in the world,establishment of an efficient genetic transformation system is critical for its improvement.In the current study,several elite corn lines were tested for suitability of Agrobacterium tumefaciens-mediated transformation by using immature embryos as explants.Infection ability and efficiency of transformation of A.tumefaciens sp.strains EHA105 and LBA4404,different heat treatment times of immature embryos before infection,influence of L-cysteine addition in co-cultivation medium after transformation,and how different ways of selection and cultivation influence the efficiency of transformation were compared.Glyphosate-resistant gene 2mG2-EPSPS was transformed into several typical maize genotypes including 78599,Zong 31 and BA,under the optimum conditions.Results showed that the hypervirulent Agrobacterium tumefaciens sp.strain EHA105 was more infectious than LBA4404.Inclusion of L-cysteine（100 mg L-1） in co-cultivation medium,and heating of the immature embryos for 3 min prior to infection led to a significant increase in the transformation efficiency.Growth in resting medium for 4-10 d and delaying selection was beneficial to the survival of resistant calli.During induction of germination,adding a high concentration of 6-BA（5 mg L-1） and a low concentration of 2,4-D（0.2 mg L-1） to regeneration medium significantly enhanced germination percentage.Using the optimized transformation procedure,more than 800 transgenic plants were obtained from 78599,Zong 31 and BA.By spraying herbicide glyphosate on leaves of transgenic lines,we identified 66 primary glyphosate-resistant plants.The transformation efficiency was 8.2%.PCR and Southern-blot analyses confirmed the integration of the transgenes in the maize genome.展开更多
The major royal jelly proteins(MRJPs)are the central constituents responsible for the specific activities of royal jelly.Here MRJPs via oral administration daily for 45 consecutive days were evaluated the effects on t...The major royal jelly proteins(MRJPs)are the central constituents responsible for the specific activities of royal jelly.Here MRJPs via oral administration daily for 45 consecutive days were evaluated the effects on the reproductive parameters in immature female mice(FM).Neonatal FM were divided into four groups fed MRJPs with doses of 0,125,250 and 500 mg/kg/body weight(M125,M250 and M500).The results in M125,M250 and M500 showed that the times of estrus were accelerated by 10.7%,15.5%and 10.7%,the secondary follicles number were increased by 50.7%,78.8%and 38.6%,the Graafian follicles were increased by 600.0%and 774.0%and 150.0%,respectively.M500 induced multi-oocyte follicles.The serum estradiol levels of the three groups were increased by 47.1%,64.9%and 31.1%,the action of MRJPs raising hormone secretion level is mainly via upregulating expression of ERˇgene.Antioxidant parameters of ovarian tissue showed that the malondialdehyde levels in M125 and M250 were decreased,the superoxide dismutase activities and glutathione peroxidase activities in M125 and M250 were increased.In conclusion,MRJPs may accelerate onset of puberty and promote follicular development in FM.Our findings would facilitate better understanding of the benefit effect of MRJPs as the key ingredient in royal jelly on promoting fertility performance.展开更多
The regeneration rate of wheat immature embryo varies among genotypes, howbeit many elite agriculture wheat varieties have low regeneration rates. Optimization of tissue culture conditions and attempts of adding signa...The regeneration rate of wheat immature embryo varies among genotypes, howbeit many elite agriculture wheat varieties have low regeneration rates. Optimization of tissue culture conditions and attempts of adding signal molecules are effective ways to increase plant regeneration rate. Inter-culture is one of ways that have not been investigated in plant tissue culture. Moreover, the use of arabinogalactan proteins （AGPs） and hydrogen peroxide （H202） have been reported to increase regeneration rate in a few plant species other than wheat. The current research pioneeringly uses inter-culture of immature embryos of different wheat genotypes, and also investigates impacts of AGP and H2O2 on the induction of embryogenic calli and plant regeneration. As a result, high-frequency regeneration wheat cultivars Kenong 199 （KN 199） and Xinchun 9 （XC9）, together with low-frequency regeneration wheat line Chinese Spring （CS）, presented striking increase in the induction of embryogenic calli and plant regeneration rate of CS through inter-culture strategy, up to 52.19 and 67.98%, respectively. Adding 50 to 200 mg L-1 AGP or 0.005 to 0.01‰ H2O2 to the callus induction medium, enhanced growth of embryogenic calli and plant regeneration rate in quite a few wheat genotypes. At 50 mg L-1 AGP application level in callus induction medium plant regeneration rates of 8.49,409.06 and 283.16% were achieved for Jimai 22 （JM22）, Jingdong 18 （JD18） and Yangmai 18 （YM18）, respectively; whereas at 100 mg L-1 AGP level, CS （105.44%）, Chuannong 16 （CN16） （80.60%） and Ningchun 4 （NC4） （62.87%） acted the best. Moreover CS （79.05%）, JM22 （7.55%）, CN16 （101.87%）, YM18 （365.56%）, Yangmai 20 （YM20） （10.48%）, and CB301 （187.40%） were more responsive to 0.005 %o of H2O2, and NC4 （35.37%） obtained the highest shoot regeneration rates at 0.01%o of H2O2. Overall, these two methods, inter-culture and AGP （or H2O2） application, can be further applied to wheat transgenic research.展开更多
This research uses the immature embryos of inbred maize lines(GSH9901,Hi01,Hi02,and Chang 7-2)as receptor materials to establish the callus induction system.These inbred lines provide the receptor materials for the ge...This research uses the immature embryos of inbred maize lines(GSH9901,Hi01,Hi02,and Chang 7-2)as receptor materials to establish the callus induction system.These inbred lines provide the receptor materials for the genetic regeneration of maize and the verification of the genetic functions of maize.The factor experiment and orthogonal experiments were used to investigate the impacts of different genotypes,immature embryo size,shield orientation,2,4-D concentration,proline concentration,and folic acid concentration on the induction rate of embryogenic callus tissue.A sensitivity experiment testing glyphosate(Bar)and an antibiotic(Cefotaxime sodium)were also conducted.The results indicate that the immature embryos of inbred maize line GSH9901 were the most effective for callus tissue induction,and the immature embryos with a length of 1.6-2.0 mm produce the best result.The upward shield face is more successful for the formation of induced callus.Using orthogonal analysis,we found that the optimal combination for the induction system was A_(3)(2,4-D concentration 0.25 mg mL^(-1)),B_(1)C_(3)(proline concentration 0.8 mg mL^(-1)),and D 2(folate Concentration 0.5 mg mL^(-1))and the induction rate reached 84%.We found that cold storage at 4℃ for 1 d is more conducive for the formation of embryogenic callus than the other treatments tested.The sensitivity experiment for callus tissue screening revealed the critical concentration of glyphosate to be 10 mg ml^(-1),and the critical concentration of antibiotic is 250 mg ml^(-1).Using this combination of glyphosate and antibiotic resulted in regenerated plants.This study established the optimal conditions for immature embryo callus tissue induction in maize.展开更多
文摘BACKGROUND Immature ovarian teratoma is a rare and aggressive neoplasm that affects young women.This report is the first to describe the development of immature teratoma after ovarian cystectomy for mature teratoma of the ovary in an adolescent female with a family history of ovarian teratoma.CASE SUMMARY A 16-year-old girl who had undergone bilateral ovarian cystectomy for mature teratomas 3 years ago showed bilateral adnexal tumors during her regular ultrasonography follow-up every 6 months.She received laparoscopic bilateral ovarian cystectomy,and final histopathology showed grade-1 immature teratoma of the left ovary and mature teratoma of the right ovary.Laparoscopic left salpingo-oophorectomy and staging procedures were performed again.Her mother,maternal aunt,and maternal grandmother had also received surgeries for mature ovarian teratomas.CONCLUSION It is important to have guidance on management of patient and family members with familial ovarian teratomas.
文摘Common or bread wheat ( Triticum aestivum L., AABBDD, 2n=42) originated ca. 8 000 years ago from hybridization of tetraploid wheat ( Triticum turgidum L., AABB, 2n=28) and diploid Aegilops tauschii Coss. (DD, 2n=14). An essential prerequisite for this evolutionary step is that the natural hybrids between tetraploid wheat and diploid Aegilops tauschii can produce relatively many filled seeds which germinated well. In this study, without special techniques, e.g. immature embryo culture, out of 22 Ae. tauschii accessions, the genotype AS60 produced relatively many filled seeds which germinated well. The seed germination percentages in the crosses of Ae. tauschii ×tetraploid wheat, tetraploid wheat× Ae. tauschii and Ae. tauschii ×common wheat were, respectively, 50.0%, 57.1% and 45.5%. It seems that Ae. tauschii accession AS60 has a unique genotype which facilitate hybrid seed development and viability, and which meets with the prerequisite for wheat evolutionary. Furthermore, the significance of this finding for common wheat improvement and evolution was discussed.
文摘In this experiment, floral development from tissue culture of bread wheat (Triticum aestivum L.) was investigated. Immature embryos of 45 wheat cultivars were cultured, and 11.1% of the genotypes regenerated floral organs from the calli near the bases of the green buds or plantlets regenerated. The floral buds were morphologically incomplete with the appearances of unisexual pistillate flowers which were naked, clustered with normal ovaries and exuberant feather-like stigmas, but without stamens, paleas, lemmas and glumes. The histological examination showed that the pistils originated from the meristematic cells near the green buds or plantlets, and the clustered pistils were formed by secondary pistillate regeneration. The development of the feather-like structures was earlier than that of the ovules. Biovule developed from an ovary besides normal uniovule. Statistical analysis by X 2 test for independency demonstrated highly significant difference of flower regeneration among the tested genotypes. Wheat cultivar YA-1 revealed higher percentage (44.4%) than other genotypes, and the response could well be repeated in different years. It was indicated that the floral regeneration of immature embryo explants of YA-1 is relatively stable. The frequency of floral regeneration was mainly regulated by the components in the subculture media, compared with the response of the dedifferentiation media, despite the obviously different components involving basal medium type, inorganic Fe2+ concentration and plant growth regulators. The results suggested the combination of 6-benzylaminopurine, alpha-naphthalene acetic-acid and doubled inorganic Fe2+ might be more beneficial to inducing the floral development than that of 2,4-D and normal inorganic Fe2+ concentration in subculture medium. However, both immature inflorescence and mature embryo, as cultured explants of YA-1, did not regenerate any flower organs. It is believed that the immature embryo culture of YA-1 can be used to establish ideal experimental system for the study of floral developmental mechanism in wheat.
文摘The line of T240-6 was selected from 32 SC 2 lines of immature embryo culture of T240 (common wheat (Triticum aestivum L.)× Wheat-Haynaldia villosa (L.) Schur. 6D/6V substitution line) through powdery mildew characterization, glutamate oxaloacetate transaminase (GOT) enzyme and gliadin (Gli) analyses and in situ hybridization. All of the individual plants resistant to powdery mildew lacked the locus of GOT at 6VL arm (GOT-V 2) and had gliadin locus at 6VS arm (Gli-V 2) of Haynaldia villosa. All the plants resistant to powdery mildew had one or two telocentric chromosomes that did not pair with wheat chromosomes but paired between themselves. T240-6 was identified as a telocentric line through in situ hybridization.
基金Supported by Natural Science Foundation of Guangxi Province(Guikezi 0991096)~~
文摘[Objective] The aim was to explore the conditions of high frequency induction of embryonic callus and plant regeneration of maize. [Method] Immature embryos of maize inbred lines were used as explants to study the effect of different 2,4-D concentrations on the induction of callus,the effect of different 6-BA concentrations on the differentiation of test-tube plantlet,as well as the effect of different IBA concentrations on the rooting of test-tube plantlet. [Result] 2,4-D showed obvious effect on the induction of inducement rate of maize,and the optimum induction medium was：N6 ＋ 2 mg/L of 2,4-D ＋ 500 mg/L of CH ＋ 500 mg/L of Pro ＋10 mg/L of AgNO3; the optimum differentiation medium was：N6 ＋ 0.5 mg/L of BA ＋ 500 mg/L of Pro; the optimum for the rooting of test-tube plantlet was 1/2 MS ＋ 0.5 mg/L of IBA. [Conclusion] The study had provided basis for the genetic transformation of maize.
基金The project was supported by "Education Grant" of National Natural SciencesFoundation of China (No. 108)
文摘Immature stages of 3 species of the tribe Ozophorini from China are described for the first time, they are Bryanellocoris orientalis Hidaka (3rd^5th instars), Primierus longispinus Zheng (4th^5th instars) and Vertomannus brevicollum Zheng (4th^5th instars). Abrief study on fine structure of pale spots on head, pronotum and abdomen of 5th instar nymph of Bryanellocoris orientalis Hidaka was also given.
文摘In this paper, the crawler, second instar female and male, prepupa, pupa and adult male of Cryptococcus ulmi are described and illustrated for the first time; adult female is redescribed; a key to all stages is given and the biology is briefly repor ted.
基金the Major State Basic Research Programs of the People’s Republic of China(No.2001CB409806).
文摘According to the study on the peripheral orogenic belts of the Junggar basin and combined with the interpretation of geophysical data, this paper points out that there is an Early Paleozoic basement of immature continental crust in the Junggar area, which is mainly composed of Neoproterozoic-Ordovician oceanic crust and weakly metamorphosed covering sedimentary rocks. The Late Paleozoic tectonism and mineralization were developed on the basement of the Early Paleozoic immature continental crust. The Junggar metallogenic province is dominated by Cr, Cu, Ni and Au mineralization. Those large and medium-scale deposits are mainly distributed along the deep faults and particularly near the ophiolitic melange zones, and formed in the Late Paleozoic with the peak of mineralization occurring in the Carboniferous-Permian post-collisional stage. The intrusions related to Cu, Ni and Au mineralization generally have low Is, and positive εNd(t) values. The δ34S values of the ore deposits are mostly near zero, and the lead isotopes are mostly of normal lead. All these indicate that the ore-forming material comes either directly from the mantle-derived magma (for chromite and Cu-Ni deposits) or from recirculation of the basement material of the Early Paleozoic immature crust (for most Cu and Au deposits).
基金supported by the Hunan Provincial Innovation Foundation for Postgraduate in China,No.CX2014B099(to XH)the Science Foundation of Hunan Provincial Education Department of China,No.11C1264(to FJD),13C958(to XH)
文摘The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volume and promote proliferation and differentiation of neural stem cells in the hippocampus and lateral ventricles. However, there is a lack of studies on whether total saponins of Panax notoginsertg have potential benefits on immature neuroblasts in the olfactory bulb following ischemia and reperfusion. This study established a rat model of global cerebral ischemia and reperfusion using four-vessel occlusion. Rats were administered total sa- ponins of Panax notoginseng at 75 mg/kg intraperitoneally 30 minutes after ischemia then once a day, for either 7 or 14 days. Total saponins of Panax notoginseng enhanced the number of dou- blecortin （DCX）＋ neural progenitor ceils and increased co-localization of DCX with neuronal nuclei and phosphorylated cAMP response element-binding/DCX＋ neural progenitor cells in the olfactory bulb at 7 and 14 days post ischemia. These findings indicate that following global brain ischemia/reperfusion, total saponins of Panax notoginseng promote differentiation of DCX＋ cells expressing immature neuroblasts in the olfactory bulb and the underlying mechanism is related to the activation of the signaling pathway of cyclic adenosine monophosphate response element binding protein.
文摘Revascularization of immature necrotic teeth is a reliable treatment alternative to conventional apexogenesis or apexification. In case 1, a 12-year-old boy had his necrotic, immature mandibular left second premolar treated with a revascularization technique. At a 24-month follow-up, periapical radiolucency had disappeared and thickening of the root wall was observed. In cases 2 and 3, a lO-year-old boy had his necrotic, immature, bilateral mandibular second premolars treated with the same modality. At 48-month （in case 2） and 42-month （in case 3） follow-ups, loss of periapical radiolucencies and increases in the root wall thickness were also observed.
基金supported by the National Natural Science Foundation of China (30971776)the National Transgenic Specialized Research Program of China (2008ZX08010-004)
文摘Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues often become severely necrotic after co-cultivation with Agrobacterium, which is one of the major obstacles in gene delivery. In this study, wheat varieties CB037, Kenong 199, Xinchun 9, Lunxuan 987, and Shi 4185 showed desirable culture potential or high infection ability in Agrobacterium-mediated transformation. Similarly, optimal regeneration conditions were determined by testing their ability to inhibit the cell necrosis and cell death phenotype. Two auxins of 2,4-dichlorophenoxyacetic acid （2,4-D） and 3,6-dichloro-o-anisic acid （dicamba） were evaluated for highly significant effect on both callus and plantlet production, although they were genotype-dependent in wheat. Substitution of 2,4-D by dicamba enhanced the growth and regeneration ability of callus from the immature embryos of most genotypes tested. The callus growth state couldn’t be modified by adding antioxidants such as ascorbic acid, cysteine, and silver nitrate or organic additives such as thiamine HCl and asparagine to the media. On the contrary, the best tissue statement and plant regeneration was achieved by employing the media containing the simplest MS （Murashige and Skoog） components and dicamba without organic components and vitamins. Thereby, our results are thought to inhibit wheat cell necrosis effectively and suggested to be used for more wheat genotypes.
基金supported by the Academic Research fund of Hoseo University in 2011-0069
文摘Plants of the genus Taraxacum, commonly known as dandelions, are used to treat breast cancer in traditional folk medicine. However, their use has mainly been based on empirical findings without sufficient scientific evidence. Therefore, we hypothesized that dandelions would behave as a Selective estrogen receptor modulator （SERM） and be effective as hormone replacement therapy （HRT） in the postmenopausal women. In the present study, in vitro assay systems, including cell proliferation assay, reporter gene assay, and RT-PCR to evaluate the mRNA expression of estrogen-related genes （pS2 and progesterone receptor, PR）, were performed in human breast cancer cells. Dandelion ethanol extract （DEE） significantly increased cell proliferation and estrogen response element （ERE）-driven luciferase activity. DEE significantly induced the expression of estrogen related genes such as pS2 and PR, which was inhibited by tamoxifen at 1 gmol L-1. These results indicated that DEE could induce estrogenic activities mediated by a classical estrogen receptor pathway. In addition, immature rat uterotrophic assay was carried out to identify estrogenic activity of DEE in vivo. The lowest concentration of DEE slightly increased the uterine wet weight, but there was no significant effect with the highest concentration of DEE. The results demonstrate the potential estrogenic activities of DEE, providing scientific evidence supporting their use in traditional medicine.
基金the National Natural Science Foundation of China(30971776)the Transgenic Major Projects,Ministry of Agriculture of China(2011ZX08010-004)
文摘The immature embryos （IEs） of wheat are the most widely used tissues for in vitro culture and genetic transformation due to its high regeneration competency. However, this explant can only be maintained in 4℃ daily cooler for a short period time for its use in plant tissue culture or transformation experiments. This study aimed to investigate the effects of environmental temperature, cryopreservation storage temperature, and heat shock culture （HSC） temperature on the regeneration frequency of wheat IEs. Results indicated that environmental temperature significantly affected the induction of embryonic calli. The optimum total accumulated temperature （TAT） during the time of anthesis and sampling for regeneration of these tissues was around 280℃ for spring wheat type cv. CB037 and approximately 300℃ for winter wheat type cv. Kenong 199. Regeneration ability obviously declined when the highest environmental temperature was over 35℃ for 1 d or a high temperature between 30 and 33℃ lasted for 5 d during anthesis and sampling. This finding was verified by culturing the freshly isolated IEs under different temperatures from 29 to 37℃ in different controlled growth incubators for 5 d; the IEs almost completely lost regeneration ability when the temperature rose to 37℃. Cryopreservation of-20℃ caused the wheat samples lost ability of producing callus or embryonic callus in a few days, and cryopreservation of-10℃ more than 10 d made the regeneration potential of the tissues dramatically declined. Comparatively, the temperature that best maintained high regeneration ability was -5℃, at which the materials can be maintained for around 1 mon. In addition, the preservation of the immature samples at -5 or -10℃ inhibited the direct germination of the IEs, avoiding the embryo axis removing process. Our results are useful for ensuring that field collection and cryopreservation of the wheat IEs are done correctly to enable tissue culture and genetic transformation.
基金financially supported by the earmarked fund for China Agriculture Research System (CARS-36)the Hunan Provincial Natural Science Foundation of China (2018JJ2176 and 2018JJ3219)
文摘MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically(P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased(P<0.001). In addition, the mRNA expression of miR-10b was negatively(P<0.01) correlated with DAZAP1 mRNA expression(r=–0.550). In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells. DAZAP1 mRNA and protein expressions were both regulated(P<0.05) by miR-10b. In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted(P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8(CCK-8) assay and the 5-Ethynyl-2′-deoxyuridine(EdU) assay. However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect(P>0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted(P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene. Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene.
基金Military Medical Research Projects Fund of China (No. CLZ11JA25)
文摘·AIM: To study the role of immature dendritic cells (imDCs) on immune tolerance in rat penetrating keratoplasty (PKP) in high -risk eyes and to investigate the mechanism of immune hyporesponsiveness induced by donor-derived imDCs. ·METHODS: Seventy-five SD rats (recipient) and 39 Wistar rats (donor) were randomly divided into 3 groups: control, imDC and mature dendritic cell (mDC) group respectively. Using a model of orthotopic corneal transplantation in which allografts were placed in neovascularized high -risk eyes of recipient rat. Corneal neovascularization was induced by alkaline burn in the central cornea of recipient rat. Recipients in imDC group or mDC group were injected donor bone marrow-derived imDCs or mDCs of 1 ×10 6 respectively 1 week before corneal transplantation tail vein. Control rat received the same volume of PBS. In each group, 16 recipients were kept for determination of survival time and other 9 recipients were executed on day 3, 7 and 14 after transplantation. Cornea was harvested for hematoxylin eosin staining and acute rejection evaluation, Western blot was used to detect the expression level of Foxp3. ·RESULTS: The mean survival time of imDC group was significantly longer than that of control and mDC groups (all 【0.05). The expression level of Foxp3 on CD4 + CD25 + T cells of imDC group (2.24 ±0.18) was significantly higher than that in the control (1.68 ±0.09) and mDC groups (1.46±0.13) (all 【0.05).·CONCLUSION: Donor -derived imDC is an effective treatment in inducing immune hyporesponsiveness in rat PKP. The mechanism of immune tolerance induced by imDC might be inhibit T lymphocytes responsiveness by regulatory T cells. ·
基金grants from Jilin Bureau of Science and Technology(No. 20030430)Traditional Chinese Medicine and Drug Adminsitration of Jilin Provinece(No. 2004079)
文摘BACKGROUND： α-asarone and acrous gramineus have been shown to play a necessary function in enhancing the reactivity and convulsant threshold to electric stimulation of immature rats. They have also been shown to effectively suppress epileptic seizures induced by pentylenetetrazol in young rats. However, the mechanisms for these roles have been still unclear. OBJECTIVE： To observe the effects in immature rats of acrous gramineus and α -asarone on apoptosis of hippocampal neurons after epileptic seizure at the protein level, and to analyze the mechanism for these effects. DESIGN： A randomized controlled animal experiment. SETTINGS： Department of Pediatrics, First Hospital of Jilin University; Department of Histology and Embryology, Norman Bethune Medical School of Jilin University; Department of Internal Medicine, Children＇s Hospital of Changchun City; Department of Neurology, First Clinical Hospital affiliated to Harbin Medical University. MATERIALS： Fifty 3-week old Wistar rats, 34-40 g, irrespective of gender, were provided by Gaoxin Research Center of Medical Animal Experiment, Changchun. The animals were treated according to the animal ethical standards. The following chemicals were used for this study： acrous gramineus powders or infusion （Batch No, 0307113, Tianjiang Medicine Company Limited, Jiangyin）, α-asarone tablets （Batch No. 030219, Tianwei Pharmaceutical Factory, Shenyang）, and phenobarbital sodium tablets （Batch No. 020608, Xinya Medicine Company Limited, Shanghai）. The animals were divided into five groups randomly. First, ten rats were chosen as the normal controls. The remaining rats were treated with i.p. injections of pentylenetetrazol to stimulate an epileptic model. METHODS： The experiments were performed at the Neurological Laboratory of the First Hospital of Jilin University between October and December 2004. The rats were treated with i.p. injections of pentylenetetrazol （60 mg/kg） to establish an epileptic model. According to Racine＇ s standard, animals that reached stage 4 and 5 were chosen and randomly divided into 4 groups： model group, phenobarbital sodium, acrous gramineus, and a-asarone group. The normal control group was treated with an i.p. injection of physiological saline （0.5 mL）. After modeling, the model groups were intragastrically administrated 0.5 mL saline. The phenobarbital sodium, acrous gramineus, and α-asarone groups were intragastrically administrated 18 mg/kg/d phenobarbital sodium, 2 350 mg/kg/d acrous gramineus and 29 mg/kg/d α-asarone, respectively. The course of treatment was twice a day for 7 days. The normal group received intragastric administration of 0.5 mL saline at the same time. The rats were sacrificed and brain sections were prepared for light microscopy and electron microscopy. MAIN OUTCOME MEASURES： （1） Pathological changes of CA1 and CA3 hippocampal region neurons were observed by light microscopy and electronic microscopy. （2） Neuronal apoptosis in the CA1 and CA3 region was measured by TUNEL staining. （3） Bcl-2 and Bax expression in CA1 and CA3 region neurons was detected by immunohistochemistry and a ratio of Bcl-2/Bax was calculated. RESULTS： All 50 immature rats were included in the final analysis. （1） Pathological changes of CA1 and CA3 region hippocampal neurons： there were different pathological changes in all groups other than the normal control group. The number of damaged neurons in the model group was highest. The phenobarbital sodium, acrous gramineus, and α -asarone group exhibited different degrees of improvement. （2） Neuronal apoptosis in the CA1 and CA3 regions： there were less TUNEL-positive cells in the CA1 and CA3 regions in the normal control group. One week after PTZ-induced seizure, numerous TUNEL-positive cells were detected in the CA1 and CA3 regions in the remaining four groups. There was a significant difference between the normal control group and the remaining four groups （t = 12.089-19.162, P 〈 0.0 1）. The number of TUNEL-positive cells was less in the phenobarbital sodium, acrous gramineus, and α-asarone groups compared to the model group （t = 4.707-5.268, P 〈 0.01）. （3）Bcl-2 and Bax expression of neurons in the CA1 and CA3 regions： The number of Bcl-2- and Bax-positive cells was less in the normal control group. The Bax-positive cells exhibited a normal shape and had large round nuclei that were predominant. One week after PTZ-induced epilepsy, the number of Bcl-2- and Bax-positive cells in the CA1 and CA2 regions was significantly increased in the remaining four groups compared to the normal control group （t = 11.606-27.042, P 〈 0.01）. The Bax-positive cells exhibited a reduced size and nuclear pyknosis was predominant. However, there was no significant difference among the four groups （P 〉 0.05）. The number of Bcl-2-positive cells in the phenobarbital sodium, acrous gramineus, and a-asarone groups were significantly increased compared to the model group （t = 4.051-6.404, P 〈 0.01）. However, the number of Bax-positive cells was not significantly different among the four groups. The ratio of Bcl-2 to Bax expression was approximately 6.0 in the normal controls, 0.7 in the model group, and 1.0 in the remaining three groups. CONCLUSION： Acrous gramineus and a-asarone increased Bcl-2 expression and decreased Bax expression, and also reduced the number of apoptotic hippocampal neurons during PTZ-induced epileptic seizures in immature rats.
基金supported by the National Key Project of transgenic varieties breeding(2009ZX08003-003B)the Light of West Talent Training Project of China(2010-2011)the Project of Sichuan Province Finance Genetic Engineering,China(2011JYGC01-002)
文摘Since maize is one of the most important cereal crops in the world,establishment of an efficient genetic transformation system is critical for its improvement.In the current study,several elite corn lines were tested for suitability of Agrobacterium tumefaciens-mediated transformation by using immature embryos as explants.Infection ability and efficiency of transformation of A.tumefaciens sp.strains EHA105 and LBA4404,different heat treatment times of immature embryos before infection,influence of L-cysteine addition in co-cultivation medium after transformation,and how different ways of selection and cultivation influence the efficiency of transformation were compared.Glyphosate-resistant gene 2mG2-EPSPS was transformed into several typical maize genotypes including 78599,Zong 31 and BA,under the optimum conditions.Results showed that the hypervirulent Agrobacterium tumefaciens sp.strain EHA105 was more infectious than LBA4404.Inclusion of L-cysteine（100 mg L-1） in co-cultivation medium,and heating of the immature embryos for 3 min prior to infection led to a significant increase in the transformation efficiency.Growth in resting medium for 4-10 d and delaying selection was beneficial to the survival of resistant calli.During induction of germination,adding a high concentration of 6-BA（5 mg L-1） and a low concentration of 2,4-D（0.2 mg L-1） to regeneration medium significantly enhanced germination percentage.Using the optimized transformation procedure,more than 800 transgenic plants were obtained from 78599,Zong 31 and BA.By spraying herbicide glyphosate on leaves of transgenic lines,we identified 66 primary glyphosate-resistant plants.The transformation efficiency was 8.2%.PCR and Southern-blot analyses confirmed the integration of the transgenes in the maize genome.
基金The authors are grateful to Dr.Quanwei Wei from Nanjing Agricultural University,Nanjing,China for his technical assistance.This work was supported by the National Natural Science Foundation of China(no.31271848)。
文摘The major royal jelly proteins(MRJPs)are the central constituents responsible for the specific activities of royal jelly.Here MRJPs via oral administration daily for 45 consecutive days were evaluated the effects on the reproductive parameters in immature female mice(FM).Neonatal FM were divided into four groups fed MRJPs with doses of 0,125,250 and 500 mg/kg/body weight(M125,M250 and M500).The results in M125,M250 and M500 showed that the times of estrus were accelerated by 10.7%,15.5%and 10.7%,the secondary follicles number were increased by 50.7%,78.8%and 38.6%,the Graafian follicles were increased by 600.0%and 774.0%and 150.0%,respectively.M500 induced multi-oocyte follicles.The serum estradiol levels of the three groups were increased by 47.1%,64.9%and 31.1%,the action of MRJPs raising hormone secretion level is mainly via upregulating expression of ERˇgene.Antioxidant parameters of ovarian tissue showed that the malondialdehyde levels in M125 and M250 were decreased,the superoxide dismutase activities and glutathione peroxidase activities in M125 and M250 were increased.In conclusion,MRJPs may accelerate onset of puberty and promote follicular development in FM.Our findings would facilitate better understanding of the benefit effect of MRJPs as the key ingredient in royal jelly on promoting fertility performance.
基金financially supported in part by the National Key Project for Tansgenic Study, Ministry of Agriculture of China(2011ZX08010-004)
文摘The regeneration rate of wheat immature embryo varies among genotypes, howbeit many elite agriculture wheat varieties have low regeneration rates. Optimization of tissue culture conditions and attempts of adding signal molecules are effective ways to increase plant regeneration rate. Inter-culture is one of ways that have not been investigated in plant tissue culture. Moreover, the use of arabinogalactan proteins （AGPs） and hydrogen peroxide （H202） have been reported to increase regeneration rate in a few plant species other than wheat. The current research pioneeringly uses inter-culture of immature embryos of different wheat genotypes, and also investigates impacts of AGP and H2O2 on the induction of embryogenic calli and plant regeneration. As a result, high-frequency regeneration wheat cultivars Kenong 199 （KN 199） and Xinchun 9 （XC9）, together with low-frequency regeneration wheat line Chinese Spring （CS）, presented striking increase in the induction of embryogenic calli and plant regeneration rate of CS through inter-culture strategy, up to 52.19 and 67.98%, respectively. Adding 50 to 200 mg L-1 AGP or 0.005 to 0.01‰ H2O2 to the callus induction medium, enhanced growth of embryogenic calli and plant regeneration rate in quite a few wheat genotypes. At 50 mg L-1 AGP application level in callus induction medium plant regeneration rates of 8.49,409.06 and 283.16% were achieved for Jimai 22 （JM22）, Jingdong 18 （JD18） and Yangmai 18 （YM18）, respectively; whereas at 100 mg L-1 AGP level, CS （105.44%）, Chuannong 16 （CN16） （80.60%） and Ningchun 4 （NC4） （62.87%） acted the best. Moreover CS （79.05%）, JM22 （7.55%）, CN16 （101.87%）, YM18 （365.56%）, Yangmai 20 （YM20） （10.48%）, and CB301 （187.40%） were more responsive to 0.005 %o of H2O2, and NC4 （35.37%） obtained the highest shoot regeneration rates at 0.01%o of H2O2. Overall, these two methods, inter-culture and AGP （or H2O2） application, can be further applied to wheat transgenic research.
基金This research was funded by Jilin province science and technology research projects(20170204005NY)Jilin Science and Technology Development Plan Major Science and Technology R&D Project(20180201029NY)+2 种基金Jilin Province Science and Technology Development Plan Project(20190802012ZG)Jilin Province Natural Science Foundation(20190201168JC)a thirteenth five-year plan for the Education Department of Jilin Province(JJKH20180661KJ)were jointly funded.
文摘This research uses the immature embryos of inbred maize lines(GSH9901,Hi01,Hi02,and Chang 7-2)as receptor materials to establish the callus induction system.These inbred lines provide the receptor materials for the genetic regeneration of maize and the verification of the genetic functions of maize.The factor experiment and orthogonal experiments were used to investigate the impacts of different genotypes,immature embryo size,shield orientation,2,4-D concentration,proline concentration,and folic acid concentration on the induction rate of embryogenic callus tissue.A sensitivity experiment testing glyphosate(Bar)and an antibiotic(Cefotaxime sodium)were also conducted.The results indicate that the immature embryos of inbred maize line GSH9901 were the most effective for callus tissue induction,and the immature embryos with a length of 1.6-2.0 mm produce the best result.The upward shield face is more successful for the formation of induced callus.Using orthogonal analysis,we found that the optimal combination for the induction system was A_(3)(2,4-D concentration 0.25 mg mL^(-1)),B_(1)C_(3)(proline concentration 0.8 mg mL^(-1)),and D 2(folate Concentration 0.5 mg mL^(-1))and the induction rate reached 84%.We found that cold storage at 4℃ for 1 d is more conducive for the formation of embryogenic callus than the other treatments tested.The sensitivity experiment for callus tissue screening revealed the critical concentration of glyphosate to be 10 mg ml^(-1),and the critical concentration of antibiotic is 250 mg ml^(-1).Using this combination of glyphosate and antibiotic resulted in regenerated plants.This study established the optimal conditions for immature embryo callus tissue induction in maize.
